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Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDTVitaminK Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my summer vacation (long)Reply-VitaminK > > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the > beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence. > > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands, > organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane > surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we > have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing > them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes > generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins. > > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the > kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming > biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve > the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of > antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved > using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > > > > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves. > > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be > treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes. > > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends' > children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too > hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > > > Interfase by Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1 > capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > > > > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a > maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > > > > Children 16+ and adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin > K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. > > > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica. > > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies. If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED! > The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large > quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps > from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. > > > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which > seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in > the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to > hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms, which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off > the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be > acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located > inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis, > resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. > > > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the > various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has > been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and > regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes. > > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling > once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in > the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION > > > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the > kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live > microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used. > > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes > some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was > slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more > responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our > cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less > trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer. > > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with. > > > > > > > >

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How do you know you have biofilms?CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Friday, September 9, 2011 6:35 AMSubject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDTVitaminK Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK > > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the > beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence. > > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands, > organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane > surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we > have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing > them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes > generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins. > > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the > kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming > biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve > the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of > antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved > using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > > > > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves. > > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes. > > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends' > children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too > hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > > > Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1 > capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > > > > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a > maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > > > > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin > K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. > > > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica. > > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED! > The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large > quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps > from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which > seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in > the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to > hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off > the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be > acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located > inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. > > > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the > various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has > been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and > regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes. > > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling > once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION > > > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the > kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live > microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used. > > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes > some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was > slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more > responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our > cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less > trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer. > > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with. > > > > > > > >

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I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

 

How do you know you have biofilms?Cecilia

From: " Goldstein@... " <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

 

Yes.  True.  L.From: " Aandraya Da Silva " <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

 

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing " biofilm protocols " assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other " biofilm protocols " in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book " Healing Lyme " by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was " yes " which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Understanding BiofilmsAuthor: Amy Proal26MAY2008As humans, our environment consistently exposes us to a variety of dangers. Tornadoes, lightning, flooding and hurricanes can all hamper our survival. Not to mention the fact that most of us can encounter swerving cars or ill-intentioned people at any given moment.Biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of materialThousands of years ago, humans realized that they could better survive a dangerous world if they formed into communities, particularly communities consisting of people with different talents. They realized that a community is far more likely to survive through division of labor– one person makes food, another gathers resources, still another protects the community against invaders. Working together in this manner requires communication and cooperation.Inhabitants of a community live in close proximity and create various forms of shelter in order to protect themselves from external threats. We build houses that protect our families and larger buildings that protect the entire community. Grouping together inside places of shelter is a logical way to enhance survival.With the above in mind, it should come as no surprise that the pathogens we harbor are seldom found as single entities. Although the pathogens that cause acute infection are generally free-floating bacteria – also referred to as planktonic bacteria – those chronic bacterial forms that stick around for decades long ago evolved ways to join together into communities. Why? Because by doing so, they are better able to combat the cells of our immune system bent upon destroying them.It turns out that a vast number of the pathogens we harbor are grouped into communities called biofilms. In an article titled “Bacterial Biofilms: A Common Cause of Persistent Infections,†JW Costerton at the Center for Biofilm Engineering in Montana defines a bacterial biofilm as “a structured community of bacterial cells enclosed in a self-produced polymeric matrix and adherent to an inert or living surface.â€[1] In layman’s terms, that means that bacteria can join together on essentially any surface and start to form a protective matrix around their group. The matrix is made of polymers – substances composed of molecules with repeating structural units that are connected by chemical bonds.According to the Center for Biofilm Engineering at Montana State University, biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of material – such as metals, plastics, soil particles, medical implant materials and, most significantly, human or animal tissue. The first bacterial colonists to adhere to a surface initially do so by inducing weak, reversible bonds called van der Waals forces. If the colonists are not immediately separated from the surface, they can anchor themselves more permanently using cell adhesion molecules, proteins on their surfaces that bind other cells in a process called cell adhesion.A biofilm in the gut.These bacterial pioneers facilitate the arrival of other pathogens by providing more diverse adhesion sites. They also begin to build the matrix that holds the biofilm together. If there are species that are unable to attach to a surface on their own, they are often able to anchor themselves to the matrix or directly to earlier colonists.During colonization, things start to get interesting. Multiple studies have shown that during the time a biofilm is being created, the pathogens inside it can communicate with each other thanks to a phenomenon called quorum sensing. Although the mechanisms behind quorum sensing are not fully understood, the phenomenon allows a single-celled bacterium to perceive how many other bacteria are in close proximity. If a bacterium can sense that it is surrounded by a dense population of other pathogens, it is more inclined to join them and contribute to the formation of a biofilm.Bacteria that engage in quorum sensing communicate their presence by emitting chemical messages that their fellow infectious agents are able to recognize. When the messages grow strong enough, the bacteria respond en masse, behaving as a group. Quorum sensing can occur within a single bacterial species as well as between diverse species, and can regulate a host of different processes, essentially serving as a simple communication network. A variety of different molecules can be used as signals.“Disease-causing bacteria talk to each other with a chemical vocabulary,†says Doug Hibbins of Princeton University. A graduate student in the lab of Princeton University microbiologist Dr. Bonnie Bassler, Hibbins was part of a research effort which shed light on how the bacteria that cause cholera form biofilms and communicate via quorum sensing.[2]“Forming a biofilm is one of the crucial steps in cholera’s progression,†states Bassler. “They [bacteria] cover themselves in a sort of goop that’s a shield against antibiotics, allowing them to grow rapidly. When they sense there are enough of them, they try to leave the body.â€Although cholera bacteria use the intestines as a breeding ground, after enough biofilms have formed, planktonic bacteria inside the biofilm seek to leave the body in order to infect a new host. It didn’t take long for Bassler and team to realize that the bacteria inside cholera biofilms must signal each other in order to communicate that it’s time for the colony to stop reproducing and focus instead on leaving the body.“We generically understood that bacteria talk to each other with quorum sensing, but we didn’t know the specific chemical words that cholera uses,†Bassler said.Then Higgins isolated the CAI-1 – a chemical which occurs naturally in cholera. Another graduate student figured out how to make the molecule in the laboratory. By moderating the level of CAI-1 in contact with cholera bacteria, Higgins was successfully able to chemically control cholera’s behavior in lab tests. His team eventually confirmed that when CAI-1 is absent, cholera bacteria attach in biofilms to their current host. But when the bacteria detect enough of the chemical, they stop making biofilms and releasing toxins, perceiving that it is time to leave the body instead. Thus, CAI-1 may very well be the single molecule that allow the bacteria inside a cholera biofilm to communicate. Although it is likely that the bacteria in a cholera biofilm may communicate with other signals besides CAI-1, the study is a good example of the fact that signaling molecules serve a key role in determining the state of a biofilm.Sessile cells in a biofilm “talk†to each other via quorum sensing to build microcolonies and to keep water channels open.Similarly, researchers at the University of Iowa (several of whom are now at the University of Washington) have spent the last decade identifying the molecules that allow the bacterial species P. aeruginosa to form biofilms in the lungs of patients with cystic fibrosis.[3] Although the P. auruginosa isolated from the lungs of patients with cystic fibrosis looks like a biofilm and acts like a biofilm, up until recently, there were no objective tests available to confirm that the bacterial species did indeed form biofilms in the lungs of patients with the disease, nor was there a way to tell what proportion of P. aeruginosa in the lungs were actually in biofilm mode.“We needed a way to show that the P. auruginosa in cystic fibrosis lungs was communicating like a biofilm. That could tell us about the P. auruginosalifestyle,†said Pradeep Singh, M.D., a lead author on the study who is now at the University of Washington.Singh and his colleagues finally discovered that P. aeruginosa uses one of two particular quorum-sensing molecules to initiate the formation of biofilms. In November 1999, his research team screened the entire bacterial genome, identifying 39 genes that are strongly controlled by the quorum-sensing system.In a 2000 study published in Nature, Singh and colleagues developed a sensitive test which shows P. auruginosa from cystic fibrosis lungs produces the telltale, quorum-sensing molecules that are the signals for biofilm formation.[3]It turns out that P. aerugnosa secretes two signaling molecules, one that is long, and another that is short. Using the new test, the team was able to show that planktonic forms of P. aeruginosa produce more long signaling molecules. Alternately, when they tested the P. aeruginosa strains isolated from the lungs of patients with cystic fibrosis (which were in biofilm form), all of the strains produced the signaling molecules, but in the opposite ratio – more short than long.Interestingly, when the biofilm strains of P. aeruginosa were separated in broth into individual bacterial forms, they reverted to producing more long signal molecules than short ones. Does this mean that a change in signaling molecular length can indicate whether bacteria remain as planktonic forms or develop into biofilms?To find out, the team took the bacteria from the broth and made them grow as a biofilm again. Sure enough, those strains of bacteria in biofilm form produced more short signal molecules than long.“The fact that the P. aeruginosa in [the lungs of cystic fibrosis patients] is making the signals in the ratios that we see tells us that there is a biofilm and that most of the P. aeruginosa in the lung is in the biofilm state,†states Greenberg, another member of the research team. He believes that the findings allow for a clear biochemical definition of whether bacteria are in a biofilm. Techniques similar to those used by his group will likely be used to determine the properties of other biofilm signaling molecules.DevelopmentOnce colonization has begun, the biofilm grows through a combination of cell division and recruitment. The final stage of biofilm formation is known as development and is the stage in which the biofilm is established and may only change in shape and size. This development of a biofilm allows for the cells inside to become more resistant to antibiotics administered in a standard fashion. In fact, depending on the organism and type of antimicrobial and experimental system, biofilm bacteria can be up to a thousand times more resistant to antimicrobial stress than free-swimming bacteria of the same species.Biofilms grow slowly, in diverse locations, and biofilm infections are often slow to produce overt symptoms. However, biofilm bacteria can move in numerous ways that allow them to easily infect new tissues. Biofilms may move collectively, by rippling or rolling across the surface, or by detaching in clumps. Sometimes, in a dispersal strategy referred to as “swarming/seedingâ€, a biofilm colony differentiates to form an outer “wall†of stationary bacteria, while the inner region of the biofilm “liquefiesâ€, allowing planktonic cells to “swim†out of the biofilm and leave behind a hollow mound.[4]Biofilm bacteria can move in numerous ways: Collectively, by rippling or rolling across the surface, or by detaching in clumps. Individually, through a “swarming and seeding†dispersal.Research on the molecular and genetic basis of biofilm development has made it clear that when cells switch from planktonic to community mode, they also undergo a shift in behavior that involves alterations in the activity of numerous genes. There is evidence that specific genes must be transcribed during the attachment phase of biofilm development. In many cases, the activation of these genes is required for synthesis of the extracellular matrix that protects the pathogens inside.According to Costerton, the genes that allow a biofilm to develop are activated after enough cells attach to a solid surface. “Thus, it appears that attachment itself is what stimulates synthesis of the extracellular matrix in which the sessile bacteria are embedded,†states the molecular biologist. “This notion– that bacteria have a sense of touch that enables detection of a surface and the expression of specific genes– is in itself an exciting area of research…â€[1]Certain characteristics may also facilitate the ability of some bacteria to form biofilms. Scientists at the Department of Microbiology and Molecular Genetics, Harvard Medical School, performed a study in which they created a “mutant†form of the bacterial species P. aeguinosa (PA).[5] The mutants lacked genes that code for hair-like appendages called pili. Interestingly, the mutants were unable to form biofilms. Since the pili of PA are involved in a type of surface-associated motility called twitching, the team hypothesized this twitching might be required for the aggregation of cells into the microcolonies that subsequently form a stable biofilm.Once a biofilm has officially formed, it often contains channels in which nutrients can circulate. Cells in different regions of a biofilm also exhibit different patterns of gene expression. Because biofilms often develop their own metabolism, they are sometimes compared to the tissues of higher organisms, in which closely packed cells work together and create a network in which minerals can flow.“There is a perception that single-celled organisms are asocial, but that is misguided,†said Andre Levchenko, assistant professor of biomedical engineering in s Hopkins University’s Whiting School of Engineering and an affiliate of the University’s Institute for NanoBioTechnology. “When bacteria are under stress—which is the story of their lives—they team up and form this collective called a biofilm. If you look at naturally occurring biofilms, they have very complicated architecture. They are like cities with channels for nutrients to go in and waste to go out.â€[6]The biofilm life cycle in three steps: attachment, growth of colonies (development), and periodic detachment of planktonic cells.Understanding how such cooperation among pathogens evolves and is maintained represents one of evolutionary biology’s thorniest problems. This stems from the reality that, in nature, freeloading cheats inevitably evolve to exploit any cooperative group that doesn’t defend itself, leading to the breakdown of cooperation. So what causes the bacteria in a biofilm to contribute to and share resources rather than steal them? Recently, Dr. Brockhurst of the University of Liverpool and colleagues at the Université Montpellier and the University of Oxford conducted several studies in an effort to understand why the bacteria in a biofilm cooperate and share resources rather than horde them.[7]The team took a closer look at P. fluorescens biofilms, which are formed when individual cells overproduce a polymer that sticks the cells together, allowing the colonization of liquid surfaces. While production of the polymer is metabolically costly to individual cells, the biofilm group benefits from the increased access to oxygen that surface colonization provides. Yet, evolutionarily speaking, such a setup allows possible “cheaters†to enter the biofilm. Such cheats can take advantage of the protective matrix while failing to contribute energy to actually building the matrix. If too many “cheaters†enter a biofilm, it will weaken and eventually break apart.After several years of study, Brockhurst and team realized that the short-term evolution of diversity within a biofilm is a major factor in how successfully its members cooperate. The team found that once inside a biofilm, P. fluorescensdifferentiates into various forms, each of which uses different nutrient resources. The fact that these “diverse cooperators†don’t all compete for the same chemicals and nutrients substantially reduces competition for resources within the biofilm.When the team manipulated diversity within experimental biofilms, they found that diverse biofilms contained fewer “cheaters†and produced larger groups than non-diverse biofilms.Levchenko and team used this device to observe bacteria growing in cramped conditions.Similarly, this year, researchers from s Hopkins; Virginia Tech; the University of California, San Diego; and Lund University in Sweden recently released the results of a study which found that once bacteria cooperate and form a biofilm, packing tightly together further enhances their survival.[6]The team created a new device in order to observe the behavior of E. coli bacteria forced to grow in the cramped conditions. The device, which allows scientists to use extremely small volumes of cells in solution, contains a series of tiny chambers of various shapes and sizes that keep the bacteria uniformly suspended in a culture medium.Not surprisingly, the cramped bacteria in the device began to form a biofilm. The team captured the development of the biofilm on video, and were able to observe the gradual self-organization and eventual construction of bacterial biofilms over a 24-hour period.First, Andre Levchenko and Hojung Cho of s Hopkins recorded the behavior of single layers of E. coli cells using real-time microscopy. “We were surprised to find that cells growing in chambers of all sorts of shapes gradually organized themselves into highly regular structures,†Levchenko said.Dr. Levchenko of s Hopkins and Hojung Cho, a biomedical engineering doctoral studentFurther observations using microscopy revealed that the longer the packed cell population resided in the chambers, the more ordered the biofilm structure became. As the cells in the biofilm became more ordered and tightly packed, the biofilm became harder and harder to penetrate.Levchenko also noted that rod-shaped E. colithat were too short or too long typically did not organize well into the dense, circular main hub of the biofilm. Instead, the bacteria of odd shapes or highly disordered groups of cells were found on the edges of the biofilm, where they formed sharp corners.Nodes of relapsing infection?Researchers often note that, once biofilms are established, planktonic bacteria may periodically leave the biofilm on their own. When they do, they can rapidly multiply and disperse.According to Costerton, there is a natural pattern of programmed detachment of planktonic cells from biofilms. This means that biofilms can act as what Costerton refers to as “niduses†of acute infection. Because the bacteria in a biofilm are protected by a matrix, the host immune system is less likely to mount a response to their presence.[1]But if planktonic bacteria are periodically released from the biofilms, each time single bacterial forms enter the tissues, the immune system suddenly becomes aware of their presence. It may proceed to mount an inflammatory response that leads to heightened symptoms. Thus, the periodic release of planktonic bacteria from some biofilms may be what causes many chronic relapsing infections.Planktonic bacteria are periodically released from a biofilmAs R. Parsek of Northwestern University describes in a 2003 paper in the Annual Review of Microbiology, any pathogen that survives in a chronic form benefits by keeping the host alive.[8] After all, if a chronic bacterial form simply kills its host, it will no longer have a place to live. So according to Parsek, chronic infection often results in a “disease stalemate†where bacteria of moderate virulence are somewhat contained by the defenses of the host. The infectious agents never actually kill the host, but the host is never able to fully kill the invading pathogens either.Parsek believes that the optimal way for bacteria to survive under such circumstances is in a biofilm, stating that “Increasing evidence suggests that the biofilm mode of growth may play a key role in both of these adaptations. Biofilm growth increases the resistance of bacteria to killing and may make organisms less conspicuous to the immune system… ultimately this moderation of virulence may serve the bacteria’s interest by increasing the longevity of the host.â€The acceptance of biofilms as infectious entitiesAnton van Leeuwenhoek.Perhaps because many biofilms are sufficiently thick to be visible to the naked eye, the microbial communities were among the first to be studied by early microbiologists. Anton van Leeuwenhoek scraped the plaque biofilm from his teeth and observed what he described as the “animalculi†inside them under his primitive microscope. However, according to Costerton and team at the Center for Biofilm Research at Montana State University, it was not until the 1970s that scientists began to appreciate that bacteria in the biofilm mode of existence constitute such a major component of the bacterial biomass in most environments. Then, it was not until the 1980s and 1990s that scientists truly began to understand how elaborately organized a bacterial biofilm community can be.[1]As Kolter, professor of microbiology and molecular genetics at Harvard Medical School, and one of the first scientists to study how biofilms developstates, “At first, however, studying biofilms was a radical departure from previous work.â€Like most microbial geneticists, Kolter had been trained in the tradition dating back to Koch and Louis Pasteur, namely that bacteriology is best conducted by studying pure strains of planktonic bacteria. “While this was a tremendous advance for modern microbiology, it also distracted microbiologists from a more organismic view of bacteria, Kolter adds, “Certainly we felt that pure, planktonic cultures were the only way to work. Yet in nature bacteria don’t live like that,†he says. “In fact, most of them occur in mixed, surface-dwelling communities.â€Although research on biofilms has surged over the past few decades, the majority of biofilm research to date has focused on external biofilms, or those that form on various surfaces in our natural environment.Over the past years, as scientists developed better tools to analyze external biofilms, they quickly discovered that biofilms can cause a wide range of problems in industrial environments. For example, biofilms can develop on the interiors of pipes, which can lead to clogging and corrosion. Biofilms on floors and counters can make sanitation difficult in food preparation areas.Since biofilms have the ability to clog pipes, watersheds, storage areas, and contaminate food products, large companies with facilities that are negatively impacted by their presence have naturally taken an interest in supporting biofilm research, particularly research that specifies how biofilms can be eliminated.This means that many recent advances in biofilm detection have resulted from collaborations between microbial ecologists, environmental engineers, and mathematicians. This research has generated new analytical tools that help scientists identify biofilms.Biofilm in a swamp gas reactor.For example, the Canadian company FAS International Ltd. has justcreated an endoluminal brush, which will be launched this spring. Physicians can use the brush to obtain samples from the interior of catheters. Samples taken from catheters can be sent to a lab, where researchers determine if biofilms are present in the sample. If biofilms are detected, the catheter is immediately replaced, since the insertion of catheters with biofilms can cause the patient to suffer from numerous infections, some of which are potentially life threatening.Scientists now realize that biofilms are not just composed of bacteria. Nearly every species of microorganism – including viruses, fungi, and Archaea – have mechanisms by which they can adhere to surfaces and to each other. Furthermore, it is now understood that biofilms are extremely diverse. For example, upward of 300 different species of bacteria can inhabit the biofilms that form dental plaque.[9]Furthermore, biofilms have been found literally everywhere in nature, to the point where any mainstream microbiologist would acknowledge that their presence is ubiquitous. They can be found on rocks and pebbles at the bottom of most streams or rivers and often form on the surface of stagnant pools of water. In fact, biofilms are important components of food chains in rivers and streams and are grazed upon by the aquatic invertebrates upon which many fish feed. Biofilms even grow in the hot, acidic pools at Yellowstone National Park and on glaciers in Antarctica.Biofilm in acidic pools at Yellowstone National Park.It is also now understood that the biofilm mode of existence has been around for millenia. For example, filamentous biofilms have been identified in the 3.2-billion-year-old deep-sea hydrothermal rocks of the Pilbara Craton, Australia. According to a 2004 article in Nature Reviews Microbiology, “Biofilm formation appears early in the fossil record (approximately 3.25 billion years ago) and is common throughout a diverse range of organisms in both the Archaea and Bacteria lineages. It is evident that biofilm formation is an ancient and integral component of the prokaryotic life cycle, and is a key factor for survival in diverse environments.â€[10]Biofilms and diseaseThe fact that external biofilms are ubiquitous raises the question – if biofilms can form on essentially every surface in our external environments, can they do the same inside the human body? The answer seems to be yes, and over the past few years, research on internal biofilms has finally started to pick up pace. After all, it’s easy for biofilm researchers to see that the human body, with its wide range of moist surfaces and mucosal tissue, is an excellent place for biofilms to thrive. Not to mention the fact that those bacteria which join a biofilm have a significantly greater chance of evading the battery of immune system cells that more easily attack planktonic forms.Many would argue that research on internal biofilms has been largely neglected, despite the fact that bacterial biofilms seem to have great potential for causing human disease.Common sites of biofilm infection. One biofilm reach the bloodstream they can spread to any moist surface of the human body. Stoodley of the Center for Biofilm Engineering at Montana State University, attributes much of the lag in studying biofilms to the difficulties of working with heterogeneous biofilms compared with homogeneous planktonic populations. In a 2004 paper in Nature Reviews, the molecular biologist describes many reasons why biofilms are extremely difficult to culture, such as the fact that the diffusion of liquid through a biofilm and the fluid forces acting on a biofilm must be carefully calculated if it is to be cultured correctly. According to Stoodley, the need to master such difficult laboratory techniques has deterred many scientists from attempting to work with biofilms. [10]Also, since much of the technology needed to detect internal biofilms was created at the same time as the sequencing of the human genome, interest in biofilm bacteria, and the research grants that would accompany such interest, have been largely diverted to projects with a decidedly genetic focus. However, since genetic research has failed to uncover the cause of any of the common chronic diseases, biofilms are finally – just over the past few years – being studied more intensely, and being given the credit they deserve as serious infectious entities, capable of causing a wide array of chronic illnesses.In just a short period of time, researchers studying internal biofilms have already pegged them as the cause of numerous chronic infections and diseases, and the list of illnesses attributed to these bacterial colonies continues to grow rapidly.According to a recent public statement from the National Institutes of Health, more than 65% of all microbial infections are caused by biofilms. This number might seem high, but according to Kim of the Department of Chemical and Biological Engineering at Tufts University, “If one recalls that such common infections as urinary tract infections (caused by E. coli and other pathogens), catheter infections (caused by Staphylococcus aureus and other gram-positive pathogens), child middle-ear infections (caused by Haemophilus influenzae, for example), common dental plaque formation, and gingivitis, all of which are caused by biofilms, are hard to treat or frequently relapsing, this figure appears realistic.â€[11]Hundreds of microbial biofilm colonize the human mouth, causing tooth decay and gum disease.As mentions, perhaps the most well-studied biofilms are those that make up what is commonly referred to as dental plaque. “Plaque is a biofilm on the surfaces of the teeth,†states Parsek. “This accumulation of microorganisms subject the teeth and gingival tissues to high concentrations of bacterial metabolites which results in dental disease.â€[12]It has also recently been shown that biofilms are present on the removed tissue of 80% of patients undergoing surgery for chronic sinusitis. According to Parsek, biofilms may also cause osteomyelitis, a disease in which the bones and bone marrow become infected. This is supported by the fact that microscopy studies have shown biofilm formation on infected bone surfaces from humans and experimental animal models. Parsek also implicates biofilms in chronic prostatitis since microscopy studies have also documented biofilms on the surface of the prostatic duct. Microbes that colonize vaginal tissue and tampon fibers can also form into biofilms, causing inflammation and disease such as Toxic Shock Syndrome.Biofilms also cause the formation of kidney stones. The stones cause disease by obstructing urine flow and by producing inflammation and recurrent infection that can lead to kidney failure. Approximately 15%–20% of kidney stones occur in the setting of urinary tract infection. According to Parsek, these stones are produced by the interplay between infecting bacteria and mineral substrates derived from the urine. This interaction results in a complex biofilm composed of bacteria, bacterial exoproducts, and mineralized stone material.Microbes that colonize vaginal tissue and tampon fibers can become pathogenic, causing inflammation and disease such as Toxic Shock Syndrome.Perhaps the first hint of the role of bacteria in these stones came in 1938 when Hellstrom examined stones passed by his patients and found bacteria embedded deep inside them. Microscopic analysis of stones removed from infected patients has revealed features that characterize biofilm growth. For one thing, bacteria on the surface and inside the stones are organized in microcolonies and surrounded by a matrix composed of crystallized (struvite) minerals.Then there’s endocarditis, a disease that involves inflammation of the inner layers of the heart. The primary infectious lesion in endocarditis is a complex biofilm composed of both bacterial and host components that is located on a cardiac valve. This biofilm, known as a vegetation, causes disease by three basic mechanisms. First, the vegetation physically disrupts valve function, causing leakage when the valve is closed and inducing turbulence and diminished flow when the valve is open. Second, the vegetation provides a source for near-continuous infection of the bloodstream that persists even during antibiotic treatment. This causes recurrent fever, chronic systemic inflammation, and other infections. Third, pieces of the infected vegetation can break off and be carried to a terminal point in the circulation where they block the flow of blood (a process known as embolization). The brain, kidney, and extremities are particularly vulnerable to the effects of embolization.A variety of pathogenic biofims are also commonly found on medical devices such as joint prostheses and heart valves. According to Parsek, electron microscopy of the surfaces of medical devices that have been foci of device-related infections shows the presence of large numbers of slime-encased bacteria. Tissues taken from non-device-related chronic infections also show the presence of biofilm bacteria surrounded by an exopolysaccharide matrix. These biofilm infections may be caused by a single species or by a mixture of species of bacteria or fungi.According to Dr. Patel of the Mayo Clinic, individuals with prosthetic joints are often oblivious to the fact that their prosthetic joints harbor biofilm infections.[13]Cells of Staphylococcus epidermidis causing devastating disease as they grow on the cuff at a mechanical heart valve.“When people think of infection, they may think of fever or pus coming out of a wound,†explains Dr. Patel. “However, this is not the case with prosthetic joint infection. Patients will often experience pain, but not other symptoms usually associated with infection. Often what happens is that the bacteria that cause infection on prosthetic joints are the same as bacteria that live harmlessly on our skin. However, on a prosthetic joint they can stick, grow and cause problems over the long term. Many of these bacteria would not infect the joint were it not for the prosthesis.â€Biofilms also cause Leptospirosis, a serious but neglected emerging disease that infects humans through contaminated water. New research published in the May issue of the journal Microbiology shows for the first time how bacteria that cause the disease survive in the environment.Leptospirosis is a major public health problem in southeast Asia and South America, with over 500,000 severe cases every year. Between 5% and 20% of these cases are fatal. Rats and other mammals carry the disease-causing pathogen Leptospira interrogans in their kidneys. When they urinate, they contaminate surface water with the bacteria, which can survive in the environment for long periods.“This led us to see if the bacteria build a protective casing around themselves for protection,†said Professor Mathieu Picardeau from the Institut Pasteur in Paris, France. [14]Previously, scientists believed the bacteria were planktonic. But Professor Picardeau and his team have shown that L. interrogans can make biofilms, which could be one of the main factors controlling survival and disease transmission. “90% of the species of Leptospira we tested could form biofilms. It takes L. interrogans an average of 20 days to make a biofilm,†says Picardeau.Biofilms have also been implicated in a wide array of veterinary diseases. For example, researchers at the Virginia-land Regional College of Veterinary Medicine at Virginia Tech were just awarded a grant from the United States Department of Agriculture to study the role biofilms play in the development of Bovine Respiratory Disease Complex (BRDC). If biofilms play a role in bovine respiratory disease, it’s likely only a matter of time before they will be established as a cause of human respiratory diseases as well.When the immune response is compromised, Pseudomonas aeruginosabiofilms are able to colonize the alveoli, and to form biofilms.As mentioned previously, infection by the bacterium Pseudomonas aeruginosa (P. aeruginosa) is the main cause of death among patients with cystic fibrosis. Pseudomonas is able to set up permanent residence in the lungs of patients with cystic fibrosis where, if you ask most mainstream researchers, it is impossible to kill. Eventually, chronic inflammation produced by the immune system in response to Pseudomonas destroys the lung and causes respiratory failure. In the permanent infection phase, P. aeruginosa biofilms are thought to be present in the airway, although much about the infection pathogenesis remains unclear.[15]Cystic fibrosis is caused by mutations in the proteins of channels that regulates chloride. How abnormal chloride channel protein leads to biofilm infection remains hotly debated. It is clear, however, that cystic fibrosis patients manifest some kind of host-defense defect localized to the airway surface. Somehow this leads to a debilitating biofilm infection.Biofilms have the potential to cause a tremendous array of infections and diseasesBecause internal pathogenic biofilm research comprises such a new field of study, the infections described above almost certainly represent just the tip of the iceberg when it comes to the number of chronic diseases and infections currently caused by biofilms.For example, it wasn’t until July of 2006 that researchers realized that the majority of ear infections are caused by biofilm bacteria. These infections, which can be either acute or chronic, are referred to collectively as otitis media (OM). They are the most common illness for which children visit a physician, receive antibiotics, or undergo surgery in the United States.There are two subtypes of chronic OM. Recurrent OM (ROM) is diagnosed when children suffer repeated infections over a span of time and during which clinical evidence of the disease resolves between episodes. Chronic OM with effusion is diagnosed when children have persistent fluid in the ears that lasts for months in the absence of any other symptoms except conductive hearing loss.It took over ten years for researchers to realize that otitis media is caused by biofilms. Finally, in 2002, Drs. Ehrlich and J. Post, an Allegheny General Hospital pediatric ear specialist and medical director of the Center for Genomic Sciences, published the first animal evidence of biofilms in the middle ear in the Journal of the American Medical Association, setting the stage for further clinical investigation.In a subsequent study, Ehrlich and Post obtained middle ear mucosa – or membrane tissue – biopsies from children undergoing a procedure for otitis. The team gathered uninfected mucosal biopsies from children and adults undergoing cochlear implantation as a control.[16]Using advanced confocal laser scanning microscopy, Luanne Hall Stoodley, Ph.D. and her ASRI colleagues obtained three dimensional images of the biopsies and evaluated them for biofilm morphology using generic stains and species-specific probes for Haemophilus influenzae, Streptococcus pneumoniaeand Moraxella catarrhalis. Effusions, when present, were also evaluated for evidence of pathogen specific nucleic acid sequences (indicating presence of live bacteria).The study found mucosal biofilms in the middle ears of 46/50 children (92%) with both forms of otitis. Biofilms were not observed in eight control middle ear mucosa specimens obtained from cochlear implant patients.Otitis media, or inflammation of the inner ear, is caused by biofilm.In fact, all of the children in the study who suffered from chronic otitis media tested positive for biofilms in the middle ear, even those who were asymptomatic, causing Erlich to conclude that, “It appears that in many cases recurrent disease stems not from re-infection as was previously thought and which forms the basis for conventional treatment, but from a persistent biofilm.â€He went on to state that the discovery of biofilms in the setting of chronic otitis media represented “a landmark evolution in the medical community’s understanding about a disease that afflicts millions of children world-wide each year and further endorses the emerging biofilm paradigm of chronic infectious disease.â€The emerging biofilm paradigm of chronic disease refers to a new movement in which researchers such as Ehrlich are calling for a tremendous shift in the way the medical community views bacterial biofilms. Those scientists who support an emerging biofilm paradigm of chronic disease feel that biofilm research is of utmost importance because of the fact that the infectious entities have the potential to cause so many forms of chronic disease. The Marshall Pathogenesis is an important part of this paradigm shift.It was also just last year that researchers realized that biofilms cause most infections associated with contact lens use. In 2006, Bausch & Lomb withdrew its ReNu with MoistureLoc contact lens solution because a high proportion of corneal infections were associated with it. It wasn’t long before researchers at the University Hospitals Case Medical Center found that the infections were caused by biofilms. [17]“Once they live in that type of state [a biofilm], the cells become resistant to lens solutions and immune to the body’s own defense system,†said Mahmoud A. Ghannoum, Ph.D, senior investigator of the study. “This study should alert contact lens wearers to the importance of proper care for contact lenses to protect against potentially virulent eye infections,†he said.It turns out that the biofilms detected by Ghannoum and team were composed of fungi, particularly a species called Fusarium. His team also discovered that the strain of fungus (with the catchy name, ATCC 36031) used for testing the effectiveness of lens care solutions is a strain that does not produce biofilms as the clinical fungal strains do. ReNu contact solution, therefore, was effective in the laboratory, but failed when faced with strains in real-world situations.Fungal biofilm can form in contact lens solution leading to potentially virulent eye infectionsUnfortunately, Ghannoum and team were not able to create a method to target and destroy the fungal biofilms that plague users of ReNu and some other contact lens solutions.Then there’s Dr. Randall Wolcott who just recently discovered and confirmed that the sludge covering diabetic wounds is largely made up of biofilms. Whereas before Wolcott’s work such limbs generally had to be amputated, now that they have been correctly linked to biofilms, measures such as those described in thisinterview can be taken to stop the spread of infection and save the limb. Wolcott has finally been given a grant by the National Institutes of Health to further study chronic biofilms and wound development.Dr. Garth and the Medical Biofilm Laboratory team at Montana State University are also researching wounds and biofilms. Their latest article and an image showing wound biofilm was featured on the cover of the January-February 2008 issue of Wound Repair and Regeneration.[18]Biofilm bacteria and chronic inflammatory diseaseIn just a few short years, the potential of biofilms to cause debilitating chronic infections has become so clear that there is little doubt that biofilms are part of the pathogenic mix or “pea soup†that cause most or all chronic “autoimmune†and inflammatory diseases.In fact, thanks, in large part, to the research of biomedical researcher Dr. Trevor Marshall, it is now increasingly understood that chronic inflammatory diseases result from infection with a large microbiota of chronic biofilm and L-form bacteria (collectively called the Th1 pathogens).[19][20] The microbiota is thought to be comprised of numerous bacterial species, some of which have yet to be discovered. However, most of the pathogens that cause inflammatory disease have one thing in common – they have all developed ways to evade the immune system and persist as chronic forms that the body is unable to eliminate naturally.Some L-form bacteria are able to evade the immune system because, long ago, they evolved the ability to reside inside macrophages, the very white bloods cells of the immune system that are supposed to kill invading pathogens. Upon formation, L-form bacteria also lose their cell walls, which makes them impervious to components of the immune response that detect invading pathogens by identifying the proteins on their cell walls. The fact that L-form bacteria lack cell walls also means that the beta-lactam antibiotics, which work by targeting the bacterial cell wall, are completely ineffective at killing them.[21]Clearly, transforming into the L-form offers any pathogen a survival advantage. But among those pathogens not in an L-form state, joining a biofilm is just as likely to enhance their ability to evade the immune system. Once enough chronic pathogens have grouped together and formed a stable community with a strong protective matrix, they are likely able to reside in any area of the body, causing the host to suffer from chronic symptoms that are both mental and physical in nature.Biofilm researchers will also tell you that, not surprisingly, biofilms form with greater ease in an immunocompromised host. Marshall’s research has made it clear that many of the Th1 pathogens are capable of creating substances that bind and inactivate the Vitamin D Receptor – a fundamental receptor of the body that controls the activity of the innate immune system, or the body’s first line of defense against intracellular infection.[22]Diagram of the Vitamin D Receptor and capnine.Thus, as patients accumulate a greater number of the Th1 pathogens, more and more of the chronic bacterial forms create substances capable of disabling the VDR. This causes a snowball effect, in which the patient becomes increasingly immunocompromised as they acquire a larger bacterial load.For one thing, it’s possible that many of the bacteria that survive inside biofilms are capable of creating VDR blocking substances. Thus, the formation of biofilms may contribute to immune dysfunction. Conversely, as patients acquire L-form bacteria and other persistent bacterial forms capable of creating VDR-blocking substances, it becomes exceptionally easy for biofilms to form on any tissue surface of the human body.Thus, patients who begin to acquire L-form bacteria almost always fall victim to biofilm infections as well, since it is all too easy for pathogens to group together into a biofilm when the immune system isn’t working up to par.To date, there is also no strict criteria that separate L-form bacteria from biofilm bacteria or any other chronic pathogenic forms. This means that L-form bacteria may also form into biofilms, and by doing so enter a mode of survival that makes them truly impervious to the immune system. Some L-form bacteria may not form complete biofilms, yet may still possess the ability to surround themselves in a protective matrix. Under these circumstances one might say they are in a “biofilm-like†state.Marshall often refers to the pathogens that cause inflammatory disease as an intraphgocytic, metagenomic microbiota of bacteria, terms which suggest that most chronic bacterial forms possess properties of both L-form and biofilm bacteria. Intraphagocytic refers to the fact that the pathogens can be found inside the cells of the immune system. The term metagenomic indicates that there are a tremendous number of different species of these chronic bacterial forms. Finally, microbiota refers to the fact that biofilm communities sustain their pathogenic activity.For example, when observed under a darkfield microscope, L-form bacteria are often encased in protective biofilm sheaths. If the blood containing the pathogens are aged overnight, the bacterial colonies reach a point where they expand and burst out of the cell, causing the cell to burst as well. Then they extend as huge, long biofilm tubules, which are presumably helping the pathogens spread to other cells. The tubules also help spread bacterial DNA to neighboring cells.Clearly, there is a great need for more research on how different chronic bacterial forms interact. To date, L-form researchers have essentially focused soley on the L-form, while failing to investigate how frequently the wall-less pathogens form into biofilms or become parts of biofilm communities together with bacteria with cell walls. Conversely, most biofilm researchers are intently studying the biofilm mode of growth without considering the presence of L-form bacteria. So, it will likely take several years before we will be better able to understand probable overlaps between the lifestyles of L-form and biofilm bacteria.Anyone who is skeptical about the fact that biofilms likely form a large percentage of the microbiota that cause inflammatory disease should consider many of the recent studies that have linked established biofilm infections to a higher risk for multiple forms of chronic inflammatory disease. Take, for example, studies that have found a link between periodontal disease and several major inflammatory conditions. A 1989 article published in British Medical Journal showed a correlation between dental disease and systemic disease (stroke, heart disease, diabetes). After correcting for age, exercise, diet, smoking, weight, blood cholesterol level, alcohol use and health care, people who had periodontal disease had a significantly higher incidence of heart disease, stroke and premature death. More recently, these results were confirmed in studies in the United States, Canada, Great Britain, Sweden, and Germany. The effects are striking. For example, researchers from the Canadian Health Bureau found that people with periodontal disease had a two times higher risk of dying from cardiovascular disease.[23]Dental plaque as seen under a scanning electron microcroscope.Since we know that periodontal disease is caused by biofilm bacteria, the most logical explanation for the fact that people with dental problems are much more likely to suffer from heart disease and stroke is that the biofilms in their mouths have gradually spread to the moist surfaces of their circulatory systems. Or perhaps if the bacteria in periodontal biofilms create VDR binding substances, their ability to slow innate immune function allows new biofilms (and L-form bacteria as well) to more easily form and infect the heart and blood vessels. Conversely, systemic infection with VDR blocking biofilm bacteria is also likely to weaken immune defenses in the gums and facilitate periodontal disease.In fact, it appears that biofilm bacteria in the mouth also facilitate the formation of biofilm and L-form bacteria in the brain. Just last year, researchers at Vasant Hirani at University College London released the results of a study which found that elderly people who have lost their teeth are at more than three-fold greater risk of memory problems and dementia.[24]At the moment, Autoimmunity Research Foundation does not have the resources to culture biofilms from patients on the treatment and, even if they did, current methods for culturing internal biofilms remain unreliable. According to Stoodley, “The lack of standard methods for growing, quantifying and testing biofilms in continuous culture results in incalculable variability between laboratory systems. Biofilm microbiology is complex and not well represented by flask cultures. Although homogeneity allows statistical enumeration, the extent to which it reflects the real, less orderly world is questionable.â€[10]How else do we acquire biofilm bacteria?As discussed thus far, biofilms form spontaneously as bacteria inside the human body group together. Yet people can also ingest biofilms by eating contaminated food.According to researchers at the University of Guelph in Ontario Canada, it is increasingly suspected that biofilms play an important role in contamination of meat during processing and packaging. The group warns that greater action must be taken to reduce the presence of food-borne pathogens like Escherichia coli and Listeria monocytogenes and spoilage microorganisms such as thePseudomonas species (all of which form biofilms) throughout the food processing chain to ensure the safety and shelf-life of the product. Most of these microorganisms are ubiquitous in the environment or brought into processing facilities through healthy animal carriers.Hans Blaschek of the University of Illinois has discovered that biofilms form on much of the other food products we consume as well.A biofilm on a piece of lettuce“If you could see a piece of celery that’s been magnified 10,000 times, you’d know what the scientists fighting foodborne pathogens are up against,†says Blaschek.“It’s like looking at a moonscape, full of craters and crevices. And many of the pathogens that cause foodborne illness, such as Shigella, E. coli,and Listeria, make sticky, sugary biofilms that get down in these crevices, stick like glue, and hang on like crazy.â€According to Blaschek, the problem faced by produce suppliers can be a triple whammy. “If you’re unlucky enough to be dealing with a pathogen–and the pathogen has the additional attribute of being able to form biofilm—and you’re dealing with a food product that’s minimally processed, well, you’re triply unlucky,†the scientist said. “You may be able to scrub the organism off the surface, but the cells in these biofilms are very good at aligning themselves in the subsurface areas of produce.†, a University of Illinois food science and human nutrition professor agrees, stating,â€Once the pathogenic organism gets on the product, no amount of washing will remove it. The microbes attach to the surface of produce in a sticky biofilm, and washing just isn’t very effective.â€Biofilms can even be found in processed water. Just this month, a study was released in which researchers at the Department of Biological Sciences, at Virginia Polytechnic Institute isolated M. avium biofilm from the shower head of a woman with M. avium pulmonary disease.[25] A molecular technique called DNA fingerprinting demonstrated that M. avium isolates from the water were the same forms that were causing the woman’s respiratory illness.Effectively targeting biofilm infectionsAlthough the mainstream medical community is rapidly acknowledging the large number of diseases and infections caused by biofilms, most researchers are convinced that biofilms are difficult or impossible to destroy, particularly those cells that form the deeper layers of a thick biofilm. Most papers on biofilms state that they are resistant to antibiotics administered in a standard manner. For example, despite the fact that Ehrlich and team discovered that biofilm bacteria cause otitis media, they are unable to offer an effective solution that would actually allow for the destruction of biofilms in the ear canal. Other teams have also come up short in creating methods to break up the biofilms they implicate as the cause of numerous infections.This means patients with biofilm infections are generally told by mainstream doctors that they have an untreatable infection. In some cases, a disease-causing biofilm can be cut out of a patient’s tissues, or efforts are made to drain components of the biofilm out of the body. For example, doctors treating otitis media often treats patients with myringotomy, a surgical procedure in which small tubes are placed in the eardrum to continuously drain infectious fluid.When it comes to administering antibiotics in an effort to target biofilms, one thing is certain. Mainstream researchers have repeatedly tried to kill biofilms by giving patients high, constant doses of antibiotics. Unfortunately, when administered in high doses, the antibiotic may temporarily weaken the biofilm but is incapable of destroying it, as certain cells inevitably persist and allow the biofilm to regenerate.“You can put a patient on [a high dose] antibiotics, and it may seem that the infection has disappeared,†says Levchenko. “But in a few months, it reappears, and it is usually in an antibiotic-resistant form.â€What the vast majority of researchers working with biofilms fail to realize is that antibiotics are capable of destroying biofilms. The catch is that antibiotics are only effective against biofilms if administered in a very specific manner. Furthermore, only certain antibiotics appear to effectively target biofilms. After decades of research, much of which was derived from molecular modeling data, Marshall was the first to create an antibiotic regimen that appears to effectively target and destroy biofilms. Central to the treatment, which is called the Marshall Protocol, is the fact that biofilms and other Th1 pathogens succumb to specific bacteriostatic antibiotics taken in very low, pulsed doses. It is only when antibiotics are administered in this manner that they appear capable of fully eradicating biofilms.[19][20]In a paper entitled “The Riddle of Biofilm Resistance,†Dr. Kim of Tulane University discusses the mechanisms by which pulsed, low dose antibiotics are able to break up biofilms, while antibiotics administered in a standard manner (high, constant doses) cannot. According to , the use of pulsed, low-dose antibiotics to target biofilm bacteria is supported by observations she and her colleagues have made in the laboratory.[11]Some researchers claim that antibiotics cannot penetrate the matrix that surrounds a biofilm. But research by and other scientists has confirmed that the inability of antibiotics to penetrate the biofilm matrix is much more of an exception than a rule. According to , “In most cases involving small antimicrobial molecules, the barrier of the polysaccharide matrix should only postpone the death of cells rather than afford useful protection.â€For example, a recent study that used low concentrations of an antibiotic to killP. aeruginosa biofilm bacteria found that the majority of biofilm cells were effectively eliminated by antibiotics in a manner that did not differ much from what is observed when the same antibiotic concentrations are administered to single planktonic cells.[26]After antibiotics are applied to a biofilm, a number of cells called “persisters†are left behind.Thus, since antibiotics can generally penetrate biofilms, some other factor is responsible for the fact that they cannot be killed by standard high dose antibiotic therapy. It turns out that after antibiotics are applied to a biofilm, a number of cells called “persisters†are left behind. Persisters are simply cells that are able to survive the first onslaught of antibiotics, and if left unchecked, gradually allow the biofilm to form again. According to , persister cells form with particular ease in immunocompromised patients because the immune system is unable to help the antibiotic “mop up†all the biofilm cells it has targeted.“This simple observation suggests a new paradigm for explaining, at least in principle, the phenomenon of biofilm resistance to killing by a wide range of antimicrobials,†states . “The majority of cells in a biofilm are not necessarily more resistant to killing than planktonic cells and die rapidly when treated with [an antibiotic] that can kill slowly growing cells.â€Thus, a dose of antibiotics – particularly in immunocompromised patients – eradicates most of the biofilm population but leaves a small fraction of surviving persisters behind. Unfortunately, in the same sense that the beta-lactam antibiotics promote the formation of L-form bacteria, persister cells are actually preserved by the presence of an antibiotic that inhibits their growth. Thus, paradoxically, dosing an antibiotic in a constant, high-dose manner (in which the antibiotic is always present) helps persisters persevere.But in the case of low, pulsed dosing, where an antibiotic is administered, withdrawn, then administered again, the first application of antibiotic will eradicate the bulk of biofilm cells, leaving persister cells behind. Withdrawl of the antibiotic allows the persister population to start growing. Since administration of the antibiotic is temporarily stopped, the survival of persisters is not enhanced. This causes the persister cells to lose their phenotype (their shape and biochemical properties), meaning that they are unable to switch back into biofilm mode. A second application of the antibiotic should then completely eliminate the persister cells, which are still in planktonic mode. has found that the feasibility of a pulsed, or cyclical biofilm eradication approach depends on the rate at which persisters lose resistance to killing and regenerate new persisters. It also depends on the ability to manipulate the antibiotic concentration – something that is done quite effectively by patients on the Marshall Protocol who carefully dose their antibiotics at different levels, allowing constant variation in antibiotic concentration. Although speculates that allowing the concentration of an antibiotic to drop could potentially lead to resistance towards the antibiotic, she is quick to add that if two or more antibiotics are used to target a biofilm at one time, such resistance would not occur. Again, since the Marshall Protocol uses a total of five bacteriostatic antibiotics, usually taken two or three at a time, concerns of resistance are essentially negligible.Model of biofilm resistance based on persister survival. An initial treatment of high-dose constant antibiotic kills planktonic cells and the majority of biofilm cells. But persisters remain alive and resurrect the biofilm, causing the infection to relapse“It is entirely possible that successful cases of antimicrobial therapy of biofilm infections result from a fortuitous optimal cycling [pulsed dosing] of an antibiotic concentration that eliminated first the bulk of the biofilm and then the progeny of the persisters that began to divide,†states .’ work has been supported by other research teams. Recently, researchers at the University of Iowa found that subinhibitory (extremely low dose) concentrations of the bacteriostatic antibiotic azithromycin significantly decreased biomass and maximal thickness in both forming and established biofilms.[27] These extremely low concentrations of azithromycin inhibited biofilms in all but the most highly resistant isolates. In contrast, subinhibitory concentrations of gentamicin, which is not a bacteriostatic antibiotic, had no effect on biofilm formation. In fact, biofilms actually became resistant to gentamicin at concentrations far above the minimum inhibitory concentration.Researchers at Tulane University recently confirmed yet again that low, pulsed dosing is a superior way of targeting treatment-resistant biofilm bacteria. According to the team, who mathematically modeled the action of antibiotics on bacterial biofilms, “Exposing a biofilm to low concentration doses of an antimicrobial agent for longer time is more effective than short time dosing with high antimicrobial agent concentration.â€[28]Similarly, a bioengineer led team at the University of Washington recently created an antibiotic- containing polymer that releases antibiotic slowly onto the surface of hospital devices, such as catheters and prostheses, to reduce the risk of biofilm-related infections.“Rather than massively dosing the patient with high levels of released antibiotic, this strategy allows the release of extremely low levels of this very potent antibiotic over long periods of time,†explained Buddy Ratner, PhD, Professor and Director of the Engineered Biomaterials Program at the University of Washington, Seattle. “We calculated the amount released at the surface that would kill 100% of the bacteria entering the surface zone.â€When challenged by Dr. Leonard A. Mermel from Brown University School of Medicine on the issue that long-term use of pulsed, low-dose antibiotics might allow for increased resistance on the part of the bacteria being treated, Ratner responded, “Dr. Mermel’s concerns are, in fact, why we developed this system for [antibiotic] release. Bacteria that live through antibiotic dosing can go on to produce resistant strains. If 100% of the bacteria approaching the surface are killed, they can’t produce resistant offspring. The classical physician approach, dosing the patient systemically and heavily to rid the patient of persistent bacteria, can lead to those resistant strains. Our approach releases miniscule doses compared to what a physician would use, but releases the antibiotic where it will be optimally effective and least likely to leave antibiotic-resistant survivors.â€Although taken orally, the MP antibiotics are taken in the same manner as those administered by Ratner and team. Because they too are dosed at optimal times in extremely small doses, the chance that long-term antibiotic use might foster resistant bacteria is again, essentially negligible, especially when multiple antibiotics are typically used.Key to the ability of the Marshall Protocol to effectively target biofilm bacteria is the fact that the specific pulsed, low-dose bacteriostatic antibiotics used by the treatment are taken in conjunction with a medication called Benicar. Benicar binds and activates the Vitamin D Receptor, displacing bacterial substances and 25-D from the receptor, so that it can once again activate the innate immune system.[29] Benicar is so effective at strengthening the innate immune response that the patient’s own immune system ultimately helps destroy the biofilm weakened by pulsed, low-dose antibiotics.Thus, it is not enough for patients on the Marshall Protocol to simply take specific pulsed, low-dose antibiotics. The activity of their innate immune system must also be restored so that the cells of the immune system can actively combat biofilm bacteria, the matrix that surrounds them, and persister cells.After antibiotics are applied to a biofilm, a number of cells called “persisters†are left behindHow do we know that the Marshall Protocol effectively kills biofilm bacteria? Namely because those patients to reach the later stages of the treatment do not report symptoms associated with established biofilm diseases. Patients on the MP who once suffered from chronic ear infections (OM), chronic sinus infections, or periodontal disease find that such infections resolve over the course of treatment. Furthermore, since we now understand that biofilms almost certainly form a large part of the chronic microbiota of pathogens that cause chronic inflammatory and autoimmune diseases, the fact that patients can use the Marshall Protocol to recover from such illnesses again suggests that the treatment must be effectively allowing them to target and destroy biofilms.Because all evidence points to the fact that the MP does indeed effectively target biofilm bacteria, it is of utmost importance that people who suffer from any sort of biofilm infection start the treatment. Knowledge of the Marshall Protocol has yet to reach the cystic fibrosis community, but there is great hope that if people with the disease were to start the MP, they could destroy the P. aeruginosabiofilms that cause their untimely deaths. In the same vein, people with a wide range of infections, such as those infected with biofilm during surgery, can likely restore their health with the MP.It is to be hoped that the clinical data emerging from the Marshall Protocol study site, which shows patients recovering from biofilm-related diseases, will inspire future researchers to invest a great deal of energy into further research aimed at identifying and studying the biofilm bacteria – bacteria that almost certainly form part of the microbiota of pathogens that cause inflammatory disease. In the coming years, as the technology to detect biofilms becomes even more sophisticated, it is almost certain that a great number of biofilms will be officially detected and documented in patients with a vast array of chronic diseases.REFERENCESCosterton, J. W., , P. S., & Greenberg, E. P. (1999). Bacterial biofilms: a common cause of persistent infections. Science (New York, N.Y.), 284(5418), 1318-22. [↩] [↩] [↩] [↩]Higgins, D. A., Pomianek, M. E., Kraml, C. M., , R. K., Semmelhack, M. F., & Bassler, B. L. (2007). The major Vibrio cholerae autoinducer and its role in virulence factor production. Nature, 450(7171), 883-6. [↩]Singh, P. K., Schaefer, A. L., Parsek, M. R., Moninger, T. O., Welsh, M. J., & Greenberg, E. P. (2000). Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. Nature, 407(6805), 762-4. [↩] [↩]Stoodley, P., Purevdorj-Gage, B., & Costerton, J. W. (2005). Clinical significance of seeding dispersal in biofilms: a response. Microbiology, 151(11), 3453. [↩]O’toole, G. A., & Kolter, R. (1998). Flagellar and Twitching Motility Are Necessary for Pseudomonas Aeruginosa Biofilm Development. Molecular Microbiology, 30(2), 295-304. [↩]Cho, H., Jönsson, H., , K., Melke, P., , J. W., Jedynak, B., et al. (2007). Self-Organization in High-Density Bacterial Colonies: Efficient Crowd Control. PLoS Biology, 5(11), e302 EP -. [↩] [↩]Brockhurst, M. A., Hochberg, M. E., Bell, T., & Buckling, A. (2006). Character displacement promotes cooperation in bacterial biofilms. Current biology: CB, 16(20), 2030-4. [↩]Parsek, M. R., & Singh, P. K. (2003). Bacterial biofilms: an emerging link to disease pathogenesis. Annual review of microbiology, 57, 677-701. [↩]Kraigsley, A., Ronney, P., & Finkel, S. Hydrodynamic effects on biofilm formation. Retrieved May 28, 2008. [↩]Hall-Stoodley, L., Costerton, J. W., & Stoodley, P. (2004). Bacterial biofilms: from the Natural environment to infectious diseases. Nat Rev Micro, 2(2), 95-108. [↩] [↩] [↩], K. (2001). Riddle of biofilm resistance. Antimicrobial agents and chemotherapy, 45(4), 999-1007. [↩] [↩]Parsek, M. R., & Singh, P. K. (2003). Bacterial biofilms: an emerging link to disease pathogenesis. Annual review of microbiology, 57, 677-701. [↩]Trampuz, A., Piper, K. E., son, M. J., Hanssen, A. D., Unni, K. K., Osmon, D. R., et al. (2007). Sonication of Removed Hip and Knee Prostheses for Diagnosis of Infection. N Engl J Med, 357(7), 654-663. [↩]Ristow, P., Bourhy, P., Kerneis, S., Schmitt, C., Prevost, M., Lilenbaum, W., et al. (2008). Biofilm formation by saprophytic and pathogenic leptospires. Microbiology, 154(5), 1309-1317. [↩]Moreau-Marquis, S., Stanton, B. A., & O’Toole, G. A. (2008). Pseudomonas aeruginosa biofilm formation in the cystic fibrosis airway. Pulmonary pharmacology & therapeutics. [↩]Hall-Stoodley, L., Hu, F. Z., Gieseke, A., Nistico, L., Nguyen, D., , J., et al. (2006). Direct Detection of Bacterial Biofilms on the Middle-Ear Mucosa of Children With Chronic Otitis Media.JAMA, 296(2), 202-211. [↩]Imamura, Y., Chandra, J., Mukherjee, P. K., Lattif, A. A., Szczotka-Flynn, L. B., Pearlman, E., et al. (2008). Fusarium and Candida albicans Biofilms on Soft Contact Lenses: Model Development, Influence of Lens Type, and Susceptibility to Lens Care Solutions. Antimicrob. Agents Chemother., 52(1), 171-182. [↩], G. A., Swogger, E., Wolcott, R., Pulcini, E. D., Secor, P., Sestrich, J., et al. (2008).Biofilms in Chronic Wounds. Wound Repair and Regeneration, 16(1), 37-44. [↩]Marshall, T. G. (2006b). A New Approach to Treating Intraphagocytic CWD Bacterial Pathogens in Sarcoidosis, CFS, Lyme and other Inflammatory Diseases. [↩] [↩]Marshall, T. G., & Marshall, F. E. (2004). Sarcoidosis succumbs to antibiotics–implications for autoimmune disease. Autoimmunity reviews, 3(4), 295-300. [↩] [↩]Sr, G. J. D., & Woody, H. B. (1997). Bacterial persistence and expression of disease. Clinical Microbiology Reviews, 10(2). [↩]Marshall, T. G. (2007). Bacterial Capnine Blocks Transcription of Human Antimicrobial Peptides. Nature Precedings. [↩]on, H. I., Ellison, L. F., & , G. W. (1999). Periodontal disease and risk of fatal coronary heart and cerebrovascular diseases. Journal of cardiovascular risk, 6(1), 7-11. [↩], R., & Hirani, V. (2007). Dental Health and Cognitive Impairment in an English National Survey Population. Journal of the American Geriatrics Society, 55(9), 1410-1414. [↩]Falkinham Iii, J. O., Iseman, M. D., Haas, P. D., & Soolingen, D. V. (2008). Mycobacterium avium in a shower linked to pulmonary disease. Journal of water and health, 6(2), 209-13. [↩], K. (2001). Riddle of biofilm resistance. Antimicrobial agents and chemotherapy, 45(4), 999-1007. [↩]Starner, D et al. 2008. Subinhibitory Concentrations of Azithromycin Decrease Nontypeable Haemophilus influenzae Biofilm Formation and Diminish Established Biofilms.Antimicrobial agents and chemotherapy 52(1):137-45. [↩]Cogan, N. G., Cortez, R., & Fauci, L. (2005). Modeling physiological resistance in bacterial biofilms. Bulletin of mathematical biology, 67(4), 831-53. [↩]Marshall, T. G. (2006). VDR Nuclear Receptor Competence is the Key to Recovery from Chronic Inflammatory and Autoimmune Disease. [↩]Filed under: biofilms, featured articlesRSS feed for comments on this post37 Responses for "Understanding Biofilms"Ken Collerman May 29th, 2008 at 1:57 am1Amy,This article is a masterpiece!Amy Proal May 29th, 2008 at 9:40 am2Thanks Ken!It was a pleasure to write the article because I find the subject matter so fascinating!AmySherry June 2nd, 2008 at 4:54 pm3Is it possible to kill biofilms on our food by heating? I am having second thoughts about eating any raw vegetables…The article is well researched, well written, and very insightful.Thank you!Amy Proal June 2nd, 2008 at 5:00 pm4Hi Sherry,I’m glad you enjoyed the article.I’m not sure about the answer to your question. The foods items that seem to be most affected by biofilm contamination are raw, and can’t be heated in order to remove biofilms.I assume that if you had a food item and you heated it to high temperatures the high level of heat would have an effect on the homeostasis and potential survival of a biofilm, although I don’t know for sure.Are you on the MP? If so, you will almost certainly easily kill any biofilm bacteria that you might acquire by eating a raw vegetable before it is able to enter the bloodstream. So I wouldn’t worry about eating them.Don’t forget that until we reach the late stages of the MP, our own bodies are already filled with bacteria. They cover our hands and entire skin surface. They are already in our mouth. So coming in contact with biofilm bacteria in the mouth isn’t all that different from what we experience on a daily basis. Of course, being on the MP is key, so that an ingested biofilm will not be able to cause any harm.Best,Amy W. June 2nd, 2008 at 5:02 pm5I wonder if there’s a way to interfere with the biofilm’s chemical signaling directly. It would require much more specific diagnosis of the underlying pathogens, or else a good general cocktail.Do biofilm-creating pathogens just travel together or do they ‘learn’ how to signal one another based on shared plasmids? If the second, the plasmids themselves might be chemically attacked.Amy Proal June 2nd, 2008 at 5:30 pm6Hi ,Yes, there are research teams who are working to try to interfere with the chemical signaling in a biofilm. In fact, I think Higgins and team who are working with CAI-1 are actively seeking ways to inhibit the signal in an effort to slow biofilm growth. If they succeed, andother researchers successfully stop other biofilm signals, potential drugs that inhibit the signals they identify could form part of a larger arsenal to target biofilms. Of course, with any drug comes side effects, so people would have to weigh the pro/cons of taking drugs to interfere with biofilm signals, particularly when they can already target biofilms with just a few pulsed, low-dose antibiotics.I imagine that groups of people with specific biofilm infections (such as cystic fibrosis patients with P. auruginosa infections), would most benefit from therapies aimed at stopping chemical signals that aid the formation of biofilms.I think that the bacteria inside biofilms communicate in myriad ways, depending largely on the species present. So a lot of individual work with particular biofilms must be done to uncover ways to break up signaling pathways.I’m not sure biofilm communication has much to do with sharing plasmids. Biofilm bacteria literally emit chemical signals (not plasmids) into the surrounding environment. These signals are picked up by other nearby bacteria. Depending on the strength of the signal they react indifferent ways. For example, if a bacterium picks up a lot of other signals, it knows it is surrounded by a lot of other bacteria. That way it is able to “realize†that forming a biofilm with the nearby bacteria is a possibility. Other modes of signaling remain less clear as far as I can tell. I’m sure further research will turn up much more complex modes of action.Best, W. June 3rd, 2008 at 1:09 am7Hi Amy, you wrote;I’m not sure biofilm communication has to do with sharing plasmids. They literally emit chemical signals (not plasmids) into the surrounding environment.My point was that the bacteria had to have a common ‘language’ in order to communicate and I wondered if the key for that language was found in their chromosome or in a transmissable, extra-chromosomal unit (Kind of like giving a person a gene which lets them speak English so that you can talk to them.)There was some possibility for developing drugs that targeted plasmids, so I thought that that might be a good way to break up a biofilm.Amy Proal June 3rd, 2008 at 9:01 am8Hi ,I see. It possible that some genes are embedded within plasmids and others within the genome. We’re dealing with a lot of different genes here so they may be in diverse locations. Hopefully future researchers projects will address this issue….Dr.Abbas Ubaid Al_Janabi June 16th, 2008 at 8:29 am9Thanks for this best artical about the biofilms,I am finsed my Ph.D.Microbial Biofilms and I havemany papears in this field ,So, Plz keep attach with me amy.All the best YoursDr. Abbas Al_JanabiCollege of Medicine, University of Al_Anbar,Ramadi,IraqAmy Proal June 16th, 2008 at 10:05 am10Hi Dr. Abbas,I’m glad to hear that a biofilm expert enjoyed my article. Congrats on getting your PhD in biofilm-related research. I think you are pursuing a very interesting course of study that will be extremely relevant to the future of medicine.Do you have a link to any of your published papers? I’d enjoy looking over them…Best,AmyBeth August 19th, 2008 at 11:38 pm11AmyWe are doing a Nursing Grand Rounds which includes a segment on oral care. I would like to use some of your pictures. Can I have permission to do so?Thank you for your consideration.BethAmy Proal August 20th, 2008 at 7:50 am12Hi Beth,Many of these images are from the Center for Biofilm Engineering in Montana. One of the missions of CBE is to promote the understanding of biofilms. The Center has a page on its website explaining acceptable reuse.Best,Amy Rifkin, MD September 8th, 2008 at 5:22 pm13Amy: Terrific article. I’ve been asked to give a talk on biofilms for our new Masters in Biotechnology course and I’d love to be able to use some of your slides (proper acknowledgement of course). Would that be possible. Thanks. GRAmy Proal September 9th, 2008 at 1:54 am14Hi ,Of course, you are welcome to use my content for the purposes of a presentation. Please note, as per comment #12, that when it comes to the images, you’re going to want to attribute the CBE in Montana.If you haven’t done so already, you may also want to look at my 87-minute videoIntroduction to the Marshall Protocol.Best,AmyAnusha December 2nd, 2008 at 7:12 pm15Hi Amy,Thank you for this detailed report on biofilms. I am suspicious of biofilm contamination in my airway epithelial cell culture. I see a network of floating dead/dying epithelial cells on the top layer of cell culture medium and a translucent jelly-like strands that dont seem to be fungi. The culture is an established immortalized cell line derived from a patient’s lung biopsy. Is there any way to test for persistant contamination? Any stain or culture technique to detect presence of biofilms?Thank you!Amy Proal December 3rd, 2008 at 1:05 pm16Hi Anusha,There are tests that detect biofilms but they are not necessarily available in a standard lab. In order to test you samples, I think you would have to contact a researcher working with biofilms and ask them for help and guidance.You could start by contacting the Center for Biofilm Engineering at Montana State University. It’s likely that someone on their staff could tell you more about biofilm testing and who to contact to do the research.Personally, I think there’s a great group of biofilm researchers at the University of Washington. Drs. Parsek, Singh, Harwood and others. You could look them up and send them an email with your question. I’m not sure if they would respond but they might give you feedback.Best,AmyShari Gold April 14th, 2009 at 5:44 am17Hi Amy,I just re-read this article. It is such a great, well researched article, written in a very direct and easy to understand manner. I had a few quick questions. I am familiar with biofilms contaminating artificial surfaces within the human body, but wondered if biofilms colonize regular joints, not altered by artificial replacements. In particular, shoulder and hips, wrists and fingers – could this be what is going on in something like RA too? Also, what about your nerves – say, peripheral nerves? Is this something they might colonize as well? Reason I ask is I do have peripheral neuropathy in my back. Neuropathy is something I know that resolves on the MP – although it is something that takes a bit longer. Could this be because much of what might be affecting that signal transduction in the nerve is stopped by biofilm inflammation somewhere from the nerve root to the muscle? In me, my neuropathy is getting much better and I can definitely see the waxing and waning of symptoms as I increase abx and ramp different abx combos. Based on what I just re-read tonight, that might seem a plausible explanation for the physical phenomenon I am experiencing – killing a bunch of bacteria, one layer at a time. Also, as noted in the patients w/ cystic fibrosis or chronic sinus infected patients, as I have gotten further into phase 2, I have had a lot of sinus drainage and much nasty stuff coming up from lower respiratory, like I have a constant cold, but it isn’t “infection†in the clinical way, just sputum breaking up constantly, might this be evidence of biofilm existence in these places? This is truly a fascinating area of study.Thanks for you insight.Best,Shari Albert April 14th, 2009 at 8:48 am18Hi Shari,Amy is away, so you’re stuck with me. : )I don’t think there’s anything unique about non-human surfaces that leads to formation of biofilm. You may be shocked to learn that most people manually remove biofilm from themselves every day, sometimes twice a day. This odd ritual? Teeth brushing! Teeth brushing is nothing more than the act of manually removing biofilm from the surface of one’s teeth.But, it would be very un-bascterialike for bacteria to confine themselves to prosthetic hip joints and teeth. If you think about it, the only reason we don’t brush every other part of our body is that we can’t – regrettably, we cannot brush our kidney, liver, nerves, lungs, etc.Do biofilm colonize nerves? They have had hundreds of thousands of years to figure it out. I would say absolutely. One of the limiting factors as far as progress on the MP is concerned is blood flow to a region. I remember hearing that nerve cells don’t get as much blood as other areas. That’s not to the say that neuropathy doesn’t resolve, only that the process may take longer. However, the waxing and waning of your neuropathy symptoms with your antibiotics supports the idea that these symptoms will resolve.As for your theory on signal transduction, I’m sure it’s involved, but I’m afraid I couldn’t say how.Also, I think you’re on target when it comes to sinus drainage – bacteria are involved – although I’m not sure we have any good idea on the relative proportion of different forms of bacteria by region of the body: L-forms, intracellular pathogens, biofilm, etc.Best, May 13th, 2009 at 4:49 pm19Hi Amy,thanks for the article and research… In your research, did you happen to find any effective alternative medicines for removal of biofilms? or energy medicine? Ondamed has been used successfully now in Europe for 15+ yrs in chronic lyme….Amy Proal May 15th, 2009 at 11:38 am20Hi ,Unfortunately I do not know of any substance that can target internal biofilms. External biofilms – or those that grow outside the body – are a different story. Those can be treated with several chemicals, however they are chemicals that would be toxic if swallowed.Actually, some studies show that a substance called Lactoferrin can reduce biofilm growth. However I believe more research is needed to confirm its effects on biofilm proliferation and to make sure it doesn’t interfere with other pathways in the body when taken.Does Ondamed cause patients to experience a bacterial die-off reaction? I ask, because if it doesn’t cause an exacerbation in disease symptoms then it’s not causing bacteria to be killed. We know this because a rise in symptoms generated by bacterial death cannot be avoided when the MP medications begin to stimulate the immune response to target biofilm species.If Ondamed makes patients feel better instead of worse, it is probably just a palliative treatment option. What it may actually do is slow the immune response. While this would lower the inflammation associated with bacterial death it wouldn’t actually kill biofilm bacteria and would be ineffective in the long run. Clearly Lyme is still a big problem in Europe so unfortunately I suspect that Ondamed falls into this latter category.I have not researched energy medicine and biofilms enough to comment on whether it would help with their elimination. What I can say is that the MP seems quite effective at targeting biofilm species! Essentially 100% of patients to start the treatment have experienced immunopathology or bacterial die-off and these reactions remain strong and steady during much of treatment. Then, during later stages of treatment, we have patients reporting improvement from many known biofilm-related infections. So I would say the MP is certainly your best bet if you are trying to target biofilm. I encourage you not to worry about taking the MP medications as they have an excellent safety profiles and are likely safer than taking herbs or other “natural†supplements whose properties have not been investigated at the molecular level.This article discusses the safety of the MP meds:http://bacteriality.com/2008/02/23/misconceptions/#2Hope this helps!Amy W. May 15th, 2009 at 12:17 pm21 – Xylitol and Serrapeptase both seem pretty safe and have activity against biofilms, though I’m not sure how long xylitol lasts or how well it works inside the body. Xylitol is a sugar alcohol used as a sweetner and in some toothpastes because of its flavor and effect on oral biofilms. Serrapeptase is an anti-inflammatory and blood thinner used routinely in Germany as an aspirin substitute since it doesn’t interfere with K1 (it doesn’t promote heart disease via arterial calcification like Aspirin does) or seem to cause the liver problems that salyciliates like Aspirin do. Both seem very safe in proper dosages. It may be hard to tell if Serrapeptase caused a bacterial dieoff reaction since it’s also an anti-inflammatory but it seems like one of the more promising candidates for attacking internal biofilms.Amy Proal May 15th, 2009 at 12:57 pm22Hi ,Yes, Xylitol has been proposed to curb biofilm growth. It’s in my toothpaste and mouthwash (I use a brand of tooth products called Biotene which is designed to target biofilm bacteria). It’s also some chewing gum. However I have not found any studies that show it’s effective at targeting biofilm other than those on the teeth and I’m not really sure if it even works as intended.When I googled Serapeptase, I read that “Serrapeptase is an enzyme that is produced in the intestines of silk worms to break down cocoon walls. This enzyme is proving to be an alternative to the non-steroidal anti-inflammatory agents (NSAIDs) traditionally used to treat rheumatoid arthritis and osteoarthritis.â€NSAIDs are completely contraindicated for use for MP patients because they palliate inflammation rather than actually killing the bacteria causing inflammation. So that supplement seems suspect to me and I would certainly never take it.I cannot emphasize enough the risk of taking supplements when most of the claims about how they work are just based on speculation and not on molecular data. I took way too many supplements before the MP which were supposed to kill bacteria and they did nothing but make me more ill.Best,Amy W. May 15th, 2009 at 1:35 pm23When I googled SerapeptaseIf you’re looking for hard data, why not use pubmed?Serrapeptase has the demonstrated capacity to increase antibiotic effectiveness against biofilm forming bacteria. It’s demonstratablynot simply palliative. I understand that it’s contraindicated under the Marshall protocol, but there’s still very good evidence it’s helpful and well tolerated and doesn’t just mask symptoms.linkHere’s a study showing injected serrapeptase + antibiotics is significantly more effective at clearing biofilm producing infections than antibiotics alone.We have chosen serratiopeptidase (SPEP), an extracellular metalloprotease produced by Serratia marcescens that is already widely used as an anti-inflammatory agent, and has been shown to modulate adhesin expression and to induce antibiotic sensitivity in other bacteria. Treatment of L. monocytogenes with sublethal concentrations of SPEP reduced their ability to form biofilms and to invade host cells. Zymograms of the treated cells revealed that Ami4b autolysin, internalinB, and ActA were sharply reduced. These cell-surface proteins are known to function as ligands in the interaction between these bacteria and their host cells, and our data suggest that treatment with this natural enzyme may provide a useful tool in the prevention of the initial adhesion of L. monocytogenes to the human gut.http://www.ncbi.nlm.nih.gov/pubmed/18479885?ordinalpos=1 & itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSumAntibiotic susceptibility tests on both planktonic and sessile cultures, studies on the dynamics of colonization of 10 biofilm-forming isolates, and then bioluminescence and scanning electron microscopy under seven different experimental conditions showed that serratiopeptidase greatly enhances the activity of ofloxacin on sessile cultures and can inhibit biofilm formation.http://www.ncbi.nlm.nih.gov/pubmed/8109925?ordinalpos=3 & itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSumAmy Proal May 16th, 2009 at 12:36 pm24,Thank you, I know how to use PubMed. The reason I didn’t start searching PubMed for info about Serapeptase is because it’s not a topic relevant to what I’m spending my energy on at the moment. On this site, I try to explain the MP in simple terms – the MP as it is created and the molecular data we have about how the medications interact. I already have many comments from people asking about these topics and I need/want to answer their basic questions first before I speculate about a supplement with you.You know yourself that the MP has a no supplement policy, and for a reason. If you actually started the MP, you’d realize that the combination of Olmesartan and low dose antibiotics is so powerful that our main problem is certainly not whether people are targeting enough bacteria. Our main problem is how to control the die-off reaction and keep people from killing so many bacteria in a day that they can’t tolerate the die-off symptoms. So at this point we don’t need any extra supplements to target biofilm bacteria.That’s especially true since we don’t know how serapeptase works at the molecular level. Maybe it does hinder biofilm formation somewhat but what’s to say it doesn’t interfere with any of the receptors involved in the immune response? How do we know that it wouldn’t interfere with the ability of Olmesartan to bind the VDR? We don’t know at this point and so I worry that when you mention these things people who I really believe should try the MP are going to skip the treatment and think that a supplement can help them instead – which is exactly what I don’t want them to think.Best,AmyL. Lange June 3rd, 2009 at 9:59 pm25Hi Amy –I was reading your article and enjoyed it a lot, thank you! Do you know anything about Lymes disease and if the bacteria are protected by the the concepts of biofilms? If yes, do you think that the Marshal protocol would help someone recover from the disease?Thanks in advance for your reply!-Amy Proal June 7th, 2009 at 10:05 pm26Hi ,Sorry for the delayed response. I just got back from a trip to China.Yes, it is very likely that many of the bacteria that cause Lyme disease persist in a biofilm state and there are definitely many people on the MP with Lyme disease who are recovering.Here are interviews with three of them:http://bacteriality.com/2007/09/22/interview3/http://bacteriality.com/2007/12/28/interview14/http://bacteriality.com/2008/03/31/interview19/You might also want to post on the website http://www.curemyth1.org (Th1 refers to diseases caused by bacteria, hence the name). The site is a place where people can ask questions about the MP. But once you are a member of the site, you can also communicate with other people on the MP for Lyme.If you have Lyme, I definitely hope that you start the MP. The best way to learn more about the treatment is to read as much as possible, on this site and also on the MP study site – http://www.marshallprotocol.com.I also highly recommend watching the following video which describes the MP and the science that forms its backbone in simple terms:http://bacteriality.com/2008/05/07/mpintro/Best,Amy June 7th, 2009 at 10:15 pm27Hey amy,thanks for the info:) As far as I know, ondamed therapy it is able to approach micro-organisims at a molecular level using frequencies…there’s been a ton of research to back up its effectiveness and YES you have to be careful with how much time you use b/c of the die off effect! The first time i used this therapy I did only 10 mins and was in bed for about a week! I will do some more research on the MP protocol… i hope u have a blessed week! p.s thanks ryan for the info too:)Amy Proal June 8th, 2009 at 11:09 am28Hi ,Thanks for writing. I know nothing about Ondamed. Are the papers you mention about it’s efficacy peer-reviewed? If so, I’d be interested in reading one.In the meantime, I’d like to emphasize that I wouldn’t definitely not recommend using Ondamed or any other frequency therapy or supplement along with the MP. As I mentioned to before, there is a careful balance of the immune system set up in the body by the MP meds as they target bacteria, and other therapies may interfere. Also, if any therapy causes extra bacterial die-off it could lead to intolerable immunopathology when combined with the MP meds so one must be careful.I definitely hope you look into the MP !Best,Amy June 22nd, 2009 at 1:33 pm29Amy,How do I find a doctor who will use the Marshall Protocol? I have Interstitial Cystitis and tested positive for Strep D, only after a week long broth culture. Based on what I have read, I think that the bacteria has formed a biofilm, which is why shorter cultures aren’t picking it up. My doctor doesn’t know a whole lot about biofilm, so I am trying to find a doctor who does, so that I can get treatment.Thanks in advance for any help!Amy Proal June 24th, 2009 at 12:01 pm30Hi ,I’m sorry about your illness and positive strep cultures. In my opinion you are correct – it is very likely that many of the bacteria making you ill are in a biofilm state and cannot be kill by conventional therapies. On the other hand, the MP appears to effectively break up biofilms and I think you would definitely benefit from the treatment.I’m not sure where you live. However the best way to find a doctor in your area is to post at the following website:http://www.curemyth1.org (Th1 refers to diseases caused by bacteria, hence the name). The patient advocates on the site, who are volunteers, can likely provide you with a list of doctors that administer the MP in your area.If that doesn’t work out I recommend showing your current doctor or a new open-minded doctor in your area Dr. Marshall and team’s latest peer-reviewed papers and presentations. See if then you can convince them to prescribe you the necessary MP meds. The publications etc. can be found here:http://mpkb.org/doku.php#publications_presentationsThis article also give more tips on finding an MP doctor.http://mpkb.org/doku.php/home:starting:physician:findingGood luck and take care!AmyJD Bear August 8th, 2009 at 11:06 am31Hello Amy,I have spent some time perusing your website and I complement you on your good work. As such, it is all the more important to not mar this good work by using inappropriate terminology. Specifically, the term “biofilm†refers to a community of bacteria surrounded by *Extracellular* Polymeric Substances. A tubule, or filament, or “protective sheath†formed by an L-form bacteria may be a “film†and it is no doubt biological, but it is NOT a “biofilm†as commonly defined. It is a disservice to conflate these two different phenomena. Whatever is going on with L-form bacteria deserves its own terminology so as not to introduce unnecessary confusion into the field.Best Regards,-JDBAmy Proal August 8th, 2009 at 12:09 pm32Hi JDB,Thanks for your comment. I’m not sure exactly what statement I’ve made that you are referring to. Nevertheless, I don’t see why L-form bacteria could not be able to form a biofilm. Bacteria in the L-form are simply in a different part of the microbial lifecycle and have temporarily lost their cell walls. Is a cell wall necessary to become part of biofilm community? I have not seen research along those lines.This being said why wouldn’t L-form bacteria also form into communities in order to better protect themselves from the immune response etc? How do we know that a biofilm cannot be composed of both bacteria with cell walls and bacteria without cell walls?So I don’t really think we need to come up with completely different terminology to describe bacteria in the L-form seeing as their characteristics are generally very similar to those of their walled counterparts and they interact with classical forms of bacteria very closely in vivo.Best,Amy August 26th, 2009 at 7:02 am33Dear Amy,Thanks for this article. And I hope what you do next is to study Borrelia and biofilms. Leptospira and Treponema denticola are known to be able to persist inside biofilms. But there has not been published anything on Borrelia yet. No major journal has anything on it. It would meen so much to so many if you’d get that out…Amy Proal August 28th, 2009 at 11:45 pm34Hi ,Thank you for your interest in our research. Borrelia is definitely a very hardy pathogen that we would expect would be able to survive in biofilms. Over the course of the next few years, Autoimmunity Research Foundation, which is the organization I work with, is hoping to run in which we will look at the DNA of the infected cells in patients with chronic disease. This study is still in the planning stages, but, in time, could eventually better help us understand how Borrelia survive in the body and in a biofilm-like environment.Meanwhile, there is research suggesting that the low pulsed manner with which antibiotics are administered is much more effective at targeting bacteria in biofilms than standard regular doses of antibiotics or antibiotics taken in an IV form. Our own clinical data also shows people reporting improvement and recovery from Lyme disease thanks to therapy with the Marshall Protocol. So, if you think you may harbor Borrelia, and you want to target Borrelia in the biofilm state, looking into the MP may be a worthwhile option.Best,Amygregg zulauf September 12th, 2009 at 4:16 pm35I was impressed by the article in ‘Discover Magazine’ about biofilms (link below). Anecdotal evidence of xylitol breaking down certain biofilm structures was interesting to me.This article suggests that xylitol may work internally and not only in the mouth. I am interested in your reaction to this article.Regards, Gregghttp://discovermagazine.com/2009/jul-aug/17-slime-city-germs-talk-each-other-plan-attacks/article_view?b_start:int=2 & -C= Albert September 14th, 2009 at 12:47 pm36Hi Gregg,Interesting. One of the principles in that article is Randall Wolcott. If you do a Google search for Randall Wolcott, the #1 hit is Amy’s interview with him:http://bacteriality.com/2008/04/13/wolcott/So, yes indeed we have heard about Xylitol before.However, the problem with Marshall Protocol patients isn’t generating bacterial die-off but controlling the reaction. That said, it may be worth giving those few patients who are non-responders xylitol and seeing if that can lead to bacterial killing. This might be an avenue we will research in time….Best, September 30th, 2009 at 8:57 pm37I’ve been reading that EDTA has been used to erradicate Biofilm??Also, some claim Noni and Grapefruit seed extract works….any comments?NEWS FLASHApril 4, 2009:, Milk consumption tied to Parkinson’s diseaseMarch 21, 2009:, Hey there, how’s your Kineosphaeram holding up?Peer-Reviewed PapersAutoimmune disease in the era of the metagenome (PDF)Vitamin D: the alternative hypothesis (PDF)Dysregulation of the Vitamin D Nuclear Receptor may contribute to the higher prevalence of some autoimmune diseases in women (PDF)Vitamin D metabolites as clinical markers in autoimmune and chronic disease (PDF)Amy's Conference PresentationsMetagenomic symbiosis between bacterial and viral pathogens in autoimmune diseaseNotes from the 2009 International Congress of AntibodiesNotes from the 2008 International Congress on AutoimmunityFeatured ArticlesUpdate on tone and other issuesWhy patients with chronic disease are disaffected and how online social networks meet their needsSun-blocking culture among the ChineseSecond-guessing the consensus on vitamin DTravels, papers, and more… an updateThree days at the J. Craig Venter InstituteThe bacteria boom – implications of the Human Microbiome ProjectUnderstanding BiofilmsInterview with Dr. Randall Wolcott, bacterial biofilm wound specialistInsights into horizontal gene transfer: conversations with Dr. Gogarten and Dr. LakeVoices of reason in the vitamin D debateInterview with evolutionary biologist EwaldWhat can medical research learn from the open source software movement?Interview with Dr. Greg Blaney: MP physicianBacteria and cancer: an interview with Dr. Alan CantwellInterview with Nadya Markova: L-form ExpertGerald Domingue: Pioneer of Atypical BacteriaA History of Cell Wall Deficient Bacteria: A Selection of Researchers Who Have Worked with the L-formPatient InterviewsInterview with Gene – sarcoidosis, bladder cancerInterview with Bonnie B – lupus, Sjogren’s SyndromeInterview with Eastlund – diabetes, sarcoidosis, irritable bowel syndromeInterview with Roy P. – sarcoidosis, rheumatoid arthritisInterview with de Jager: chronic fatigue syndrome, multiple chemical sensitivityInterview with Ken L. – Post Treatment Lyme Disease Syndrome (PTLDS)Interview with Doreen V. – autism, ADHD depression, severe anxiety, CFSInterview with Jane -Aoki: Neurosarcoidosis, systemic sarcoidosis; spasticity, myasthenia, CNS dysfunction, joint pain, pulmonary, splenic and cardiac involvement.Interview with Melinda Stiles – Lyme, Irritable bowel syndrome/colitis, Radiculitis (inflammation of the nerve roots)Interview with Freddie Ash – Sarcoidosis of the heart, coronary artery disease, atrial fibrillationInterview with P. Bear R.N. – Chronic Borreliosis (“Lymeâ€), MCS (multiple chemical sensitivities), Chronic Spinal Inflammation, Peripheral NeuropathyInterview with Sherry Cook – Sarcoidosis, Cat Scratch Fever, Restless Leg SyndromeInterview with Leesa Shanahan – Sarcoidosis (Heerfordt’s Syndrome), UveitisInterview with Mirek Wozga – sarcoidosisInterview with Albert – CFS, depression, food sensitivitiesInterview with Carole – Sarcoidosis, fibromyalgia, CFSInterview with Shirley J. (Saj) – SarcoidosisInterview with Robyn – Lyme, myoclonusInterview with Sue Andorn – Lyme, BabesiaInterview with Ival Meyer – Arthritis, dyslexiaInterview with Guss Wilkinson – Sarcoidosis, psoriasis, insomniaAbout Amy ProalAmy Proal graduated from town University in 2005 with a degree in biology. While at town, she wrote her senior thesis on Chronic Fatigue Syndrome and the Marshall Protocol.Amy has spoken at several international conferences and authored several peer-reviewed papers on the intersection of bacteria and chronic disease.If you have questions about the MP, please visitCureMyTh1.org where volunteer patient advocates will answer your questions. Another good resource is the MP Knowledge Base, which is scheduled to be completed within the next year.Categories Select Category aging (1) biofilms (3) cancer (2) cardiovascular disease (1) cognitive dysfunction (3) conferences and trainings (7) diet (4) familial aggregation (3) featured articles (18) history (3) horizontal gene transfer (2) interview (doctor/researcher) (5) interview (patient) (21) L-form bacteria (6) marshall protocol (21) medical research (5) mental illness (2) microbiome (3) News Flash (38) obesity (2) personal (2) presentations (3) statins (1) Uncategorized (1) videos (4) vitamin d (12) RSS FeedsPostsCommentsCopyright © 2009 - Bacteriality — Exploring Chronic DiseaseThis site uses a modified version of Illacrimo Theme created by Design DiseaseFrom: Goldstein@...To: bird mites Sent: Friday, September 9, 2011 8:28:49 AMSubject: Re: Good summary about biofilms

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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http://www.youtube.com/watch?v=AmvgOfIN_8cIf you have time, watch this doctor/researcher talking about her own experience with Lyme disease; she did cancer research and now does Lyme disease research, particularly into biofilms. Dr. Eva Sapi--From: Goldstein@...To: bird mites Sent: Friday, September 9, 2011 8:32:43 AMSubject: Re: Good summary about biofilms

Understanding BiofilmsAuthor: Amy Proal26MAY2008As humans, our environment consistently exposes us to a variety of dangers. Tornadoes, lightning, flooding and hurricanes can all hamper our survival. Not to mention the fact that most of us can encounter swerving cars or ill-intentioned people at any given moment.Biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of materialThousands of years ago, humans realized that they could better survive a dangerous world if they formed into communities, particularly communities consisting of people with different talents. They realized that a community is far more likely to survive through division of labor– one person makes food, another gathers resources, still another protects the community against invaders. Working together in this manner requires communication and cooperation.Inhabitants of a community live in close proximity and create various forms of shelter in order to protect themselves from external threats. We build houses that protect our families and larger buildings that protect the entire community. Grouping together inside places of shelter is a logical way to enhance survival.With the above in mind, it should come as no surprise that the pathogens we harbor are seldom found as single entities. Although the pathogens that cause acute infection are generally free-floating bacteria – also referred to as planktonic bacteria – those chronic bacterial forms that stick around for decades long ago evolved ways to join together into communities. Why? Because by doing so, they are better able to combat the cells of our immune system bent upon destroying them.It turns out that a vast number of the pathogens we harbor are grouped into communities called biofilms. In an article titled “Bacterial Biofilms: A Common Cause of Persistent Infections,†JW Costerton at the Center for Biofilm Engineering in Montana defines a bacterial biofilm as “a structured community of bacterial cells enclosed in a self-produced polymeric matrix and adherent to an inert or living surface.â€[1] In layman’s terms, that means that bacteria can join together on essentially any surface and start to form a protective matrix around their group. The matrix is made of polymers – substances composed of molecules with repeating structural units that are connected by chemical bonds.According to the Center for Biofilm Engineering at Montana State University, biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of material – such as metals, plastics, soil particles, medical implant materials and, most significantly, human or animal tissue. The first bacterial colonists to adhere to a surface initially do so by inducing weak, reversible bonds called van der Waals forces. If the colonists are not immediately separated from the surface, they can anchor themselves more permanently using cell adhesion molecules, proteins on their surfaces that bind other cells in a process called cell adhesion.A biofilm in the gut.These bacterial pioneers facilitate the arrival of other pathogens by providing more diverse adhesion sites. They also begin to build the matrix that holds the biofilm together. If there are species that are unable to attach to a surface on their own, they are often able to anchor themselves to the matrix or directly to earlier colonists.During colonization, things start to get interesting. Multiple studies have shown that during the time a biofilm is being created, the pathogens inside it can communicate with each other thanks to a phenomenon called quorum sensing. Although the mechanisms behind quorum sensing are not fully understood, the phenomenon allows a single-celled bacterium to perceive how many other bacteria are in close proximity. If a bacterium can sense that it is surrounded by a dense population of other pathogens, it is more inclined to join them and contribute to the formation of a biofilm.Bacteria that engage in quorum sensing communicate their presence by emitting chemical messages that their fellow infectious agents are able to recognize. When the messages grow strong enough, the bacteria respond en masse, behaving as a group. Quorum sensing can occur within a single bacterial species as well as between diverse species, and can regulate a host of different processes, essentially serving as a simple communication network. A variety of different molecules can be used as signals.“Disease-causing bacteria talk to each other with a chemical vocabulary,†says Doug Hibbins of Princeton University. A graduate student in the lab of Princeton University microbiologist Dr. Bonnie Bassler, Hibbins was part of a research effort which shed light on how the bacteria that cause cholera form biofilms and communicate via quorum sensing.[2]“Forming a biofilm is one of the crucial steps in cholera’s progression,†states Bassler. “They [bacteria] cover themselves in a sort of goop that’s a shield against antibiotics, allowing them to grow rapidly. When they sense there are enough of them, they try to leave the body.â€Although cholera bacteria use the intestines as a breeding ground, after enough biofilms have formed, planktonic bacteria inside the biofilm seek to leave the body in order to infect a new host. It didn’t take long for Bassler and team to realize that the bacteria inside cholera biofilms must signal each other in order to communicate that it’s time for the colony to stop reproducing and focus instead on leaving the body.“We generically understood that bacteria talk to each other with quorum sensing, but we didn’t know the specific chemical words that cholera uses,†Bassler said.Then Higgins isolated the CAI-1 – a chemical which occurs naturally in cholera. Another graduate student figured out how to make the molecule in the laboratory. By moderating the level of CAI-1 in contact with cholera bacteria, Higgins was successfully able to chemically control cholera’s behavior in lab tests. His team eventually confirmed that when CAI-1 is absent, cholera bacteria attach in biofilms to their current host. But when the bacteria detect enough of the chemical, they stop making biofilms and releasing toxins, perceiving that it is time to leave the body instead. Thus, CAI-1 may very well be the single molecule that allow the bacteria inside a cholera biofilm to communicate. Although it is likely that the bacteria in a cholera biofilm may communicate with other signals besides CAI-1, the study is a good example of the fact that signaling molecules serve a key role in determining the state of a biofilm.Sessile cells in a biofilm “talk†to each other via quorum sensing to build microcolonies and to keep water channels open.Similarly, researchers at the University of Iowa (several of whom are now at the University of Washington) have spent the last decade identifying the molecules that allow the bacterial species P. aeruginosa to form biofilms in the lungs of patients with cystic fibrosis.[3] Although the P. auruginosa isolated from the lungs of patients with cystic fibrosis looks like a biofilm and acts like a biofilm, up until recently, there were no objective tests available to confirm that the bacterial species did indeed form biofilms in the lungs of patients with the disease, nor was there a way to tell what proportion of P. aeruginosa in the lungs were actually in biofilm mode.“We needed a way to show that the P. auruginosa in cystic fibrosis lungs was communicating like a biofilm. That could tell us about the P. auruginosalifestyle,†said Pradeep Singh, M.D., a lead author on the study who is now at the University of Washington.Singh and his colleagues finally discovered that P. aeruginosa uses one of two particular quorum-sensing molecules to initiate the formation of biofilms. In November 1999, his research team screened the entire bacterial genome, identifying 39 genes that are strongly controlled by the quorum-sensing system.In a 2000 study published in Nature, Singh and colleagues developed a sensitive test which shows P. auruginosa from cystic fibrosis lungs produces the telltale, quorum-sensing molecules that are the signals for biofilm formation.[3]It turns out that P. aerugnosa secretes two signaling molecules, one that is long, and another that is short. Using the new test, the team was able to show that planktonic forms of P. aeruginosa produce more long signaling molecules. Alternately, when they tested the P. aeruginosa strains isolated from the lungs of patients with cystic fibrosis (which were in biofilm form), all of the strains produced the signaling molecules, but in the opposite ratio – more short than long.Interestingly, when the biofilm strains of P. aeruginosa were separated in broth into individual bacterial forms, they reverted to producing more long signal molecules than short ones. Does this mean that a change in signaling molecular length can indicate whether bacteria remain as planktonic forms or develop into biofilms?To find out, the team took the bacteria from the broth and made them grow as a biofilm again. Sure enough, those strains of bacteria in biofilm form produced more short signal molecules than long.“The fact that the P. aeruginosa in [the lungs of cystic fibrosis patients] is making the signals in the ratios that we see tells us that there is a biofilm and that most of the P. aeruginosa in the lung is in the biofilm state,†states Greenberg, another member of the research team. He believes that the findings allow for a clear biochemical definition of whether bacteria are in a biofilm. Techniques similar to those used by his group will likely be used to determine the properties of other biofilm signaling molecules.DevelopmentOnce colonization has begun, the biofilm grows through a combination of cell division and recruitment. The final stage of biofilm formation is known as development and is the stage in which the biofilm is established and may only change in shape and size. This development of a biofilm allows for the cells inside to become more resistant to antibiotics administered in a standard fashion. In fact, depending on the organism and type of antimicrobial and experimental system, biofilm bacteria can be up to a thousand times more resistant to antimicrobial stress than free-swimming bacteria of the same species.Biofilms grow slowly, in diverse locations, and biofilm infections are often slow to produce overt symptoms. However, biofilm bacteria can move in numerous ways that allow them to easily infect new tissues. Biofilms may move collectively, by rippling or rolling across the surface, or by detaching in clumps. Sometimes, in a dispersal strategy referred to as “swarming/seedingâ€, a biofilm colony differentiates to form an outer “wall†of stationary bacteria, while the inner region of the biofilm “liquefiesâ€, allowing planktonic cells to “swim†out of the biofilm and leave behind a hollow mound.[4]Biofilm bacteria can move in numerous ways: Collectively, by rippling or rolling across the surface, or by detaching in clumps. Individually, through a “swarming and seeding†dispersal.Research on the molecular and genetic basis of biofilm development has made it clear that when cells switch from planktonic to community mode, they also undergo a shift in behavior that involves alterations in the activity of numerous genes. There is evidence that specific genes must be transcribed during the attachment phase of biofilm development. In many cases, the activation of these genes is required for synthesis of the extracellular matrix that protects the pathogens inside.According to Costerton, the genes that allow a biofilm to develop are activated after enough cells attach to a solid surface. “Thus, it appears that attachment itself is what stimulates synthesis of the extracellular matrix in which the sessile bacteria are embedded,†states the molecular biologist. “This notion– that bacteria have a sense of touch that enables detection of a surface and the expression of specific genes– is in itself an exciting area of research…â€[1]Certain characteristics may also facilitate the ability of some bacteria to form biofilms. Scientists at the Department of Microbiology and Molecular Genetics, Harvard Medical School, performed a study in which they created a “mutant†form of the bacterial species P. aeguinosa (PA).[5] The mutants lacked genes that code for hair-like appendages called pili. Interestingly, the mutants were unable to form biofilms. Since the pili of PA are involved in a type of surface-associated motility called twitching, the team hypothesized this twitching might be required for the aggregation of cells into the microcolonies that subsequently form a stable biofilm.Once a biofilm has officially formed, it often contains channels in which nutrients can circulate. Cells in different regions of a biofilm also exhibit different patterns of gene expression. Because biofilms often develop their own metabolism, they are sometimes compared to the tissues of higher organisms, in which closely packed cells work together and create a network in which minerals can flow.“There is a perception that single-celled organisms are asocial, but that is misguided,†said Andre Levchenko, assistant professor of biomedical engineering in s Hopkins University’s Whiting School of Engineering and an affiliate of the University’s Institute for NanoBioTechnology. “When bacteria are under stress—which is the story of their lives—they team up and form this collective called a biofilm. If you look at naturally occurring biofilms, they have very complicated architecture. They are like cities with channels for nutrients to go in and waste to go out.â€[6]The biofilm life cycle in three steps: attachment, growth of colonies (development), and periodic detachment of planktonic cells.Understanding how such cooperation among pathogens evolves and is maintained represents one of evolutionary biology’s thorniest problems. This stems from the reality that, in nature, freeloading cheats inevitably evolve to exploit any cooperative group that doesn’t defend itself, leading to the breakdown of cooperation. So what causes the bacteria in a biofilm to contribute to and share resources rather than steal them? Recently, Dr. Brockhurst of the University of Liverpool and colleagues at the Université Montpellier and the University of Oxford conducted several studies in an effort to understand why the bacteria in a biofilm cooperate and share resources rather than horde them.[7]The team took a closer look at P. fluorescens biofilms, which are formed when individual cells overproduce a polymer that sticks the cells together, allowing the colonization of liquid surfaces. While production of the polymer is metabolically costly to individual cells, the biofilm group benefits from the increased access to oxygen that surface colonization provides. Yet, evolutionarily speaking, such a setup allows possible “cheaters†to enter the biofilm. Such cheats can take advantage of the protective matrix while failing to contribute energy to actually building the matrix. If too many “cheaters†enter a biofilm, it will weaken and eventually break apart.After several years of study, Brockhurst and team realized that the short-term evolution of diversity within a biofilm is a major factor in how successfully its members cooperate. The team found that once inside a biofilm, P. fluorescensdifferentiates into various forms, each of which uses different nutrient resources. The fact that these “diverse cooperators†don’t all compete for the same chemicals and nutrients substantially reduces competition for resources within the biofilm.When the team manipulated diversity within experimental biofilms, they found that diverse biofilms contained fewer “cheaters†and produced larger groups than non-diverse biofilms.Levchenko and team used this device to observe bacteria growing in cramped conditions.Similarly, this year, researchers from s Hopkins; Virginia Tech; the University of California, San Diego; and Lund University in Sweden recently released the results of a study which found that once bacteria cooperate and form a biofilm, packing tightly together further enhances their survival.[6]The team created a new device in order to observe the behavior of E. coli bacteria forced to grow in the cramped conditions. The device, which allows scientists to use extremely small volumes of cells in solution, contains a series of tiny chambers of various shapes and sizes that keep the bacteria uniformly suspended in a culture medium.Not surprisingly, the cramped bacteria in the device began to form a biofilm. The team captured the development of the biofilm on video, and were able to observe the gradual self-organization and eventual construction of bacterial biofilms over a 24-hour period.First, Andre Levchenko and Hojung Cho of s Hopkins recorded the behavior of single layers of E. coli cells using real-time microscopy. “We were surprised to find that cells growing in chambers of all sorts of shapes gradually organized themselves into highly regular structures,†Levchenko said.Dr. Levchenko of s Hopkins and Hojung Cho, a biomedical engineering doctoral studentFurther observations using microscopy revealed that the longer the packed cell population resided in the chambers, the more ordered the biofilm structure became. As the cells in the biofilm became more ordered and tightly packed, the biofilm became harder and harder to penetrate.Levchenko also noted that rod-shaped E. colithat were too short or too long typically did not organize well into the dense, circular main hub of the biofilm. Instead, the bacteria of odd shapes or highly disordered groups of cells were found on the edges of the biofilm, where they formed sharp corners.Nodes of relapsing infection?Researchers often note that, once biofilms are established, planktonic bacteria may periodically leave the biofilm on their own. When they do, they can rapidly multiply and disperse.According to Costerton, there is a natural pattern of programmed detachment of planktonic cells from biofilms. This means that biofilms can act as what Costerton refers to as “niduses†of acute infection. Because the bacteria in a biofilm are protected by a matrix, the host immune system is less likely to mount a response to their presence.[1]But if planktonic bacteria are periodically released from the biofilms, each time single bacterial forms enter the tissues, the immune system suddenly becomes aware of their presence. It may proceed to mount an inflammatory response that leads to heightened symptoms. Thus, the periodic release of planktonic bacteria from some biofilms may be what causes many chronic relapsing infections.Planktonic bacteria are periodically released from a biofilmAs R. Parsek of Northwestern University describes in a 2003 paper in the Annual Review of Microbiology, any pathogen that survives in a chronic form benefits by keeping the host alive.[8] After all, if a chronic bacterial form simply kills its host, it will no longer have a place to live. So according to Parsek, chronic infection often results in a “disease stalemate†where bacteria of moderate virulence are somewhat contained by the defenses of the host. The infectious agents never actually kill the host, but the host is never able to fully kill the invading pathogens either.Parsek believes that the optimal way for bacteria to survive under such circumstances is in a biofilm, stating that “Increasing evidence suggests that the biofilm mode of growth may play a key role in both of these adaptations. Biofilm growth increases the resistance of bacteria to killing and may make organisms less conspicuous to the immune system… ultimately this moderation of virulence may serve the bacteria’s interest by increasing the longevity of the host.â€The acceptance of biofilms as infectious entitiesAnton van Leeuwenhoek.Perhaps because many biofilms are sufficiently thick to be visible to the naked eye, the microbial communities were among the first to be studied by early microbiologists. Anton van Leeuwenhoek scraped the plaque biofilm from his teeth and observed what he described as the “animalculi†inside them under his primitive microscope. However, according to Costerton and team at the Center for Biofilm Research at Montana State University, it was not until the 1970s that scientists began to appreciate that bacteria in the biofilm mode of existence constitute such a major component of the bacterial biomass in most environments. Then, it was not until the 1980s and 1990s that scientists truly began to understand how elaborately organized a bacterial biofilm community can be.[1]As Kolter, professor of microbiology and molecular genetics at Harvard Medical School, and one of the first scientists to study how biofilms developstates, “At first, however, studying biofilms was a radical departure from previous work.â€Like most microbial geneticists, Kolter had been trained in the tradition dating back to Koch and Louis Pasteur, namely that bacteriology is best conducted by studying pure strains of planktonic bacteria. “While this was a tremendous advance for modern microbiology, it also distracted microbiologists from a more organismic view of bacteria, Kolter adds, “Certainly we felt that pure, planktonic cultures were the only way to work. Yet in nature bacteria don’t live like that,†he says. “In fact, most of them occur in mixed, surface-dwelling communities.â€Although research on biofilms has surged over the past few decades, the majority of biofilm research to date has focused on external biofilms, or those that form on various surfaces in our natural environment.Over the past years, as scientists developed better tools to analyze external biofilms, they quickly discovered that biofilms can cause a wide range of problems in industrial environments. For example, biofilms can develop on the interiors of pipes, which can lead to clogging and corrosion. Biofilms on floors and counters can make sanitation difficult in food preparation areas.Since biofilms have the ability to clog pipes, watersheds, storage areas, and contaminate food products, large companies with facilities that are negatively impacted by their presence have naturally taken an interest in supporting biofilm research, particularly research that specifies how biofilms can be eliminated.This means that many recent advances in biofilm detection have resulted from collaborations between microbial ecologists, environmental engineers, and mathematicians. This research has generated new analytical tools that help scientists identify biofilms.Biofilm in a swamp gas reactor.For example, the Canadian company FAS International Ltd. has justcreated an endoluminal brush, which will be launched this spring. Physicians can use the brush to obtain samples from the interior of catheters. Samples taken from catheters can be sent to a lab, where researchers determine if biofilms are present in the sample. If biofilms are detected, the catheter is immediately replaced, since the insertion of catheters with biofilms can cause the patient to suffer from numerous infections, some of which are potentially life threatening.Scientists now realize that biofilms are not just composed of bacteria. Nearly every species of microorganism – including viruses, fungi, and Archaea – have mechanisms by which they can adhere to surfaces and to each other. Furthermore, it is now understood that biofilms are extremely diverse. For example, upward of 300 different species of bacteria can inhabit the biofilms that form dental plaque.[9]Furthermore, biofilms have been found literally everywhere in nature, to the point where any mainstream microbiologist would acknowledge that their presence is ubiquitous. They can be found on rocks and pebbles at the bottom of most streams or rivers and often form on the surface of stagnant pools of water. In fact, biofilms are important components of food chains in rivers and streams and are grazed upon by the aquatic invertebrates upon which many fish feed. Biofilms even grow in the hot, acidic pools at Yellowstone National Park and on glaciers in Antarctica.Biofilm in acidic pools at Yellowstone National Park.It is also now understood that the biofilm mode of existence has been around for millenia. For example, filamentous biofilms have been identified in the 3.2-billion-year-old deep-sea hydrothermal rocks of the Pilbara Craton, Australia. According to a 2004 article in Nature Reviews Microbiology, “Biofilm formation appears early in the fossil record (approximately 3.25 billion years ago) and is common throughout a diverse range of organisms in both the Archaea and Bacteria lineages. It is evident that biofilm formation is an ancient and integral component of the prokaryotic life cycle, and is a key factor for survival in diverse environments.â€[10]Biofilms and diseaseThe fact that external biofilms are ubiquitous raises the question – if biofilms can form on essentially every surface in our external environments, can they do the same inside the human body? The answer seems to be yes, and over the past few years, research on internal biofilms has finally started to pick up pace. After all, it’s easy for biofilm researchers to see that the human body, with its wide range of moist surfaces and mucosal tissue, is an excellent place for biofilms to thrive. Not to mention the fact that those bacteria which join a biofilm have a significantly greater chance of evading the battery of immune system cells that more easily attack planktonic forms.Many would argue that research on internal biofilms has been largely neglected, despite the fact that bacterial biofilms seem to have great potential for causing human disease.Common sites of biofilm infection. One biofilm reach the bloodstream they can spread to any moist surface of the human body. Stoodley of the Center for Biofilm Engineering at Montana State University, attributes much of the lag in studying biofilms to the difficulties of working with heterogeneous biofilms compared with homogeneous planktonic populations. In a 2004 paper in Nature Reviews, the molecular biologist describes many reasons why biofilms are extremely difficult to culture, such as the fact that the diffusion of liquid through a biofilm and the fluid forces acting on a biofilm must be carefully calculated if it is to be cultured correctly. According to Stoodley, the need to master such difficult laboratory techniques has deterred many scientists from attempting to work with biofilms. [10]Also, since much of the technology needed to detect internal biofilms was created at the same time as the sequencing of the human genome, interest in biofilm bacteria, and the research grants that would accompany such interest, have been largely diverted to projects with a decidedly genetic focus. However, since genetic research has failed to uncover the cause of any of the common chronic diseases, biofilms are finally – just over the past few years – being studied more intensely, and being given the credit they deserve as serious infectious entities, capable of causing a wide array of chronic illnesses.In just a short period of time, researchers studying internal biofilms have already pegged them as the cause of numerous chronic infections and diseases, and the list of illnesses attributed to these bacterial colonies continues to grow rapidly.According to a recent public statement from the National Institutes of Health, more than 65% of all microbial infections are caused by biofilms. This number might seem high, but according to Kim of the Department of Chemical and Biological Engineering at Tufts University, “If one recalls that such common infections as urinary tract infections (caused by E. coli and other pathogens), catheter infections (caused by Staphylococcus aureus and other gram-positive pathogens), child middle-ear infections (caused by Haemophilus influenzae, for example), common dental plaque formation, and gingivitis, all of which are caused by biofilms, are hard to treat or frequently relapsing, this figure appears realistic.â€[11]Hundreds of microbial biofilm colonize the human mouth, causing tooth decay and gum disease.As mentions, perhaps the most well-studied biofilms are those that make up what is commonly referred to as dental plaque. “Plaque is a biofilm on the surfaces of the teeth,†states Parsek. “This accumulation of microorganisms subject the teeth and gingival tissues to high concentrations of bacterial metabolites which results in dental disease.â€[12]It has also recently been shown that biofilms are present on the removed tissue of 80% of patients undergoing surgery for chronic sinusitis. According to Parsek, biofilms may also cause osteomyelitis, a disease in which the bones and bone marrow become infected. This is supported by the fact that microscopy studies have shown biofilm formation on infected bone surfaces from humans and experimental animal models. Parsek also implicates biofilms in chronic prostatitis since microscopy studies have also documented biofilms on the surface of the prostatic duct. Microbes that colonize vaginal tissue and tampon fibers can also form into biofilms, causing inflammation and disease such as Toxic Shock Syndrome.Biofilms also cause the formation of kidney stones. The stones cause disease by obstructing urine flow and by producing inflammation and recurrent infection that can lead to kidney failure. Approximately 15%–20% of kidney stones occur in the setting of urinary tract infection. According to Parsek, these stones are produced by the interplay between infecting bacteria and mineral substrates derived from the urine. This interaction results in a complex biofilm composed of bacteria, bacterial exoproducts, and mineralized stone material.Microbes that colonize vaginal tissue and tampon fibers can become pathogenic, causing inflammation and disease such as Toxic Shock Syndrome.Perhaps the first hint of the role of bacteria in these stones came in 1938 when Hellstrom examined stones passed by his patients and found bacteria embedded deep inside them. Microscopic analysis of stones removed from infected patients has revealed features that characterize biofilm growth. For one thing, bacteria on the surface and inside the stones are organized in microcolonies and surrounded by a matrix composed of crystallized (struvite) minerals.Then there’s endocarditis, a disease that involves inflammation of the inner layers of the heart. The primary infectious lesion in endocarditis is a complex biofilm composed of both bacterial and host components that is located on a cardiac valve. This biofilm, known as a vegetation, causes disease by three basic mechanisms. First, the vegetation physically disrupts valve function, causing leakage when the valve is closed and inducing turbulence and diminished flow when the valve is open. Second, the vegetation provides a source for near-continuous infection of the bloodstream that persists even during antibiotic treatment. This causes recurrent fever, chronic systemic inflammation, and other infections. Third, pieces of the infected vegetation can break off and be carried to a terminal point in the circulation where they block the flow of blood (a process known as embolization). The brain, kidney, and extremities are particularly vulnerable to the effects of embolization.A variety of pathogenic biofims are also commonly found on medical devices such as joint prostheses and heart valves. According to Parsek, electron microscopy of the surfaces of medical devices that have been foci of device-related infections shows the presence of large numbers of slime-encased bacteria. Tissues taken from non-device-related chronic infections also show the presence of biofilm bacteria surrounded by an exopolysaccharide matrix. These biofilm infections may be caused by a single species or by a mixture of species of bacteria or fungi.According to Dr. Patel of the Mayo Clinic, individuals with prosthetic joints are often oblivious to the fact that their prosthetic joints harbor biofilm infections.[13]Cells of Staphylococcus epidermidis causing devastating disease as they grow on the cuff at a mechanical heart valve.“When people think of infection, they may think of fever or pus coming out of a wound,†explains Dr. Patel. “However, this is not the case with prosthetic joint infection. Patients will often experience pain, but not other symptoms usually associated with infection. Often what happens is that the bacteria that cause infection on prosthetic joints are the same as bacteria that live harmlessly on our skin. However, on a prosthetic joint they can stick, grow and cause problems over the long term. Many of these bacteria would not infect the joint were it not for the prosthesis.â€Biofilms also cause Leptospirosis, a serious but neglected emerging disease that infects humans through contaminated water. New research published in the May issue of the journal Microbiology shows for the first time how bacteria that cause the disease survive in the environment.Leptospirosis is a major public health problem in southeast Asia and South America, with over 500,000 severe cases every year. Between 5% and 20% of these cases are fatal. Rats and other mammals carry the disease-causing pathogen Leptospira interrogans in their kidneys. When they urinate, they contaminate surface water with the bacteria, which can survive in the environment for long periods.“This led us to see if the bacteria build a protective casing around themselves for protection,†said Professor Mathieu Picardeau from the Institut Pasteur in Paris, France. [14]Previously, scientists believed the bacteria were planktonic. But Professor Picardeau and his team have shown that L. interrogans can make biofilms, which could be one of the main factors controlling survival and disease transmission. “90% of the species of Leptospira we tested could form biofilms. It takes L. interrogans an average of 20 days to make a biofilm,†says Picardeau.Biofilms have also been implicated in a wide array of veterinary diseases. For example, researchers at the Virginia-land Regional College of Veterinary Medicine at Virginia Tech were just awarded a grant from the United States Department of Agriculture to study the role biofilms play in the development of Bovine Respiratory Disease Complex (BRDC). If biofilms play a role in bovine respiratory disease, it’s likely only a matter of time before they will be established as a cause of human respiratory diseases as well.When the immune response is compromised, Pseudomonas aeruginosabiofilms are able to colonize the alveoli, and to form biofilms.As mentioned previously, infection by the bacterium Pseudomonas aeruginosa (P. aeruginosa) is the main cause of death among patients with cystic fibrosis. Pseudomonas is able to set up permanent residence in the lungs of patients with cystic fibrosis where, if you ask most mainstream researchers, it is impossible to kill. Eventually, chronic inflammation produced by the immune system in response to Pseudomonas destroys the lung and causes respiratory failure. In the permanent infection phase, P. aeruginosa biofilms are thought to be present in the airway, although much about the infection pathogenesis remains unclear.[15]Cystic fibrosis is caused by mutations in the proteins of channels that regulates chloride. How abnormal chloride channel protein leads to biofilm infection remains hotly debated. It is clear, however, that cystic fibrosis patients manifest some kind of host-defense defect localized to the airway surface. Somehow this leads to a debilitating biofilm infection.Biofilms have the potential to cause a tremendous array of infections and diseasesBecause internal pathogenic biofilm research comprises such a new field of study, the infections described above almost certainly represent just the tip of the iceberg when it comes to the number of chronic diseases and infections currently caused by biofilms.For example, it wasn’t until July of 2006 that researchers realized that the majority of ear infections are caused by biofilm bacteria. These infections, which can be either acute or chronic, are referred to collectively as otitis media (OM). They are the most common illness for which children visit a physician, receive antibiotics, or undergo surgery in the United States.There are two subtypes of chronic OM. Recurrent OM (ROM) is diagnosed when children suffer repeated infections over a span of time and during which clinical evidence of the disease resolves between episodes. Chronic OM with effusion is diagnosed when children have persistent fluid in the ears that lasts for months in the absence of any other symptoms except conductive hearing loss.It took over ten years for researchers to realize that otitis media is caused by biofilms. Finally, in 2002, Drs. Ehrlich and J. Post, an Allegheny General Hospital pediatric ear specialist and medical director of the Center for Genomic Sciences, published the first animal evidence of biofilms in the middle ear in the Journal of the American Medical Association, setting the stage for further clinical investigation.In a subsequent study, Ehrlich and Post obtained middle ear mucosa – or membrane tissue – biopsies from children undergoing a procedure for otitis. The team gathered uninfected mucosal biopsies from children and adults undergoing cochlear implantation as a control.[16]Using advanced confocal laser scanning microscopy, Luanne Hall Stoodley, Ph.D. and her ASRI colleagues obtained three dimensional images of the biopsies and evaluated them for biofilm morphology using generic stains and species-specific probes for Haemophilus influenzae, Streptococcus pneumoniaeand Moraxella catarrhalis. Effusions, when present, were also evaluated for evidence of pathogen specific nucleic acid sequences (indicating presence of live bacteria).The study found mucosal biofilms in the middle ears of 46/50 children (92%) with both forms of otitis. Biofilms were not observed in eight control middle ear mucosa specimens obtained from cochlear implant patients.Otitis media, or inflammation of the inner ear, is caused by biofilm.In fact, all of the children in the study who suffered from chronic otitis media tested positive for biofilms in the middle ear, even those who were asymptomatic, causing Erlich to conclude that, “It appears that in many cases recurrent disease stems not from re-infection as was previously thought and which forms the basis for conventional treatment, but from a persistent biofilm.â€He went on to state that the discovery of biofilms in the setting of chronic otitis media represented “a landmark evolution in the medical community’s understanding about a disease that afflicts millions of children world-wide each year and further endorses the emerging biofilm paradigm of chronic infectious disease.â€The emerging biofilm paradigm of chronic disease refers to a new movement in which researchers such as Ehrlich are calling for a tremendous shift in the way the medical community views bacterial biofilms. Those scientists who support an emerging biofilm paradigm of chronic disease feel that biofilm research is of utmost importance because of the fact that the infectious entities have the potential to cause so many forms of chronic disease. The Marshall Pathogenesis is an important part of this paradigm shift.It was also just last year that researchers realized that biofilms cause most infections associated with contact lens use. In 2006, Bausch & Lomb withdrew its ReNu with MoistureLoc contact lens solution because a high proportion of corneal infections were associated with it. It wasn’t long before researchers at the University Hospitals Case Medical Center found that the infections were caused by biofilms. [17]“Once they live in that type of state [a biofilm], the cells become resistant to lens solutions and immune to the body’s own defense system,†said Mahmoud A. Ghannoum, Ph.D, senior investigator of the study. “This study should alert contact lens wearers to the importance of proper care for contact lenses to protect against potentially virulent eye infections,†he said.It turns out that the biofilms detected by Ghannoum and team were composed of fungi, particularly a species called Fusarium. His team also discovered that the strain of fungus (with the catchy name, ATCC 36031) used for testing the effectiveness of lens care solutions is a strain that does not produce biofilms as the clinical fungal strains do. ReNu contact solution, therefore, was effective in the laboratory, but failed when faced with strains in real-world situations.Fungal biofilm can form in contact lens solution leading to potentially virulent eye infectionsUnfortunately, Ghannoum and team were not able to create a method to target and destroy the fungal biofilms that plague users of ReNu and some other contact lens solutions.Then there’s Dr. Randall Wolcott who just recently discovered and confirmed that the sludge covering diabetic wounds is largely made up of biofilms. Whereas before Wolcott’s work such limbs generally had to be amputated, now that they have been correctly linked to biofilms, measures such as those described in thisinterview can be taken to stop the spread of infection and save the limb. Wolcott has finally been given a grant by the National Institutes of Health to further study chronic biofilms and wound development.Dr. Garth and the Medical Biofilm Laboratory team at Montana State University are also researching wounds and biofilms. Their latest article and an image showing wound biofilm was featured on the cover of the January-February 2008 issue of Wound Repair and Regeneration.[18]Biofilm bacteria and chronic inflammatory diseaseIn just a few short years, the potential of biofilms to cause debilitating chronic infections has become so clear that there is little doubt that biofilms are part of the pathogenic mix or “pea soup†that cause most or all chronic “autoimmune†and inflammatory diseases.In fact, thanks, in large part, to the research of biomedical researcher Dr. Trevor Marshall, it is now increasingly understood that chronic inflammatory diseases result from infection with a large microbiota of chronic biofilm and L-form bacteria (collectively called the Th1 pathogens).[19][20] The microbiota is thought to be comprised of numerous bacterial species, some of which have yet to be discovered. However, most of the pathogens that cause inflammatory disease have one thing in common – they have all developed ways to evade the immune system and persist as chronic forms that the body is unable to eliminate naturally.Some L-form bacteria are able to evade the immune system because, long ago, they evolved the ability to reside inside macrophages, the very white bloods cells of the immune system that are supposed to kill invading pathogens. Upon formation, L-form bacteria also lose their cell walls, which makes them impervious to components of the immune response that detect invading pathogens by identifying the proteins on their cell walls. The fact that L-form bacteria lack cell walls also means that the beta-lactam antibiotics, which work by targeting the bacterial cell wall, are completely ineffective at killing them.[21]Clearly, transforming into the L-form offers any pathogen a survival advantage. But among those pathogens not in an L-form state, joining a biofilm is just as likely to enhance their ability to evade the immune system. Once enough chronic pathogens have grouped together and formed a stable community with a strong protective matrix, they are likely able to reside in any area of the body, causing the host to suffer from chronic symptoms that are both mental and physical in nature.Biofilm researchers will also tell you that, not surprisingly, biofilms form with greater ease in an immunocompromised host. Marshall’s research has made it clear that many of the Th1 pathogens are capable of creating substances that bind and inactivate the Vitamin D Receptor – a fundamental receptor of the body that controls the activity of the innate immune system, or the body’s first line of defense against intracellular infection.[22]Diagram of the Vitamin D Receptor and capnine.Thus, as patients accumulate a greater number of the Th1 pathogens, more and more of the chronic bacterial forms create substances capable of disabling the VDR. This causes a snowball effect, in which the patient becomes increasingly immunocompromised as they acquire a larger bacterial load.For one thing, it’s possible that many of the bacteria that survive inside biofilms are capable of creating VDR blocking substances. Thus, the formation of biofilms may contribute to immune dysfunction. Conversely, as patients acquire L-form bacteria and other persistent bacterial forms capable of creating VDR-blocking substances, it becomes exceptionally easy for biofilms to form on any tissue surface of the human body.Thus, patients who begin to acquire L-form bacteria almost always fall victim to biofilm infections as well, since it is all too easy for pathogens to group together into a biofilm when the immune system isn’t working up to par.To date, there is also no strict criteria that separate L-form bacteria from biofilm bacteria or any other chronic pathogenic forms. This means that L-form bacteria may also form into biofilms, and by doing so enter a mode of survival that makes them truly impervious to the immune system. Some L-form bacteria may not form complete biofilms, yet may still possess the ability to surround themselves in a protective matrix. Under these circumstances one might say they are in a “biofilm-like†state.Marshall often refers to the pathogens that cause inflammatory disease as an intraphgocytic, metagenomic microbiota of bacteria, terms which suggest that most chronic bacterial forms possess properties of both L-form and biofilm bacteria. Intraphagocytic refers to the fact that the pathogens can be found inside the cells of the immune system. The term metagenomic indicates that there are a tremendous number of different species of these chronic bacterial forms. Finally, microbiota refers to the fact that biofilm communities sustain their pathogenic activity.For example, when observed under a darkfield microscope, L-form bacteria are often encased in protective biofilm sheaths. If the blood containing the pathogens are aged overnight, the bacterial colonies reach a point where they expand and burst out of the cell, causing the cell to burst as well. Then they extend as huge, long biofilm tubules, which are presumably helping the pathogens spread to other cells. The tubules also help spread bacterial DNA to neighboring cells.Clearly, there is a great need for more research on how different chronic bacterial forms interact. To date, L-form researchers have essentially focused soley on the L-form, while failing to investigate how frequently the wall-less pathogens form into biofilms or become parts of biofilm communities together with bacteria with cell walls. Conversely, most biofilm researchers are intently studying the biofilm mode of growth without considering the presence of L-form bacteria. So, it will likely take several years before we will be better able to understand probable overlaps between the lifestyles of L-form and biofilm bacteria.Anyone who is skeptical about the fact that biofilms likely form a large percentage of the microbiota that cause inflammatory disease should consider many of the recent studies that have linked established biofilm infections to a higher risk for multiple forms of chronic inflammatory disease. Take, for example, studies that have found a link between periodontal disease and several major inflammatory conditions. A 1989 article published in British Medical Journal showed a correlation between dental disease and systemic disease (stroke, heart disease, diabetes). After correcting for age, exercise, diet, smoking, weight, blood cholesterol level, alcohol use and health care, people who had periodontal disease had a significantly higher incidence of heart disease, stroke and premature death. More recently, these results were confirmed in studies in the United States, Canada, Great Britain, Sweden, and Germany. The effects are striking. For example, researchers from the Canadian Health Bureau found that people with periodontal disease had a two times higher risk of dying from cardiovascular disease.[23]Dental plaque as seen under a scanning electron microcroscope.Since we know that periodontal disease is caused by biofilm bacteria, the most logical explanation for the fact that people with dental problems are much more likely to suffer from heart disease and stroke is that the biofilms in their mouths have gradually spread to the moist surfaces of their circulatory systems. Or perhaps if the bacteria in periodontal biofilms create VDR binding substances, their ability to slow innate immune function allows new biofilms (and L-form bacteria as well) to more easily form and infect the heart and blood vessels. Conversely, systemic infection with VDR blocking biofilm bacteria is also likely to weaken immune defenses in the gums and facilitate periodontal disease.In fact, it appears that biofilm bacteria in the mouth also facilitate the formation of biofilm and L-form bacteria in the brain. Just last year, researchers at Vasant Hirani at University College London released the results of a study which found that elderly people who have lost their teeth are at more than three-fold greater risk of memory problems and dementia.[24]At the moment, Autoimmunity Research Foundation does not have the resources to culture biofilms from patients on the treatment and, even if they did, current methods for culturing internal biofilms remain unreliable. According to Stoodley, “The lack of standard methods for growing, quantifying and testing biofilms in continuous culture results in incalculable variability between laboratory systems. Biofilm microbiology is complex and not well represented by flask cultures. Although homogeneity allows statistical enumeration, the extent to which it reflects the real, less orderly world is questionable.â€[10]How else do we acquire biofilm bacteria?As discussed thus far, biofilms form spontaneously as bacteria inside the human body group together. Yet people can also ingest biofilms by eating contaminated food.According to researchers at the University of Guelph in Ontario Canada, it is increasingly suspected that biofilms play an important role in contamination of meat during processing and packaging. The group warns that greater action must be taken to reduce the presence of food-borne pathogens like Escherichia coli and Listeria monocytogenes and spoilage microorganisms such as thePseudomonas species (all of which form biofilms) throughout the food processing chain to ensure the safety and shelf-life of the product. Most of these microorganisms are ubiquitous in the environment or brought into processing facilities through healthy animal carriers.Hans Blaschek of the University of Illinois has discovered that biofilms form on much of the other food products we consume as well.A biofilm on a piece of lettuce“If you could see a piece of celery that’s been magnified 10,000 times, you’d know what the scientists fighting foodborne pathogens are up against,†says Blaschek.“It’s like looking at a moonscape, full of craters and crevices. And many of the pathogens that cause foodborne illness, such as Shigella, E. coli,and Listeria, make sticky, sugary biofilms that get down in these crevices, stick like glue, and hang on like crazy.â€According to Blaschek, the problem faced by produce suppliers can be a triple whammy. “If you’re unlucky enough to be dealing with a pathogen–and the pathogen has the additional attribute of being able to form biofilm—and you’re dealing with a food product that’s minimally processed, well, you’re triply unlucky,†the scientist said. “You may be able to scrub the organism off the surface, but the cells in these biofilms are very good at aligning themselves in the subsurface areas of produce.†, a University of Illinois food science and human nutrition professor agrees, stating,â€Once the pathogenic organism gets on the product, no amount of washing will remove it. The microbes attach to the surface of produce in a sticky biofilm, and washing just isn’t very effective.â€Biofilms can even be found in processed water. Just this month, a study was released in which researchers at the Department of Biological Sciences, at Virginia Polytechnic Institute isolated M. avium biofilm from the shower head of a woman with M. avium pulmonary disease.[25] A molecular technique called DNA fingerprinting demonstrated that M. avium isolates from the water were the same forms that were causing the woman’s respiratory illness.Effectively targeting biofilm infectionsAlthough the mainstream medical community is rapidly acknowledging the large number of diseases and infections caused by biofilms, most researchers are convinced that biofilms are difficult or impossible to destroy, particularly those cells that form the deeper layers of a thick biofilm. Most papers on biofilms state that they are resistant to antibiotics administered in a standard manner. For example, despite the fact that Ehrlich and team discovered that biofilm bacteria cause otitis media, they are unable to offer an effective solution that would actually allow for the destruction of biofilms in the ear canal. Other teams have also come up short in creating methods to break up the biofilms they implicate as the cause of numerous infections.This means patients with biofilm infections are generally told by mainstream doctors that they have an untreatable infection. In some cases, a disease-causing biofilm can be cut out of a patient’s tissues, or efforts are made to drain components of the biofilm out of the body. For example, doctors treating otitis media often treats patients with myringotomy, a surgical procedure in which small tubes are placed in the eardrum to continuously drain infectious fluid.When it comes to administering antibiotics in an effort to target biofilms, one thing is certain. Mainstream researchers have repeatedly tried to kill biofilms by giving patients high, constant doses of antibiotics. Unfortunately, when administered in high doses, the antibiotic may temporarily weaken the biofilm but is incapable of destroying it, as certain cells inevitably persist and allow the biofilm to regenerate.“You can put a patient on [a high dose] antibiotics, and it may seem that the infection has disappeared,†says Levchenko. “But in a few months, it reappears, and it is usually in an antibiotic-resistant form.â€What the vast majority of researchers working with biofilms fail to realize is that antibiotics are capable of destroying biofilms. The catch is that antibiotics are only effective against biofilms if administered in a very specific manner. Furthermore, only certain antibiotics appear to effectively target biofilms. After decades of research, much of which was derived from molecular modeling data, Marshall was the first to create an antibiotic regimen that appears to effectively target and destroy biofilms. Central to the treatment, which is called the Marshall Protocol, is the fact that biofilms and other Th1 pathogens succumb to specific bacteriostatic antibiotics taken in very low, pulsed doses. It is only when antibiotics are administered in this manner that they appear capable of fully eradicating biofilms.[19][20]In a paper entitled “The Riddle of Biofilm Resistance,†Dr. Kim of Tulane University discusses the mechanisms by which pulsed, low dose antibiotics are able to break up biofilms, while antibiotics administered in a standard manner (high, constant doses) cannot. According to , the use of pulsed, low-dose antibiotics to target biofilm bacteria is supported by observations she and her colleagues have made in the laboratory.[11]Some researchers claim that antibiotics cannot penetrate the matrix that surrounds a biofilm. But research by and other scientists has confirmed that the inability of antibiotics to penetrate the biofilm matrix is much more of an exception than a rule. According to , “In most cases involving small antimicrobial molecules, the barrier of the polysaccharide matrix should only postpone the death of cells rather than afford useful protection.â€For example, a recent study that used low concentrations of an antibiotic to killP. aeruginosa biofilm bacteria found that the majority of biofilm cells were effectively eliminated by antibiotics in a manne

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Wow, what a fascinating article. Thanks .RegardsKrysOn 9 September 2011 10:32, <Goldstein@...> wrote:

 

Understanding BiofilmsAuthor: Amy Proal

26MAY2008

As humans, our environment consistently exposes us to a variety of dangers. Tornadoes, lightning, flooding and hurricanes can all hamper our survival. Not to mention the fact that most of us can encounter swerving cars or ill-intentioned people at any given moment.

Biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of materialThousands of years ago, humans realized that they could better survive a dangerous world if they formed into communities, particularly communities consisting of people with different talents. They realized that a community is far more likely to survive through division of labor– one person makes food, another gathers resources, still another protects the community against invaders. Working together in this manner requires communication and cooperation.

Inhabitants of a community live in close proximity and create various forms of shelter in order to protect themselves from external threats. We build houses that protect our families and larger buildings that protect the entire community. Grouping together inside places of shelter is a logical way to enhance survival.

With the above in mind, it should come as no surprise that the pathogens we harbor are seldom found as single entities. Although the pathogens that cause acute infection are generally free-floating bacteria – also referred to as planktonic bacteria – those chronic bacterial forms that stick around for decades long ago evolved ways to join together into communities. Why? Because by doing so, they are better able to combat the cells of our immune system bent upon destroying them.

It turns out that a vast number of the pathogens we harbor are grouped into communities called biofilms. In an article titled “Bacterial Biofilms: A Common Cause of Persistent Infections,†JW Costerton at the Center for Biofilm Engineering in Montana defines a bacterial biofilm as “a structured community of bacterial cells enclosed in a self-produced polymeric matrix and adherent to an inert or living surface.â€[1] In layman’s terms, that means that bacteria can join together on essentially any surface and start to form a protective matrix around their group. The matrix is made of polymers – substances composed of molecules with repeating structural units that are connected by chemical bonds.

According to the Center for Biofilm Engineering at Montana State University, biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of material – such as metals, plastics, soil particles, medical implant materials and, most significantly, human or animal tissue. The first bacterial colonists to adhere to a surface initially do so by inducing weak, reversible bonds called van der Waals forces. If the colonists are not immediately separated from the surface, they can anchor themselves more permanently using cell adhesion molecules, proteins on their surfaces that bind other cells in a process called cell adhesion.

A biofilm in the gut.These bacterial pioneers facilitate the arrival of other pathogens by providing more diverse adhesion sites. They also begin to build the matrix that holds the biofilm together. If there are species that are unable to attach to a surface on their own, they are often able to anchor themselves to the matrix or directly to earlier colonists.

During colonization, things start to get interesting. Multiple studies have shown that during the time a biofilm is being created, the pathogens inside it can communicate with each other thanks to a phenomenon called quorum sensing. Although the mechanisms behind quorum sensing are not fully understood, the phenomenon allows a single-celled bacterium to perceive how many other bacteria are in close proximity. If a bacterium can sense that it is surrounded by a dense population of other pathogens, it is more inclined to join them and contribute to the formation of a biofilm.

Bacteria that engage in quorum sensing communicate their presence by emitting chemical messages that their fellow infectious agents are able to recognize. When the messages grow strong enough, the bacteria respond en masse, behaving as a group. Quorum sensing can occur within a single bacterial species as well as between diverse species, and can regulate a host of different processes, essentially serving as a simple communication network. A variety of different molecules can be used as signals.

“Disease-causing bacteria talk to each other with a chemical vocabulary,†says Doug Hibbins of Princeton University. A graduate student in the lab of Princeton University microbiologist Dr. Bonnie Bassler, Hibbins was part of a research effort which shed light on how the bacteria that cause cholera form biofilms and communicate via quorum sensing.[2]

“Forming a biofilm is one of the crucial steps in cholera’s progression,†states Bassler. “They [bacteria] cover themselves in a sort of goop that’s a shield against antibiotics, allowing them to grow rapidly. When they sense there are enough of them, they try to leave the body.â€

Although cholera bacteria use the intestines as a breeding ground, after enough biofilms have formed, planktonic bacteria inside the biofilm seek to leave the body in order to infect a new host. It didn’t take long for Bassler and team to realize that the bacteria inside cholera biofilms must signal each other in order to communicate that it’s time for the colony to stop reproducing and focus instead on leaving the body.

“We generically understood that bacteria talk to each other with quorum sensing, but we didn’t know the specific chemical words that cholera uses,†Bassler said.Then Higgins isolated the CAI-1 – a chemical which occurs naturally in cholera. Another graduate student figured out how to make the molecule in the laboratory. By moderating the level of CAI-1 in contact with cholera bacteria, Higgins was successfully able to chemically control cholera’s behavior in lab tests. His team eventually confirmed that when CAI-1 is absent, cholera bacteria attach in biofilms to their current host. But when the bacteria detect enough of the chemical, they stop making biofilms and releasing toxins, perceiving that it is time to leave the body instead. Thus, CAI-1 may very well be the single molecule that allow the bacteria inside a cholera biofilm to communicate. Although it is likely that the bacteria in a cholera biofilm may communicate with other signals besides CAI-1, the study is a good example of the fact that signaling molecules serve a key role in determining the state of a biofilm.

Sessile cells in a biofilm “talk†to each other via quorum sensing to build microcolonies and to keep water channels open.Similarly, researchers at the University of Iowa (several of whom are now at the University of Washington) have spent the last decade identifying the molecules that allow the bacterial species P. aeruginosa to form biofilms in the lungs of patients with cystic fibrosis.[3] Although the P. auruginosa isolated from the lungs of patients with cystic fibrosis looks like a biofilm and acts like a biofilm, up until recently, there were no objective tests available to confirm that the bacterial species did indeed form biofilms in the lungs of patients with the disease, nor was there a way to tell what proportion of P. aeruginosa in the lungs were actually in biofilm mode.

“We needed a way to show that the P. auruginosa in cystic fibrosis lungs was communicating like a biofilm. That could tell us about the P. auruginosalifestyle,†said Pradeep Singh, M.D., a lead author on the study who is now at the University of Washington.

Singh and his colleagues finally discovered that P. aeruginosa uses one of two particular quorum-sensing molecules to initiate the formation of biofilms. In November 1999, his research team screened the entire bacterial genome, identifying 39 genes that are strongly controlled by the quorum-sensing system.

In a 2000 study published in Nature, Singh and colleagues developed a sensitive test which shows P. auruginosa from cystic fibrosis lungs produces the telltale, quorum-sensing molecules that are the signals for biofilm formation.[3]

It turns out that P. aerugnosa secretes two signaling molecules, one that is long, and another that is short. Using the new test, the team was able to show that planktonic forms of P. aeruginosa produce more long signaling molecules. Alternately, when they tested the P. aeruginosa strains isolated from the lungs of patients with cystic fibrosis (which were in biofilm form), all of the strains produced the signaling molecules, but in the opposite ratio – more short than long.

Interestingly, when the biofilm strains of P. aeruginosa were separated in broth into individual bacterial forms, they reverted to producing more long signal molecules than short ones. Does this mean that a change in signaling molecular length can indicate whether bacteria remain as planktonic forms or develop into biofilms?

To find out, the team took the bacteria from the broth and made them grow as a biofilm again. Sure enough, those strains of bacteria in biofilm form produced more short signal molecules than long.

“The fact that the P. aeruginosa in [the lungs of cystic fibrosis patients] is making the signals in the ratios that we see tells us that there is a biofilm and that most of the P. aeruginosa in the lung is in the biofilm state,†states Greenberg, another member of the research team. He believes that the findings allow for a clear biochemical definition of whether bacteria are in a biofilm. Techniques similar to those used by his group will likely be used to determine the properties of other biofilm signaling molecules.

DevelopmentOnce colonization has begun, the biofilm grows through a combination of cell division and recruitment. The final stage of biofilm formation is known as development and is the stage in which the biofilm is established and may only change in shape and size. This development of a biofilm allows for the cells inside to become more resistant to antibiotics administered in a standard fashion. In fact, depending on the organism and type of antimicrobial and experimental system, biofilm bacteria can be up to a thousand times more resistant to antimicrobial stress than free-swimming bacteria of the same species.

Biofilms grow slowly, in diverse locations, and biofilm infections are often slow to produce overt symptoms. However, biofilm bacteria can move in numerous ways that allow them to easily infect new tissues. Biofilms may move collectively, by rippling or rolling across the surface, or by detaching in clumps. Sometimes, in a dispersal strategy referred to as “swarming/seedingâ€, a biofilm colony differentiates to form an outer “wall†of stationary bacteria, while the inner region of the biofilm “liquefiesâ€, allowing planktonic cells to “swim†out of the biofilm and leave behind a hollow mound.[4]

Biofilm bacteria can move in numerous ways: Collectively, by rippling or rolling across the surface, or by detaching in clumps. Individually, through a “swarming and seeding†dispersal.

Research on the molecular and genetic basis of biofilm development has made it clear that when cells switch from planktonic to community mode, they also undergo a shift in behavior that involves alterations in the activity of numerous genes. There is evidence that specific genes must be transcribed during the attachment phase of biofilm development. In many cases, the activation of these genes is required for synthesis of the extracellular matrix that protects the pathogens inside.

According to Costerton, the genes that allow a biofilm to develop are activated after enough cells attach to a solid surface. “Thus, it appears that attachment itself is what stimulates synthesis of the extracellular matrix in which the sessile bacteria are embedded,†states the molecular biologist. “This notion– that bacteria have a sense of touch that enables detection of a surface and the expression of specific genes– is in itself an exciting area of research…â€[1]

Certain characteristics may also facilitate the ability of some bacteria to form biofilms. Scientists at the Department of Microbiology and Molecular Genetics, Harvard Medical School, performed a study in which they created a “mutant†form of the bacterial species P. aeguinosa (PA).[5] The mutants lacked genes that code for hair-like appendages called pili. Interestingly, the mutants were unable to form biofilms. Since the pili of PA are involved in a type of surface-associated motility called twitching, the team hypothesized this twitching might be required for the aggregation of cells into the microcolonies that subsequently form a stable biofilm.

Once a biofilm has officially formed, it often contains channels in which nutrients can circulate. Cells in different regions of a biofilm also exhibit different patterns of gene expression. Because biofilms often develop their own metabolism, they are sometimes compared to the tissues of higher organisms, in which closely packed cells work together and create a network in which minerals can flow.

“There is a perception that single-celled organisms are asocial, but that is misguided,†said Andre Levchenko, assistant professor of biomedical engineering in s Hopkins University’s Whiting School of Engineering and an affiliate of the University’s Institute for NanoBioTechnology. “When bacteria are under stress—which is the story of their lives—they team up and form this collective called a biofilm. If you look at naturally occurring biofilms, they have very complicated architecture. They are like cities with channels for nutrients to go in and waste to go out.â€[6]

The biofilm life cycle in three steps: attachment, growth of colonies (development), and periodic detachment of planktonic cells.Understanding how such cooperation among pathogens evolves and is maintained represents one of evolutionary biology’s thorniest problems. This stems from the reality that, in nature, freeloading cheats inevitably evolve to exploit any cooperative group that doesn’t defend itself, leading to the breakdown of cooperation. So what causes the bacteria in a biofilm to contribute to and share resources rather than steal them? Recently, Dr. Brockhurst of the University of Liverpool and colleagues at the Université Montpellier and the University of Oxford conducted several studies in an effort to understand why the bacteria in a biofilm cooperate and share resources rather than horde them.[7]

The team took a closer look at P. fluorescens biofilms, which are formed when individual cells overproduce a polymer that sticks the cells together, allowing the colonization of liquid surfaces. While production of the polymer is metabolically costly to individual cells, the biofilm group benefits from the increased access to oxygen that surface colonization provides. Yet, evolutionarily speaking, such a setup allows possible “cheaters†to enter the biofilm. Such cheats can take advantage of the protective matrix while failing to contribute energy to actually building the matrix. If too many “cheaters†enter a biofilm, it will weaken and eventually break apart.

After several years of study, Brockhurst and team realized that the short-term evolution of diversity within a biofilm is a major factor in how successfully its members cooperate. The team found that once inside a biofilm, P. fluorescensdifferentiates into various forms, each of which uses different nutrient resources. The fact that these “diverse cooperators†don’t all compete for the same chemicals and nutrients substantially reduces competition for resources within the biofilm.

When the team manipulated diversity within experimental biofilms, they found that diverse biofilms contained fewer “cheaters†and produced larger groups than non-diverse biofilms.

Levchenko and team used this device to observe bacteria growing in cramped conditions.Similarly, this year, researchers from s Hopkins; Virginia Tech; the University of California, San Diego; and Lund University in Sweden recently released the results of a study which found that once bacteria cooperate and form a biofilm, packing tightly together further enhances their survival.[6]

The team created a new device in order to observe the behavior of E. coli bacteria forced to grow in the cramped conditions. The device, which allows scientists to use extremely small volumes of cells in solution, contains a series of tiny chambers of various shapes and sizes that keep the bacteria uniformly suspended in a culture medium.

Not surprisingly, the cramped bacteria in the device began to form a biofilm. The team captured the development of the biofilm on video, and were able to observe the gradual self-organization and eventual construction of bacterial biofilms over a 24-hour period.

First, Andre Levchenko and Hojung Cho of s Hopkins recorded the behavior of single layers of E. coli cells using real-time microscopy. “We were surprised to find that cells growing in chambers of all sorts of shapes gradually organized themselves into highly regular structures,†Levchenko said.

Dr. Levchenko of s Hopkins and Hojung Cho, a biomedical engineering doctoral studentFurther observations using microscopy revealed that the longer the packed cell population resided in the chambers, the more ordered the biofilm structure became. As the cells in the biofilm became more ordered and tightly packed, the biofilm became harder and harder to penetrate.

Levchenko also noted that rod-shaped E. colithat were too short or too long typically did not organize well into the dense, circular main hub of the biofilm. Instead, the bacteria of odd shapes or highly disordered groups of cells were found on the edges of the biofilm, where they formed sharp corners.

Nodes of relapsing infection?Researchers often note that, once biofilms are established, planktonic bacteria may periodically leave the biofilm on their own. When they do, they can rapidly multiply and disperse.

According to Costerton, there is a natural pattern of programmed detachment of planktonic cells from biofilms. This means that biofilms can act as what Costerton refers to as “niduses†of acute infection. Because the bacteria in a biofilm are protected by a matrix, the host immune system is less likely to mount a response to their presence.[1]

But if planktonic bacteria are periodically released from the biofilms, each time single bacterial forms enter the tissues, the immune system suddenly becomes aware of their presence. It may proceed to mount an inflammatory response that leads to heightened symptoms. Thus, the periodic release of planktonic bacteria from some biofilms may be what causes many chronic relapsing infections.

Planktonic bacteria are periodically released from a biofilmAs R. Parsek of Northwestern University describes in a 2003 paper in the Annual Review of Microbiology, any pathogen that survives in a chronic form benefits by keeping the host alive.[8] After all, if a chronic bacterial form simply kills its host, it will no longer have a place to live. So according to Parsek, chronic infection often results in a “disease stalemate†where bacteria of moderate virulence are somewhat contained by the defenses of the host. The infectious agents never actually kill the host, but the host is never able to fully kill the invading pathogens either.

Parsek believes that the optimal way for bacteria to survive under such circumstances is in a biofilm, stating that “Increasing evidence suggests that the biofilm mode of growth may play a key role in both of these adaptations. Biofilm growth increases the resistance of bacteria to killing and may make organisms less conspicuous to the immune system… ultimately this moderation of virulence may serve the bacteria’s interest by increasing the longevity of the host.â€

The acceptance of biofilms as infectious entities

Anton van Leeuwenhoek.Perhaps because many biofilms are sufficiently thick to be visible to the naked eye, the microbial communities were among the first to be studied by early microbiologists. Anton van Leeuwenhoek scraped the plaque biofilm from his teeth and observed what he described as the “animalculi†inside them under his primitive microscope. However, according to Costerton and team at the Center for Biofilm Research at Montana State University, it was not until the 1970s that scientists began to appreciate that bacteria in the biofilm mode of existence constitute such a major component of the bacterial biomass in most environments. Then, it was not until the 1980s and 1990s that scientists truly began to understand how elaborately organized a bacterial biofilm community can be.[1]

As Kolter, professor of microbiology and molecular genetics at Harvard Medical School, and one of the first scientists to study how biofilms developstates, “At first, however, studying biofilms was a radical departure from previous work.â€

Like most microbial geneticists, Kolter had been trained in the tradition dating back to Koch and Louis Pasteur, namely that bacteriology is best conducted by studying pure strains of planktonic bacteria. “While this was a tremendous advance for modern microbiology, it also distracted microbiologists from a more organismic view of bacteria, Kolter adds, “Certainly we felt that pure, planktonic cultures were the only way to work. Yet in nature bacteria don’t live like that,†he says. “In fact, most of them occur in mixed, surface-dwelling communities.â€

Although research on biofilms has surged over the past few decades, the majority of biofilm research to date has focused on external biofilms, or those that form on various surfaces in our natural environment.

Over the past years, as scientists developed better tools to analyze external biofilms, they quickly discovered that biofilms can cause a wide range of problems in industrial environments. For example, biofilms can develop on the interiors of pipes, which can lead to clogging and corrosion. Biofilms on floors and counters can make sanitation difficult in food preparation areas.

Since biofilms have the ability to clog pipes, watersheds, storage areas, and contaminate food products, large companies with facilities that are negatively impacted by their presence have naturally taken an interest in supporting biofilm research, particularly research that specifies how biofilms can be eliminated.

This means that many recent advances in biofilm detection have resulted from collaborations between microbial ecologists, environmental engineers, and mathematicians. This research has generated new analytical tools that help scientists identify biofilms.

Biofilm in a swamp gas reactor.For example, the Canadian company FAS International Ltd. has justcreated an endoluminal brush, which will be launched this spring. Physicians can use the brush to obtain samples from the interior of catheters. Samples taken from catheters can be sent to a lab, where researchers determine if biofilms are present in the sample. If biofilms are detected, the catheter is immediately replaced, since the insertion of catheters with biofilms can cause the patient to suffer from numerous infections, some of which are potentially life threatening.

Scientists now realize that biofilms are not just composed of bacteria. Nearly every species of microorganism – including viruses, fungi, and Archaea – have mechanisms by which they can adhere to surfaces and to each other. Furthermore, it is now understood that biofilms are extremely diverse. For example, upward of 300 different species of bacteria can inhabit the biofilms that form dental plaque.[9]

Furthermore, biofilms have been found literally everywhere in nature, to the point where any mainstream microbiologist would acknowledge that their presence is ubiquitous. They can be found on rocks and pebbles at the bottom of most streams or rivers and often form on the surface of stagnant pools of water. In fact, biofilms are important components of food chains in rivers and streams and are grazed upon by the aquatic invertebrates upon which many fish feed. Biofilms even grow in the hot, acidic pools at Yellowstone National Park and on glaciers in Antarctica.

Biofilm in acidic pools at Yellowstone National Park.It is also now understood that the biofilm mode of existence has been around for millenia. For example, filamentous biofilms have been identified in the 3.2-billion-year-old deep-sea hydrothermal rocks of the Pilbara Craton, Australia. According to a 2004 article in Nature Reviews Microbiology, “Biofilm formation appears early in the fossil record (approximately 3.25 billion years ago) and is common throughout a diverse range of organisms in both the Archaea and Bacteria lineages. It is evident that biofilm formation is an ancient and integral component of the prokaryotic life cycle, and is a key factor for survival in diverse environments.â€[10]

Biofilms and diseaseThe fact that external biofilms are ubiquitous raises the question – if biofilms can form on essentially every surface in our external environments, can they do the same inside the human body? The answer seems to be yes, and over the past few years, research on internal biofilms has finally started to pick up pace. After all, it’s easy for biofilm researchers to see that the human body, with its wide range of moist surfaces and mucosal tissue, is an excellent place for biofilms to thrive. Not to mention the fact that those bacteria which join a biofilm have a significantly greater chance of evading the battery of immune system cells that more easily attack planktonic forms.

Many would argue that research on internal biofilms has been largely neglected, despite the fact that bacterial biofilms seem to have great potential for causing human disease.

Common sites of biofilm infection. One biofilm reach the bloodstream they can spread to any moist surface of the human body. Stoodley of the Center for Biofilm Engineering at Montana State University, attributes much of the lag in studying biofilms to the difficulties of working with heterogeneous biofilms compared with homogeneous planktonic populations. In a 2004 paper in Nature Reviews, the molecular biologist describes many reasons why biofilms are extremely difficult to culture, such as the fact that the diffusion of liquid through a biofilm and the fluid forces acting on a biofilm must be carefully calculated if it is to be cultured correctly. According to Stoodley, the need to master such difficult laboratory techniques has deterred many scientists from attempting to work with biofilms. [10]

Also, since much of the technology needed to detect internal biofilms was created at the same time as the sequencing of the human genome, interest in biofilm bacteria, and the research grants that would accompany such interest, have been largely diverted to projects with a decidedly genetic focus. However, since genetic research has failed to uncover the cause of any of the common chronic diseases, biofilms are finally – just over the past few years – being studied more intensely, and being given the credit they deserve as serious infectious entities, capable of causing a wide array of chronic illnesses.

In just a short period of time, researchers studying internal biofilms have already pegged them as the cause of numerous chronic infections and diseases, and the list of illnesses attributed to these bacterial colonies continues to grow rapidly.

According to a recent public statement from the National Institutes of Health, more than 65% of all microbial infections are caused by biofilms. This number might seem high, but according to Kim of the Department of Chemical and Biological Engineering at Tufts University, “If one recalls that such common infections as urinary tract infections (caused by E. coli and other pathogens), catheter infections (caused by Staphylococcus aureus and other gram-positive pathogens), child middle-ear infections (caused by Haemophilus influenzae, for example), common dental plaque formation, and gingivitis, all of which are caused by biofilms, are hard to treat or frequently relapsing, this figure appears realistic.â€[11]

Hundreds of microbial biofilm colonize the human mouth, causing tooth decay and gum disease.As mentions, perhaps the most well-studied biofilms are those that make up what is commonly referred to as dental plaque. “Plaque is a biofilm on the surfaces of the teeth,†states Parsek. “This accumulation of microorganisms subject the teeth and gingival tissues to high concentrations of bacterial metabolites which results in dental disease.â€[12]

It has also recently been shown that biofilms are present on the removed tissue of 80% of patients undergoing surgery for chronic sinusitis. According to Parsek, biofilms may also cause osteomyelitis, a disease in which the bones and bone marrow become infected. This is supported by the fact that microscopy studies have shown biofilm formation on infected bone surfaces from humans and experimental animal models. Parsek also implicates biofilms in chronic prostatitis since microscopy studies have also documented biofilms on the surface of the prostatic duct. Microbes that colonize vaginal tissue and tampon fibers can also form into biofilms, causing inflammation and disease such as Toxic Shock Syndrome.

Biofilms also cause the formation of kidney stones. The stones cause disease by obstructing urine flow and by producing inflammation and recurrent infection that can lead to kidney failure. Approximately 15%–20% of kidney stones occur in the setting of urinary tract infection. According to Parsek, these stones are produced by the interplay between infecting bacteria and mineral substrates derived from the urine. This interaction results in a complex biofilm composed of bacteria, bacterial exoproducts, and mineralized stone material.

Microbes that colonize vaginal tissue and tampon fibers can become pathogenic, causing inflammation and disease such as Toxic Shock Syndrome.Perhaps the first hint of the role of bacteria in these stones came in 1938 when Hellstrom examined stones passed by his patients and found bacteria embedded deep inside them. Microscopic analysis of stones removed from infected patients has revealed features that characterize biofilm growth. For one thing, bacteria on the surface and inside the stones are organized in microcolonies and surrounded by a matrix composed of crystallized (struvite) minerals.

Then there’s endocarditis, a disease that involves inflammation of the inner layers of the heart. The primary infectious lesion in endocarditis is a complex biofilm composed of both bacterial and host components that is located on a cardiac valve. This biofilm, known as a vegetation, causes disease by three basic mechanisms. First, the vegetation physically disrupts valve function, causing leakage when the valve is closed and inducing turbulence and diminished flow when the valve is open. Second, the vegetation provides a source for near-continuous infection of the bloodstream that persists even during antibiotic treatment. This causes recurrent fever, chronic systemic inflammation, and other infections. Third, pieces of the infected vegetation can break off and be carried to a terminal point in the circulation where they block the flow of blood (a process known as embolization). The brain, kidney, and extremities are particularly vulnerable to the effects of embolization.

A variety of pathogenic biofims are also commonly found on medical devices such as joint prostheses and heart valves. According to Parsek, electron microscopy of the surfaces of medical devices that have been foci of device-related infections shows the presence of large numbers of slime-encased bacteria. Tissues taken from non-device-related chronic infections also show the presence of biofilm bacteria surrounded by an exopolysaccharide matrix. These biofilm infections may be caused by a single species or by a mixture of species of bacteria or fungi.

According to Dr. Patel of the Mayo Clinic, individuals with prosthetic joints are often oblivious to the fact that their prosthetic joints harbor biofilm infections.[13]

Cells of Staphylococcus epidermidis causing devastating disease as they grow on the cuff at a mechanical heart valve.“When people think of infection, they may think of fever or pus coming out of a wound,†explains Dr. Patel. “However, this is not the case with prosthetic joint infection. Patients will often experience pain, but not other symptoms usually associated with infection. Often what happens is that the bacteria that cause infection on prosthetic joints are the same as bacteria that live harmlessly on our skin. However, on a prosthetic joint they can stick, grow and cause problems over the long term. Many of these bacteria would not infect the joint were it not for the prosthesis.â€

Biofilms also cause Leptospirosis, a serious but neglected emerging disease that infects humans through contaminated water. New research published in the May issue of the journal Microbiology shows for the first time how bacteria that cause the disease survive in the environment.

Leptospirosis is a major public health problem in southeast Asia and South America, with over 500,000 severe cases every year. Between 5% and 20% of these cases are fatal. Rats and other mammals carry the disease-causing pathogen Leptospira interrogans in their kidneys. When they urinate, they contaminate surface water with the bacteria, which can survive in the environment for long periods.

“This led us to see if the bacteria build a protective casing around themselves for protection,†said Professor Mathieu Picardeau from the Institut Pasteur in Paris, France. [14]

Previously, scientists believed the bacteria were planktonic. But Professor Picardeau and his team have shown that L. interrogans can make biofilms, which could be one of the main factors controlling survival and disease transmission. “90% of the species of Leptospira we tested could form biofilms. It takes L. interrogans an average of 20 days to make a biofilm,†says Picardeau.

Biofilms have also been implicated in a wide array of veterinary diseases. For example, researchers at the Virginia-land Regional College of Veterinary Medicine at Virginia Tech were just awarded a grant from the United States Department of Agriculture to study the role biofilms play in the development of Bovine Respiratory Disease Complex (BRDC). If biofilms play a role in bovine respiratory disease, it’s likely only a matter of time before they will be established as a cause of human respiratory diseases as well.

When the immune response is compromised, Pseudomonas aeruginosabiofilms are able to colonize the alveoli, and to form biofilms.As mentioned previously, infection by the bacterium Pseudomonas aeruginosa (P. aeruginosa) is the main cause of death among patients with cystic fibrosis. Pseudomonas is able to set up permanent residence in the lungs of patients with cystic fibrosis where, if you ask most mainstream researchers, it is impossible to kill. Eventually, chronic inflammation produced by the immune system in response to Pseudomonas destroys the lung and causes respiratory failure. In the permanent infection phase, P. aeruginosa biofilms are thought to be present in the airway, although much about the infection pathogenesis remains unclear.[15]

Cystic fibrosis is caused by mutations in the proteins of channels that regulates chloride. How abnormal chloride channel protein leads to biofilm infection remains hotly debated. It is clear, however, that cystic fibrosis patients manifest some kind of host-defense defect localized to the airway surface. Somehow this leads to a debilitating biofilm infection.

Biofilms have the potential to cause a tremendous array of infections and diseasesBecause internal pathogenic biofilm research comprises such a new field of study, the infections described above almost certainly represent just the tip of the iceberg when it comes to the number of chronic diseases and infections currently caused by biofilms.

For example, it wasn’t until July of 2006 that researchers realized that the majority of ear infections are caused by biofilm bacteria. These infections, which can be either acute or chronic, are referred to collectively as otitis media (OM). They are the most common illness for which children visit a physician, receive antibiotics, or undergo surgery in the United States.

There are two subtypes of chronic OM. Recurrent OM (ROM) is diagnosed when children suffer repeated infections over a span of time and during which clinical evidence of the disease resolves between episodes. Chronic OM with effusion is diagnosed when children have persistent fluid in the ears that lasts for months in the absence of any other symptoms except conductive hearing loss.

It took over ten years for researchers to realize that otitis media is caused by biofilms. Finally, in 2002, Drs. Ehrlich and J. Post, an Allegheny General Hospital pediatric ear specialist and medical director of the Center for Genomic Sciences, published the first animal evidence of biofilms in the middle ear in the Journal of the American Medical Association, setting the stage for further clinical investigation.

In a subsequent study, Ehrlich and Post obtained middle ear mucosa – or membrane tissue – biopsies from children undergoing a procedure for otitis. The team gathered uninfected mucosal biopsies from children and adults undergoing cochlear implantation as a control.[16]

Using advanced confocal laser scanning microscopy, Luanne Hall Stoodley, Ph.D. and her ASRI colleagues obtained three dimensional images of the biopsies and evaluated them for biofilm morphology using generic stains and species-specific probes for Haemophilus influenzae, Streptococcus pneumoniaeand Moraxella catarrhalis. Effusions, when present, were also evaluated for evidence of pathogen specific nucleic acid sequences (indicating presence of live bacteria).

The study found mucosal biofilms in the middle ears of 46/50 children (92%) with both forms of otitis. Biofilms were not observed in eight control middle ear mucosa specimens obtained from cochlear implant patients.

Otitis media, or inflammation of the inner ear, is caused by biofilm.In fact, all of the children in the study who suffered from chronic otitis media tested positive for biofilms in the middle ear, even those who were asymptomatic, causing Erlich to conclude that, “It appears that in many cases recurrent disease stems not from re-infection as was previously thought and which forms the basis for conventional treatment, but from a persistent biofilm.â€

He went on to state that the discovery of biofilms in the setting of chronic otitis media represented “a landmark evolution in the medical community’s understanding about a disease that afflicts millions of children world-wide each year and further endorses the emerging biofilm paradigm of chronic infectious disease.â€

The emerging biofilm paradigm of chronic disease refers to a new movement in which researchers such as Ehrlich are calling for a tremendous shift in the way the medical community views bacterial biofilms. Those scientists who support an emerging biofilm paradigm of chronic disease feel that biofilm research is of utmost importance because of the fact that the infectious entities have the potential to cause so many forms of chronic disease. The Marshall Pathogenesis is an important part of this paradigm shift.

It was also just last year that researchers realized that biofilms cause most infections associated with contact lens use. In 2006, Bausch & Lomb withdrew its ReNu with MoistureLoc contact lens solution because a high proportion of corneal infections were associated with it. It wasn’t long before researchers at the University Hospitals Case Medical Center found that the infections were caused by biofilms. [17]

“Once they live in that type of state [a biofilm], the cells become resistant to lens solutions and immune to the body’s own defense system,†said Mahmoud A. Ghannoum, Ph.D, senior investigator of the study. “This study should alert contact lens wearers to the importance of proper care for contact lenses to protect against potentially virulent eye infections,†he said.

It turns out that the biofilms detected by Ghannoum and team were composed of fungi, particularly a species called Fusarium. His team also discovered that the strain of fungus (with the catchy name, ATCC 36031) used for testing the effectiveness of lens care solutions is a strain that does not produce biofilms as the clinical fungal strains do. ReNu contact solution, therefore, was effective in the laboratory, but failed when faced with strains in real-world situations.

Fungal biofilm can form in contact lens solution leading to potentially virulent eye infectionsUnfortunately, Ghannoum and team were not able to create a method to target and destroy the fungal biofilms that plague users of ReNu and some other contact lens solutions.

Then there’s Dr. Randall Wolcott who just recently discovered and confirmed that the sludge covering diabetic wounds is largely made up of biofilms. Whereas before Wolcott’s work such limbs generally had to be amputated, now that they have been correctly linked to biofilms, measures such as those described in thisinterview can be taken to stop the spread of infection and save the limb. Wolcott has finally been given a grant by the National Institutes of Health to further study chronic biofilms and wound development.

Dr. Garth and the Medical Biofilm Laboratory team at Montana State University are also researching wounds and biofilms. Their latest article and an image showing wound biofilm was featured on the cover of the January-February 2008 issue of Wound Repair and Regeneration.[18]

Biofilm bacteria and chronic inflammatory diseaseIn just a few short years, the potential of biofilms to cause debilitating chronic infections has become so clear that there is little doubt that biofilms are part of the pathogenic mix or “pea soup†that cause most or all chronic “autoimmune†and inflammatory diseases.

In fact, thanks, in large part, to the research of biomedical researcher Dr. Trevor Marshall, it is now increasingly understood that chronic inflammatory diseases result from infection with a large microbiota of chronic biofilm and L-form bacteria (collectively called the Th1 pathogens).[19][20] The microbiota is thought to be comprised of numerous bacterial species, some of which have yet to be discovered. However, most of the pathogens that cause inflammatory disease have one thing in common – they have all developed ways to evade the immune system and persist as chronic forms that the body is unable to eliminate naturally.

Some L-form bacteria are able to evade the immune system because, long ago, they evolved the ability to reside inside macrophages, the very white bloods cells of the immune system that are supposed to kill invading pathogens. Upon formation, L-form bacteria also lose their cell walls, which makes them impervious to components of the immune response that detect invading pathogens by identifying the proteins on their cell walls. The fact that L-form bacteria lack cell walls also means that the beta-lactam antibiotics, which work by targeting the bacterial cell wall, are completely ineffective at killing them.[21]

Clearly, transforming into the L-form offers any pathogen a survival advantage. But among those pathogens not in an L-form state, joining a biofilm is just as likely to enhance their ability to evade the immune system. Once enough chronic pathogens have grouped together and formed a stable community with a strong protective matrix, they are likely able to reside in any area of the body, causing the host to suffer from chronic symptoms that are both mental and physical in nature.

Biofilm researchers will also tell you that, not surprisingly, biofilms form with greater ease in an immunocompromised host. Marshall’s research has made it clear that many of the Th1 pathogens are capable of creating substances that bind and inactivate the Vitamin D Receptor – a fundamental receptor of the body that controls the activity of the innate immune system, or the body’s first line of defense against intracellular infection.[22]

Diagram of the Vitamin D Receptor and capnine.Thus, as patients accumulate a greater number of the Th1 pathogens, more and more of the chronic bacterial forms create substances capable of disabling the VDR. This causes a snowball effect, in which the patient becomes increasingly immunocompromised as they acquire a larger bacterial load.

For one thing, it’s possible that many of the bacteria that survive inside biofilms are capable of creating VDR blocking substances. Thus, the formation of biofilms may contribute to immune dysfunction. Conversely, as patients acquire L-form bacteria and other persistent bacterial forms capable of creating VDR-blocking substances, it becomes exceptionally easy for biofilms to form on any tissue surface of the human body.

Thus, patients who begin to acquire L-form bacteria almost always fall victim to biofilm infections as well, since it is all too easy for pathogens to group together into a biofilm when the immune system isn’t working up to par.

To date, there is also no strict criteria that separate L-form bacteria from biofilm bacteria or any other chronic pathogenic forms. This means that L-form bacteria may also form into biofilms, and by doing so enter a mode of survival that makes them truly impervious to the immune system. Some L-form bacteria may not form complete biofilms, yet may still possess the ability to surround themselves in a protective matrix. Under these circumstances one might say they are in a “biofilm-like†state.

Marshall often refers to the pathogens that cause inflammatory disease as an intraphgocytic, metagenomic microbiota of bacteria, terms which suggest that most chronic bacterial forms possess properties of both L-form and biofilm bacteria. Intraphagocytic refers to the fact that the pathogens can be found inside the cells of the immune system. The term metagenomic indicates that there are a tremendous number of different species of these chronic bacterial forms. Finally, microbiota refers to the fact that biofilm communities sustain their pathogenic activity.

For example, when observed under a darkfield microscope, L-form bacteria are often encased in protective biofilm sheaths. If the blood containing the pathogens are aged overnight, the bacterial colonies reach a point where they expand and burst out of the cell, causing the cell to burst as well. Then they extend as huge, long biofilm tubules, which are presumably helping the pathogens spread to other cells. The tubules also help spread bacterial DNA to neighboring cells.

Clearly, there is a great need for more research on how different chronic bacterial forms interact. To date, L-form researchers have essentially focused soley on the L-form, while failing to investigate how frequently the wall-less pathogens form into biofilms or become parts of biofilm communities together with bacteria with cell walls. Conversely, most biofilm researchers are intently studying the biofilm mode of growth without considering the presence of L-form bacteria. So, it will likely take several years before we will be better able to understand probable overlaps between the lifestyles of L-form and biofilm bacteria.

Anyone who is skeptical about the fact that biofilms likely form a large percentage of the microbiota that cause inflammatory disease should consider many of the recent studies that have linked established biofilm infections to a higher risk for multiple forms of chronic inflammatory disease. Take, for example, studies that have found a link between periodontal disease and several major inflammatory conditions. A 1989 article published in British Medical Journal showed a correlation between dental disease and systemic disease (stroke, heart disease, diabetes). After correcting for age, exercise, diet, smoking, weight, blood cholesterol level, alcohol use and health care, people who had periodontal disease had a significantly higher incidence of heart disease, stroke and premature death. More recently, these results were confirmed in studies in the United States, Canada, Great Britain, Sweden, and Germany. The effects are striking. For example, researchers from the Canadian Health Bureau found that people with periodontal disease had a two times higher risk of dying from cardiovascular disease.[23]

Dental plaque as seen under a scanning electron microcroscope.Since we know that periodontal disease is caused by biofilm bacteria, the most logical explanation for the fact that people with dental problems are much more likely to suffer from heart disease and stroke is that the biofilms in their mouths have gradually spread to the moist surfaces of their circulatory systems. Or perhaps if the bacteria in periodontal biofilms create VDR binding substances, their ability to slow innate immune function allows new biofilms (and L-form bacteria as well) to more easily form and infect the heart and blood vessels. Conversely, systemic infection with VDR blocking biofilm bacteria is also likely to weaken immune defenses in the gums and facilitate periodontal disease.

In fact, it appears that biofilm bacteria in the mouth also facilitate the formation of biofilm and L-form bacteria in the brain. Just last year, researchers at Vasant Hirani at University College London released the results of a study which found that elderly people who have lost their teeth are at more than three-fold greater risk of memory problems and dementia.[24]

At the moment, Autoimmunity Research Foundation does not have the resources to culture biofilms from patients on the treatment and, even if they did, current methods for culturing internal biofilms remain unreliable. According to Stoodley, “The lack of standard methods for growing, quantifying and testing biofilms in continuous culture results in incalculable variability between laboratory systems. Biofilm microbiology is complex and not well represented by flask cultures. Although homogeneity allows statistical enumeration, the extent to which it reflects the real, less orderly world is questionable.â€[10]

How else do we acquire biofilm bacteria?As discussed thus far, biofilms form spontaneously as bacteria inside the human body group together. Yet people can also ingest biofilms by eating contaminated food.

According to researchers at the University of Guelph in Ontario Canada, it is increasingly suspected that biofilms play an important role in contamination of meat during processing and packaging. The group warns that greater action must be taken to reduce the presence of food-borne pathogens like Escherichia coli and Listeria monocytogenes and spoilage microorganisms such as thePseudomonas species (all of which form biofilms) throughout the food processing chain to ensure the safety and shelf-life of the product. Most of these microorganisms are ubiquitous in the environment or brought into processing facilities through healthy animal carriers.

Hans Blaschek of the University of Illinois has discovered that biofilms form on much of the other food products we consume as well.

A biofilm on a piece of lettuce“If you could see a piece of celery that’s been magnified 10,000 times, you’d know what the scientists fighting foodborne pathogens are up against,†says Blaschek.

“It’s like looking at a moonscape, full of craters and crevices. And many of the pathogens that cause foodborne illness, such as Shigella, E. coli,and Listeria, make sticky, sugary biofilms that get down in these crevices, stick like glue, and hang on like crazy.â€

According to Blaschek, the problem faced by produce suppliers can be a triple whammy. “If you’re unlucky enough to be dealing with a pathogen–and the pathogen has the additional attribute of being able to form biofilm—and you’re dealing with a food product that’s minimally processed, well, you’re triply unlucky,†the scientist said. “You may be able to scrub the organism off the surface, but the cells in these biofilms are very good at aligning themselves in the subsurface areas of produce.â€

, a University of Illinois food science and human nutrition professor agrees, stating,â€Once the pathogenic organism gets on the product, no amount of washing will remove it. The microbes attach to the surface of produce in a sticky biofilm, and washing just isn’t very effective.â€

Biofilms can even be found in processed water. Just this month, a study was released in which researchers at the Department of Biological Sciences, at Virginia Polytechnic Institute isolated M. avium biofilm from the shower head of a woman with M. avium pulmonary disease.[25] A molecular technique called DNA fingerprinting demonstrated that M. avium isolates from the water were the same forms that were causing the woman’s respiratory illness.

Effectively targeting biofilm infectionsAlthough the mainstream medical community is rapidly acknowledging the large number of diseases and infections caused by biofilms, most researchers are convinced that biofilms are difficult or impossible to destroy, particularly those cells that form the deeper layers of a thick biofilm. Most papers on biofilms state that they are resistant to antibiotics administered in a standard manner. For example, despite the fact that Ehrlich and team discovered that biofilm bacteria cause otitis media, they are unable to offer an effective solution that would actually allow for the destruction of biofilms in the ear canal. Other teams have also come up short in creating methods to break up the biofilms they implicate as the cause of numerous infections.

This means patients with biofilm infections are generally told by mainstream doctors that they have an untreatable infection. In some cases, a disease-causing biofilm can be cut out of a patient’s tissues, or efforts are made to drain components of the biofilm out of the body. For example, doctors treating otitis media often treats patients with myringotomy, a surgical procedure in which small tubes are placed in the eardrum to continuously drain infectious fluid.

When it comes to administering antibiotics in an effort to target biofilms, one thing is certain. Mainstream researchers have repeatedly tried to kill biofilms by giving patients high, constant doses of antibiotics. Unfortunately, when administered in high doses, the antibiotic may temporarily weaken the biofilm but is incapable of destroying it, as certain cells inevitably persist and allow the biofilm to regenerate.

“You can put a patient on [a high dose] antibiotics, and it may seem that the infection has disappeared,â€Â says Levchenko. “But in a few months, it reappears, and it is usually in an antibiotic-resistant form.â€

What the vast majority of researchers working with biofilms fail to realize is that antibiotics are capable of destroying biofilms. The catch is that antibiotics are only effective against biofilms if administered in a very specific manner. Furthermore, only certain antibiotics appear to effectively target biofilms. After decades of research, much of which was derived from molecular modeling data, Marshall was the first to create an antibiotic regimen that appears to effectively target and destroy biofilms. Central to the treatment, which is called the Marshall Protocol, is the fact that biofilms and other Th1 pathogens succumb to specific bacteriostatic antibiotics taken in very low, pulsed doses. It is only when antibiotics are administered in this manner that they appear capable of fully eradicating biofilms.[19][20]

In a paper entitled “The Riddle of Biofilm Resistance,†Dr. Kim of Tulane University discusses the mechanisms by which pulsed, low dose antibiotics are able to break up biofilms, while antibiotics administered in a standard manner (high, constant doses) cannot. According to , the use of pulsed, low-dose antibiotics to target biofilm bacteria is supported by observations she and her colleagues have made in the laboratory.[11]

Some researchers claim that antibiotics cannot penetrate the matrix that surrounds a biofilm. But research by and other scientists has confirmed that the inability of antibiotics to penetrate the biofilm matrix is much more of an exception than a rule. According to , “In most cases involving small antimicrobial molecules, the barrier of the polysaccharide matrix should only postpone the death of cells rather than afford useful protection.â€

For example, a recent study that used low concentrations of an antibiotic to killP. aeruginosa biofilm bacteria found that the majority of biofilm cells were effectively eliminated by antibiotics in a manner that did not differ much from what is observed when the same antibiotic concentrations are administered to single planktonic cells.[26]

After antibiotics are applied to a biofilm, a number of cells called “persisters†are left behind.Thus, since antibiotics can generally penetrate biofilms, some other factor is responsible for the fact that they cannot be killed by standard high dose antibiotic therapy. It turns out that after antibiotics are applied to a biofilm, a number of cells called “persisters†are left behind. Persisters are simply cells that are able to survive the first onslaught of antibiotics, and if left unchecked, gradually allow the biofilm to form again. According to , persister cells form with particular ease in immunocompromised patients because the immune system is unable to help the antibiotic “mop up†all the biofilm cells it has targeted.

“This simple observation suggests a new paradigm for explaining, at least in principle, the phenomenon of biofilm resistance to killing by a wide range of antimicrobials,†states . “The majority of cells in a biofilm are not necessarily more resistant to killing than planktonic cells and die rapidly when treated with [an antibiotic] that can kill slowly growing cells.â€

Thus, a dose of antibiotics – particularly in immunocompromised patients – eradicates most of the biofilm population but leaves a small fraction of surviving persisters behind. Unfortunately, in the same sense that the beta-lactam antibiotics promote the formation of L-form bacteria, persister cells are actually preserved by the presence of an antibiotic that inhibits their growth. Thus, paradoxically, dosing an antibiotic in a constant, high-dose manner (in which the antibiotic is always present) helps persisters persevere.

But in the case of low, pulsed dosing, where an antibiotic is administered, withdrawn, then administered again, the first application of antibiotic will eradicate the bulk of biofilm cells, leaving persister cells behind. Withdrawl of the antibiotic allows the persister population to start growing. Since administration of the antibiotic is temporarily stopped, the survival of persisters is not enhanced. This causes the persister cells to lose their phenotype (their shape and biochemical properties), meaning that they are unable to switch back into biofilm mode. A second application of the antibiotic should then completely eliminate the persister cells, which are still in planktonic mode.

has found that the feasibility of a pulsed, or cyclical biofilm eradication approach depends on the rate at which persisters lose resistance to killing and regenerate new persisters. It also depends on the ability to manipulate the antibiotic concentration – something that is done quite effectively by patients on the Marshall Protocol who carefully dose their antibiotics at different levels, allowing constant variation in antibiotic concentration. Although speculates that allowing the concentration of an antibiotic to drop could potentially lead to resistance towards the antibiotic, she is quick to add that if two or more antibiotics are used to target a biofilm at one time, such resistance would not occur. Again, since the Marshall Protocol uses a total of five bacteriostatic antibiotics, usually taken two or three at a time, concerns of resistance are essentially negligible.

Model of biofilm resistance based on persister survival. An initial treatment of high-dose constant antibiotic kills planktonic cells and the majority of biofilm cells. But persisters remain alive and resurrect the biofilm, causing the infection to relapse

“It is entirely possible that successful cases of antimicrobial therapy of biofilm infections result from a fortuitous optimal cycling [pulsed dosing] of an antibiotic concentration that eliminated first the bulk of the biofilm and then the progeny of the persisters that began to divide,†states .

’ work has been supported by other research teams. Recently, researchers at the University of Iowa found that subinhibitory (extremely low dose) concentrations of the bacteriostatic antibiotic azithromycin significantly decreased biomass and maximal thickness in both forming and established biofilms.[27] These extremely low concentrations of azithromycin inhibited biofilms in all but the most highly resistant isolates. In contrast, subinhibitory concentrations of gentamicin, which is not a bacteriostatic antibiotic, had no effect on biofilm formation. In fact, biofilms actually became resistant to gentamicin at concentrations far above the minimum inhibitory concentration.

Researchers at Tulane University recently confirmed yet again that low, pulsed dosing is a superior way of targeting treatment-resistant biofilm bacteria. According to the team, who mathematically modeled the action of antibiotics on bacterial biofilms, “Exposing a biofilm to low concentration doses of an antimicrobial agent for longer time is more effective than short time dosing with high antimicrobial agent concentration.â€[28]

Similarly, a bioengineer led team at the University of Washington recently created an antibiotic- containing polymer that releases antibiotic slowly onto the surface of hospital devices, such as catheters and prostheses, to reduce the risk of biofilm-related infections.

“Rather than massively dosing the patient with high levels of released antibiotic, this strategy allows the release of extremely low levels of this very potent antibiotic over long periods of time,â€Â explained Buddy Ratner, PhD, Professor and Director of the Engineered Biomaterials Program at the University of Washington, Seattle. “We calculated the amount released at the surface that would kill 100% of the bacteria entering the surface zone.â€

When challenged by Dr. Leonard A. Mermel from Brown University School of Medicine on the issue that long-term use of pulsed, low-dose antibiotics might allow for increased resistance on the part of the bacteria being treated, Ratner responded, “Dr. Mermel’s concerns are, in fact, why we developed this system for [antibiotic] release. Bacteria that live through antibiotic dosing can go on to produce resistant strains. If 100% of the bacteria approaching the surface are killed, they can’t produce resistant offspring. The classical physician approach, dosing the patient systemically and heavily to rid the patient of persistent bacteria, can lead to those resistant strains. Our approach releases miniscule doses compared to what a physician would use, but releases the antibiotic where it will be optimally effective and least likely to leave antibiotic-resistant survivors.â€

Although taken orally, the MP antibiotics are taken in the same manner as those administered by Ratner and team. Because they too are dosed at optimal times in extremely small doses, the chance that long-term antibiotic use might foster resistant bacteria is again, essentially negligible, especially when multiple antibiotics are typically used.

Key to the ability of the Marshall Protocol to effectively target biofilm bacteria is the fact that the specific pulsed, low-dose bacteriostatic antibiotics used by the treatment are taken in conjunction with a medication called Benicar. Benicar binds and activates the Vitamin D Receptor, displacing bacterial substances and 25-D from the receptor, so that it can once again activate the innate immune system.[29] Benicar is so effective at strengthening the innate immune response that the patient’s own immune system ultimately helps destroy the biofilm weakened by pulsed, low-dose antibiotics.

Thus, it is not enough for patients on the Marshall Protocol to simply take specific pulsed, low-dose antibiotics. The activity of their innate immune system must also be restored so that the cells of the immune system can actively combat biofilm bacteria, the matrix that surrounds them, and persister cells.

After antibiotics are applied to a biofilm, a number of cells called “persisters†are left behindHow do we know that the Marshall Protocol effectively kills biofilm bacteria? Namely because those patients to reach the later stages of the treatment do not report symptoms associated with established biofilm diseases. Patients on the MP who once suffered from chronic ear infections (OM), chronic sinus infections, or periodontal disease find that such infections resolve over the course of treatment. Furthermore, since we now understand that biofilms almost certainly form a large part of the chronic microbiota of pathogens that cause chronic inflammatory and autoimmune diseases, the fact that patients can use the Marshall Protocol to recover from such illnesses again suggests that the treatment must be effectively allowing them to target and destroy biofilms.

Because all evidence points to the fact that the MP does indeed effectively target biofilm bacteria, it is of utmost importance that people who suffer from any sort of biofilm infection start the treatment. Knowledge of the Marshall Protocol has yet to reach the cystic fibrosis community, but there is great hope that if people with the disease were to start the MP, they could destroy the P. aeruginosabiofilms that cause their untimely deaths. In the same vein, people with a wide range of infections, such as those infected with biofilm during surgery, can likely restore their health with the MP.

It is to be hoped that the clinical data emerging from the Marshall Protocol study site, which shows patients recovering from biofilm-related diseases, will inspire future researchers to invest a great deal of energy into further research aimed at identifying and studying the biofilm bacteria – bacteria that almost certainly form part of the microbiota of pathogens that cause inflammatory disease. In the coming years, as the technology to detect biofilms becomes even more sophisticated, it is almost certain that a great number of biofilms will be officially detected and documented in patients with a vast array of chronic diseases.

REFERENCESCosterton, J. W., , P. S., & Greenberg, E. P. (1999). Bacterial biofilms: a common cause of persistent infections. Science (New York, N.Y.), 284(5418), 1318-22. [↩] [↩] [↩] [↩]

Higgins, D. A., Pomianek, M. E., Kraml, C. M., , R. K., Semmelhack, M. F., & Bassler, B. L. (2007). The major Vibrio cholerae autoinducer and its role in virulence factor production. Nature, 450(7171), 883-6. [↩]

Singh, P. K., Schaefer, A. L., Parsek, M. R., Moninger, T. O., Welsh, M. J., & Greenberg, E. P. (2000). Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. Nature, 407(6805), 762-4. [↩] [↩]

Stoodley, P., Purevdorj-Gage, B., & Costerton, J. W. (2005). Clinical significance of seeding dispersal in biofilms: a response. Microbiology, 151(11), 3453. [↩]

O’toole, G. A., & Kolter, R. (1998). Flagellar and Twitching Motility Are Necessary for Pseudomonas Aeruginosa Biofilm Development. Molecular Microbiology, 30(2), 295-304. [↩]

Cho, H., Jönsson, H., , K., Melke, P., , J. W., Jedynak, B., et al. (2007). Self-Organization in High-Density Bacterial Colonies: Efficient Crowd Control. PLoS Biology, 5(11), e302 EP -. [↩] [↩]

Brockhurst, M. A., Hochberg, M. E., Bell, T., & Buckling, A. (2006). Character displacement promotes cooperation in bacterial biofilms. Current biology: CB, 16(20), 2030-4. [↩]

Parsek, M. R., & Singh, P. K. (2003). Bacterial biofilms: an emerging link to disease pathogenesis. Annual review of microbiology, 57, 677-701. [↩]

Kraigsley, A., Ronney, P., & Finkel, S. Hydrodynamic effects on biofilm formation. Retrieved May 28, 2008. [↩]

Hall-Stoodley, L., Costerton, J. W., & Stoodley, P. (2004). Bacterial biofilms: from the Natural environment to infectious diseases. Nat Rev Micro, 2(2), 95-108. [↩] [↩] [↩]

, K. (2001). Riddle of biofilm resistance. Antimicrobial agents and chemotherapy, 45(4), 999-1007. [↩] [↩]

Parsek, M. R., & Singh, P. K. (2003). Bacterial biofilms: an emerging link to disease pathogenesis. Annual review of microbiology, 57, 677-701. [↩]

Trampuz, A., Piper, K. E., son, M. J., Hanssen, A. D., Unni, K. K., Osmon, D. R., et al. (2007). Sonication of Removed Hip and Knee Prostheses for Diagnosis of Infection. N Engl J Med, 357(7), 654-663. [↩]

Ristow, P., Bourhy, P., Kerneis, S., Schmitt, C., Prevost, M., Lilenbaum, W., et al. (2008). Biofilm formation by saprophytic and pathogenic leptospires. Microbiology, 154(5), 1309-1317. [↩]

Moreau-Marquis, S., Stanton, B. A., & O’Toole, G. A. (2008). Pseudomonas aeruginosa biofilm formation in the cystic fibrosis airway. Pulmonary pharmacology & therapeutics. [↩]

Hall-Stoodley, L., Hu, F. Z., Gieseke, A., Nistico, L., Nguyen, D., , J., et al. (2006). Direct Detection of Bacterial Biofilms on the Middle-Ear Mucosa of Children With Chronic Otitis Media.JAMA, 296(2), 202-211. [↩]

Imamura, Y., Chandra, J., Mukherjee, P. K., Lattif, A. A., Szczotka-Flynn, L. B., Pearlman, E., et al. (2008). Fusarium and Candida albicans Biofilms on Soft Contact Lenses: Model Development, Influence of Lens Type, and Susceptibility to Lens Care Solutions. Antimicrob. Agents Chemother., 52(1), 171-182. [↩]

, G. A., Swogger, E., Wolcott, R., Pulcini, E. D., Secor, P., Sestrich, J., et al. (2008).Biofilms in Chronic Wounds. Wound Repair and Regeneration, 16(1), 37-44. [↩]

Marshall, T. G. (2006b). A New Approach to Treating Intraphagocytic CWD Bacterial Pathogens in Sarcoidosis, CFS, Lyme and other Inflammatory Diseases. [↩] [↩]

Marshall, T. G., & Marshall, F. E. (2004). Sarcoidosis succumbs to antibiotics–implications for autoimmune disease. Autoimmunity reviews, 3(4), 295-300. [↩] [↩]

Sr, G. J. D., & Woody, H. B. (1997). Bacterial persistence and expression of disease. Clinical Microbiology Reviews, 10(2). [↩]

Marshall, T. G. (2007). Bacterial Capnine Blocks Transcription of Human Antimicrobial Peptides. Nature Precedings. [↩]

on, H. I., Ellison, L. F., & , G. W. (1999). Periodontal disease and risk of fatal coronary heart and cerebrovascular diseases. Journal of cardiovascular risk, 6(1), 7-11. [↩]

, R., & Hirani, V. (2007). Dental Health and Cognitive Impairment in an English National Survey Population. Journal of the American Geriatrics Society, 55(9), 1410-1414. [↩]

Falkinham Iii, J. O., Iseman, M. D., Haas, P. D., & Soolingen, D. V. (2008). Mycobacterium avium in a shower linked to pulmonary disease. Journal of water and health, 6(2), 209-13. [↩]

, K. (2001). Riddle of biofilm resistance. Antimicrobial agents and chemotherapy, 45(4), 999-1007. [↩]

Starner, D et al. 2008. Subinhibitory Concentrations of Azithromycin Decrease Nontypeable Haemophilus influenzae Biofilm Formation and Diminish Established Biofilms.Antimicrobial agents and chemotherapy 52(1):137-45. [↩]

Cogan, N. G., Cortez, R., & Fauci, L. (2005). Modeling physiological resistance in bacterial biofilms. Bulletin of mathematical biology, 67(4), 831-53. [↩]

Marshall, T. G. (2006). VDR Nuclear Receptor Competence is the Key to Recovery from Chronic Inflammatory and Autoimmune Disease. [↩]

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37 Responses for " Understanding Biofilms "

Ken Collerman May 29th, 2008 at 1:57 am1

Amy,This article is a masterpiece!Amy Proal May 29th, 2008 at 9:40 am2

Thanks Ken!It was a pleasure to write the article because I find the subject matter so fascinating!AmySherry June 2nd, 2008 at 4:54 pm3

Is it possible to kill biofilms on our food by heating? I am having second thoughts about eating any raw vegetables…The article is well researched, well written, and very insightful.Thank you!

Amy Proal June 2nd, 2008 at 5:00 pm4

Hi Sherry,I’m glad you enjoyed the article.I’m not sure about the answer to your question. The foods items that seem to be most affected by biofilm contamination are raw, and can’t be heated in order to remove biofilms.

I assume that if you had a food item and you heated it to high temperatures the high level of heat would have an effect on the homeostasis and potential survival of a biofilm, although I don’t know for sure.Are you on the MP? If so, you will almost certainly easily kill any biofilm bacteria that you might acquire by eating a raw vegetable before it is able to enter the bloodstream. So I wouldn’t worry about eating them.

Don’t forget that until we reach the late stages of the MP, our own bodies are already filled with bacteria. They cover our hands and entire skin surface. They are already in our mouth. So coming in contact with biofilm bacteria in the mouth isn’t all that different from what we experience on a daily basis. Of course, being on the MP is key, so that an ingested biofilm will not be able to cause any harm.

Best,Amy W. June 2nd, 2008 at 5:02 pm5

I wonder if there’s a way to interfere with the biofilm’s chemical signaling directly. It would require much more specific diagnosis of the underlying pathogens, or else a good general cocktail.Do biofilm-creating pathogens just travel together or do they ‘learn’ how to signal one another based on shared plasmids? If the second, the plasmids themselves might be chemically attacked.

Amy Proal June 2nd, 2008 at 5:30 pm6

Hi ,Yes, there are research teams who are working to try to interfere with the chemical signaling in a biofilm. In fact, I think Higgins and team who are working with CAI-1 are actively seeking ways to inhibit the signal in an effort to slow biofilm growth. If they succeed, and

other researchers successfully stop other biofilm signals, potential drugs that inhibit the signals they identify could form part of a larger arsenal to target biofilms. Of course, with any drug comes side effects, so people would have to weigh the pro/cons of taking drugs to interfere with biofilm signals, particularly when they can already target biofilms with just a few pulsed, low-dose antibiotics.

I imagine that groups of people with specific biofilm infections (such as cystic fibrosis patients with P. auruginosa infections), would most benefit from therapies aimed at stopping chemical signals that aid the formation of biofilms.

I think that the bacteria inside biofilms communicate in myriad ways, depending largely on the species present. So a lot of individual work with particular biofilms must be done to uncover ways to break up signaling pathways.

I’m not sure biofilm communication has much to do with sharing plasmids. Biofilm bacteria literally emit chemical signals (not plasmids) into the surrounding environment. These signals are picked up by other nearby bacteria. Depending on the strength of the signal they react in

different ways. For example, if a bacterium picks up a lot of other signals, it knows it is surrounded by a lot of other bacteria. That way it is able to “realize†that forming a biofilm with the nearby bacteria is a possibility. Other modes of signaling remain less clear as far as I can tell. I’m sure further research will turn up much more complex modes of action.

Best, W. June 3rd, 2008 at 1:09 am7

Hi Amy, you wrote;I’m not sure biofilm communication has to do with sharing plasmids. They literally emit chemical signals (not plasmids) into the surrounding environment.My point was that the bacteria had to have a common ‘language’ in order to communicate and I wondered if the key for that language was found in their chromosome or in a transmissable, extra-chromosomal unit (Kind of like giving a person a gene which lets them speak English so that you can talk to them.)

There was some possibility for developing drugs that targeted plasmids, so I thought that that might be a good way to break up a biofilm.

Amy Proal June 3rd, 2008 at 9:01 am8

Hi ,I see. It possible that some genes are embedded within plasmids and others within the genome. We’re dealing with a lot of different genes here so they may be in diverse locations. Hopefully future researchers projects will address this issue….

Dr.Abbas Ubaid Al_Janabi June 16th, 2008 at 8:29 am9

Thanks for this best artical about the biofilms,I am finsed my Ph.D.Microbial Biofilms and I havemany papears in this field ,So, Plz keep attach with me amy.All the best YoursDr. Abbas Al_JanabiCollege of Medicine, University of Al_Anbar,Ramadi,Iraq

Amy Proal June 16th, 2008 at 10:05 am10

Hi Dr. Abbas,I’m glad to hear that a biofilm expert enjoyed my article. Congrats on getting your PhD in biofilm-related research. I think you are pursuing a very interesting course of study that will be extremely relevant to the future of medicine.

Do you have a link to any of your published papers? I’d enjoy looking over them…Best,AmyBeth August 19th, 2008 at 11:38 pm11

AmyWe are doing a Nursing Grand Rounds which includes a segment on oral care. I would like to use some of your pictures. Can I have permission to do so?Thank you for your consideration.Beth

Amy Proal August 20th, 2008 at 7:50 am12

Hi Beth,Many of these images are from the Center for Biofilm Engineering in Montana. One of the missions of CBE is to promote the understanding of biofilms. The Center has a page on its website explaining acceptable reuse.

Best,Amy Rifkin, MD September 8th, 2008 at 5:22 pm13

Amy: Terrific article. I’ve been asked to give a talk on biofilms for our new Masters in Biotechnology course and I’d love to be able to use some of your slides (proper acknowledgement of course). Would that be possible. Thanks. GR

Amy Proal September 9th, 2008 at 1:54 am14

Hi ,Of course, you are welcome to use my content for the purposes of a presentation. Please note, as per comment #12, that when it comes to the images, you’re going to want to attribute the CBE in Montana.

If you haven’t done so already, you may also want to look at my 87-minute videoIntroduction to the Marshall Protocol.

Best,AmyAnusha December 2nd, 2008 at 7:12 pm15

Hi Amy,Thank you for this detailed report on biofilms. I am suspicious of biofilm contamination in my airway epithelial cell culture. I see a network of floating dead/dying epithelial cells on the top layer of cell culture medium and a translucent jelly-like strands that dont seem to be fungi. The culture is an established immortalized cell line derived from a patient’s lung biopsy. Is there any way to test for persistant contamination? Any stain or culture technique to detect presence of biofilms?

Thank you!Amy Proal December 3rd, 2008 at 1:05 pm16

Hi Anusha,There are tests that detect biofilms but they are not necessarily available in a standard lab. In order to test you samples, I think you would have to contact a researcher working with biofilms and ask them for help and guidance.

You could start by contacting the Center for Biofilm Engineering at Montana State University. It’s likely that someone on their staff could tell you more about biofilm testing and who to contact to do the research.

Personally, I think there’s a great group of biofilm researchers at the University of Washington. Drs. Parsek, Singh, Harwood and others. You could look them up and send them an email with your question. I’m not sure if they would respond but they might give you feedback.

Best,AmyShari Gold April 14th, 2009 at 5:44 am17

Hi Amy,I just re-read this article. It is such a great, well researched article, written in a very direct and easy to understand manner. I had a few quick questions. I am familiar with biofilms contaminating artificial surfaces within the human body, but wondered if biofilms colonize regular joints, not altered by artificial replacements. In particular, shoulder and hips, wrists and fingers – could this be what is going on in something like RA too? Also, what about your nerves – say, peripheral nerves? Is this something they might colonize as well? Reason I ask is I do have peripheral neuropathy in my back. Neuropathy is something I know that resolves on the MP – although it is something that takes a bit longer. Could this be because much of what might be affecting that signal transduction in the nerve is stopped by biofilm inflammation somewhere from the nerve root to the muscle? In me, my neuropathy is getting much better and I can definitely see the waxing and waning of symptoms as I increase abx and ramp different abx combos. Based on what I just re-read tonight, that might seem a plausible explanation for the physical phenomenon I am experiencing – killing a bunch of bacteria, one layer at a time. Also, as noted in the patients w/ cystic fibrosis or chronic sinus infected patients, as I have gotten further into phase 2, I have had a lot of sinus drainage and much nasty stuff coming up from lower respiratory, like I have a constant cold, but it isn’t “infection†in the clinical way, just sputum breaking up constantly, might this be evidence of biofilm existence in these places? This is truly a fascinating area of study.

Thanks for you insight.Best,Shari Albert April 14th, 2009 at 8:48 am18

Hi Shari,Amy is away, so you’re stuck with me. : )I don’t think there’s anything unique about non-human surfaces that leads to formation of biofilm. You may be shocked to learn that most people manually remove biofilm from themselves every day, sometimes twice a day. This odd ritual? Teeth brushing! Teeth brushing is nothing more than the act of manually removing biofilm from the surface of one’s teeth.

But, it would be very un-bascterialike for bacteria to confine themselves to prosthetic hip joints and teeth. If you think about it, the only reason we don’t brush every other part of our body is that we can’t – regrettably, we cannot brush our kidney, liver, nerves, lungs, etc.

Do biofilm colonize nerves? They have had hundreds of thousands of years to figure it out. I would say absolutely. One of the limiting factors as far as progress on the MP is concerned is blood flow to a region. I remember hearing that nerve cells don’t get as much blood as other areas. That’s not to the say that neuropathy doesn’t resolve, only that the process may take longer. However, the waxing and waning of your neuropathy symptoms with your antibiotics supports the idea that these symptoms will resolve.

As for your theory on signal transduction, I’m sure it’s involved, but I’m afraid I couldn’t say how.Also, I think you’re on target when it comes to sinus drainage – bacteria are involved – although I’m not sure we have any good idea on the relative proportion of different forms of bacteria by region of the body: L-forms, intracellular pathogens, biofilm, etc.

Best, May 13th, 2009 at 4:49 pm19

Hi Amy,thanks for the article and research… In your research, did you happen to find any effective alternative medicines for removal of biofilms? or energy medicine? Ondamed has been used successfully now in Europe for 15+ yrs in chronic lyme….

Amy Proal May 15th, 2009 at 11:38 am20

Hi ,Unfortunately I do not know of any substance that can target internal biofilms. External biofilms – or those that grow outside the body – are a different story. Those can be treated with several chemicals, however they are chemicals that would be toxic if swallowed.

Actually, some studies show that a substance called Lactoferrin can reduce biofilm growth. However I believe more research is needed to confirm its effects on biofilm proliferation and to make sure it doesn’t interfere with other pathways in the body when taken.

Does Ondamed cause patients to experience a bacterial die-off reaction? I ask, because if it doesn’t cause an exacerbation in disease symptoms then it’s not causing bacteria to be killed. We know this because a rise in symptoms generated by bacterial death cannot be avoided when the MP medications begin to stimulate the immune response to target biofilm species.

If Ondamed makes patients feel better instead of worse, it is probably just a palliative treatment option. What it may actually do is slow the immune response. While this would lower the inflammation associated with bacterial death it wouldn’t actually kill biofilm bacteria and would be ineffective in the long run. Clearly Lyme is still a big problem in Europe so unfortunately I suspect that Ondamed falls into this latter category.

I have not researched energy medicine and biofilms enough to comment on whether it would help with their elimination. What I can say is that the MP seems quite effective at targeting biofilm species! Essentially 100% of patients to start the treatment have experienced immunopathology or bacterial die-off and these reactions remain strong and steady during much of treatment. Then, during later stages of treatment, we have patients reporting improvement from many known biofilm-related infections. So I would say the MP is certainly your best bet if you are trying to target biofilm. I encourage you not to worry about taking the MP medications as they have an excellent safety profiles and are likely safer than taking herbs or other “natural†supplements whose properties have not been investigated at the molecular level.

This article discusses the safety of the MP meds:http://bacteriality.com/2008/02/23/misconceptions/#2

Hope this helps!Amy W. May 15th, 2009 at 12:17 pm21

– Xylitol and Serrapeptase both seem pretty safe and have activity against biofilms, though I’m not sure how long xylitol lasts or how well it works inside the body. Xylitol is a sugar alcohol used as a sweetner and in some toothpastes because of its flavor and effect on oral biofilms. Serrapeptase is an anti-inflammatory and blood thinner used routinely in Germany as an aspirin substitute since it doesn’t interfere with K1 (it doesn’t promote heart disease via arterial calcification like Aspirin does) or seem to cause the liver problems that salyciliates like Aspirin do. Both seem very safe in proper dosages. It may be hard to tell if Serrapeptase caused a bacterial dieoff reaction since it’s also an anti-inflammatory but it seems like one of the more promising candidates for attacking internal biofilms.

Amy Proal May 15th, 2009 at 12:57 pm22

Hi ,Yes, Xylitol has been proposed to curb biofilm growth. It’s in my toothpaste and mouthwash (I use a brand of tooth products called Biotene which is designed to target biofilm bacteria). It’s also some chewing gum. However I have not found any studies that show it’s effective at targeting biofilm other than those on the teeth and I’m not really sure if it even works as intended.

When I googled Serapeptase, I read that “Serrapeptase is an enzyme that is produced in the intestines of silk worms to break down cocoon walls. This enzyme is proving to be an alternative to the non-steroidal anti-inflammatory agents (NSAIDs) traditionally used to treat rheumatoid arthritis and osteoarthritis.â€

NSAIDs are completely contraindicated for use for MP patients because they palliate inflammation rather than actually killing the bacteria causing inflammation. So that supplement seems suspect to me and I would certainly never take it.

I cannot emphasize enough the risk of taking supplements when most of the claims about how they work are just based on speculation and not on molecular data. I took way too many supplements before the MP which were supposed to kill bacteria and they did nothing but make me more ill.

Best,Amy W. May 15th, 2009 at 1:35 pm23

When I googled SerapeptaseIf you’re looking for hard data, why not use pubmed?Serrapeptase has the demonstrated capacity to increase antibiotic effectiveness against biofilm forming bacteria. It’s demonstratablynot simply palliative. I understand that it’s contraindicated under the Marshall protocol, but there’s still very good evidence it’s helpful and well tolerated and doesn’t just mask symptoms.

link

Here’s a study showing injected serrapeptase + antibiotics is significantly more effective at clearing biofilm producing infections than antibiotics alone.We have chosen serratiopeptidase (SPEP), an extracellular metalloprotease produced by Serratia marcescens that is already widely used as an anti-inflammatory agent, and has been shown to modulate adhesin expression and to induce antibiotic sensitivity in other bacteria. Treatment of L. monocytogenes with sublethal concentrations of SPEP reduced their ability to form biofilms and to invade host cells. Zymograms of the treated cells revealed that Ami4b autolysin, internalinB, and ActA were sharply reduced. These cell-surface proteins are known to function as ligands in the interaction between these bacteria and their host cells, and our data suggest that treatment with this natural enzyme may provide a useful tool in the prevention of the initial adhesion of L. monocytogenes to the human gut.

http://www.ncbi.nlm.nih.gov/pubmed/18479885?ordinalpos=1 & itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum

Antibiotic susceptibility tests on both planktonic and sessile cultures, studies on the dynamics of colonization of 10 biofilm-forming isolates, and then bioluminescence and scanning electron microscopy under seven different experimental conditions showed that serratiopeptidase greatly enhances the activity of ofloxacin on sessile cultures and can inhibit biofilm formation.

http://www.ncbi.nlm.nih.gov/pubmed/8109925?ordinalpos=3 & itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum

Amy Proal May 16th, 2009 at 12:36 pm24

,Thank you, I know how to use PubMed. The reason I didn’t start searching PubMed for info about Serapeptase is because it’s not a topic relevant to what I’m spending my energy on at the moment. On this site, I try to explain the MP in simple terms – the MP as it is created and the molecular data we have about how the medications interact. I already have many comments from people asking about these topics and I need/want to answer their basic questions first before I speculate about a supplement with you.

You know yourself that the MP has a no supplement policy, and for a reason. If you actually started the MP, you’d realize that the combination of Olmesartan and low dose antibiotics is so powerful that our main problem is certainly not whether people are targeting enough bacteria. Our main problem is how to control the die-off reaction and keep people from killing so many bacteria in a day that they can’t tolerate the die-off symptoms. So at this point we don’t need any extra supplements to target biofilm bacteria.

That’s especially true since we don’t know how serapeptase works at the molecular level. Maybe it does hinder biofilm formation somewhat but what’s to say it doesn’t interfere with any of the receptors involved in the immune response? How do we know that it wouldn’t interfere with the ability of Olmesartan to bind the VDR? We don’t know at this point and so I worry that when you mention these things people who I really believe should try the MP are going to skip the treatment and think that a supplement can help them instead – which is exactly what I don’t want them to think.

Best,AmyL. Lange June 3rd, 2009 at 9:59 pm25

Hi Amy –I was reading your article and enjoyed it a lot, thank you! Do you know anything about Lymes disease and if the bacteria are protected by the the concepts of biofilms? If yes, do you think that the Marshal protocol would help someone recover from the disease?

Thanks in advance for your reply!-Amy Proal June 7th, 2009 at 10:05 pm26

Hi ,Sorry for the delayed response. I just got back from a trip to China.Yes, it is very likely that many of the bacteria that cause Lyme disease persist in a biofilm state and there are definitely many people on the MP with Lyme disease who are recovering.

Here are interviews with three of them:http://bacteriality.com/2007/09/22/interview3/

http://bacteriality.com/2007/12/28/interview14/

http://bacteriality.com/2008/03/31/interview19/

You might also want to post on the website http://www.curemyth1.org (Th1 refers to diseases caused by bacteria, hence the name). The site is a place where people can ask questions about the MP. But once you are a member of the site, you can also communicate with other people on the MP for Lyme.

If you have Lyme, I definitely hope that you start the MP. The best way to learn more about the treatment is to read as much as possible, on this site and also on the MP study site – http://www.marshallprotocol.com.

I also highly recommend watching the following video which describes the MP and the science that forms its backbone in simple terms:http://bacteriality.com/2008/05/07/mpintro/

Best,Amy June 7th, 2009 at 10:15 pm27

Hey amy,thanks for the info:) As far as I know, ondamed therapy it is able to approach micro-organisims at a molecular level using frequencies…there’s been a ton of research to back up its effectiveness and YES you have to be careful with how much time you use b/c of the die off effect! The first time i used this therapy I did only 10 mins and was in bed for about a week! I will do some more research on the MP protocol… i hope u have a blessed week!

p.s thanks ryan for the info too:)Amy Proal June 8th, 2009 at 11:09 am28

Hi ,Thanks for writing. I know nothing about Ondamed. Are the papers you mention about it’s efficacy peer-reviewed? If so, I’d be interested in reading one.In the meantime, I’d like to emphasize that I wouldn’t definitely not recommend using Ondamed or any other frequency therapy or supplement along with the MP. As I mentioned to before, there is a careful balance of the immune system set up in the body by the MP meds as they target bacteria, and other therapies may interfere. Also, if any therapy causes extra bacterial die-off it could lead to intolerable immunopathology when combined with the MP meds so one must be careful.

I definitely hope you look into the MP !Best,Amy June 22nd, 2009 at 1:33 pm29

Amy,How do I find a doctor who will use the Marshall Protocol? I have Interstitial Cystitis and tested positive for Strep D, only after a week long broth culture. Based on what I have read, I think that the bacteria has formed a biofilm, which is why shorter cultures aren’t picking it up. My doctor doesn’t know a whole lot about biofilm, so I am trying to find a doctor who does, so that I can get treatment.

Thanks in advance for any help!Amy Proal June 24th, 2009 at 12:01 pm30

Hi ,I’m sorry about your illness and positive strep cultures. In my opinion you are correct – it is very likely that many of the bacteria making you ill are in a biofilm state and cannot be kill by conventional therapies. On the other hand, the MP appears to effectively break up biofilms and I think you would definitely benefit from the treatment.

I’m not sure where you live. However the best way to find a doctor in your area is to post at the following website:http://www.curemyth1.org (Th1 refers to diseases caused by bacteria, hence the name). The patient advocates on the site, who are volunteers, can likely provide you with a list of doctors that administer the MP in your area.

If that doesn’t work out I recommend showing your current doctor or a new open-minded doctor in your area Dr. Marshall and team’s latest peer-reviewed papers and presentations. See if then you can convince them to prescribe you the necessary MP meds. The publications etc. can be found here:

http://mpkb.org/doku.php#publications_presentations

This article also give more tips on finding an MP doctor.http://mpkb.org/doku.php/home:starting:physician:finding

Good luck and take care!AmyJD Bear August 8th, 2009 at 11:06 am31

Hello Amy,I have spent some time perusing your website and I complement you on your good work. As such, it is all the more important to not mar this good work by using inappropriate terminology. Specifically, the term “biofilm†refers to a community of bacteria surrounded by *Extracellular* Polymeric Substances. A tubule, or filament, or “protective sheath†formed by an L-form bacteria may be a “film†and it is no doubt biological, but it is NOT a “biofilm†as commonly defined. It is a disservice to conflate these two different phenomena. Whatever is going on with L-form bacteria deserves its own terminology so as not to introduce unnecessary confusion into the field.

Best Regards,-JDBAmy Proal August 8th, 2009 at 12:09 pm32

Hi JDB,Thanks for your comment. I’m not sure exactly what statement I’ve made that you are referring to. Nevertheless, I don’t see why L-form bacteria could not be able to form a biofilm. Bacteria in the L-form are simply in a different part of the microbial lifecycle and have temporarily lost their cell walls. Is a cell wall necessary to become part of biofilm community? I have not seen research along those lines.

This being said why wouldn’t L-form bacteria also form into communities in order to better protect themselves from the immune response etc? How do we know that a biofilm cannot be composed of both bacteria with cell walls and bacteria without cell walls?

So I don’t really think we need to come up with completely different terminology to describe bacteria in the L-form seeing as their characteristics are generally very similar to those of their walled counterparts and they interact with classical forms of bacteria very closely in vivo.

Best,Amy  August 26th, 2009 at 7:02 am33

Dear Amy,Thanks for this article. And I hope what you do next is to study Borrelia and biofilms. Leptospira and Treponema denticola are known to be able to persist inside biofilms. But there has not been published anything on Borrelia yet. No major journal has anything on it. It would meen so much to so many if you’d get that out…

Amy Proal August 28th, 2009 at 11:45 pm34

Hi ,Thank you for your interest in our research. Borrelia is definitely a very hardy pathogen that we would expect would be able to survive in biofilms. Over the course of the next few years, Autoimmunity Research Foundation, which is the organization I work with, is hoping to run in which we will look at the DNA of the infected cells in patients with chronic disease. This study is still in the planning stages, but, in time, could eventually better help us understand how Borrelia survive in the body and in a biofilm-like environment.

Meanwhile, there is research suggesting that the low pulsed manner with which antibiotics are administered is much more effective at targeting bacteria in biofilms than standard regular doses of antibiotics or antibiotics taken in an IV form. Our own clinical data also shows people reporting improvement and recovery from Lyme disease thanks to therapy with the Marshall Protocol. So, if you think you may harbor Borrelia, and you want to target Borrelia in the biofilm state, looking into the MP may be a worthwhile option.

Best,Amygregg zulauf September 12th, 2009 at 4:16 pm35

I was impressed by the article in ‘Discover Magazine’ about biofilms (link below). Anecdotal evidence of xylitol breaking down certain biofilm structures was interesting to me.This article suggests that xylitol may work internally and not only in the mouth. I am interested in your reaction to this article.

Regards, Gregghttp://discovermagazine.com/2009/jul-aug/17-slime-city-germs-talk-each-other-plan-attacks/article_view?b_start:int=2 & -C=

Albert September 14th, 2009 at 12:47 pm36

Hi Gregg,Interesting. One of the principles in that article is Randall Wolcott. If you do a Google search for Randall Wolcott, the #1 hit is Amy’s interview with him:http://bacteriality.com/2008/04/13/wolcott/

So, yes indeed we have heard about Xylitol before.However, the problem with Marshall Protocol patients isn’t generating bacterial die-off but controlling the reaction. That said, it may be worth giving those few patients who are non-responders xylitol and seeing if that can lead to bacterial killing. This might be an avenue we will research in time….

Best, September 30th, 2009 at 8:57 pm37

I’ve been reading that EDTA has been used to erradicate Biofilm??Also, some claim Noni and Grapefruit seed extract works….any comments?

NEWS FLASH

April 4, 2009:,  Milk consumption tied to Parkinson’s disease

March 21, 2009:,  Hey there, how’s your Kineosphaeram holding up?

Peer-Reviewed Papers

Autoimmune disease in the era of the metagenome (PDF)

Vitamin D: the alternative hypothesis (PDF)

Dysregulation of the Vitamin D Nuclear Receptor may contribute to the higher prevalence of some autoimmune diseases in women (PDF)

Vitamin D metabolites as clinical markers in autoimmune and chronic disease (PDF)

Amy's Conference Presentations

Metagenomic symbiosis between bacterial and viral pathogens in autoimmune disease

Notes from the 2009 International Congress of Antibodies

Notes from the 2008 International Congress on Autoimmunity

Featured Articles

Update on tone and other issues

Why patients with chronic disease are disaffected and how online social networks meet their needs

Sun-blocking culture among the Chinese

Second-guessing the consensus on vitamin D

Travels, papers, and more… an update

Three days at the J. Craig Venter Institute

The bacteria boom – implications of the Human Microbiome Project

Understanding Biofilms

Interview with Dr. Randall Wolcott, bacterial biofilm wound specialist

Insights into horizontal gene transfer: conversations with Dr. Gogarten and Dr. Lake

Voices of reason in the vitamin D debate

Interview with evolutionary biologist Ewald

What can medical research learn from the open source software movement?

Interview with Dr. Greg Blaney: MP physician

Bacteria and cancer: an interview with Dr. Alan Cantwell

Interview with Nadya Markova: L-form Expert

Gerald Domingue: Pioneer of Atypical Bacteria

A History of Cell Wall Deficient Bacteria: A Selection of Researchers Who Have Worked with the L-form

Patient Interviews

Interview with Gene – sarcoidosis, bladder cancer

Interview with Bonnie B – lupus, Sjogren’s Syndrome

Interview with Eastlund – diabetes, sarcoidosis, irritable bowel syndrome

Interview with Roy P. – sarcoidosis, rheumatoid arthritis

Interview with de Jager: chronic fatigue syndrome, multiple chemical sensitivity

Interview with Ken L. – Post Treatment Lyme Disease Syndrome (PTLDS)

Interview with Doreen V. – autism, ADHD depression, severe anxiety, CFS

Interview with Jane -Aoki: Neurosarcoidosis, systemic sarcoidosis; spasticity, myasthenia, CNS dysfunction, joint pain, pulmonary, splenic and cardiac involvement.

Interview with Melinda Stiles – Lyme, Irritable bowel syndrome/colitis, Radiculitis (inflammation of the nerve roots)

Interview with Freddie Ash – Sarcoidosis of the heart, coronary artery disease, atrial fibrillation

Interview with P. Bear R.N. – Chronic Borreliosis (“Lymeâ€), MCS (multiple chemical sensitivities), Chronic Spinal Inflammation, Peripheral Neuropathy

Interview with Sherry Cook – Sarcoidosis, Cat Scratch Fever, Restless Leg Syndrome

Interview with Leesa Shanahan – Sarcoidosis (Heerfordt’s Syndrome), Uveitis

Interview with Mirek Wozga – sarcoidosis

Interview with Albert – CFS, depression, food sensitivities

Interview with Carole – Sarcoidosis, fibromyalgia, CFS

Interview with Shirley J. (Saj) – Sarcoidosis

Interview with Robyn – Lyme, myoclonus

Interview with Sue Andorn – Lyme, Babesia

Interview with Ival Meyer – Arthritis, dyslexia

Interview with Guss Wilkinson – Sarcoidosis, psoriasis, insomnia

About Amy Proal

Amy Proal graduated from town University in 2005 with a degree in biology. While at town, she wrote her senior thesis on Chronic Fatigue Syndrome and the Marshall Protocol.

Amy has spoken at several international conferences and authored several peer-reviewed papers on the intersection of bacteria and chronic disease.

If you have questions about the MP, please visitCureMyTh1.org where volunteer patient advocates will answer your questions. Another good resource is the MP Knowledge Base, which is scheduled to be completed within the next year.

CategoriesSelect Categoryaging  (1)biofilms  (3)cancer  (2)cardiovascular disease  (1)cognitive dysfunction  (3)conferences and trainings  (7)diet  (4)familial aggregation  (3)featured articles  (18)history  (3)horizontal gene transfer  (2)interview (doctor/researcher)  (5)interview (patient)  (21)L-form bacteria  (6)marshall protocol  (21)medical research  (5)mental illness  (2)microbiome  (3)News Flash  (38)obesity  (2)personal  (2)presentations  (3)statins  (1)Uncategorized  (1)videos  (4)vitamin d  (12)

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Copyright © 2009 - Bacteriality — Exploring Chronic DiseaseThis site uses a modified version of Illacrimo Theme created by Design Disease

From: Goldstein@...To: bird mites

Sent: Friday, September 9, 2011 8:28:49 AMSubject: Re: Good summary about biofilms

 

Hi Cecilia,Two things, first, you are right.  I don't think it is good to use Hibiclens long term.  When we finish these bottles, I think that will be the end of that.  Normally have not used antibacterial soaps here... Dr. told us to use it.  But, what do they know?  Right?  Sometimes they give incorrect or bad advice. 

Secondly about biofilms.  I can just speak from experience.  At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin.  Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair.  Husband has it too.  Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm.  I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and new ones appear.  I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy.  Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created  by Lyme organisms and other organisms together.  I think there is some research being done on this.  I'll see if I can locate anything on it.  Maybe Aandraya knows of something too.

From: " Krys Brennand " <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

 

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

 

How do you know you have biofilms?Cecilia

From: " Goldstein@... " <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

 

Yes.  True.  L.From: " Aandraya Da Silva " <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

 

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing " biofilm protocols " assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other " biofilm protocols " in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book " Healing Lyme " by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was " yes " which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out. AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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This was a good article if you have time to read it in its entirety. A couple of things that I did not know that I learned:1. After antibiotics (all) there are bacteria they have referred to in science as persisters. This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.2. Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc. Much like a family or civilization. They have their own language; they are intelligent.3. Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums. The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear. 4. One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth. As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body. My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.5. Research is picking up on the study of biofilms. The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.6. Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7. Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi. Maybe there is more information out about that... I'll look. So much to be gleaned from this article. From: "Aandraya" <aandraya@...>bird mites Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out. AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned. 

I found the theory, in the last section, about using low level antibiotics in " pulses "  to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing. 

 Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a " language " & being " intelligent " could be misleading. I suppose it depends how we define " intelligent " . We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to. 

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

 

This was a good article if you have time to read it in its entirety.  A couple of things that I did not know that I learned:1.  After antibiotics (all) there are bacteria they have referred to in science as persisters.  This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2.  Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc.  Much like a family or civilization.  They have their own language; they are intelligent.

3.  Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums.  The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.  

4.  One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth.  As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body.  My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5.  Research is picking up on the study of biofilms.  The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6.  Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7.  Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi.  Maybe there is more information out about that... I'll look.  

So much to be gleaned from this article.  From: " Aandraya " <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

 

I had that film too big time.  I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked.  I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same.  Either way, treating my chronic infections including the antifungals is knocking it out.  

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

 

Hi Cecilia,Two things, first, you are right.  I don't think it is good to use Hibiclens long term.  When we finish these bottles, I think that will be the end of that.  Normally have not used antibacterial soaps here... Dr. told us to use it.  But, what do they know?  Right?  Sometimes they give incorrect or bad advice. 

Secondly about biofilms.  I can just speak from experience.  At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin.  Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair.  Husband has it too.  Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm.  I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear.  I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy.  Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created  by Lyme organisms and other organisms together.  I think there is some research being done on this.  I'll see if I can locate anything on it.  Maybe Aandraya knows of something too.

From: " Krys Brennand " <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

 

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

 

How do you know you have biofilms?Cecilia

From: " Goldstein@... " <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

 

Yes.  True.  L.From: " Aandraya Da Silva " <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

 

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing " biofilm protocols " assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other " biofilm protocols " in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book " Healing Lyme " by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was " yes " which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Krys-With chronic Lyme and coinfections low dose abx are not recommended because the microbes can build up resistance more easily. It's better to hit them with therapeutic doses. I've tried a lot of strategies, what's worked best in my case is multiple antimicrobials given in high doses long term. That's where I'm at now. The treatment is no picnic but I am progressing.Aandraya On Sep 9, 2011, at 1:32 PM, Krys Brennand <krys109uk@...> wrote:

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned.

I found the theory, in the last section, about using low level antibiotics in "pulses" to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing.

Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a "language" & being "intelligent" could be misleading. I suppose it depends how we define "intelligent". We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to.

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

This was a good article if you have time to read it in its entirety. A couple of things that I did not know that I learned:1. After antibiotics (all) there are bacteria they have referred to in science as persisters. This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2. Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc. Much like a family or civilization. They have their own language; they are intelligent.

3. Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums. The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.

4. One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth. As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body. My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5. Research is picking up on the study of biofilms. The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6. Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7. Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi. Maybe there is more information out about that... I'll look.

So much to be gleaned from this article. From: "Aandraya" <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out.

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice.

Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.

From: "Krys Brennand" <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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The LLD I went to wanted to pulse the antibiotics. I think a lot of LLDs do pulse antibiotics. I agree with you about intelligence and language. In a broad sense maybe it could be thought of that way. They do seem to learn and adapt. I know what you mean about what we learned in college feel somewhat obsolete. Sometimes it feels like things are really speeding up, including science and technology. I don't know how people keep up with it. Just seems near impossible. This is one of the reasons why I like to be around young people - they are like sponges... pick things up so much more quickly than I ever did.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 11:32:21 AMSubject: Re: Good summary about biofilms

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned.

I found the theory, in the last section, about using low level antibiotics in "pulses" to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing.

Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a "language" & being "intelligent" could be misleading. I suppose it depends how we define "intelligent". We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to.

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

This was a good article if you have time to read it in its entirety. A couple of things that I did not know that I learned:1. After antibiotics (all) there are bacteria they have referred to in science as persisters. This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2. Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc. Much like a family or civilization. They have their own language; they are intelligent.

3. Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums. The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.

4. One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth. As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body. My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5. Research is picking up on the study of biofilms. The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6. Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7. Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi. Maybe there is more information out about that... I'll look.

So much to be gleaned from this article. From: "Aandraya" <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out.

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice.

Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.

From: "Krys Brennand" <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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We tend to think that the microbes live randomly in the body, but they don't. They live in organized colonies in biofilms. Some Biofilm dissolvers I know of: neem, Rizols-ozonated plant oils, enzymes, and I know there are many others. Also, chelating agents like EDTA break them down- chelates calcium and magnesium which is an important part of the biofilm's skeleton. It's a long slow process breaking down the biofilms. We all have them in our tissues, but healthy people have an immune system that doesn't allow the microbes to proliferate to the point they makes us sick like they do in us.I have used multi enzymes, Nattokinase, EDTA and rizol Gamma from biopure to break down biofilms. Aandraya On Sep 9, 2011, at 12:58 PM, Goldstein@... wrote:

This was a good article if you have time to read it in its entirety. A couple of things that I did not know that I learned:1. After antibiotics (all) there are bacteria they have referred to in science as persisters. This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.2. Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc. Much like a family or civilization. They have their own language; they are intelligent.3. Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums. The fact that the infections come back year after year

is a signal that there are persisters and there is biofilm in the inner ear. 4. One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth. As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body. My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.5. Research is picking up on the study of biofilms. The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.6. Biofilm is made up of a complex city of

workers, including fungi, viruses and bacteria.7. Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi. Maybe there is more information out about that... I'll look. So much to be gleaned from this article. From: "Aandraya" <aandraya@...>bird mites Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out. AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away

and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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I don't doubt that a big part of this is mold. If it is a biofilm, it would contain mold and other things too. L.From: "Aandraya" <aandraya@...>bird mites Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out. AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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I see what you are saying.  

Really, leaving out the added complications of biofilm & quorum sensing, every time we give antibiotics we are, in effect, breeding for resistance.

I don't think the writer was talking about Lyme per sé.  The cells, which they called " persisters " are particularly interesting. If what is written is found to be the case then one would imagine pulsing antibiotics even in high doses could work better than continuous long term high doses?

On 9 September 2011 13:39, Aandraya <aandraya@...> wrote:

 

Krys-With chronic Lyme and coinfections low dose abx are not recommended because the microbes can build up resistance more easily.  It's better to hit them with therapeutic doses.  I've tried a lot of strategies, what's worked best in my case is multiple antimicrobials given in high doses long term.  That's where I'm at now.  The treatment is no picnic but I am progressing.

Aandraya On Sep 9, 2011, at 1:32 PM, Krys Brennand <krys109uk@...> wrote:

 

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned. 

I found the theory, in the last section, about using low level antibiotics in " pulses "  to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing. 

 Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a " language " & being " intelligent " could be misleading. I suppose it depends how we define " intelligent " . We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to. 

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

 

This was a good article if you have time to read it in its entirety.  A couple of things that I did not know that I learned:1.  After antibiotics (all) there are bacteria they have referred to in science as persisters.  This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2.  Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc.  Much like a family or civilization.  They have their own language; they are intelligent.

3.  Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums.  The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.  

4.  One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth.  As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body.  My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5.  Research is picking up on the study of biofilms.  The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6.  Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7.  Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi.  Maybe there is more information out about that... I'll look.  

So much to be gleaned from this article.  From: " Aandraya " <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

 

I had that film too big time.  I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked.  I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same.  Either way, treating my chronic infections including the antifungals is knocking it out.  

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

 

Hi Cecilia,Two things, first, you are right.  I don't think it is good to use Hibiclens long term.  When we finish these bottles, I think that will be the end of that.  Normally have not used antibacterial soaps here... Dr. told us to use it.  But, what do they know?  Right?  Sometimes they give incorrect or bad advice. 

Secondly about biofilms.  I can just speak from experience.  At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin.  Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair.  Husband has it too.  Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm.  I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear.  I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy.  Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created  by Lyme organisms and other organisms together.  I think there is some research being done on this.  I'll see if I can locate anything on it.  Maybe Aandraya knows of something too.

From: " Krys Brennand " <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

 

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

 

How do you know you have biofilms?Cecilia

From: " Goldstein@... " <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

 

Yes.  True.  L.From: " Aandraya Da Silva " <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

 

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing " biofilm protocols " assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other " biofilm protocols " in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book " Healing Lyme " by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was " yes " which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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I agree Krys. I wish my doctor had pushed that idea of pulsing harder towards me. Suppose I could still do it, but must get my stomach back in balance first and I want to get a liver enzyme test before I really go for it. I've requested testing from my doctor today. L.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 1:07:43 PMSubject: Re: Good summary about biofilms

I see what you are saying.

Really, leaving out the added complications of biofilm & quorum sensing, every time we give antibiotics we are, in effect, breeding for resistance.

I don't think the writer was talking about Lyme per sé. The cells, which they called "persisters" are particularly interesting. If what is written is found to be the case then one would imagine pulsing antibiotics even in high doses could work better than continuous long term high doses?

On 9 September 2011 13:39, Aandraya <aandraya@...> wrote:

Krys-With chronic Lyme and coinfections low dose abx are not recommended because the microbes can build up resistance more easily. It's better to hit them with therapeutic doses. I've tried a lot of strategies, what's worked best in my case is multiple antimicrobials given in high doses long term. That's where I'm at now. The treatment is no picnic but I am progressing.

Aandraya On Sep 9, 2011, at 1:32 PM, Krys Brennand <krys109uk@...> wrote:

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned.

I found the theory, in the last section, about using low level antibiotics in "pulses" to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing.

Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a "language" & being "intelligent" could be misleading. I suppose it depends how we define "intelligent". We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to.

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

This was a good article if you have time to read it in its entirety. A couple of things that I did not know that I learned:1. After antibiotics (all) there are bacteria they have referred to in science as persisters. This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2. Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc. Much like a family or civilization. They have their own language; they are intelligent.

3. Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums. The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.

4. One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth. As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body. My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5. Research is picking up on the study of biofilms. The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6. Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7. Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi. Maybe there is more information out about that... I'll look.

So much to be gleaned from this article. From: "Aandraya" <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out.

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice.

Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.

From: "Krys Brennand" <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Absolutely they seem to " learn " & adapt; evolution in progress. Just hope we mammals can keep one step ahead. ;-PIt's amazing how far knowledge of biology has moved on in the last 20-30 years, & it's accelerating. LOL. Even simple stuff like classification....it gets to the stage that the kids are teaching us. And that's good really, shows they're learning more than us LOL. 

KrysOn 9 September 2011 13:44, <Goldstein@...> wrote:

 

The LLD I went to wanted to pulse the antibiotics.  I think a lot of LLDs do pulse antibiotics.  I agree with you about intelligence and language.  In a broad sense maybe it could be thought of that way.  They do seem to learn and adapt.  

I know what you mean about what we learned in college feel somewhat obsolete.  Sometimes it feels like things are really speeding up, including science and technology.  I don't know how people keep up with it. Just seems near impossible.  This is one of the reasons why I like to be around young people - they are like sponges... pick things up so much more quickly than I ever did.

From: " Krys Brennand " <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 11:32:21 AMSubject: Re: Good summary about biofilms

 

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned. 

I found the theory, in the last section, about using low level antibiotics in " pulses "  to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing. 

 Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a " language " & being " intelligent " could be misleading. I suppose it depends how we define " intelligent " . We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to. 

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

 

This was a good article if you have time to read it in its entirety.  A couple of things that I did not know that I learned:1.  After antibiotics (all) there are bacteria they have referred to in science as persisters.  This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2.  Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc.  Much like a family or civilization.  They have their own language; they are intelligent.

3.  Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums.  The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.  

4.  One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth.  As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body.  My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5.  Research is picking up on the study of biofilms.  The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6.  Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7.  Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi.  Maybe there is more information out about that... I'll look.  

So much to be gleaned from this article.  From: " Aandraya " <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

 

I had that film too big time.  I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked.  I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same.  Either way, treating my chronic infections including the antifungals is knocking it out.  

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

 

Hi Cecilia,Two things, first, you are right.  I don't think it is good to use Hibiclens long term.  When we finish these bottles, I think that will be the end of that.  Normally have not used antibacterial soaps here... Dr. told us to use it.  But, what do they know?  Right?  Sometimes they give incorrect or bad advice. 

Secondly about biofilms.  I can just speak from experience.  At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin.  Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair.  Husband has it too.  Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm.  I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear.  I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy.  Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created  by Lyme organisms and other organisms together.  I think there is some research being done on this.  I'll see if I can locate anything on it.  Maybe Aandraya knows of something too.

From: " Krys Brennand " <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

 

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

 

How do you know you have biofilms?Cecilia

From: " Goldstein@... " <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

 

Yes.  True.  L.From: " Aandraya Da Silva " <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

 

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing " biofilm protocols " assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other " biofilm protocols " in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book " Healing Lyme " by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was " yes " which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Could be, though it hasn't been so in my case. Depends on what you consider pulsing, I've been on a variety of antimicrobial cocktails. It's all trial and error. What's important is I'm getting better and getting multiple infections under control.On Sep 9, 2011, at 3:07 PM, Krys Brennand <krys109uk@...> wrote:

I see what you are saying.

Really, leaving out the added complications of biofilm & quorum sensing, every time we give antibiotics we are, in effect, breeding for resistance.

I don't think the writer was talking about Lyme per sé. The cells, which they called "persisters" are particularly interesting. If what is written is found to be the case then one would imagine pulsing antibiotics even in high doses could work better than continuous long term high doses?

On 9 September 2011 13:39, Aandraya <aandraya@...> wrote:

Krys-With chronic Lyme and coinfections low dose abx are not recommended because the microbes can build up resistance more easily. It's better to hit them with therapeutic doses. I've tried a lot of strategies, what's worked best in my case is multiple antimicrobials given in high doses long term. That's where I'm at now. The treatment is no picnic but I am progressing.

Aandraya On Sep 9, 2011, at 1:32 PM, Krys Brennand <krys109uk@...> wrote:

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned.

I found the theory, in the last section, about using low level antibiotics in "pulses" to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing.

Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a "language" & being "intelligent" could be misleading. I suppose it depends how we define "intelligent". We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to.

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

This was a good article if you have time to read it in its entirety. A couple of things that I did not know that I learned:1. After antibiotics (all) there are bacteria they have referred to in science as persisters. This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2. Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc. Much like a family or civilization. They have their own language; they are intelligent.

3. Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums. The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.

4. One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth. As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body. My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5. Research is picking up on the study of biofilms. The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6. Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7. Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi. Maybe there is more information out about that... I'll look.

So much to be gleaned from this article. From: "Aandraya" <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out.

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice.

Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.

From: "Krys Brennand" <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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I've just spotted, there was more of the article which had been cut off. Apparently it does apply to Lyme. Here are the last paragraphs: has found that the feasibility of a pulsed, or cyclical biofilm eradication approach depends on the rate at which persisters lose resistance to killing and regenerate new persisters. It also depends on the ability to manipulate the antibiotic concentration – something that is done quite effectively by patients on the Marshall Protocol who carefully dose their antibiotics at different levels, allowing constant variation in antibiotic concentration. Although speculates that allowing the concentration of an antibiotic to drop could potentially lead to resistance towards the antibiotic, she is quick to add that if two or more antibiotics are used to target a biofilm at one time, such resistance would not occur. Again, since the Marshall Protocol uses a total of five bacteriostatic antibiotics, usually taken two or three at a time, concerns of resistance are essentially negligible.

Model of biofilm resistance based on persister survival. An initial treatment of high-dose constant antibiotic kills planktonic cells and the majority of biofilm cells. But persisters remain alive and resurrect the biofilm, causing the infection to relapse

“It is entirely possible that successful cases of antimicrobial therapy of biofilm infections result from a fortuitous optimal cycling [pulsed dosing] of an antibiotic concentration that eliminated first the bulk of the biofilm and then the progeny of the persisters that began to divide,” states .

’ work has been supported by other research teams. Recently, researchers at the University of Iowa found that subinhibitory (extremely low dose) concentrations of the bacteriostatic antibiotic azithromycin significantly decreased biomass and maximal thickness in both forming and established biofilms.[27] These extremely low concentrations of azithromycin inhibited biofilms in all but the most highly resistant isolates. In contrast, subinhibitory concentrations of gentamicin, which is not a bacteriostatic antibiotic, had no effect on biofilm formation. In fact, biofilms actually became resistant to gentamicin at concentrations far above the minimum inhibitory concentration.

Researchers at Tulane University recently confirmed yet again that low, pulsed dosing is a superior way of targeting treatment-resistant biofilm bacteria. According to the team, who mathematically modeled the action of antibiotics on bacterial biofilms, “Exposing a biofilm to low concentration doses of an antimicrobial agent for longer time is more effective than short time dosing with high antimicrobial agent concentration.”[28]

Similarly, a bioengineer led team at the University of Washington recently created an antibiotic- containing polymer that releases antibiotic slowly onto the surface of hospital devices, such as catheters and prostheses, to reduce the risk of biofilm-related infections.

“Rather than massively dosing the patient with high levels of released antibiotic, this strategy allows the release of extremely low levels of this very potent antibiotic over long periods of time,” explained Buddy Ratner, PhD, Professor and Director of the Engineered Biomaterials Program at the University of Washington, Seattle. “We calculated the amount released at the surface that would kill 100% of the bacteria entering the surface zone.”

When challenged by Dr. Leonard A. Mermel from Brown University School of Medicine on the issue that long-term use of pulsed, low-dose antibiotics might allow for increased resistance on the part of the bacteria being treated, Ratner responded, “Dr. Mermel’s concerns are, in fact, why we developed this system for [antibiotic] release. Bacteria that live through antibiotic dosing can go on to produce resistant strains. If 100% of the bacteria approaching the surface are killed, they can’t produce resistant offspring. The classical physician approach, dosing the patient systemically and heavily to rid the patient of persistent bacteria, can lead to those resistant strains. Our approach releases miniscule doses compared to what a physician would use, but releases the antibiotic where it will be optimally effective and least likely to leave antibiotic-resistant survivors.”

Although taken orally, the MP antibiotics are taken in the same manner as those administered by Ratner and team. Because they too are dosed at optimal times in extremely small doses, the chance that long-term antibiotic use might foster resistant bacteria is again, essentially negligible, especially when multiple antibiotics are typically used.

Key to the ability of the Marshall Protocol to effectively target biofilm bacteria is the fact that the specific pulsed, low-dose bacteriostatic antibiotics used by the treatment are taken in conjunction with a medication called Benicar. Benicar binds and activates the Vitamin D Receptor, displacing bacterial substances and 25-D from the receptor, so that it can once again activate the innate immune system.[29] Benicar is so effective at strengthening the innate immune response that the patient’s own immune system ultimately helps destroy the biofilm weakened by pulsed, low-dose antibiotics.

Thus, it is not enough for patients on the Marshall Protocol to simply take specific pulsed, low-dose antibiotics. The activity of their innate immune system must also be restored so that the cells of the immune system can actively combat biofilm bacteria, the matrix that surrounds them, and persister cells.

After antibiotics are applied to a biofilm, a number of cells called “persisters” are left behindHow do we know that the Marshall Protocol effectively kills biofilm bacteria? Namely because those patients to reach the later stages of the treatment do not report symptoms associated with established biofilm diseases. Patients on the MP who once suffered from chronic ear infections (OM), chronic sinus infections, or periodontal disease find that such infections resolve over the course of treatment. Furthermore, since we now understand that biofilms almost certainly form a large part of the chronic microbiota of pathogens that cause chronic inflammatory and autoimmune diseases, the fact that patients can use the Marshall Protocol to recover from such illnesses again suggests that the treatment must be effectively allowing them to target and destroy biofilms.

Because all evidence points to the fact that the MP does indeed effectively target biofilm bacteria, it is of utmost importance that people who suffer from any sort of biofilm infection start the treatment. Knowledge of the Marshall Protocol has yet to reach the cystic fibrosis community, but there is great hope that if people with the disease were to start the MP, they could destroy the P. aeruginosabiofilms that cause their untimely deaths. In the same vein, people with a wide range of infections, such as those infected with biofilm during surgery, can likely restore their health with the MP.

It is to be hoped that the clinical data emerging from the Marshall Protocol study site, which shows patients recovering from biofilm-related diseases, will inspire future researchers to invest a great deal of energy into further research aimed at identifying and studying the biofilm bacteria – bacteria that almost certainly form part of the microbiota of pathogens that cause inflammatory disease. In the coming years, as the technology to detect biofilms becomes even more sophisticated, it is almost certain that a great number of biofilms will be officially detected and documented in patients with a vast array of chronic diseases.

REFERENCESOn 9 September 2011 15:07, Krys Brennand <krys109uk@...> wrote:

I see what you are saying.  

Really, leaving out the added complications of biofilm & quorum sensing, every time we give antibiotics we are, in effect, breeding for resistance.

I don't think the writer was talking about Lyme per sé.  The cells, which they called " persisters " are particularly interesting. If what is written is found to be the case then one would imagine pulsing antibiotics even in high doses could work better than continuous long term high doses?

On 9 September 2011 13:39, Aandraya <aandraya@...> wrote:

 

Krys-With chronic Lyme and coinfections low dose abx are not recommended because the microbes can build up resistance more easily.  It's better to hit them with therapeutic doses.  I've tried a lot of strategies, what's worked best in my case is multiple antimicrobials given in high doses long term.  That's where I'm at now.  The treatment is no picnic but I am progressing.

Aandraya On Sep 9, 2011, at 1:32 PM, Krys Brennand <krys109uk@...> wrote:

 

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned. 

I found the theory, in the last section, about using low level antibiotics in " pulses "  to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing. 

 Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.I wonder whether the description of the quorum sensing bacteria as using a " language " & being " intelligent " could be misleading. I suppose it depends how we define " intelligent " . We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to. 

All the best,KrysOn 9 September 2011 12:58, <Goldstein@...> wrote:

 

This was a good article if you have time to read it in its entirety.  A couple of things that I did not know that I learned:1.  After antibiotics (all) there are bacteria they have referred to in science as persisters.  This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2.  Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc.  Much like a family or civilization.  They have their own language; they are intelligent.

3.  Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums.  The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.  

4.  One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth.  As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body.  My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5.  Research is picking up on the study of biofilms.  The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6.  Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.7.  Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi.  Maybe there is more information out about that... I'll look.  

So much to be gleaned from this article.  From: " Aandraya " <aandraya@...>bird mites

Sent: Friday, September 9, 2011 10:27:53 AMSubject: Re: Good summary about biofilms

 

I had that film too big time.  I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked.  I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same.  Either way, treating my chronic infections including the antifungals is knocking it out.  

AandrayaOn Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

 

Hi Cecilia,Two things, first, you are right.  I don't think it is good to use Hibiclens long term.  When we finish these bottles, I think that will be the end of that.  Normally have not used antibacterial soaps here... Dr. told us to use it.  But, what do they know?  Right?  Sometimes they give incorrect or bad advice. 

Secondly about biofilms.  I can just speak from experience.  At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin.  Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair.  Husband has it too.  Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm.  I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and

new ones appear.  I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy.  Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created  by Lyme organisms and other organisms together.  I think there is some research being done on this.  I'll see if I can locate anything on it.  Maybe Aandraya knows of something too.

From: " Krys Brennand " <krys109uk@...>bird mites

Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

 

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

 

How do you know you have biofilms?Cecilia

From: " Goldstein@... " <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

 

Yes.  True.  L.From: " Aandraya Da Silva " <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

 

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing " biofilm protocols " assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other " biofilm protocols " in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book " Healing Lyme " by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was " yes " which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Yes, I'm very familiar with her work. Her latest recommendation is Tinidazol for breaking up the cyst form of Borrelia, I want to try it when I finish treating Babesia and Bartonella.I forgot earlier- Xylitol and Lactoferrin for disintegrating Biofilm. I used to mix it together and use it on my skin lesions to clean them out, and I've taken it internally as well. I use pure Xylitol for dental plaque.On Sep 9, 2011, at 10:39 AM, Goldstein@... wrote:

http://www.youtube.com/watch?v=AmvgOfIN_8cIf you have time, watch this doctor/researcher talking about her own experience with Lyme disease; she did cancer research and now does Lyme disease research, particularly into biofilms. Dr. Eva Sapi--From: Goldstein@...To: bird mites Sent: Friday, September 9, 2011 8:32:43 AMSubject: Re: Good summary about biofilms

Understanding BiofilmsAuthor: Amy Proal26MAY2008As humans, our environment consistently exposes us to a variety of dangers. Tornadoes, lightning, flooding and hurricanes can all hamper our survival. Not to mention the fact that most of us can encounter swerving cars or ill-intentioned people at any given moment.Biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of materialThousands of years ago, humans realized that they could better survive a dangerous world if they formed into communities, particularly communities consisting of people with different talents. They realized that a community is far more likely to survive through division of labor– one person makes food, another gathers resources, still another protects the community against invaders. Working together in this manner requires communication and cooperation.Inhabitants of a community live in close proximity and create various forms of shelter in order to protect themselves from external threats. We build houses that protect our families and larger buildings that protect the entire community. Grouping together inside places of shelter is a logical way to enhance survival.With the above in mind, it should come as no surprise that the pathogens we harbor are seldom found as single entities. Although the pathogens that cause acute infection are generally free-floating bacteria – also referred to as planktonic bacteria – those chronic bacterial forms that stick around for decades long ago evolved ways to join together into communities. Why? Because by doing so, they are better able to combat the cells of our immune system bent upon destroying them.It turns out that a vast number of the pathogens we harbor are grouped into communities called biofilms. In an article titled “Bacterial Biofilms: A Common Cause of Persistent Infections,†JW Costerton at the Center for Biofilm Engineering in Montana defines a bacterial biofilm as “a structured community of bacterial cells enclosed in a self-produced polymeric matrix and adherent to an inert or living surface.â€[1] In layman’s terms, that means that bacteria can join together on essentially any surface and start to form a protective matrix around their group. The matrix is made of polymers – substances composed of molecules with repeating structural units that are connected by chemical bonds.According to the Center for Biofilm Engineering at Montana State University, biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of material – such as metals, plastics, soil particles, medical implant materials and, most significantly, human or animal tissue. The first bacterial colonists to adhere to a surface initially do so by inducing weak, reversible bonds called van der Waals forces. If the colonists are not immediately separated from the surface, they can anchor themselves more permanently using cell adhesion molecules, proteins on their surfaces that bind other cells in a process called cell adhesion.A biofilm in the gut.These bacterial pioneers facilitate the arrival of other pathogens by providing more diverse adhesion sites. They also begin to build the matrix that holds the biofilm together. If there are species that are unable to attach to a surface on their own, they are often able to anchor themselves to the matrix or directly to earlier colonists.During colonization, things start to get interesting. Multiple studies have shown that during the time a biofilm is being created, the pathogens inside it can communicate with each other thanks to a phenomenon called quorum sensing. Although the mechanisms behind quorum sensing are not fully understood, the phenomenon allows a single-celled bacterium to perceive how many other bacteria are in close proximity. If a bacterium can sense that it is surrounded by a dense population of other pathogens, it is more inclined to join them and contribute to the formation of a biofilm.Bacteria that engage in quorum sensing communicate their presence by emitting chemical messages that their fellow infectious agents are able to recognize. When the messages grow strong enough, the bacteria respond en masse, behaving as a group. Quorum sensing can occur within a single bacterial species as well as between diverse species, and can regulate a host of different processes, essentially serving as a simple communication network. A variety of different molecules can be used as signals.“Disease-causing bacteria talk to each other with a chemical vocabulary,†says Doug Hibbins of Princeton University. A graduate student in the lab of Princeton University microbiologist Dr. Bonnie Bassler, Hibbins was part of a research effort which shed light on how the bacteria that cause cholera form biofilms and communicate via quorum sensing.[2]“Forming a biofilm is one of the crucial steps in cholera’s progression,†states Bassler. “They [bacteria] cover themselves in a sort of goop that’s a shield against antibiotics, allowing them to grow rapidly. When they sense there are enough of them, they try to leave the body.â€Although cholera bacteria use the intestines as a breeding ground, after enough biofilms have formed, planktonic bacteria inside the biofilm seek to leave the body in order to infect a new host. It didn’t take long for Bassler and team to realize that the bacteria inside cholera biofilms must signal each other in order to communicate that it’s time for the colony to stop reproducing and focus instead on leaving the body.“We generically understood that bacteria talk to each other with quorum sensing, but we didn’t know the specific chemical words that cholera uses,†Bassler said.Then Higgins isolated the CAI-1 – a chemical which occurs naturally in cholera. Another graduate student figured out how to make the molecule in the laboratory. By moderating the level of CAI-1 in contact with cholera bacteria, Higgins was successfully able to chemically control cholera’s behavior in lab tests. His team eventually confirmed that when CAI-1 is absent, cholera bacteria attach in biofilms to their current host. But when the bacteria detect enough of the chemical, they stop making biofilms and releasing toxins, perceiving that it is time to leave the body instead. Thus, CAI-1 may very well be the single molecule that allow the bacteria inside a cholera biofilm to communicate. Although it is likely that the bacteria in a cholera biofilm may communicate with other signals besides CAI-1, the study is a good example of the fact that signaling molecules serve a key role in determining the state of a biofilm.Sessile cells in a biofilm “talk†to each other via quorum sensing to build microcolonies and to keep water channels open.Similarly, researchers at the University of Iowa (several of whom are now at the University of Washington) have spent the last decade identifying the molecules that allow the bacterial species P. aeruginosa to form biofilms in the lungs of patients with cystic fibrosis.[3] Although the P. auruginosa isolated from the lungs of patients with cystic fibrosis looks like a biofilm and acts like a biofilm, up until recently, there were no objective tests available to confirm that the bacterial species did indeed form biofilms in the lungs of patients with the disease, nor was there a way to tell what proportion of P. aeruginosa in the lungs were actually in biofilm mode.“We needed a way to show that the P. auruginosa in cystic fibrosis lungs was communicating like a biofilm. That could tell us about the P. auruginosalifestyle,†said Pradeep Singh, M.D., a lead author on the study who is now at the University of Washington.Singh and his colleagues finally discovered that P. aeruginosa uses one of two particular quorum-sensing molecules to initiate the formation of biofilms. In November 1999, his research team screened the entire bacterial genome, identifying 39 genes that are strongly controlled by the quorum-sensing system.In a 2000 study published in Nature, Singh and colleagues developed a sensitive test which shows P. auruginosa from cystic fibrosis lungs produces the telltale, quorum-sensing molecules that are the signals for biofilm formation.[3]It turns out that P. aerugnosa secretes two signaling molecules, one that is long, and another that is short. Using the new test, the team was able to show that planktonic forms of P. aeruginosa produce more long signaling molecules. Alternately, when they tested the P. aeruginosa strains isolated from the lungs of patients with cystic fibrosis (which were in biofilm form), all of the strains produced the signaling molecules, but in the opposite ratio – more short than long.Interestingly, when the biofilm strains of P. aeruginosa were separated in broth into individual bacterial forms, they reverted to producing more long signal molecules than short ones. Does this mean that a change in signaling molecular length can indicate whether bacteria remain as planktonic forms or develop into biofilms?To find out, the team took the bacteria from the broth and made them grow as a biofilm again. Sure enough, those strains of bacteria in biofilm form produced more short signal molecules than long.“The fact that the P. aeruginosa in [the lungs of cystic fibrosis patients] is making the signals in the ratios that we see tells us that there is a biofilm and that most of the P. aeruginosa in the lung is in the biofilm state,†states Greenberg, another member of the research team. He believes that the findings allow for a clear biochemical definition of whether bacteria are in a biofilm. Techniques similar to those used by his group will likely be used to determine the properties of other biofilm signaling molecules.DevelopmentOnce colonization has begun, the biofilm grows through a combination of cell division and recruitment. The final stage of biofilm formation is known as development and is the stage in which the biofilm is established and may only change in shape and size. This development of a biofilm allows for the cells inside to become more resistant to antibiotics administered in a standard fashion. In fact, depending on the organism and type of antimicrobial and experimental system, biofilm bacteria can be up to a thousand times more resistant to antimicrobial stress than free-swimming bacteria of the same species.Biofilms grow slowly, in diverse locations, and biofilm infections are often slow to produce overt symptoms. However, biofilm bacteria can move in numerous ways that allow them to easily infect new tissues. Biofilms may move collectively, by rippling or rolling across the surface, or by detaching in clumps. Sometimes, in a dispersal strategy referred to as “swarming/seedingâ€, a biofilm colony differentiates to form an outer “wall†of stationary bacteria, while the inner region of the biofilm “liquefiesâ€, allowing planktonic cells to “swim†out of the biofilm and leave behind a hollow mound.[4]Biofilm bacteria can move in numerous ways: Collectively, by rippling or rolling across the surface, or by detaching in clumps. Individually, through a “swarming and seeding†dispersal.Research on the molecular and genetic basis of biofilm development has made it clear that when cells switch from planktonic to community mode, they also undergo a shift in behavior that involves alterations in the activity of numerous genes. There is evidence that specific genes must be transcribed during the attachment phase of biofilm development. In many cases, the activation of these genes is required for synthesis of the extracellular matrix that protects the pathogens inside.According to Costerton, the genes that allow a biofilm to develop are activated after enough cells attach to a solid surface. “Thus, it appears that attachment itself is what stimulates synthesis of the extracellular matrix in which the sessile bacteria are embedded,†states the molecular biologist. “This notion– that bacteria have a sense of touch that enables detection of a surface and the expression of specific genes– is in itself an exciting area of research…â€[1]Certain characteristics may also facilitate the ability of some bacteria to form biofilms. Scientists at the Department of Microbiology and Molecular Genetics, Harvard Medical School, performed a study in which they created a “mutant†form of the bacterial species P. aeguinosa (PA).[5] The mutants lacked genes that code for hair-like appendages called pili. Interestingly, the mutants were unable to form biofilms. Since the pili of PA are involved in a type of surface-associated motility called twitching, the team hypothesized this twitching might be required for the aggregation of cells into the microcolonies that subsequently form a stable biofilm.Once a biofilm has officially formed, it often contains channels in which nutrients can circulate. Cells in different regions of a biofilm also exhibit different patterns of gene expression. Because biofilms often develop their own metabolism, they are sometimes compared to the tissues of higher organisms, in which closely packed cells work together and create a network in which minerals can flow.“There is a perception that single-celled organisms are asocial, but that is misguided,†said Andre Levchenko, assistant professor of biomedical engineering in s Hopkins University’s Whiting School of Engineering and an affiliate of the University’s Institute for NanoBioTechnology. “When bacteria are under stress—which is the story of their lives—they team up and form this collective called a biofilm. If you look at naturally occurring biofilms, they have very complicated architecture. They are like cities with channels for nutrients to go in and waste to go out.â€[6]The biofilm life cycle in three steps: attachment, growth of colonies (development), and periodic detachment of planktonic cells.Understanding how such cooperation among pathogens evolves and is maintained represents one of evolutionary biology’s thorniest problems. This stems from the reality that, in nature, freeloading cheats inevitably evolve to exploit any cooperative group that doesn’t defend itself, leading to the breakdown of cooperation. So what causes the bacteria in a biofilm to contribute to and share resources rather than steal them? Recently, Dr. Brockhurst of the University of Liverpool and colleagues at the Université Montpellier and the University of Oxford conducted several studies in an effort to understand why the bacteria in a biofilm cooperate and share resources rather than horde them.[7]The team took a closer look at P. fluorescens biofilms, which are formed when individual cells overproduce a polymer that sticks the cells together, allowing the colonization of liquid surfaces. While production of the polymer is metabolically costly to individual cells, the biofilm group benefits from the increased access to oxygen that surface colonization provides. Yet, evolutionarily speaking, such a setup allows possible “cheaters†to enter the biofilm. Such cheats can take advantage of the protective matrix while failing to contribute energy to actually building the matrix. If too many “cheaters†enter a biofilm, it will weaken and eventually break apart.After several years of study, Brockhurst and team realized that the short-term evolution of diversity within a biofilm is a major factor in how successfully its members cooperate. The team found that once inside a biofilm, P. fluorescensdifferentiates into various forms, each of which uses different nutrient resources. The fact that these “diverse cooperators†don’t all compete for the same chemicals and nutrients substantially reduces competition for resources within the biofilm.When the team manipulated diversity within experimental biofilms, they found that diverse biofilms contained fewer “cheaters†and produced larger groups than non-diverse biofilms.Levchenko and team used this device to observe bacteria growing in cramped conditions.Similarly, this year, researchers from s Hopkins; Virginia Tech; the University of California, San Diego; and Lund University in Sweden recently released the results of a study which found that once bacteria cooperate and form a biofilm, packing tightly together further enhances their survival.[6]The team created a new device in order to observe the behavior of E. coli bacteria forced to grow in the cramped conditions. The device, which allows scientists to use extremely small volumes of cells in solution, contains a series of tiny chambers of various shapes and sizes that keep the bacteria uniformly suspended in a culture medium.Not surprisingly, the cramped bacteria in the device began to form a biofilm. The team captured the development of the biofilm on video, and were able to observe the gradual self-organization and eventual construction of bacterial biofilms over a 24-hour period.First, Andre Levchenko and Hojung Cho of s Hopkins recorded the behavior of single layers of E. coli cells using real-time microscopy. “We were surprised to find that cells growing in chambers of all sorts of shapes gradually organized themselves into highly regular structures,†Levchenko said.Dr. Levchenko of s Hopkins and Hojung Cho, a biomedical engineering doctoral studentFurther observations using microscopy revealed that the longer the packed cell population resided in the chambers, the more ordered the biofilm structure became. As the cells in the biofilm became more ordered and tightly packed, the biofilm became harder and harder to penetrate.Levchenko also noted that rod-shaped E. colithat were too short or too long typically did not organize well into the dense, circular main hub of the biofilm. Instead, the bacteria of odd shapes or highly disordered groups of cells were found on the edges of the biofilm, where they formed sharp corners.Nodes of relapsing infection?Researchers often note that, once biofilms are established, planktonic bacteria may periodically leave the biofilm on their own. When they do, they can rapidly multiply and disperse.According to Costerton, there is a natural pattern of programmed detachment of planktonic cells from biofilms. This means that biofilms can act as what Costerton refers to as “niduses†of acute infection. Because the bacteria in a biofilm are protected by a matrix, the host immune system is less likely to mount a response to their presence.[1]But if planktonic bacteria are periodically released from the biofilms, each time single bacterial forms enter the tissues, the immune system suddenly becomes aware of their presence. It may proceed to mount an inflammatory response that leads to heightened symptoms. Thus, the periodic release of planktonic bacteria from some biofilms may be what causes many chronic relapsing infections.Planktonic bacteria are periodically released from a biofilmAs R. Parsek of Northwestern University describes in a 2003 paper in the Annual Review of Microbiology, any pathogen that survives in a chronic form benefits by keeping the host alive.[8] After all, if a chronic bacterial form simply kills its host, it will no longer have a place to live. So according to Parsek, chronic infection often results in a “disease stalemate†where bacteria of moderate virulence are somewhat contained by the defenses of the host. The infectious agents never actually kill the host, but the host is never able to fully kill the invading pathogens either.Parsek believes that the optimal way for bacteria to survive under such circumstances is in a biofilm, stating that “Increasing evidence suggests that the biofilm mode of growth may play a key role in both of these adaptations. Biofilm growth increases the resistance of bacteria to killing and may make organisms less conspicuous to the immune system… ultimately this moderation of virulence may serve the bacteria’s interest by increasing the longevity of the host.â€The acceptance of biofilms as infectious entitiesAnton van Leeuwenhoek.Perhaps because many biofilms are sufficiently thick to be visible to the naked eye, the microbial communities were among the first to be studied by early microbiologists. Anton van Leeuwenhoek scraped the plaque biofilm from his teeth and observed what he described as the “animalculi†inside them under his primitive microscope. However, according to Costerton and team at the Center for Biofilm Research at Montana State University, it was not until the 1970s that scientists began to appreciate that bacteria in the biofilm mode of existence constitute such a major component of the bacterial biomass in most environments. Then, it was not until the 1980s and 1990s that scientists truly began to understand how elaborately organized a bacterial biofilm community can be.[1]As Kolter, professor of microbiology and molecular genetics at Harvard Medical School, and one of the first scientists to study how biofilms developstates, “At first, however, studying biofilms was a radical departure from previous work.â€Like most microbial geneticists, Kolter had been trained in the tradition dating back to Koch and Louis Pasteur, namely that bacteriology is best conducted by studying pure strains of planktonic bacteria. “While this was a tremendous advance for modern microbiology, it also distracted microbiologists from a more organismic view of bacteria, Kolter adds, “Certainly we felt that pure, planktonic cultures were the only way to work. Yet in nature bacteria don’t live like that,†he says. “In fact, most of them occur in mixed, surface-dwelling communities.â€Although research on biofilms has surged over the past few decades, the majority of biofilm research to date has focused on external biofilms, or those that form on various surfaces in our natural environment.Over the past years, as scientists developed better tools to analyze external biofilms, they quickly discovered that biofilms can cause a wide range of problems in industrial environments. For example, biofilms can develop on the interiors of pipes, which can lead to clogging and corrosion. Biofilms on floors and counters can make sanitation difficult in food preparation areas.Since biofilms have the ability to clog pipes, watersheds, storage areas, and contaminate food products, large companies with facilities that are negatively impacted by their presence have naturally taken an interest in supporting biofilm research, particularly research that specifies how biofilms can be eliminated.This means that many recent advances in biofilm detection have resulted from collaborations between microbial ecologists, environmental engineers, and mathematicians. This research has generated new analytical tools that help scientists identify biofilms.Biofilm in a swamp gas reactor.For example, the Canadian company FAS International Ltd. has justcreated an endoluminal brush, which will be launched this spring. Physicians can use the brush to obtain samples from the interior of catheters. Samples taken from catheters can be sent to a lab, where researchers determine if biofilms are present in the sample. If biofilms are detected, the catheter is immediately replaced, since the insertion of catheters with biofilms can cause the patient to suffer from numerous infections, some of which are potentially life threatening.Scientists now realize that biofilms are not just composed of bacteria. Nearly every species of microorganism – including viruses, fungi, and Archaea – have mechanisms by which they can adhere to surfaces and to each other. Furthermore, it is now understood that biofilms are extremely diverse. For example, upward of 300 different species of bacteria can inhabit the biofilms that form dental plaque.[9]Furthermore, biofilms have been found literally everywhere in nature, to the point where any mainstream microbiologist would acknowledge that their presence is ubiquitous. They can be found on rocks and pebbles at the bottom of most streams or rivers and often form on the surface of stagnant pools of water. In fact, biofilms are important components of food chains in rivers and streams and are grazed upon by the aquatic invertebrates upon which many fish feed. Biofilms even grow in the hot, acidic pools at Yellowstone National Park and on glaciers in Antarctica.Biofilm in acidic pools at Yellowstone National Park.It is also now understood that the biofilm mode of existence has been around for millenia. For example, filamentous biofilms have been identified in the 3.2-billion-year-old deep-sea hydrothermal rocks of the Pilbara Craton, Australia. According to a 2004 article in Nature Reviews Microbiology, “Biofilm formation appears early in the fossil record (approximately 3.25 billion years ago) and is common throughout a diverse range of organisms in both the Archaea and Bacteria lineages. It is evident that biofilm formation is an ancient and integral component of the prokaryotic life cycle, and is a key factor for survival in diverse environments.â€[10]Biofilms and diseaseThe fact that external biofilms are ubiquitous raises the question – if biofilms can form on essentially every surface in our external environments, can they do the same inside the human body? The answer seems to be yes, and over the past few years, research on internal biofilms has finally started to pick up pace. After all, it’s easy for biofilm researchers to see that the human body, with its wide range of moist surfaces and mucosal tissue, is an excellent place for biofilms to thrive. Not to mention the fact that those bacteria which join a biofilm have a significantly greater chance of evading the battery of immune system cells that more easily attack planktonic forms.Many would argue that research on internal biofilms has been largely neglected, despite the fact that bacterial biofilms seem to have great potential for causing human disease.Common sites of biofilm infection. One biofilm reach the bloodstream they can spread to any moist surface of the human body. Stoodley of the Center for Biofilm Engineering at Montana State University, attributes much of the lag in studying biofilms to the difficulties of working with heterogeneous biofilms compared with homogeneous planktonic populations. In a 2004 paper in Nature Reviews, the molecular biologist describes many reasons why biofilms are extremely difficult to culture, such as the fact that the diffusion of liquid through a biofilm and the fluid forces acting on a biofilm must be carefully calculated if it is to be cultured correctly. According to Stoodley, the need to master such difficult laboratory techniques has deterred many scientists from attempting to work with biofilms. [10]Also, since much of the technology needed to detect internal biofilms was created at the same time as the sequencing of the human genome, interest in biofilm bacteria, and the research grants that would accompany such interest, have been largely diverted to projects with a decidedly genetic focus. However, since genetic research has failed to uncover the cause of any of the common chronic diseases, biofilms are finally – just over the past few years – being studied more intensely, and being given the credit they deserve as serious infectious entities, capable of causing a wide array of chronic illnesses.In just a short period of time, researchers studying internal biofilms have already pegged them as the cause of numerous chronic infections and diseases, and the list of illnesses attributed to these bacterial colonies continues to grow rapidly.According to a recent public statement from the National Institutes of Health, more than 65% of all microbial infections are caused by biofilms. This number might seem high, but according to Kim of the Department of Chemical and Biological Engineering at Tufts University, “If one recalls that such common infections as urinary tract infections (caused by E. coli and other pathogens), catheter infections (caused by Staphylococcus aureus and other gram-positive pathogens), child middle-ear infections (caused by Haemophilus influenzae, for example), common dental plaque formation, and gingivitis, all of which are caused by biofilms, are hard to treat or frequently relapsing, this figure appears realistic.â€[11]Hundreds of microbial biofilm colonize the human mouth, causing tooth decay and gum disease.As mentions, perhaps the most well-studied biofilms are those that make up what is commonly referred to as dental plaque. “Plaque is a biofilm on the surfaces of the teeth,†states Parsek. “This accumulation of microorganisms subject the teeth and gingival tissues to high concentrations of bacterial metabolites which results in dental disease.â€[12]It has also recently been shown that biofilms are present on the removed tissue of 80% of patients undergoing surgery for chronic sinusitis. According to Parsek, biofilms may also cause osteomyelitis, a disease in which the bones and bone marrow become infected. This is supported by the fact that microscopy studies have shown biofilm formation on infected bone surfaces from humans and experimental animal models. Parsek also implicates biofilms in chronic prostatitis since microscopy studies have also documented biofilms on the surface of the prostatic duct. Microbes that colonize vaginal tissue and tampon fibers can also form into biofilms, causing inflammation and disease such as Toxic Shock Syndrome.Biofilms also cause the formation of kidney stones. The stones cause disease by obstructing urine flow and by producing inflammation and recurrent infection that can lead to kidney failure. Approximately 15%–20% of kidney stones occur in the setting of urinary tract infection. According to Parsek, these stones are produced by the interplay between infecting bacteria and mineral substrates derived from the urine. This interaction results in a complex biofilm composed of bacteria, bacterial exoproducts, and mineralized stone material.Microbes that colonize vaginal tissue and tampon fibers can become pathogenic, causing inflammation and disease such as Toxic Shock Syndrome.Perhaps the first hint of the role of bacteria in these stones came in 1938 when Hellstrom examined stones passed by his patients and found bacteria embedded deep inside them. Microscopic analysis of stones removed from infected patients has revealed features that characterize biofilm growth. For one thing, bacteria on the surface and inside the stones are organized in microcolonies and surrounded by a matrix composed of crystallized (struvite) minerals.Then there’s endocarditis, a disease that involves inflammation of the inner layers of the heart. The primary infectious lesion in endocarditis is a complex biofilm composed of both bacterial and host components that is located on a cardiac valve. This biofilm, known as a vegetation, causes disease by three basic mechanisms. First, the vegetation physically disrupts valve function, causing leakage when the valve is closed and inducing turbulence and diminished flow when the valve is open. Second, the vegetation provides a source for near-continuous infection of the bloodstream that persists even during antibiotic treatment. This causes recurrent fever, chronic systemic inflammation, and other infections. Third, pieces of the infected vegetation can break off and be carried to a terminal point in the circulation where they block the flow of blood (a process known as embolization). The brain, kidney, and extremities are particularly vulnerable to the effects of embolization.A variety of pathogenic biofims are also commonly found on medical devices such as joint prostheses and heart valves. According to Parsek, electron microscopy of the surfaces of medical devices that have been foci of device-related infections shows the presence of large numbers of slime-encased bacteria. Tissues taken from non-device-related chronic infections also show the presence of biofilm bacteria surrounded by an exopolysaccharide matrix. These biofilm infections may be caused by a single species or by a mixture of species of bacteria or fungi.According to Dr. Patel of the Mayo Clinic, individuals with prosthetic joints are often oblivious to the fact that their prosthetic joints harbor biofilm infections.[13]Cells of Staphylococcus epidermidis causing devastating disease as they grow on the cuff at a mechanical heart valve.“When people think of infection, they may think of fever or pus coming out of a wound,†explains Dr. Patel. “However, this is not the case with prosthetic joint infection. Patients will often experience pain, but not other symptoms usually associated with infection. Often what happens is that the bacteria that cause infection on prosthetic joints are the same as bacteria that live harmlessly on our skin. However, on a prosthetic joint they can stick, grow and cause problems over the long term. Many of these bacteria would not infect the joint were it not for the prosthesis.â€Biofilms also cause Leptospirosis, a serious but neglected emerging disease that infects humans through contaminated water. New research published in the May issue of the journal Microbiology shows for the first time how bacteria that cause the disease survive in the environment.Leptospirosis is a major public health problem in southeast Asia and South America, with over 500,000 severe cases every year. Between 5% and 20% of these cases are fatal. Rats and other mammals carry the disease-causing pathogen Leptospira interrogans in their kidneys. When they urinate, they contaminate surface water with the bacteria, which can survive in the environment for long periods.“This led us to see if the bacteria build a protective casing around themselves for protection,†said Professor Mathieu Picardeau from the Institut Pasteur in Paris, France. [14]Previously, scientists believed the bacteria were planktonic. But Professor Picardeau and his team have shown that L. interrogans can make biofilms, which could be one of the main factors controlling survival and disease transmission. “90% of the species of Leptospira we tested could form biofilms. It takes L. interrogans an average of 20 days to make a biofilm,†says Picardeau.Biofilms have also been implicated in a wide array of veterinary diseases. For example, researchers at the Virginia-land Regional College of Veterinary Medicine at Virginia Tech were just awarded a grant from the United States Department of Agriculture to study the role biofilms play in the development of Bovine Respiratory Disease Complex (BRDC). If biofilms play a role in bovine respiratory disease, it’s likely only a matter of time before they will be established as a cause of human respiratory diseases as well.When the immune response is compromised, Pseudomonas aeruginosabiofilms are able to colonize the alveoli, and to form biofilms.As mentioned previously, infection by the bacterium Pseudomonas aeruginosa (P. aeruginosa) is the main cause of death among patients with cystic fibrosis. Pseudomonas is able to set up permanent residence in the lungs of patients with cystic fibrosis where, if you ask most mainstream researchers, it is impossible to kill. Eventually, chronic inflammation produced by the immune system in response to Pseudomonas destroys the lung and causes respiratory failure. In the permanent infection phase, P. aeruginosa biofilms are thought to be present in the airway, although much about the infection pathogenesis remains unclear.[15]Cystic fibrosis is caused by mutations in the proteins of channels that regulates chloride. How abnormal chloride channel protein leads to biofilm infection remains hotly debated. It is clear, however, that cystic fibrosis patients manifest some kind of host-defense defect localized to the airway surface. Somehow this leads to a debilitating biofilm infection.Biofilms have the potential to cause a tremendous array of infections and diseasesBecause internal pathogenic biofilm research comprises such a new field of study, the infections described above almost certainly represent just the tip of the iceberg when it comes to the number of chronic diseases and infections currently caused by biofilms.For example, it wasn’t until July of 2006 that researchers realized that the majority of ear infections are caused by biofilm bacteria. These infections, which can be either acute or chronic, are referred to collectively as otitis media (OM). They are the most common illness for which children visit a physician, receive antibiotics, or undergo surgery in the United States.There are two subtypes of chronic OM. Recurrent OM (ROM) is diagnosed when children suffer repeated infections over a span of time and during which clinical evidence of the disease resolves between episodes. Chronic OM with effusion is diagnosed when children have persistent fluid in the ears that lasts for months in the absence of any other symptoms except conductive hearing loss.It took over ten years for researchers to realize that otitis media is caused by biofilms. Finally, in 2002, Drs. Ehrlich and J. Post, an Allegheny General Hospital pediatric ear specialist and medical director of the Center for Genomic Sciences, published the first animal evidence of biofilms in the middle ear in the Journal of the American Medical Association, setting the stage for further clinical investigation.In a subsequent study, Ehrlich and Post obtained middle ear mucosa – or membrane tissue – biopsies from children undergoing a procedure for otitis. The team gathered uninfected mucosal biopsies from children and adults undergoing cochlear implantation as a control.[16]Using advanced confocal laser scanning microscopy, Luanne Hall Stoodley, Ph.D. and her ASRI colleagues obtained three dimensional images of the biopsies and evaluated them for biofilm morphology using generic stains and species-specific probes for Haemophilus influenzae, Streptococcus pneumoniaeand Moraxella catarrhalis. Effusions, when present, were also evaluated for evidence of pathogen specific nucleic acid sequences (indicating presence of live bacteria).The study found mucosal biofilms in the middle ears of 46/50 children (92%) with both forms of otitis. Biofilms were not observed in eight control middle ear mucosa specimens obtained from cochlear implant patients.Otitis media, or inflammation of the inner ear, is caused by biofilm.In fact, all of the children in the study who suffered from chronic otitis media tested positive for biofilms in the middle ear, even those who were asymptomatic, causing Erlich to conclude that, “It appears that in many cases recurrent disease stems not from re-infection as was previously thought and which forms the basis for conventional treatment, but from a persistent biofilm.â€He went on to state that the discovery of biofilms in the setting of chronic otitis media represented “a landmark evolution in the medical community’s understanding about a disease that afflicts millions of children world-wide each year and further endorses the emerging biofilm paradigm of chronic infectious disease.â€The emerging biofilm paradigm of chronic disease refers to a new movement in which researchers such as Ehrlich are calling for a tremendous shift in the way the medical community views bacterial biofilms. Those scientists who support an emerging biofilm paradigm of chronic disease feel that biofilm research is of utmost importance because of the fact that the infectious entities have the potential to cause so many forms of chronic disease. The Marshall Pathogenesis is an important part of this paradigm shift.It was also just last year that researchers realized that biofilms cause most infections associated with contact lens use. In 2006, Bausch & Lomb withdrew its ReNu with MoistureLoc contact lens solution because a high proportion of corneal infections were associated with it. It wasn’t long before researchers at the University Hospitals Case Medical Center found that the infections were caused by biofilms. [17]“Once they live in that type of state [a biofilm], the cells become resistant to lens solutions and immune to the body’s own defense system,†said Mahmoud A. Ghannoum, Ph.D, senior investigator of the study. “This study should alert contact lens wearers to the importance of proper care for contact lenses to protect against potentially virulent eye infections,†he said.It turns out that the biofilms detected by Ghannoum and team were composed of fungi, particularly a species called Fusarium. His team also discovered that the strain of fungus (with the catchy name, ATCC 36031) used for testing the effectiveness of lens care solutions is a strain that does not produce biofilms as the clinical fungal strains do. ReNu contact solution, therefore, was effective in the laboratory, but failed when faced with strains in real-world situations.Fungal biofilm can form in contact lens solution leading to potentially virulent eye infectionsUnfortunately, Ghannoum and team were not able to create a method to target and destroy the fungal biofilms that plague users of ReNu and some other contact lens solutions.Then there’s Dr. Randall Wolcott who just recently discovered and confirmed that the sludge covering diabetic wounds is largely made up of biofilms. Whereas before Wolcott’s work such limbs generally had to be amputated, now that they have been correctly linked to biofilms, measures such as those described in thisinterview can be taken to stop the spread of infection and save the limb. Wolcott has finally been given a grant by the National Institutes of Health to further study chronic biofilms and wound development.Dr. Garth and the Medical Biofilm Laboratory team at Montana State University are also researching wounds and biofilms. Their latest article and an image showing wound biofilm was featured on the cover of the January-February 2008 issue of Wound Repair and Regeneration.[18]Biofilm bacteria and chronic inflammatory diseaseIn just a few short years, the potential of biofilms to cause debilitating chronic infections has become so clear that there is little doubt that biofilms are part of the pathogenic mix or “pea soup†that cause most or all chronic “autoimmune†and inflammatory diseases.In fact, thanks, in large part, to the research of biomedical researcher Dr. Trevor Marshall, it is now increasingly understood that chronic inflammatory diseases result from infection with a large microbiota of chronic biofilm and L-form bacteria (collectively called the Th1 pathogens).[19][20] The microbiota is thought to be comprised of numerous bacterial species, some of which have yet to be discovered. However, most of the pathogens that cause inflammatory disease have one thing in common – they have all developed ways to evade the immune system and persist as chronic forms that the body is unable to eliminate naturally.Some L-form bacteria are able to evade the immune system because, long ago, they evolved the ability to reside inside macrophages, the very white bloods cells of the immune system that are supposed to kill invading pathogens. Upon formation, L-form bacteria also lose their cell walls, which makes them impervious to components of the immune response that detect invading pathogens by identifying the proteins on their cell walls. The fact that L-form bacteria lack cell walls also means that the beta-lactam antibiotics, which work by targeting the bacterial cell wall, are completely ineffective at killing them.[21]Clearly, transforming into the L-form offers any pathogen a survival advantage. But among those pathogens not in an L-form state, joining a biofilm is just as likely to enhance their ability to evade the immune system. Once enough chronic pathogens have grouped together and formed a stable community with a strong protective matrix, they are likely able to reside in any area of the body, causing the host to suffer from chronic symptoms that are both mental and physical in nature.Biofilm researchers will also tell you that, not surprisingly, biofilms form with greater ease in an immunocompromised host. Marshall’s research has made it clear that many of the Th1 pathogens are capable of creating substances that bind and inactivate the Vitamin D Receptor – a fundamental receptor of the body that controls the activity of the innate immune system, or the body’s first line of defense against intracellular infection.[22]Diagram of the Vitamin D Receptor and capnine.Thus, as patients accumulate a greater number of the Th1 pathogens, more and more of the chronic bacterial forms create substances capable of disabling the VDR. This causes a snowball effect, in which the patient becomes increasingly immunocompromised as they acquire a larger bacterial load.For one thing, it’s possible that many of the bacteria that survive inside biofilms are capable of creating VDR blocking substances. Thus, the formation of biofilms may contribute to immune dysfunction. Conversely, as patients acquire L-form bacteria and other persistent bacterial forms capable of creating VDR-blocking substances, it becomes exceptionally easy for biofilms to form on any tissue surface of the human body.Thus, patients who begin to acquire L-form bacteria almost always fall victim to biofilm infections as well, since it is all too easy for pathogens to group together into a biofilm when the immune system isn’t working up to par.To date, there is also no strict criteria that separate L-form bacteria from biofilm bacteria or any other chronic pathogenic forms. This means that L-form bacteria may also form into biofilms, and by doing so enter a mode of survival that makes them truly impervious to the immune system. Some L-form bacteria may not form complete biofilms, yet may still possess the ability to surround themselves in a protective matrix. Under these circumstances one might say they are in a “biofilm-like†state.Marshall often refers to the pathogens that cause inflammatory disease as an intraphgocytic, metagenomic microbiota of bacteria, terms which suggest that most chronic bacterial forms possess properties of both L-form and biofilm bacteria. Intraphagocytic refers to the fact that the pathogens can be found inside the cells of the immune system. The term metagenomic indicates that there are a tremendous number of different species of these chronic bacterial forms. Finally, microbiota refers to the fact that biofilm communities sustain their pathogenic activity.For example, when observed under a darkfield microscope, L-form bacteria are often encased in protective biofilm sheaths. If the blood containing the pathogens are aged overnight, the bacterial colonies reach a point where they expand and burst out of the cell, causing the cell to burst as well. Then they extend as huge, long biofilm tubules, which are presumably helping the pathogens spread to other cells. The tubules also help spread bacterial DNA to neighboring cells.Clearly, there is a great need for more research on how different chronic bacterial forms interact. To date, L-form researchers have essentially focused soley on the L-form, while failing to investigate how frequently the wall-less pathogens form into biofilms or become parts of biofilm communities together with bacteria with cell walls. Conversely, most biofilm researchers are intently studying the biofilm mode of growth without considering the presence of L-form bacteria. So, it will likely take several years before we will be better able to understand probable overlaps between the lifestyles of L-form and biofilm bacteria.Anyone who is skeptical about the fact that biofilms likely form a large percentage of the microbiota that cause inflammatory disease should consider many of the recent studies that have linked established biofilm infections to a higher risk for multiple forms of chronic inflammatory disease. Take, for example, studies that have found a link between periodontal disease and several major inflammatory conditions. A 1989 article published in British Medical Journal showed a correlation between dental disease and systemic disease (stroke, heart disease, diabetes). After correcting for age, exercise, diet, smoking, weight, blood cholesterol level, alcohol use and health care, people who had periodontal disease had a significantly higher incidence of heart disease, stroke and premature death. More recently, these results were confirmed in studies in the United States, Canada, Great Britain, Sweden, and Germany. The effects are striking. For example, researchers from the Canadian Health Bureau found that people with periodontal disease had a two times higher risk of dying from cardiovascular disease.[23]Dental plaque as seen under a scanning electron microcroscope.Since we know that periodontal disease is caused by biofilm bacteria, the most logical explanation for the fact that people with dental problems are much more likely to suffer from heart disease and stroke is that the biofilms in their mouths have gradually spread to the moist surfaces of their circulatory systems. Or perhaps if the bacteria in periodontal biofilms create VDR binding substances, their ability to slow innate immune function allows new biofilms (and L-form bacteria as well) to more easily form and infect the heart and blood vessels. Conversely, systemic infection with VDR blocking biofilm bacteria is also likely to weaken immune defenses in the gums and facilitate periodontal disease.In fact, it appears that biofilm bacteria in the mouth also facilitate the formation of biofilm and L-form bacteria in the brain. Just last year, researchers at Vasant Hirani at University College London released the results of a study which found that elderly people who have lost their teeth are at more than three-fold greater risk of memory problems and dementia.[24]At the moment, Autoimmunity Research Foundation does not have the resources to culture biofilms from patients on the treatment and, even if they did, current methods for culturing internal biofilms remain unreliable. According to Stoodley, “The lack of standard methods for growing, quantifying and testing biofilms in continuous culture results in incalculable variability between laboratory systems. Biofilm microbiology is complex and not well represented by flask cultures. Although homogeneity allows statistical enumeration, the extent to which it reflects the real, less orderly world is questionable.â€[10]How else do we acquire biofilm bacteria?As discussed thus far, biofilms form spontaneously as bacteria inside the human body group together. Yet people can also ingest biofilms by eating contaminated food.According to researchers at the University of Guelph in Ontario Canada, it is increasingly suspected that biofilms play an important role in contamination of meat during processing and packaging. The group warns that greater action must be taken to reduce the presence of food-borne pathogens like Escherichia coli and Listeria monocytogenes and spoilage microorganisms such as thePseudomonas species (all of which form biofilms) throughout the food processing chain to ensure the safety and shelf-life of the product. Most of these microorganisms are ubiquitous in the environment or brought into processing facilities through healthy animal carriers.Hans Blaschek of the University of Illinois has discovered that biofilms form on much of the other food products we consume as well.A biofilm on a piece of lettuce“If you could see a piece of celery that’s been magnified 10,000 times, you’d know what the scientists fighting foodborne pathogens are up against,†says Blaschek.“It’s like looking at a moonscape, full of craters and crevices. And many of the pathogens that cause foodborne illness, such as Shigella, E. coli,and Listeria, make sticky, sugary biofilms that get down in these crevices, stick like glue, and hang on like crazy.â€According to Blaschek, the problem faced by produce suppliers can be a triple whammy. “If you’re unlucky enough to be dealing with a pathogen–and the pathogen has the additional attribute of being able to form biofilm—and you’re dealing with a food product that’s minimally processed, well, you’re triply unlucky,†the scientist said. “You may be able to scrub the organism off the surface, but the cells in these biofilms are very good at aligning themselves in the subsurface areas of produce.†, a University of Illinois food science and human nutrition professor agrees, stating,â€Once the pathogenic organism gets on the product, no amount of washing will remove it. The microbes attach to the surface of produce in a sticky biofilm, and washing just isn’t very effective.â€Biofilms can even be found in processed water. Just this month, a study was released in which researchers at the Department of Biological Sciences, at Virginia Polytechnic Institute isolated M. avium biofilm from the shower head of a woman with M. avium pulmonary disease.[25] A molecular technique called DNA fingerprinting demonstrated that M. avium isolates from the water were the same forms that were causing the woman’s respiratory illness.Effectively targeting biofilm infectionsAlthough the mainstream medical community is rapidly acknowledging the large number of diseases and infections caused by biofilms, most researchers are convinced that biofilms are difficult or impossible to destroy, particularly those cells that form the deeper layers of a thick biofilm. Most papers on biofilms state that they are resistant to antibiotics administered in a standard manner. For example, despite the fact that Ehrlich and team discovered that biofilm bacteria cause otitis media, they are unable to offer an effective solution that would actually allow for the destruction of biofilms in the ear canal. Other teams have also come up short in creating methods to break up the biofilms they implicate as the cause of numerous infections.This means patients with biofilm infections are generally told by mainstream doctors that they have an untreatable infection. In some cases, a disease-causing biofilm can be cut out of a patient’s tissues, or efforts are made to drain components of the biofilm out of the body. For example, doctors treating otitis media often treats patients with myringotomy, a surgical procedure in which small tubes are placed in the eardrum to continuously drain infectious fluid.When it comes to administering antibiotics in an effort to target biofilms, one thing is certain. Mainstream researchers have repeatedly tried to kill biofilms by giving patients high, constant doses of antibiotics. Unfortunately, when administered in high doses, the antibiotic may temporarily weaken the biofilm but is incapable of destroying it, as certain cells inevitably persist and allow the biofilm to regenerate.“You can put a patient on [a high dose] antibiotics, and it may seem that the infection has disappeared,†says Levchenko. “But in a few months, it reappears, and it is usually in an antibiotic-resistant form.â€What the vast majority of researchers working with biofilms fail to realize is that antibiotics are capable of destroying biofilms. The catch is that antibiotics are only effective against biofilms if administered in a very specific manner. Furthermore, only certain antibiotics appear to effectively target biofilms. After decades of research, much of which was derived from molecular modeling data, Marshall was the first to create an antibiotic regimen that appears to effectively target and destroy biofilms. Central to the treatment, which is called the Marshall Protocol, is the fact that biofilms and other Th1 pathogens succumb to specific bacteriostatic antibiotics taken in very low, pulsed doses. It is only when antibiotics are administered in this manner that they appear capable of fully eradicating biofilms.[19][20]In a paper entitled “The Riddle of Biofilm Resistance,†Dr. Kim of Tulane University discusses the mechanisms by which pulsed, low dose antibiotics are able to break up biofilms, while antibiotics administered in a standard manner (high, constant doses) cannot. According to , the use of pulsed, low-dose antibiotics to target biofilm bacteria is supported by observations she and her colleagues have made in the laboratory.[11]Some researchers claim that antibiotics cannot penetrate the matrix that surrounds a biofilm. But research by and other scientists has confirmed that the inability of antibiotics to penetrate the biofilm matrix is much more of an exception than a rule. According to , “In most cases involving small antimicrobial molecules, the barrier of the polysaccharide matrix should only postpone the death of cells rather than afford useful protection.â€For example, a recent study that used low concentrations of an antibiotic to killP. aeruginosa biofilm bacteria found that the majority of biofilm cells were effectively eliminated by antibiotics in a manne

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We all have them to some extent. When pathogens are present they can get out of control like in the case of Lyme and coinfections.On Sep 9, 2011, at 2:45 AM, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Friday, September 9, 2011 6:35 AMSubject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDTVitaminK Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK > > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the > beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence. > > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands, > organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane > surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we > have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing > them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes > generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins. > > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the > kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming > biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve > the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of > antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved > using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > > > > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves. > > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes. > > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends' > children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too > hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > > > Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1 > capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > > > > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a > maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > > > > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin > K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. > > > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica. > > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED! > The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large > quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps > from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which > seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in > the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to > hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off > the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be > acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located > inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. > > > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the > various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has > been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and > regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes. > > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling > once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION > > > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the > kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live > microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used. > > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes > some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was > slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more > responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our > cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less > trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer. > > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with. > > > > > > > >

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HI, !Thanks for the info. Then I know we don´t have biofilm in hair or on body (only plaque as Krys explained:)Are you doing the bleach baths? Does it help you?Take care, !CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Friday, September 9, 2011 5:28 PMSubject: Re: Good summary about biofilms

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled

things that pop up, then crust over and go away and new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Great article , now I know a lot about biofilms..:)CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Friday, September 9, 2011 5:32 PMSubject: Re: Good summary about biofilms

Understanding BiofilmsAuthor: Amy Proal26MAY2008As humans, our environment consistently exposes us to a variety of dangers. Tornadoes, lightning, flooding and hurricanes can all hamper our survival. Not to mention the fact that most of us can encounter swerving cars or ill-intentioned people at any given moment.Biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of materialThousands of years ago, humans realized that they could better survive a dangerous world if they formed into communities, particularly communities consisting of people with different talents. They realized that a community is far more likely to survive through division of labor– one person makes food, another gathers resources, still another protects the community against invaders. Working together in this manner requires communication and cooperation.Inhabitants of a community live in close proximity and create various forms of shelter in order to protect themselves from external threats. We build houses that protect our

families and larger buildings that protect the entire community. Grouping together inside places of shelter is a logical way to enhance survival.With the above in mind, it should come as no surprise that the pathogens we harbor are seldom found as single entities. Although the pathogens that cause acute infection are generally free-floating bacteria – also referred to as planktonic bacteria – those chronic bacterial forms that stick around for decades long ago evolved ways to join together into communities. Why? Because by doing so, they are better able to combat the cells of our immune system bent upon destroying them.It turns out that a vast number of the pathogens we harbor are grouped into communities called biofilms. In an article titled “Bacterial Biofilms: A Common Cause of Persistent Infections,†JW Costerton at the Center for

Biofilm Engineering in Montana defines a bacterial biofilm as “a structured community of bacterial cells enclosed in a self-produced polymeric matrix and adherent to an inert or living surface.â€[1] In layman’s terms, that means that bacteria can join together on essentially any surface and start to

form a protective matrix around their group. The matrix is made of polymers – substances composed of molecules with repeating structural units that are connected by chemical bonds.According to the Center for Biofilm Engineering at Montana State University, biofilms form when bacteria adhere to surfaces in aqueous environments and begin to excrete a slimy, glue-like substance that can anchor them to all kinds of material – such as metals, plastics, soil particles, medical implant materials and, most significantly, human or animal tissue. The first bacterial colonists to adhere to a surface initially do so by inducing weak, reversible bonds called van der Waals forces. If

the colonists are not immediately separated from the surface, they can anchor themselves more permanently using cell adhesion molecules, proteins on their surfaces that bind other cells in a process called cell adhesion.A biofilm in the gut.These bacterial pioneers facilitate the arrival of other pathogens by providing more diverse adhesion sites. They also begin to build the matrix that holds the biofilm together. If there are species that are unable to attach to a surface on their own, they are often able to anchor themselves to the

matrix or directly to earlier colonists.During colonization, things start to get interesting. Multiple studies have shown that during the time a biofilm is being created, the pathogens inside it can communicate with each other thanks to a phenomenon called quorum sensing. Although the mechanisms behind quorum sensing are not fully understood, the phenomenon allows a single-celled bacterium to perceive how many other bacteria are in close proximity. If a bacterium can sense that it is surrounded by a dense population of other pathogens, it is more inclined to join them and contribute to the formation of a biofilm.Bacteria that engage in quorum sensing communicate their presence by emitting chemical messages that their fellow infectious agents are able to recognize. When the messages grow strong enough, the bacteria respond en masse, behaving as a group. Quorum sensing can occur within

a single bacterial species as well as between diverse species, and can regulate a host of different processes, essentially serving as a simple communication network. A variety of different molecules can be used as signals.“Disease-causing bacteria talk to each other with a chemical vocabulary,†says Doug Hibbins of Princeton University. A graduate student in the lab of Princeton University microbiologist Dr. Bonnie Bassler, Hibbins was part of a research effort which shed light on how the bacteria that cause cholera form biofilms and communicate via quorum sensing.[2]“Forming a biofilm is one of the crucial steps in cholera’s progression,†states Bassler. “They [bacteria] cover themselves in a sort of goop that’s a shield against antibiotics, allowing them to grow rapidly. When they sense there are enough of them, they try to leave the body.â€Although cholera bacteria use the intestines as a breeding ground, after enough biofilms have formed, planktonic bacteria inside the biofilm seek to leave the body in order to infect a new host. It didn’t take long for Bassler and team to realize that the

bacteria inside cholera biofilms must signal each other in order to communicate that it’s time for the colony to stop reproducing and focus instead on leaving the body.“We generically understood that bacteria talk to each other with quorum sensing, but we didn’t know the specific chemical words that cholera uses,†Bassler said.Then Higgins isolated the CAI-1 – a chemical which occurs naturally in cholera. Another graduate student figured out how to make the molecule in the laboratory. By moderating the level of CAI-1 in contact with cholera bacteria, Higgins was successfully able to chemically control cholera’s behavior in lab tests. His team eventually confirmed that when CAI-1 is absent, cholera bacteria attach in biofilms to their current host. But when the bacteria detect enough of the chemical, they stop making biofilms and releasing toxins, perceiving that it is time

to leave the body instead. Thus, CAI-1 may very well be the single molecule that allow the bacteria inside a cholera biofilm to communicate. Although it is likely that the bacteria in a cholera biofilm may communicate with other signals besides CAI-1, the study is a good example of the fact that signaling molecules serve a key role in determining the state of a biofilm.Sessile cells in a biofilm “talk†to each other via quorum sensing to build microcolonies and to keep water channels open.Similarly, researchers at the University

of Iowa (several of whom are now at the University of Washington) have spent the last decade identifying the molecules that allow the bacterial species P. aeruginosa to form biofilms in the lungs of patients with cystic fibrosis.[3] Although the P. auruginosa isolated from the lungs of patients with cystic fibrosis looks like a biofilm and acts like a biofilm, up until recently, there were no objective tests available to confirm that the bacterial species did indeed form biofilms in the lungs of patients with the disease, nor was there a way to tell what proportion of P. aeruginosa in the lungs were actually in biofilm mode.“We needed a way to show that the P. auruginosa in cystic fibrosis lungs was communicating like a biofilm. That could tell us about the P. auruginosalifestyle,†said Pradeep Singh, M.D., a lead author on the study who is now at the University of Washington.Singh and his colleagues finally

discovered that P. aeruginosa uses one of two particular quorum-sensing molecules to initiate the formation of biofilms. In November 1999, his research team screened the entire bacterial genome, identifying 39 genes that are strongly controlled by the quorum-sensing system.In a 2000 study published in Nature, Singh and colleagues developed a sensitive test which shows P. auruginosa from cystic fibrosis lungs produces the telltale, quorum-sensing molecules that are the signals for biofilm formation.[3]It turns out that P. aerugnosa secretes two signaling molecules, one that is long, and another that is short. Using the new test, the team was able to show that planktonic forms of P. aeruginosa produce more long signaling molecules. Alternately, when they tested the P. aeruginosa strains isolated from the lungs of patients with cystic fibrosis (which were in biofilm form), all of the strains produced the signaling molecules, but in the opposite ratio – more short than long.Interestingly, when the biofilm

strains of P. aeruginosa were separated in broth into individual bacterial forms, they reverted to producing more long signal molecules than short ones. Does this mean that a change in signaling molecular length can indicate whether bacteria remain as planktonic forms or develop into biofilms?To find out, the team took the bacteria from the broth and made them grow as a biofilm again. Sure enough, those strains of bacteria in biofilm form produced more short signal molecules than long.“The fact that the P. aeruginosa in [the lungs of cystic fibrosis patients] is making the signals in the ratios that we see tells us that there is a biofilm and that most of the P. aeruginosa in the lung is in the biofilm state,†states Greenberg, another member of the research team. He believes that the findings allow for a clear biochemical

definition of whether bacteria are in a biofilm. Techniques similar to those used by his group will likely be used to determine the properties of other biofilm signaling molecules.DevelopmentOnce colonization has begun, the biofilm grows through a combination of cell division and recruitment. The final stage of biofilm formation is known as development and is the stage in which the biofilm is established and may only change in shape and size. This development of a biofilm allows for the cells inside to become more resistant to antibiotics administered in a standard fashion. In fact, depending on the organism and type of antimicrobial and experimental system, biofilm bacteria can be up to a thousand times more resistant to antimicrobial stress than free-swimming bacteria of the same species.Biofilms grow slowly, in diverse

locations, and biofilm infections are often slow to produce overt symptoms. However, biofilm bacteria can move in numerous ways that allow them to easily infect new tissues. Biofilms may move collectively, by rippling or rolling across the surface, or by detaching in clumps. Sometimes, in a dispersal strategy referred to as “swarming/seedingâ€, a biofilm colony differentiates to form an outer “wall†of stationary bacteria, while the inner region of the biofilm “liquefiesâ€, allowing planktonic cells to “swim†out of the biofilm and leave behind a hollow mound.[4]Biofilm bacteria can move in numerous ways: Collectively, by rippling or rolling across the surface, or by detaching in clumps. Individually, through a “swarming and seeding†dispersal.Research on the molecular and genetic basis of biofilm development has made it

clear that when cells switch from planktonic to community mode, they also undergo a shift in behavior that involves alterations in the activity of numerous genes. There is evidence that specific genes must be transcribed during the attachment phase of biofilm development. In many cases, the activation of these genes is required for synthesis of the extracellular matrix that protects the pathogens inside.According to Costerton, the genes that allow a biofilm to develop are activated after enough cells attach to a solid surface. “Thus, it appears that attachment itself is what stimulates synthesis of the extracellular matrix in which the sessile bacteria are embedded,†states the molecular biologist. “This notion– that bacteria have a sense of touch that enables detection of a surface and the expression of specific genes– is in itself an exciting area of research…â€[1]Certain characteristics may also facilitate the ability of some bacteria to form biofilms. Scientists at the Department of Microbiology and Molecular Genetics, Harvard Medical School, performed a study in which they created a “mutant†form of the bacterial species P.

aeguinosa (PA).[5] The mutants lacked genes that code for hair-like appendages called pili. Interestingly, the mutants were unable to form biofilms. Since the pili of PA are involved in a type of surface-associated motility called twitching, the team hypothesized this twitching might

be required for the aggregation of cells into the microcolonies that subsequently form a stable biofilm.Once a biofilm has officially formed, it often contains channels in which nutrients can circulate. Cells in different regions of a biofilm also exhibit different patterns of gene expression. Because biofilms often develop their own metabolism, they are sometimes compared to the tissues of higher organisms, in which closely packed cells work together and create a network in which minerals can flow.“There is a perception that single-celled organisms are asocial, but that is misguided,†said Andre Levchenko, assistant professor of biomedical engineering in s Hopkins University’s Whiting School of Engineering and an affiliate of the University’s Institute for NanoBioTechnology. “When bacteria are under stress—which is the story of their lives—they team up and form this

collective called a biofilm. If you look at naturally occurring biofilms, they have very complicated architecture. They are like cities with channels for nutrients to go in and waste to go out.â€[6]The biofilm life cycle in three steps: attachment, growth of colonies (development), and periodic detachment of planktonic cells.Understanding how such cooperation among pathogens evolves and is maintained represents one of evolutionary biology’s thorniest problems. This stems from the reality that, in nature, freeloading cheats inevitably evolve to exploit any cooperative group that doesn’t defend itself, leading to the breakdown of cooperation. So what causes the bacteria in a biofilm to contribute to and share resources rather than steal them? Recently, Dr. Brockhurst of the University of

Liverpool and colleagues at the Université Montpellier and the University of Oxford conducted several studies in an effort to understand why the bacteria in a biofilm cooperate and share resources rather than horde them.[7]The team took a closer look at P.

fluorescens biofilms, which are formed when individual cells overproduce a polymer that sticks the cells together, allowing the colonization of liquid surfaces. While production of the polymer is metabolically costly to individual cells, the biofilm group benefits from the increased access to oxygen that surface colonization provides. Yet, evolutionarily speaking, such a setup allows possible “cheaters†to enter the biofilm. Such cheats can take advantage of the protective matrix while failing to contribute energy to actually building the matrix. If too many “cheaters†enter a biofilm, it will weaken and eventually break apart.After several years of study, Brockhurst and team realized that the short-term evolution of diversity within a biofilm is a major factor in how successfully its members cooperate. The team found that once inside a biofilm, P. fluorescensdifferentiates into various

forms, each of which uses different nutrient resources. The fact that these “diverse cooperators†don’t all compete for the same chemicals and nutrients substantially reduces competition for resources within the biofilm.When the team manipulated diversity within experimental biofilms, they found that diverse biofilms contained fewer “cheaters†and produced larger groups than non-diverse biofilms.Levchenko and team used this device to observe bacteria growing in cramped conditions.Similarly, this year, researchers from s Hopkins; Virginia Tech; the University of California, San Diego; and Lund University in Sweden recently released the results of a study which found that once bacteria cooperate and form a biofilm, packing tightly together further enhances their survival.[6]The team created a new device in order to observe the behavior of E. coli bacteria forced to grow in the cramped conditions. The device, which allows scientists to use extremely small volumes of cells in solution, contains a series of tiny chambers of various shapes and sizes that keep the bacteria uniformly suspended in a culture medium.Not surprisingly, the cramped bacteria in the device began to form a biofilm. The team captured the development of the biofilm on video, and were able to observe the gradual self-organization and eventual construction of bacterial biofilms over a 24-hour period.First, Andre Levchenko and Hojung Cho of s Hopkins recorded the behavior of single layers of E.

coli cells using real-time microscopy. “We were surprised to find that cells growing in chambers of all sorts of shapes gradually organized themselves into highly regular structures,†Levchenko said.Dr. Levchenko of s Hopkins and Hojung Cho, a biomedical engineering doctoral studentFurther observations using microscopy revealed that the longer the packed cell population resided in the chambers, the more ordered the biofilm structure became. As the cells in the biofilm became more ordered and tightly packed, the biofilm

became harder and harder to penetrate.Levchenko also noted that rod-shaped E. colithat were too short or too long typically did not organize well into the dense, circular main hub of the biofilm. Instead, the bacteria of odd shapes or highly disordered groups of cells were found on the edges of the biofilm, where they formed sharp corners.Nodes of relapsing infection?Researchers often note that, once biofilms are established, planktonic bacteria may periodically leave the biofilm on their own. When they do, they can rapidly multiply and disperse.According to Costerton, there is a natural pattern of programmed detachment of planktonic cells from biofilms. This means that biofilms can act as what Costerton refers to as “niduses†of acute infection. Because the bacteria in a

biofilm are protected by a matrix, the host immune system is less likely to mount a response to their presence.[1]But if planktonic bacteria are periodically released from the biofilms, each time single bacterial forms enter the tissues, the immune system suddenly becomes

aware of their presence. It may proceed to mount an inflammatory response that leads to heightened symptoms. Thus, the periodic release of planktonic bacteria from some biofilms may be what causes many chronic relapsing infections.Planktonic bacteria are periodically released from a biofilmAs R. Parsek of Northwestern University describes in a 2003 paper in the Annual Review of Microbiology, any pathogen that survives in a chronic form benefits by keeping the host alive.[8] After all, if a chronic bacterial form simply kills its host, it will no longer have a place to live. So according to Parsek, chronic infection often results in a “disease stalemate†where bacteria of moderate virulence are somewhat contained by the defenses of the host. The infectious agents never actually kill the host, but the host is never able to

fully kill the invading pathogens either.Parsek believes that the optimal way for bacteria to survive under such circumstances is in a biofilm, stating that “Increasing evidence suggests that the biofilm mode of growth may play a key role in both of these adaptations. Biofilm growth increases the resistance of bacteria to killing and may make organisms less conspicuous to the immune system… ultimately this moderation of virulence may serve the bacteria’s interest by increasing the longevity of the host.â€The acceptance of biofilms as infectious entitiesAnton van Leeuwenhoek.Perhaps because many biofilms are sufficiently thick to be visible to the naked eye, the microbial communities were among the first to be studied by early microbiologists. Anton van Leeuwenhoek scraped the plaque biofilm from his teeth and observed what he described as the “animalculi†inside them under his primitive microscope. However, according to Costerton and team at the Center for Biofilm Research at Montana State University, it was not until the 1970s that scientists began to appreciate that bacteria in the biofilm mode of existence constitute such a major component of the bacterial biomass in most environments. Then, it was not until the 1980s and 1990s that scientists truly began to understand how elaborately organized a bacterial biofilm community can be.[1]As Kolter, professor of microbiology and molecular genetics at Harvard Medical School, and one of the first scientists to study how biofilms developstates, “At first, however, studying biofilms was a radical departure from previous work.â€Like most microbial geneticists, Kolter had been trained in the tradition dating back to Koch and Louis Pasteur, namely that bacteriology is best conducted by studying pure strains of planktonic bacteria. “While this was a tremendous advance for modern microbiology, it also distracted microbiologists from a more organismic view of bacteria, Kolter adds, “Certainly we felt that pure, planktonic cultures were the only way to work. Yet in nature bacteria don’t live like that,†he says. “In fact, most of them occur in

mixed, surface-dwelling communities.â€Although research on biofilms has surged over the past few decades, the majority of biofilm research to date has focused on external biofilms, or those that form on various surfaces in our natural environment.Over the past years, as scientists developed better tools to analyze external biofilms, they quickly discovered that biofilms can cause a wide range of problems in industrial environments. For example, biofilms can develop on the interiors of pipes, which can lead to clogging and corrosion. Biofilms on floors and counters can make sanitation difficult in food preparation areas.Since biofilms have the ability to clog pipes, watersheds, storage areas, and contaminate food products, large companies with facilities that are negatively impacted by their presence have naturally taken an interest in supporting

biofilm research, particularly research that specifies how biofilms can be eliminated.This means that many recent advances in biofilm detection have resulted from collaborations between microbial ecologists, environmental engineers, and mathematicians. This research has generated new analytical tools that help scientists identify biofilms.Biofilm in a swamp gas reactor.For example, the Canadian company FAS International Ltd. has justcreated an endoluminal brush, which will be launched this spring. Physicians can use the brush to obtain samples from the interior of catheters. Samples taken from catheters can be sent to a lab, where researchers determine if biofilms are present in the sample. If biofilms are detected, the catheter is immediately replaced, since the insertion of catheters with biofilms can cause the patient to suffer from numerous infections, some of which are potentially life threatening.Scientists now realize that biofilms are not just composed of bacteria. Nearly every species of microorganism – including viruses, fungi, and Archaea – have mechanisms by which they can adhere to

surfaces and to each other. Furthermore, it is now understood that biofilms are extremely diverse. For example, upward of 300 different species of bacteria can inhabit the biofilms that form dental plaque.[9]Furthermore, biofilms have been found literally everywhere in nature, to the point where any mainstream microbiologist would

acknowledge that their presence is ubiquitous. They can be found on rocks and pebbles at the bottom of most streams or rivers and often form on the surface of stagnant pools of water. In fact, biofilms are important components of food chains in rivers and streams and are grazed upon by the aquatic invertebrates upon which many fish feed. Biofilms even grow in the hot, acidic pools at Yellowstone National Park and on glaciers in Antarctica.Biofilm in acidic pools at Yellowstone National Park.It is also now understood that the biofilm

mode of existence has been around for millenia. For example, filamentous biofilms have been identified in the 3.2-billion-year-old deep-sea hydrothermal rocks of the Pilbara Craton, Australia. According to a 2004 article in Nature Reviews Microbiology, “Biofilm formation appears early in the fossil record (approximately 3.25 billion years ago) and is common throughout a diverse range of organisms in both the Archaea and Bacteria lineages. It is evident that biofilm formation is an ancient and integral component of the prokaryotic life cycle, and is a key factor for survival in diverse environments.â€[10]Biofilms and diseaseThe fact that external biofilms are ubiquitous raises the question – if biofilms can form on essentially every surface in our external environments, can they do the same inside the human body? The answer seems to be yes, and over the past few years, research on internal biofilms has finally started to pick up pace. After all, it’s easy for biofilm researchers to see that the human body, with its wide range of moist surfaces and mucosal tissue, is an excellent place for biofilms to thrive. Not to mention the fact that those bacteria

which join a biofilm have a significantly greater chance of evading the battery of immune system cells that more easily attack planktonic forms.Many would argue that research on internal biofilms has been largely neglected, despite the fact that bacterial biofilms seem to have great potential for causing human disease.Common sites of biofilm infection. One biofilm reach the bloodstream they can spread to any moist surface of the human body. Stoodley of the Center for Biofilm Engineering at

Montana State University, attributes much of the lag in studying biofilms to the difficulties of working with heterogeneous biofilms compared with homogeneous planktonic populations. In a 2004 paper in Nature Reviews, the molecular biologist describes many reasons why biofilms are extremely difficult to culture, such as the fact that the diffusion of liquid through a biofilm and the fluid forces acting on a biofilm must be carefully calculated if it is to be cultured correctly. According to Stoodley, the need to master such difficult laboratory techniques has deterred many scientists from attempting to work with biofilms. [10]Also, since much of the technology needed to detect internal biofilms was created at the same time as the sequencing of the human genome, interest in biofilm bacteria, and the research grants that would accompany such interest, have been largely diverted to projects with a decidedly genetic focus. However, since genetic research has failed to uncover the cause of any of the common chronic diseases, biofilms are finally – just over the past few years – being studied more intensely, and being given the credit they deserve as serious infectious entities, capable of causing a wide array of

chronic illnesses.In just a short period of time, researchers studying internal biofilms have already pegged them as the cause of numerous chronic infections and diseases, and the list of illnesses attributed to these bacterial colonies continues to grow rapidly.According to a recent public statement from the National Institutes of Health, more than 65% of all microbial infections are caused by biofilms. This number might seem high, but according to Kim of the Department of Chemical and Biological Engineering at Tufts University, “If one recalls that such common infections as urinary tract infections (caused by E. coli and other pathogens), catheter infections (caused by Staphylococcus aureus and other gram-positive pathogens), child middle-ear infections (caused by Haemophilus influenzae, for example), common dental plaque formation, and

gingivitis, all of which are caused by biofilms, are hard to treat or frequently relapsing, this figure appears realistic.â€[11]Hundreds of microbial biofilm colonize the human mouth, causing tooth decay and gum disease.As mentions, perhaps the most well-studied biofilms are those that make up what is commonly referred to as dental plaque. “Plaque is a biofilm on the surfaces of the teeth,†states Parsek. “This accumulation of microorganisms subject the teeth and gingival tissues to high concentrations of bacterial metabolites which results in dental disease.â€[12]It has also recently been shown that biofilms are present on the removed tissue of 80% of patients undergoing surgery for chronic sinusitis. According to Parsek, biofilms may also cause osteomyelitis, a disease in which the bones and bone marrow become infected. This is supported by the fact that microscopy studies have shown biofilm formation on infected bone surfaces from humans and experimental animal

models. Parsek also implicates biofilms in chronic prostatitis since microscopy studies have also documented biofilms on the surface of the prostatic duct. Microbes that colonize vaginal tissue and tampon fibers can also form into biofilms, causing inflammation and disease such as Toxic Shock Syndrome.Biofilms also cause the formation of kidney stones. The stones cause disease by obstructing urine flow and by producing inflammation and recurrent infection that can lead to kidney failure. Approximately 15%–20% of kidney stones occur in the setting of urinary tract infection. According to Parsek, these stones are produced by the interplay between infecting bacteria and mineral substrates derived from the urine. This interaction results in a complex biofilm composed of bacteria, bacterial exoproducts, and mineralized stone material.Microbes that colonize vaginal tissue and tampon fibers can become pathogenic, causing inflammation and disease such as Toxic Shock Syndrome.Perhaps the first hint of the role of bacteria in these stones came in 1938 when Hellstrom examined stones passed by his patients and found bacteria embedded deep inside them. Microscopic analysis of stones removed from infected patients has revealed features that characterize biofilm growth. For one thing, bacteria on the surface and inside the stones are organized in microcolonies and surrounded by a matrix composed of crystallized (struvite) minerals.Then there’s endocarditis, a disease that involves inflammation of the inner layers of the heart. The primary infectious lesion in endocarditis is a complex biofilm composed of both bacterial and host components that is located on a cardiac valve. This biofilm, known as a vegetation, causes disease by three basic mechanisms. First, the vegetation physically disrupts valve function, causing leakage when the valve is closed and inducing turbulence and diminished flow when the valve is open. Second, the vegetation provides a source for near-continuous infection of the bloodstream that persists even during antibiotic treatment. This causes recurrent fever, chronic systemic inflammation, and other infections. Third, pieces of the infected vegetation can break off and be carried to a terminal point in the circulation where they block the flow of blood (a process known as embolization). The brain, kidney, and extremities are

particularly vulnerable to the effects of embolization.A variety of pathogenic biofims are also commonly found on medical devices such as joint prostheses and heart valves. According to Parsek, electron microscopy of the surfaces of medical devices that have been foci of device-related infections shows the presence of large numbers of slime-encased bacteria. Tissues taken from non-device-related chronic infections also show the presence of biofilm bacteria surrounded by an exopolysaccharide matrix. These biofilm infections may be caused by a single species or by a mixture of species of bacteria or fungi.According to Dr. Patel of the Mayo Clinic, individuals with prosthetic joints are often oblivious to the fact that their prosthetic joints harbor biofilm infections.[13]Cells

of Staphylococcus epidermidis causing devastating disease as they grow on the cuff at a mechanical heart valve.“When people think of infection, they may think of fever or pus coming out of a wound,†explains Dr. Patel. “However, this is not the case with prosthetic joint infection. Patients will often experience pain, but not other symptoms usually associated with infection. Often what happens is that the bacteria that cause infection on prosthetic joints are the same as bacteria that live harmlessly on our skin. However, on a prosthetic joint they can stick, grow and cause problems over the long term. Many of these bacteria would not infect the joint were it not for the prosthesis.â€Biofilms also cause Leptospirosis, a serious but neglected emerging disease that infects humans through contaminated water. New research published in the May

issue of the journal Microbiology shows for the first time how bacteria that cause the disease survive in the environment.Leptospirosis is a major public health problem in southeast Asia and South America, with over 500,000 severe cases every year. Between 5% and 20% of these cases are fatal. Rats and other mammals carry the disease-causing pathogen Leptospira interrogans in their kidneys. When they urinate, they contaminate surface water with the bacteria, which can survive in the environment for long periods.“This led us to see if the bacteria build a protective casing around themselves for protection,†said Professor Mathieu Picardeau from the Institut Pasteur in Paris, France. [14]Previously, scientists believed the bacteria were planktonic. But Professor Picardeau and his team have shown that L. interrogans can make biofilms, which could be one of the main factors controlling survival and disease transmission. “90% of the species of Leptospira we tested could form biofilms. It takes L.

interrogans an average of 20 days to make a biofilm,†says Picardeau.Biofilms have also been implicated in a wide array of veterinary diseases. For example, researchers at the Virginia-land Regional College of Veterinary Medicine at Virginia Tech were just awarded a grant from the United States Department of Agriculture to study the role biofilms play in the development of Bovine Respiratory Disease Complex (BRDC). If biofilms play a role in bovine respiratory disease, it’s likely only a matter of time before they will be established as a cause of human respiratory diseases as well.When the immune response is compromised, Pseudomonas aeruginosabiofilms are able to colonize the alveoli, and to form biofilms.As mentioned previously, infection by the bacterium Pseudomonas aeruginosa (P. aeruginosa) is the main cause of death among patients with cystic fibrosis. Pseudomonas is able to set up permanent residence in the lungs of patients with cystic fibrosis where, if you ask most mainstream researchers, it is impossible to kill. Eventually, chronic inflammation produced by the immune system

in response to Pseudomonas destroys the lung and causes respiratory failure. In the permanent infection phase, P. aeruginosa biofilms are thought to be present in the airway, although much about the infection pathogenesis remains unclear.[15]Cystic fibrosis is caused by mutations in the proteins of channels that regulates chloride. How abnormal chloride channel protein leads to biofilm infection remains hotly debated. It is clear, however, that cystic fibrosis patients manifest some kind of host-defense defect localized to the airway surface. Somehow this leads to a debilitating biofilm infection.Biofilms have the potential to cause a tremendous array of infections and diseasesBecause internal pathogenic biofilm research comprises such a new field of study, the infections described above almost certainly represent just the tip of the iceberg when it comes to the number of chronic diseases and infections currently caused by biofilms.For example, it wasn’t until July of 2006 that researchers realized that the majority of ear infections

are caused by biofilm bacteria. These infections, which can be either acute or chronic, are referred to collectively as otitis media (OM). They are the most common illness for which children visit a physician, receive antibiotics, or undergo surgery in the United States.There are two subtypes of chronic OM. Recurrent OM (ROM) is diagnosed when children suffer repeated infections over a span of time and during which clinical evidence of the disease resolves between episodes. Chronic OM with effusion is diagnosed when children have persistent fluid in the ears that lasts for months in the absence of any other symptoms except conductive hearing loss.It took over ten years for researchers to realize that otitis media is caused by biofilms. Finally, in 2002, Drs. Ehrlich and J. Post, an Allegheny General Hospital pediatric ear specialist and medical director of the Center for

Genomic Sciences, published the first animal evidence of biofilms in the middle ear in the Journal of the American Medical Association, setting the stage for further clinical investigation.In a subsequent study, Ehrlich and Post obtained middle ear mucosa – or membrane tissue – biopsies from children undergoing a procedure for otitis. The team gathered uninfected mucosal biopsies from children and adults undergoing cochlear implantation as a control.[16]Using advanced confocal laser scanning microscopy, Luanne Hall Stoodley, Ph.D. and her ASRI colleagues obtained three dimensional images of the biopsies and evaluated them for biofilm morphology using generic stains and species-specific probes for Haemophilus influenzae, Streptococcus pneumoniaeand Moraxella catarrhalis. Effusions, when present, were also evaluated for evidence of pathogen specific nucleic acid sequences (indicating presence of live bacteria).The study found mucosal biofilms in the middle ears of 46/50 children (92%) with both forms of otitis. Biofilms were not observed in eight control middle ear mucosa

specimens obtained from cochlear implant patients.Otitis media, or inflammation of the inner ear, is caused by biofilm.In fact, all of the children in the study who suffered from chronic otitis media tested positive for biofilms in the middle ear, even those who were asymptomatic, causing Erlich to conclude that, “It appears that in many cases recurrent disease stems not from re-infection as was previously thought and which forms the basis for conventional treatment, but from a persistent biofilm.â€He went on to state that the discovery of biofilms in the setting of chronic otitis media represented “a landmark evolution in the medical community’s understanding about a disease that afflicts millions of children world-wide each year and further endorses the emerging biofilm paradigm of chronic infectious disease.â€The emerging biofilm paradigm of chronic disease refers to a new movement in which researchers such as Ehrlich are calling for a tremendous shift in the way the medical community views bacterial biofilms. Those scientists who support an

emerging biofilm paradigm of chronic disease feel that biofilm research is of utmost importance because of the fact that the infectious entities have the potential to cause so many forms of chronic disease. The Marshall Pathogenesis is an important part of this paradigm shift.It was also just last year that researchers realized that biofilms cause most infections associated with contact lens use. In 2006, Bausch & Lomb withdrew its ReNu with MoistureLoc contact lens solution because a high proportion of corneal infections were associated with it. It wasn’t long before researchers at the University Hospitals Case Medical Center found that the infections were caused by biofilms. [17]“Once they live in that type of state [a biofilm], the cells become resistant to lens solutions and immune to the body’s own defense system,†said Mahmoud A. Ghannoum, Ph.D, senior investigator of the study. “This study should alert contact lens wearers to the importance of proper care for contact lenses to protect against potentially virulent eye

infections,†he said.It turns out that the biofilms detected by Ghannoum and team were composed of fungi, particularly a species called Fusarium. His team also discovered that the strain of fungus (with the catchy name, ATCC 36031) used for testing the effectiveness of lens care solutions is a strain that does not produce biofilms as the clinical fungal strains do. ReNu contact solution, therefore, was effective in the laboratory, but failed when faced with strains in real-world situations.Fungal biofilm can form in contact lens

solution leading to potentially virulent eye infectionsUnfortunately, Ghannoum and team were not able to create a method to target and destroy the fungal biofilms that plague users of ReNu and some other contact lens solutions.Then there’s Dr. Randall Wolcott who just recently discovered and confirmed that the sludge covering diabetic wounds is largely made up of biofilms. Whereas before Wolcott’s work such limbs generally had to be amputated, now that they have been correctly linked to biofilms, measures such as those described in thisinterview can be taken to stop the spread of infection and save the limb.

Wolcott has finally been given a grant by the National Institutes of Health to further study chronic biofilms and wound development.Dr. Garth and the Medical Biofilm Laboratory team at Montana State University are also researching wounds and biofilms. Their latest article and an image showing wound biofilm was featured on the cover of the January-February 2008 issue of Wound Repair and Regeneration.[18]Biofilm bacteria and chronic inflammatory diseaseIn just a few short years, the potential of biofilms to cause debilitating chronic infections has become so clear that there is little doubt that biofilms are part of the pathogenic mix or “pea soup†that cause most or all chronic “autoimmune†and inflammatory diseases.In fact, thanks, in large part, to the research of biomedical researcher Dr. Trevor Marshall, it is now increasingly understood that chronic inflammatory diseases result from infection with a large microbiota of chronic biofilm and L-form bacteria (collectively called the Th1 pathogens).[19][20] The microbiota is thought to be comprised of numerous bacterial species, some of which have yet to be discovered. However, most of the pathogens that cause inflammatory disease have one thing in common – they have all developed ways to evade the immune system and persist as chronic forms that the body is unable to eliminate naturally.Some L-form bacteria are able to evade the immune system because, long ago, they evolved the ability to reside inside macrophages, the very white bloods cells of the immune system that are supposed to kill invading pathogens. Upon formation, L-form bacteria also lose their cell walls, which

makes them impervious to components of the immune response that detect invading pathogens by identifying the proteins on their cell walls. The fact that L-form bacteria lack cell walls also means that the beta-lactam antibiotics, which work by targeting the bacterial cell wall, are completely ineffective at killing them.[21]Clearly, transforming into the L-form offers any pathogen a survival advantage. But among those pathogens not in an L-form state, joining a biofilm is just as likely to enhance their ability to evade the immune system. Once enough chronic pathogens have grouped together and formed a stable community with a strong protective matrix, they are likely able to reside in any area of the body, causing the host to suffer from chronic symptoms that are both mental and physical in nature.Biofilm researchers will also tell you that, not surprisingly, biofilms form with greater ease in an immunocompromised host. Marshall’s research has made it clear that many of the Th1 pathogens are capable of creating substances that bind and inactivate the Vitamin D Receptor – a fundamental receptor of the body that controls the activity of the innate immune system, or the body’s first line of defense against

intracellular infection.[22]Diagram of the Vitamin D Receptor and capnine.Thus, as patients accumulate a greater number of the Th1 pathogens, more and more of the chronic bacterial forms create substances capable of disabling the VDR. This causes a snowball effect, in which the patient becomes increasingly immunocompromised as they acquire a larger bacterial load.For one thing, it’s possible that many of the bacteria that survive inside biofilms are capable of creating VDR blocking substances. Thus, the formation of biofilms may contribute to immune dysfunction. Conversely, as patients acquire L-form bacteria and other persistent bacterial forms capable of creating VDR-blocking substances, it becomes exceptionally easy for biofilms to form on any tissue surface of the human body.Thus, patients who begin to acquire

L-form bacteria almost always fall victim to biofilm infections as well, since it is all too easy for pathogens to group together into a biofilm when the immune system isn’t working up to par.To date, there is also no strict criteria that separate L-form bacteria from biofilm bacteria or any other chronic pathogenic forms. This means that L-form bacteria may also form into biofilms, and by doing so enter a mode of survival that makes them truly impervious to the immune system. Some L-form bacteria may not form complete biofilms, yet may still possess the ability to surround themselves in a protective matrix. Under these circumstances one might say they are in a “biofilm-like†state.Marshall often refers to the pathogens that cause inflammatory disease as an intraphgocytic, metagenomic microbiota of bacteria, terms which suggest that most chronic bacterial forms possess

properties of both L-form and biofilm bacteria. Intraphagocytic refers to the fact that the pathogens can be found inside the cells of the immune system. The term metagenomic indicates that there are a tremendous number of different species of these chronic bacterial forms. Finally, microbiota refers to the fact that biofilm communities sustain their pathogenic activity.For example, when observed under a darkfield microscope, L-form bacteria are often encased in protective biofilm sheaths. If the blood containing the pathogens are aged overnight, the bacterial colonies reach a point where they expand and burst out of the cell, causing the cell to burst as well. Then they extend as huge, long biofilm tubules, which are presumably helping the pathogens spread to other cells. The tubules also help spread bacterial DNA to neighboring cells.Clearly, there is a great need for more research

on how different chronic bacterial forms interact. To date, L-form researchers have essentially focused soley on the L-form, while failing to investigate how frequently the wall-less pathogens form into biofilms or become parts of biofilm communities together with bacteria with cell walls. Conversely, most biofilm researchers are intently studying the biofilm mode of growth without considering the presence of L-form bacteria. So, it will likely take several years before we will be better able to understand probable overlaps between the lifestyles of L-form and biofilm bacteria.Anyone who is skeptical about the fact that biofilms likely form a large percentage of the microbiota that cause inflammatory disease should consider many of the recent studies that have linked established biofilm infections to a higher risk for multiple forms of chronic inflammatory disease. Take, for example, studies that have found a link

between periodontal disease and several major inflammatory conditions. A 1989 article published in British Medical Journal showed a correlation between dental disease and systemic disease (stroke, heart disease, diabetes). After correcting for age, exercise, diet, smoking, weight, blood cholesterol level, alcohol use and health care, people who had periodontal disease had a significantly higher incidence of heart disease, stroke and premature death. More recently, these results were confirmed in studies in the United States, Canada, Great Britain, Sweden, and Germany. The effects are striking. For example, researchers from the Canadian Health Bureau found that people with periodontal

disease had a two times higher risk of dying from cardiovascular disease.[23]Dental plaque as seen under a scanning electron microcroscope.Since we know that periodontal disease is caused by biofilm bacteria, the most logical explanation for the fact that people with dental problems are much more likely to suffer from heart disease and stroke is that the biofilms in their mouths have gradually spread to the moist surfaces of their circulatory systems. Or perhaps if the bacteria in periodontal biofilms create VDR binding substances, their ability to slow innate immune function allows new biofilms (and L-form bacteria as well) to more easily form and infect the heart and blood vessels.

Conversely, systemic infection with VDR blocking biofilm bacteria is also likely to weaken immune defenses in the gums and facilitate periodontal disease.In fact, it appears that biofilm bacteria in the mouth also facilitate the formation of biofilm and L-form bacteria in the brain. Just last year, researchers at Vasant Hirani at University College London released the results of a study which found that elderly people who have lost their teeth are at more than three-fold greater risk of memory problems and dementia.[24]At the moment, Autoimmunity Research Foundation does not have the resources to culture biofilms from patients on the treatment and, even if they did, current methods for culturing internal biofilms remain unreliable. According to Stoodley, “The lack of standard methods for growing, quantifying and testing biofilms in continuous culture results in incalculable variability between laboratory systems. Biofilm microbiology is complex and not well represented by flask cultures. Although homogeneity allows statistical enumeration, the extent to which it reflects the real, less orderly world is questionable.â€[10]How else do we acquire biofilm bacteria?As discussed thus far, biofilms form spontaneously as bacteria inside the human body group together. Yet people can also ingest biofilms by eating contaminated food.According to researchers at the University of Guelph in Ontario Canada, it is increasingly suspected that biofilms play an important role in contamination of meat during processing and packaging. The group warns that greater action must be taken to reduce the presence of food-borne pathogens like Escherichia coli and Listeria monocytogenes and spoilage microorganisms such as thePseudomonas species (all of which form biofilms) throughout the food processing chain to ensure the safety and shelf-life of the product. Most of these microorganisms are ubiquitous in the environment or brought into processing facilities through

healthy animal carriers.Hans Blaschek of the University of Illinois has discovered that biofilms form on much of the other food products we consume as well.A biofilm on a piece of lettuce“If you could see a piece of celery

that’s been magnified 10,000 times, you’d know what the scientists fighting foodborne pathogens are up against,†says Blaschek.“It’s like looking at a moonscape, full of craters and crevices. And many of the pathogens that cause foodborne illness, such as Shigella, E. coli,and Listeria, make sticky, sugary biofilms that get down in these crevices, stick like glue, and hang on like crazy.â€According to Blaschek, the problem faced by produce suppliers can be a triple whammy. “If you’re unlucky enough to be dealing with a pathogen–and the pathogen has the additional attribute of being able to form biofilm—and you’re dealing with a food product that’s minimally processed, well, you’re triply unlucky,†the scientist said. “You may be able to scrub the organism off the surface, but the cells in these biofilms are very good at aligning

themselves in the subsurface areas of produce.†, a University of Illinois food science and human nutrition professor agrees, stating,â€Once the pathogenic organism gets on the product, no amount of washing will remove it. The microbes attach to the surface of produce in a sticky biofilm, and washing just isn’t very effective.â€Biofilms can even be found in processed water. Just this month, a study was released in which researchers at the Department of Biological Sciences, at Virginia Polytechnic Institute isolated M. avium biofilm from the shower head of a woman with M.

avium pulmonary disease.[25] A molecular technique called DNA fingerprinting demonstrated that M. avium isolates from the water were the same forms that were causing the woman’s respiratory illness.Effectively targeting biofilm infectionsAlthough the mainstream medical community is rapidly acknowledging the large number of diseases and infections caused by biofilms, most researchers are convinced that biofilms are difficult or impossible to destroy, particularly those cells that form the deeper layers of a thick biofilm. Most papers on biofilms state that they are resistant to antibiotics administered in a standard manner. For example, despite the fact that Ehrlich and team discovered that biofilm bacteria cause otitis media, they are unable to offer an effective solution that would actually allow for the destruction of biofilms in the ear canal. Other teams have also come up short in creating methods to break up the biofilms they implicate as the cause of numerous infections.This means patients with biofilm infections are generally told by mainstream doctors that they have an

untreatable infection. In some cases, a disease-causing biofilm can be cut out of a patient’s tissues, or efforts are made to drain components of the biofilm out of the body. For example, doctors treating otitis media often treats patients with myringotomy, a surgical procedure in which small tubes are placed in the eardrum to continuously drain infectious fluid.When it comes to administering antibiotics in an effort to target biofilms, one thing is certain. Mainstream researchers have repeatedly tried to kill biofilms by giving patients high, constant doses of antibiotics. Unfortunately, when administered in high doses, the antibiotic may temporarily weaken the biofilm but is incapable of destroying it, as certain cells inevitably persist and allow the biofilm to regenerate.“You can put a patient on [a high dose] antibiotics, and it may seem that the infection has

disappeared,†says Levchenko. “But in a few months, it reappears, and it is usually in an antibiotic-resistant form.â€What the vast majority of researchers working with biofilms fail to realize is that antibiotics are capable of destroying biofilms. The catch is that antibiotics are only effective against biofilms if administered in a very specific manner. Furthermore, only certain antibiotics appear to effectively target biofilms. After decades of research, much of which was derived from molecular modeling data, Marshall was the first to create an antibiotic regimen that appears to effectively target and destroy biofilms. Central to the treatment,

which is called the Marshall Protocol, is the fact that biofilms and other Th1 pathogens succumb to specific bacteriostatic antibiotics taken in very low, pulsed doses. It is only when antibiotics are administered in this manner that they appear capable of fully eradicating biofilms.[19][20]In a paper entitled “The Riddle of Biofilm Resistance,†Dr. Kim of Tulane University discusses the mechanisms by which pulsed, low dose antibiotics are able to break up biofilms, while antibiotics administered in a standard manner (high, constant doses) cannot. According to , the use of

pulsed, low-dose antibiotics to target biofilm bacteria is supported by observations she and her colleagues have made in the laboratory.[11]Some researchers claim that antibiotics cannot penetrate the matrix that surrounds a biofilm. But research by and other scientists has confirmed that the inability of antibiotics to penetrate the biofilm

matrix is much more of an exception than a rule. According to , “In most cases involving small antimicrobial molecules, the barrier of the polysaccharide matrix should only postpone the death of cells rather than afford useful protection.â€For example, a recent study that used low concentrations of an antibiotic to killP. aeruginosa biofilm bacteria found that the majority of biofilm cells were effectively eliminated by antibiotics in a manner that did not differ much from what is observed when the same antibiotic concentrations are administered to single planktonic cells.[26]After antibiotics are applied to a biofilm, a number of cells called “persisters†are left behind.Thus, since antibiotics can generally penetrate biofilms, some other factor is responsible for the fact that they cannot be killed by standard high dose antibiotic therapy. It

turns out that after antibiotics are applied to a biofilm, a number of cells called “persisters†are left behind. Persisters are simply cells that are able to survive the first onslaught of antibiotics, and if left unchecked, gradually allow the biofilm to form again. According to , persister cells form with particular ease in immunocompromised patients because the immune system is unable to help the antibiotic “mop up†all the biofilm cells it has targeted.“This simple observation suggests a new paradigm for explaining, at least in principle, the phenomenon of biofilm resistance to killing by a wide range of antimicrobials,†states . “The majority of cells in a biofilm are not necessarily more resistant to killing than planktonic cells and die rapidly when treated with [an antibiotic] that can kill slowly growing cells.â€Thus, a dose of antibiotics –

particularly in immunocompromised patients – eradicates most of the biofilm population but leaves a small fraction of surviving persisters behind. Unfortunately, in the same sense that the beta-lactam antibiotics promote the formation of L-form bacteria, persister cells are actually preserved by the presence of an antibiotic that inhibits their growth. Thus, paradoxically, dosing an antibiotic in a constant, high-dose manner (in which the antibiotic is always present) helps persisters persevere.But in the case of low, pulsed dosing, where an antibiotic is administered, withdrawn, then administered again, the first application of antibiotic will eradicate the bulk of biofilm cells, leaving persister cells behind. Withdrawl of the antibiotic allows the persister population to start growing. Since administration of the antibiotic is temporarily stopped, the survival of persisters is not enhanced. This causes the

persister cells to lose their phenotype (their shape and biochemical properties), meaning that they are unable to switch back into biofilm mode. A second application of the antibiotic should then completely eliminate the persister cells, which are still in planktonic mode. has found that the feasibility of a pulsed, or cyclical biofilm eradication approach depends on the rate at which persisters lose resistance to killing and regenerate new persisters. It also depends on the ability to manipulate the antibiotic concentration – something that is done quite effectively by patients on the Marshall Protocol who carefully dose their antibiotics at different levels, allowing constant variation in antibiotic concentration. Although speculates that allowing the concentration of an antibiotic to drop could potentially lead to resistance towards the antibiotic, she is quick to add that if two or more antibiotics are

used to target a biofilm at one time, such resistance would not occur. Again, since the Marshall Protocol uses a total of five bacteriostatic antibiotics, usually taken two or three at a time, concerns of resistance are essentially negligible.Model of biofilm resistance based on persister survival. An initial treatment of high-dose constant antibiotic kills planktonic cells and the majority of biofilm cells. But persisters remain alive and resurrect the biofilm, causing the infection to relapse“It is entirely possible that

successful cases of antimicrobial therapy of biofilm infections result from a fortuitous optimal cycling [pulsed dosing] of an antibiotic concentration that eliminated first the bulk of the biofilm and then the progeny of the persisters that began to divide,†states .’ work has been supported by other research teams. Recently, researchers at the University of Iowa found that subinhibitory (extremely low dose) concentrations of the bacteriostatic antibiotic azithromycin significantly decreased biomass and maximal thickness in both forming and established biofilms.[27] These extremely low concentrations of azithromycin inhibited biofilms in all but the most highly resistant isolates. In contrast, subinhibitory concentrations of gentamicin, which is not a bacteriostatic antibiotic, had no effect on biofilm formation. In fact, biofilms actually became resistant to gentamicin at concentrations far above the minimum inhibitory concentration.Researchers at Tulane University recently confirmed yet again that low, pulsed dosing is a superior way of targeting treatment-resistant biofilm bacteria. According to the team, who

mathematically modeled the action of antibiotics on bacterial biofilms, “Exposing a biofilm to low concentration doses of an antimicrobial agent for longer time is more effective than short time dosing with high antimicrobial agent concentration.â€[28]Similarly, a bioengineer led team at

the University of Washington recently created an antibiotic- containing polymer that releases antibiotic slowly onto the surface of hospital devices, such as catheters and prostheses, to reduce the risk of biofilm-related infections.“Rather than massively dosing the patient with high levels of released antibiotic, this strategy allows the release of extremely low levels of this very potent antibiotic over long periods of time,†explained Buddy Ratner, PhD, Professor and Director of the Engineered Biomaterials Program at the University of Washington, Seattle. “We calculated the amount released at the surface that would kill 100% of

the bacteria entering the surface zone.â€When challenged by Dr. Leonard A. Mermel from Brown University School of Medicine on the issue that long-term use of pulsed, low-dose antibiotics might allow for increased resistance on the part of the bacteria being treated, Ratner responded, “Dr. Mermel’s concerns are, in fact, why we developed this system for [antibiotic] release. Bacteria that live through antibiotic dosing can go on to produce resistant strains. If 100% of the bacteria approaching the surface are killed, they can’t produce resistant offspring. The classical physician approach, dosing the patient systemically and heavily to rid the patient of persistent bacteria, can lead to those resistant strains. Our approach releases miniscule doses compared to what a physician would use, but releases the antibiotic where it will be optimally effective and least likely to leave antibiotic-resistant

survivors.â€Although taken orally, the MP antibiotics are taken in the same manner as those administered by Ratner and team. Because they too are dosed at optimal times in extremely small doses, the chance that long-term antibiotic use might foster resistant bacteria is again, essentially negligible, especially when multiple antibiotics are typically used.Key to the ability of the Marshall Protocol to effectively target biofilm bacteria is the fact that the specific pulsed, low-dose bacteriostatic antibiotics used by the treatment are taken in conjunction with a medication called Benicar. Benicar binds and activates the Vitamin D Receptor, displacing bacterial substances and 25-D from the receptor, so that it can once again activate the innate immune system.[29] Benicar is so effective at strengthening the innate immune response that the patient’s own immune system ultimately helps destroy the biofilm weakened by pulsed, low-dose antibiotics.Thus, it is not enough for patients on the Marshall Protocol to simply take specific pulsed, low-dose antibiotics. The activity of their innate immune system must also be restored so that the cells of the immune system can actively combat

biofilm bacteria, the matrix that surrounds them, and persister cells.After antibiotics are applied to a biofilm, a number of cells called “persisters†are left behindHow do we know that the Marshall Protocol effectively kills biofilm bacteria? Namely because those patients to reach the later stages of the treatment do not report symptoms associated with established biofilm diseases. Patients on the MP who once suffered from chronic ear infections (OM), chronic sinus infections, or periodontal disease find that such infections

resolve over the course of treatment. Furthermore, since we now understand that biofilms almost certainly form a large part of the chronic microbiota of pathogens that cause chronic inflammatory and autoimmune diseases, the fact that patients can use the Marshall Protocol to recover from such illnesses again suggests that the treatment must be effectively allowing them to target and destroy biofilms.Because all evidence points to the fact that the MP does indeed effectively target biofilm bacteria, it is of utmost importance that people who suffer from any sort of biofilm infection start the treatment. Knowledge of the Marshall Protocol has yet to reach the cystic fibrosis community, but there is great hope that if people with the disease were to start the MP, they could destroy the P. aeruginosabiofilms that cause their untimely deaths. In the same vein, people with a wide range of infections, such as

those infected with biofilm during surgery, can likely restore their health with the MP.It is to be hoped that the clinical data emerging from the Marshall Protocol study site, which shows patients recovering from biofilm-related diseases, will inspire future researchers to invest a great deal of energy into further research aimed at identifying and studying the biofilm bacteria – bacteria that almost certainly form part of the microbiota of pathogens that cause inflammatory disease. In the coming years, as the technology to detect biofilms becomes even more sophisticated, it is almost certain that a great number of biofilms will be officially detected and documented in patients with a vast array of chronic diseases.REFERENCESCosterton, J. W., , P. S., & Greenberg, E. P. (1999). Bacterial biofilms: a common cause of persistent infections. Science (New York, N.Y.), 284(5418), 1318-22. [↩]

[↩] [↩] [↩]Higgins, D. A., Pomianek, M. E., Kraml, C. M., , R. K., Semmelhack, M. F., & Bassler, B. L. (2007). The major Vibrio cholerae autoinducer and its role in virulence factor production. Nature, 450(7171), 883-6. [↩]Singh, P. K., Schaefer, A. L., Parsek, M. R., Moninger, T. O., Welsh, M. J., & Greenberg, E. P. (2000). Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. Nature, 407(6805), 762-4. [↩] [↩]Stoodley, P., Purevdorj-Gage, B., & Costerton, J. W. (2005). Clinical significance of seeding dispersal in biofilms: a response. Microbiology, 151(11), 3453. [↩]O’toole, G. A., & Kolter, R. (1998). Flagellar and Twitching Motility Are Necessary for Pseudomonas Aeruginosa Biofilm Development. Molecular Microbiology, 30(2), 295-304. [↩]Cho, H., Jönsson, H., , K., Melke, P., , J. W.,

Jedynak, B., et al. (2007). Self-Organization in High-Density Bacterial Colonies: Efficient Crowd Control. PLoS Biology, 5(11), e302 EP -. [↩] [↩]Brockhurst, M. A., Hochberg, M. E., Bell, T., & Buckling, A. (2006). Character displacement promotes cooperation in bacterial biofilms. Current biology: CB, 16(20), 2030-4. [↩]Parsek, M. R., & Singh, P. K. (2003). Bacterial biofilms: an emerging link to disease pathogenesis. Annual review of microbiology, 57, 677-701. [↩]Kraigsley, A., Ronney, P., & Finkel, S. Hydrodynamic effects on biofilm formation. Retrieved May 28, 2008. [↩]Hall-Stoodley, L., Costerton, J. W., & Stoodley, P. (2004). Bacterial biofilms: from the Natural environment to infectious diseases. Nat Rev Micro, 2(2), 95-108. [↩] [↩] [↩], K. (2001). Riddle of biofilm resistance. Antimicrobial agents and chemotherapy, 45(4), 999-1007. [↩] [↩]Parsek, M. R., & Singh, P. K. (2003). Bacterial biofilms: an emerging link to disease pathogenesis. Annual review of microbiology, 57, 677-701. [↩]Trampuz, A., Piper, K. E., son, M. J., Hanssen, A. D., Unni, K. K., Osmon, D. R., et al. (2007). Sonication of Removed Hip and Knee Prostheses for Diagnosis of Infection. N Engl J Med, 357(7), 654-663. [↩]Ristow, P., Bourhy, P., Kerneis, S., Schmitt, C., Prevost, M., Lilenbaum, W., et al. (2008). Biofilm formation by saprophytic and pathogenic leptospires. Microbiology, 154(5), 1309-1317. [↩]Moreau-Marquis, S., Stanton, B. A., & O’Toole, G. A. (2008). Pseudomonas aeruginosa biofilm formation in the cystic fibrosis airway. Pulmonary pharmacology & therapeutics. [↩]Hall-Stoodley, L., Hu, F. Z., Gieseke, A., Nistico,

L., Nguyen, D., , J., et al. (2006). Direct Detection of Bacterial Biofilms on the Middle-Ear Mucosa of Children With Chronic Otitis Media.JAMA, 296(2), 202-211. [↩]Imamura, Y.,

Chandra, J., Mukherjee, P. K., Lattif, A. A., Szczotka-Flynn, L. B., Pearlman, E., et al. (2008). Fusarium and Candida albicans Biofilms on Soft Contact Lenses: Model Development, Influence of Lens Type, and Susceptibility to Lens Care Solutions. Antimicrob. Agents Chemother., 52(1), 171-182. [↩], G. A., Swogger, E., Wolcott, R., Pulcini, E. D., Secor, P., Sestrich, J., et al. (2008).Biofilms in Chronic Wounds. Wound Repair and Regeneration, 16(1), 37-44. [↩]Marshall, T. G. (2006b). A New Approach to Treating Intraphagocytic CWD Bacterial Pathogens in Sarcoidosis, CFS, Lyme and other Inflammatory Diseases. [↩] [↩]Marshall, T. G., & Marshall, F. E. (2004). Sarcoidosis succumbs to antibiotics–implications for autoimmune disease. Autoimmunity reviews, 3(4),

295-300. [↩] [↩]Sr, G. J. D., & Woody, H. B. (1997). Bacterial persistence and expression of disease. Clinical Microbiology Reviews, 10(2). [↩]Marshall, T. G. (2007). Bacterial Capnine Blocks Transcription of Human Antimicrobial Peptides. Nature Precedings. [↩]on, H. I., Ellison, L. F., & , G. W. (1999). Periodontal disease and risk of fatal coronary heart and cerebrovascular diseases. Journal of cardiovascular risk, 6(1), 7-11. [↩], R., & Hirani, V. (2007). Dental Health and Cognitive Impairment in an English National Survey Population. Journal of the American Geriatrics Society, 55(9), 1410-1414. [↩]Falkinham Iii, J. O.,

Iseman, M. D., Haas, P. D., & Soolingen, D. V. (2008). Mycobacterium avium in a shower linked to pulmonary disease. Journal of water and health, 6(2), 209-13. [↩], K. (2001). Riddle of biofilm resistance. Antimicrobial agents and chemotherapy, 45(4), 999-1007. [↩]Starner, D et al. 2008. Subinhibitory Concentrations of Azithromycin Decrease Nontypeable Haemophilus influenzae Biofilm Formation and Diminish Established Biofilms.Antimicrobial agents and chemotherapy 52(1):137-45. [↩]Cogan,

N. G., Cortez, R., & Fauci, L. (2005). Modeling physiological resistance in bacterial biofilms. Bulletin of mathematical biology, 67(4), 831-53. [↩]Marshall, T. G. (2006). VDR Nuclear Receptor Competence is the Key to Recovery from Chronic Inflammatory and Autoimmune Disease. [↩] Filed under: biofilms, featured articles RSS feed for comments on this post37 Responses for "Understanding Biofilms"Ken Collerman May 29th, 2008 at 1:57 am1Amy,This article is a masterpiece!Amy Proal May 29th, 2008 at 9:40 am2Thanks Ken!It was a pleasure to write the article because I find the subject matter so fascinating!AmySherry June 2nd, 2008 at 4:54 pm3Is it possible to kill biofilms on our food by heating? I am having second thoughts about eating any raw vegetables…The article is well researched, well written, and very insightful.Thank you!Amy Proal June 2nd, 2008 at 5:00 pm4Hi Sherry,I’m glad you enjoyed the article.I’m not sure about the answer to your question. The foods items that seem to be most affected by biofilm contamination are raw, and can’t be heated in order to remove biofilms.I assume that if you had a food item and you heated it to high temperatures the high level of heat would have an effect on the homeostasis and potential survival of a biofilm, although I don’t know for sure.Are you on the MP? If so, you will almost certainly easily kill any biofilm bacteria that you might acquire by eating a raw vegetable before it is able to enter the bloodstream. So I wouldn’t worry about eating them.Don’t forget

that until we reach the late stages of the MP, our own bodies are already filled with bacteria. They cover our hands and entire skin surface. They are already in our mouth. So coming in contact with biofilm bacteria in the mouth isn’t all that different from what we experience on a daily basis. Of course, being on the MP is key, so that an ingested biofilm will not be able to cause any harm.Best,Amy W. June 2nd, 2008 at 5:02 pm5I wonder if there’s a way to interfere with the biofilm’s chemical signaling directly. It would require much more specific diagnosis of the underlying pathogens, or else a good general cocktail.Do biofilm-creating pathogens just travel together or do they ‘learn’ how to signal one another based on shared plasmids? If the second, the plasmids themselves might be chemically attacked.Amy Proal June 2nd, 2008 at 5:30 pm6Hi ,Yes, there are research teams who are working to try to interfere with the chemical signaling in a biofilm. In fact, I think Higgins and team who are working with CAI-1 are actively seeking ways to inhibit the signal in an effort to slow biofilm growth. If they succeed, andother researchers successfully stop other biofilm signals, potential drugs that inhibit the signals they identify could form part of a larger arsenal to target biofilms. Of course, with any drug comes side effects, so people would have to weigh the pro/cons of taking drugs to interfere with biofilm signals, particularly when they can already target biofilms with just a few pulsed, low-dose antibiotics.I imagine that groups of people with

specific biofilm infections (such as cystic fibrosis patients with P. auruginosa infections), would most benefit from therapies aimed at stopping chemical signals that aid the formation of biofilms.I think that the bacteria inside biofilms communicate in myriad ways, depending largely on the species present. So a lot of individual work with particular biofilms must be done to uncover ways to break up signaling pathways.I’m not sure biofilm communication has much to do with sharing plasmids. Biofilm bacteria literally emit chemical signals (not plasmids) into the surrounding environment. These signals are picked up by other nearby bacteria. Depending on the strength of the signal they react indifferent ways. For example, if a bacterium picks up a lot of other signals, it knows it is surrounded by a lot of other bacteria. That way it is able to “realize†that forming a biofilm with the nearby bacteria is a possibility.

Other modes of signaling remain less clear as far as I can tell. I’m sure further research will turn up much more complex modes of action.Best, W. June 3rd, 2008 at 1:09 am7Hi Amy, you wrote;I’m not sure biofilm communication has to do with sharing plasmids. They literally emit chemical signals (not plasmids) into the surrounding

environment.My point was that the bacteria had to have a common ‘language’ in order to communicate and I wondered if the key for that language was found in their chromosome or in a transmissable, extra-chromosomal unit (Kind of like giving a person a gene which lets them speak English so that you can talk to them.)There was some possibility for developing drugs that targeted plasmids, so I thought that that might be a good way to break up a biofilm.Amy Proal June 3rd, 2008 at 9:01 am8Hi ,I see. It possible that some genes are embedded within plasmids and others within the genome. We’re dealing with a lot of different genes here so they may be in diverse locations. Hopefully future researchers projects will address this issue….Dr.Abbas Ubaid Al_Janabi June 16th, 2008 at 8:29 am9Thanks for this best artical about the biofilms,I am finsed my Ph.D.Microbial Biofilms and I havemany papears in this field ,So, Plz keep attach with me amy.All the best YoursDr. Abbas Al_JanabiCollege of Medicine, University of Al_Anbar,Ramadi,IraqAmy Proal June 16th, 2008 at 10:05 am10Hi Dr. Abbas,I’m glad to hear that a biofilm expert enjoyed my article. Congrats on getting your PhD in biofilm-related research. I think you are pursuing a very interesting course of study that will be extremely relevant to the future of medicine.Do you have a link to any of your published papers? I’d enjoy looking over them…Best,AmyBeth August 19th, 2008 at 11:38 pm11AmyWe are doing a Nursing Grand Rounds which includes a segment on oral care. I would like to use some of your pictures. Can I have permission to do so?Thank you for your consideration.BethAmy Proal August 20th, 2008 at 7:50 am12Hi

Beth,Many of these images are from the Center for Biofilm Engineering in Montana. One of the missions of CBE is to promote the understanding of biofilms. The Center has a page on its website explaining acceptable reuse.Best,Amy Rifkin, MD September 8th, 2008 at 5:22 pm13Amy: Terrific article. I’ve been asked to give a talk on biofilms for our new Masters in Biotechnology course and I’d love to be able to use some of your slides (proper acknowledgement of course). Would that be possible. Thanks. GRAmy Proal September 9th, 2008 at 1:54 am14Hi ,Of course, you are welcome to use my content for the purposes of a presentation. Please note, as per comment #12, that when it comes to the images, you’re going to want to attribute the CBE in Montana.If you haven’t done so already, you may also want to look at my 87-minute videoIntroduction to the Marshall Protocol.Best,AmyAnusha December 2nd, 2008 at 7:12 pm15Hi Amy,Thank you for this detailed report on biofilms. I am suspicious of biofilm contamination in my airway epithelial cell culture. I see a network of floating dead/dying epithelial cells on the top layer of cell culture medium and a translucent jelly-like strands that dont seem to be fungi. The culture is an established immortalized cell line derived from a patient’s lung biopsy. Is there any way to test for persistant contamination? Any stain or culture technique to detect presence of

biofilms?Thank you!Amy Proal December 3rd, 2008 at 1:05 pm16Hi Anusha,There are tests that detect biofilms but they are not necessarily available in a standard lab. In order to test you samples, I think you would have to contact a researcher working with biofilms and ask them for help and guidance.You could start by contacting the Center for Biofilm Engineering at Montana State University. It’s likely that someone on their staff could

tell you more about biofilm testing and who to contact to do the research.Personally, I think there’s a great group of biofilm researchers at the University of Washington. Drs. Parsek, Singh, Harwood and others. You could look them up and send them an email with your question. I’m not sure if they would respond but they might give you feedback.Best,AmyShari Gold April 14th, 2009 at 5:44 am17Hi Amy,I

just re-read this article. It is such a great, well researched article, written in a very direct and easy to understand manner. I had a few quick questions. I am familiar with biofilms contaminating artificial surfaces within the human body, but wondered if biofilms colonize regular joints, not altered by artificial replacements. In particular, shoulder and hips, wrists and fingers – could this be what is going on in something like RA too? Also, what about your nerves – say, peripheral nerves? Is this something they might colonize as well? Reason I ask is I do have peripheral neuropathy in my back. Neuropathy is something I know that resolves on the MP – although it is something that takes a bit longer. Could this be because much of what might be affecting that signal transduction in the nerve is stopped by biofilm inflammation somewhere from the nerve root to the muscle? In me, my neuropathy is getting much better and I can definitely see the

waxing and waning of symptoms as I increase abx and ramp different abx combos. Based on what I just re-read tonight, that might seem a plausible explanation for the physical phenomenon I am experiencing – killing a bunch of bacteria, one layer at a time. Also, as noted in the patients w/ cystic fibrosis or chronic sinus infected patients, as I have gotten further into phase 2, I have had a lot of sinus drainage and much nasty stuff coming up from lower respiratory, like I have a constant cold, but it isn’t “infection†in the clinical way, just sputum breaking up constantly, might this be evidence of biofilm existence in these places? This is truly a fascinating area of study.Thanks for you insight.Best,Shari Albert April 14th, 2009 at 8:48 am18Hi Shari,Amy is away, so you’re stuck with me. : )I don’t think there’s anything unique about non-human surfaces that leads to formation of biofilm. You may be shocked to learn that most people manually remove biofilm from themselves every day, sometimes twice a day. This odd ritual? Teeth brushing! Teeth brushing is nothing more than the act of manually removing biofilm from the surface of one’s teeth.But, it would be very un-bascterialike for bacteria to confine themselves to prosthetic hip joints and teeth.

If you think about it, the only reason we don’t brush every other part of our body is that we can’t – regrettably, we cannot brush our kidney, liver, nerves, lungs, etc.Do biofilm colonize nerves? They have had hundreds of thousands of years to figure it out. I would say absolutely. One of the limiting factors as far as progress on the MP is concerned is blood flow to a region. I remember hearing that nerve cells don’t get as much blood as other areas. That’s not to the say that neuropathy doesn’t resolve, only that the process may take longer. However, the waxing and waning of your neuropathy symptoms with your antibiotics supports the idea that these symptoms will resolve.As for your theory on signal transduction, I’m sure it’s involved, but I’m afraid I couldn’t say how.Also, I think you’re on target when it comes to sinus drainage – bacteria are involved – although I’m not sure we have any

good idea on the relative proportion of different forms of bacteria by region of the body: L-forms, intracellular pathogens, biofilm, etc.Best, May 13th, 2009 at 4:49 pm19Hi Amy,thanks for the article and research… In your research, did you happen to find any effective alternative medicines for removal of biofilms? or energy medicine? Ondamed has been used

successfully now in Europe for 15+ yrs in chronic lyme….Amy Proal May 15th, 2009 at 11:38 am20Hi ,Unfortunately I do not know of any substance that can target internal biofilms. External biofilms – or those that grow outside the body – are a different story. Those can be treated with several chemicals, however they are chemicals that would be toxic if swallowed.Actually, some studies show that a substance called Lactoferrin can reduce

biofilm growth. However I believe more research is needed to confirm its effects on biofilm proliferation and to make sure it doesn’t interfere with other pathways in the body when taken.Does Ondamed cause patients to experience a bacterial die-off reaction? I ask, because if it doesn’t cause an exacerbation in disease symptoms then it’s not causing bacteria to be killed. We know this because a rise in symptoms generated by bacterial death cannot be avoided when the MP medications begin to stimulate the immune response to target biofilm species.If Ondamed makes patients feel better instead of worse, it is probably just a palliative treatment option. What it may actually do is slow the immune response. While this would lower the inflammation associated with bacterial death it wouldn’t actually kill biofilm bacteria and would be ineffective in the long run. Clearly Lyme is still a big problem in Europe so unfortunately I

suspect that Ondamed falls into this latter category.I have not researched energy medicine and biofilms enough to comment on whether it would help with their elimination. What I can say is that the MP seems quite effective at targeting biofilm species! Essentially 100% of patients to start the treatment have experienced immunopathology or bacterial die-off and these reactions remain strong and steady during much of treatment. Then, during later stages of treatment, we have patients reporting improvement from many known biofilm-related infections. So I would say the MP is certainly your best bet if you are trying to target biofilm. I encourage you not to worry about taking the MP medications as they have an excellent safety profiles and are likely safer than taking herbs or other “natural†supplements whose properties have not been investigated at the molecular level.This article discusses the safety of the MP meds:http://bacteriality.com/2008/02/23/misconceptions/#2Hope this helps!Amy

W. May 15th, 2009 at 12:17 pm21 – Xylitol and Serrapeptase both seem pretty safe and have activity against

biofilms, though I’m not sure how long xylitol lasts or how well it works inside the body. Xylitol is a sugar alcohol used as a sweetner and in some toothpastes because of its flavor and effect on oral biofilms. Serrapeptase is an anti-inflammatory and blood thinner used routinely in Germany as an aspirin substitute since it doesn’t interfere with K1 (it doesn’t promote heart disease via arterial calcification like Aspirin does) or seem to cause the liver problems that salyciliates like Aspirin do. Both seem very safe in proper dosages. It may be hard to tell if Serrapeptase caused a bacterial dieoff reaction since it’s also an anti-inflammatory but it seems like one of the more promising candidates for attacking internal biofilms.Amy Proal May 15th, 2009 at 12:57 pm22Hi ,Yes, Xylitol has been proposed to curb biofilm growth. It’s in my toothpaste and mouthwash (I use a brand of tooth products called Biotene which is designed to target biofilm bacteria). It’s also some chewing gum. However I have not found any studies that show it’s effective at targeting biofilm other than those on the teeth and I’m not really sure if it even works as intended.When I googled Serapeptase, I read that “Serrapeptase is an enzyme that is produced in the intestines of silk worms to break down cocoon

walls. This enzyme is proving to be an alternative to the non-steroidal anti-inflammatory agents (NSAIDs) traditionally used to treat rheumatoid arthritis and osteoarthritis.â€NSAIDs are completely contraindicated for use for MP patients because they palliate inflammation rather than actually killing the bacteria causing inflammation. So that supplement seems suspect to me and I would certainly never take it.I cannot emphasize enough the risk of taking supplements when most of the claims about how they work are just based on speculation and not on molecular data. I took way too many supplements before the MP which were supposed to kill bacteria and they did nothing but make me more ill.Best,Amy W. May 15th, 2009 at 1:35 pm23When I googled SerapeptaseIf you’re looking for hard data, why not use pubmed?Serrapeptase has the demonstrated capacity to increase antibiotic effectiveness against biofilm forming bacteria. It’s demonstratablynot simply palliative. I understand that it’s contraindicated under the Marshall protocol, but there’s still very good evidence it’s helpful and well tolerated and doesn’t just mask symptoms.linkHere’s a study showing injected serrapeptase + antibiotics is significantly more effective at clearing biofilm producing infections than antibiotics alone.We have chosen serratiopeptidase (SPEP), an extracellular metalloprotease produced by Serratia marcescens that is already widely used as an anti-inflammatory agent, and has been shown to modulate adhesin expression and to induce antibiotic sensitivity in other bacteria. Treatment of L. monocytogenes with sublethal concentrations of SPEP reduced their ability to form biofilms and to invade host cells. Zymograms of the treated cells revealed that

Ami4b autolysin, internalinB, and ActA were sharply reduced. These cell-surface proteins are known to function as ligands in the interaction between these bacteria and their host cells, and our data suggest that treatment with this natural enzyme may provide a useful tool in the prevention of the initial adhesion of L. monocytogenes to the human gut.http://www.ncbi.nlm.nih.gov/pubmed/18479885?ordinalpos=1 & itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSumAntibiotic susceptibility tests on both planktonic and sessile

cultures, studies on the dynamics of colonization of 10 biofilm-forming isolates, and then bioluminescence and scanning electron microscopy under seven different experimental conditions showed that serratiopeptidase greatly enhances the activity of ofloxacin on sessile cultures and can inhibit biofilm formation.http://www.ncbi.nlm.nih.gov/pubmed/8109925?ordinalpos=3 & itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSumAmy Proal May 16th, 2009 at 12:36 pm24,Thank you, I know how to use PubMed. The reason I didn’t start searching PubMed for info about Serapeptase is because it’s not a topic relevant to what I’m spending my energy on at the moment. On this site, I try to explain the MP in simple terms – the MP as it is created and the molecular data we have about how the medications interact. I already have many comments from people asking about these topics and I need/want to answer their basic questions first before I speculate about a

supplement with you.You know yourself that the MP has a no supplement policy, and for a reason. If you actually started the MP, you’d realize that the combination of Olmesartan and low dose antibiotics is so powerful that our main problem is certainly not whether people are targeting enough bacteria. Our main problem is how to control the die-off reaction and keep people from killing so many bacteria in a day that they can’t tolerate the die-off symptoms. So at this point we don’t need any extra supplements to target biofilm bacteria.That’s especially true since we don’t know how serapeptase works at the molecular level. Maybe it does hinder biofilm formation somewhat but what’s to say it doesn’t interfere with any of the receptors involved in the immune response? How do we know that it wouldn’t interfere with the ability of Olmesartan to bind the VDR? We don’t know at this point and so I worry that when you

mention these things people who I really believe should try the MP are going to skip the treatment and think that a supplement can help them instead – which is exactly what I don’t want them to think.Best,AmyL. Lange June 3rd, 2009 at 9:59 pm25Hi Amy –I was reading your article and enjoyed it a lot, thank you! Do you know anything about Lymes disease and if the bacteria are protected by the the concepts of biofilms? If yes, do you

think that the Marshal protocol would help someone recover from the disease?Thanks in advance for your reply!-Amy Proal June 7th, 2009 at 10:05 pm26Hi ,Sorry for the delayed response. I just got back from a trip to China.Yes, it is very likely that many of the bacteria that cause Lyme disease persist in a biofilm state

and there are definitely many people on the MP with Lyme disease who are recovering.Here are interviews with three of them:http://bacteriality.com/2007/09/22/interview3/http://bacteriality.com/2007/12/28/interview14/http://bacteriality.com/2008/03/31/interview19/You might also want to post on the website http://www.curemyth1.org (Th1 refers to diseases caused by bacteria, hence the name). The site is a place where people can ask questions about the MP. But once you are a member of the site, you can also communicate with other people on the MP for Lyme.If you have Lyme, I definitely hope that you start the MP. The best way to learn more about the treatment is to read as much as possible, on this site and also on the MP study site – http://www.marshallprotocol.com.I also highly recommend watching the following video which describes the MP and the science that forms its backbone in simple terms:http://bacteriality.com/2008/05/07/mpintro/Best,Amy June 7th, 2009 at 10:15 pm27Hey amy,thanks for the info:) As far as I know, ondamed therapy it is able to approach micro-organisims at a molecular level using frequencies…there’s been a ton of research to back up its effectiveness and YES you have to be careful with how much time you use b/c of the die off effect! The first time i used this therapy I did only 10 mins and was in bed for about a week! I will do some more research on the MP protocol… i hope u have a blessed week! p.s thanks ryan for the info too:)Amy Proal June 8th, 2009 at 11:09 am28Hi ,Thanks for writing. I know nothing about Ondamed. Are the papers you mention about it’s efficacy peer-reviewed? If so, I’d be interested in reading one.In the meantime, I’d like to emphasize that I wouldn’t definitely not recommend using Ondamed or any other frequency therapy or supplement along with the MP. As I mentioned to before, there is a careful balance of the immune system set up

in the body by the MP meds as they target bacteria, and other therapies may interfere. Also, if any therapy causes extra bacterial die-off it could lead to intolerable immunopathology when combined with the MP meds so one must be careful.I definitely hope you look into the MP !Best,Amy June 22nd, 2009 at 1:33 pm29Amy,How do I find a doctor who will use the Marshall Protocol? I have Interstitial Cystitis and tested

positive for Strep D, only after a week long broth culture. Based on what I have read, I think that the bacteria has formed a biofilm, which is why shorter cultures aren’t picking it up. My doctor doesn’t know a whole lot about biofilm, so I am trying to find a doctor who does, so that I can get treatment.Thanks in advance for any help!Amy Proal June 24th, 2009 at 12:01 pm30Hi ,I’m sorry about your illness and

positive strep cultures. In my opinion you are correct – it is very likely that many of the bacteria making you ill are in a biofilm state and cannot be kill by conventional therapies. On the other hand, the MP appears to effectively break up biofilms and I think you would definitely benefit from the treatment.I’m not sure where you live. However the best way to find a doctor in your area is to post at the following website:http://www.curemyth1.org (Th1 refers to diseases caused by bacteria, hence the name). The patient advocates on the site, who are volunteers, can likely provide you with a list of doctors that administer the MP in your area.If that doesn’t work out I recommend

showing your current doctor or a new open-minded doctor in your area Dr. Marshall and team’s latest peer-reviewed papers and presentations. See if then you can convince them to prescribe you the necessary MP meds. The publications etc. can be found here:http://mpkb.org/doku.php#publications_presentationsThis article also give more tips on finding an MP doctor.http://mpkb.org/doku.php/home:starting:physician:findingGood luck and take care!AmyJD Bear August 8th, 2009 at 11:06 am31Hello Amy,I have spent some time perusing your website and I complement you on your good work. As such, it is all the more important to not mar this good work by using inappropriate

terminology. Specifically, the term “biofilm†refers to a community of bacteria surrounded by *Extracellular* Polymeric Substances. A tubule, or filament, or “protective sheath†formed by an L-form bacteria may be a “film†and it is no doubt biological, but it is NOT a “biofilm†as commonly defined. It is a disservice to conflate these two different phenomena. Whatever is going on with L-form bacteria deserves its own terminology so as not to introduce unnecessary confusion into the field.Best Regards,-JDBAmy Proal August 8th, 2009 at 12:09 pm32Hi JDB,Thanks for your comment. I’m not sure exactly what statement I’ve made that you are referring to. Nevertheless, I don’t see why L-form bacteria could not be able to form a biofilm. Bacteria in the L-form are simply in a different part of the microbial lifecycle and have temporarily lost their cell walls. Is a cell wall necessary to become part of biofilm community? I have not seen research along those lines.This being said why wouldn’t L-form bacteria also form into communities in order to better protect themselves from the immune response etc? How do we know that a biofilm cannot be composed of both bacteria with cell walls and bacteria without cell walls?So I don’t really think we need to

come up with completely different terminology to describe bacteria in the L-form seeing as their characteristics are generally very similar to those of their walled counterparts and they interact with classical forms of bacteria very closely in vivo.Best,Amy August 26th, 2009 at 7:02 am33Dear Amy,Thanks for this article. And I hope what you do next is to study Borrelia and biofilms. Leptospira and Treponema denticola are known to be able

to persist inside biofilms. But there has not been published anything on Borrelia yet. No major journal has anything on it. It would meen so much to so many if you’d get that out…Amy Proal August 28th, 2009 at 11:45 pm34Hi ,Thank you for your interest in our research. Borrelia is definitely a very hardy pathogen that we would expect would be able to survive in biofilms. Over the course of the next

few years, Autoimmunity Research Foundation, which is the organization I work with, is hoping to run in which we will look at the DNA of the infected cells in patients with chronic disease. This study is still in the planning stages, but, in time, could eventually better help us understand how Borrelia survive in the body and in a biofilm-like environment.Meanwhile, there is research suggesting that the low pulsed manner with which antibiotics are administered is much more effective at targeting bacteria in biofilms than standard regular doses of antibiotics or antibiotics taken in an IV form. Our own clinical data also shows people reporting improvement and recovery from Lyme disease thanks to therapy with the Marshall Protocol. So, if you think you may harbor Borrelia, and you want to target Borrelia in the biofilm state, looking into the MP may be a worthwhile option.Best,Amygregg zulauf September 12th, 2009 at 4:16 pm35I was impressed by the article in ‘Discover Magazine’ about biofilms (link below). Anecdotal evidence of xylitol breaking down certain biofilm structures was interesting to me.This article suggests that xylitol may work internally and not only in the mouth. I am interested in your reaction to this article.Regards, Gregghttp://discovermagazine.com/2009/jul-aug/17-slime-city-germs-talk-each-other-plan-attacks/article_view?b_start:int=2 & -C= Albert September 14th, 2009 at 12:47 pm36Hi

Gregg,Interesting. One of the principles in that article is Randall Wolcott. If you do a Google search for Randall Wolcott, the #1 hit is Amy’s interview with him:http://bacteriality.com/2008/04/13/wolcott/So, yes indeed we have heard about Xylitol before.However, the problem with Marshall Protocol patients isn’t generating bacterial die-off but controlling the reaction. That said, it may be worth giving those few patients who are non-responders xylitol and seeing if that can lead to bacterial killing. This might be an avenue we will research in time….Best, September 30th, 2009 at 8:57 pm37I’ve been reading that EDTA has been used to erradicate Biofilm??Also, some claim Noni and Grapefruit seed extract works….any comments?NEWS FLASHApril 4, 2009:, Milk consumption tied to Parkinson’s diseaseMarch 21, 2009:, Hey there, how’s your Kineosphaeram holding up?Peer-Reviewed PapersAutoimmune disease in the era of the metagenome (PDF)Vitamin D: the alternative hypothesis (PDF)Dysregulation of the Vitamin D Nuclear Receptor may contribute to the higher prevalence of some autoimmune diseases in women (PDF)Vitamin D metabolites as clinical markers in autoimmune and chronic disease (PDF)Amy's Conference PresentationsMetagenomic symbiosis between bacterial and viral pathogens in autoimmune diseaseNotes from the 2009 International Congress of AntibodiesNotes from the 2008 International Congress on AutoimmunityFeatured ArticlesUpdate on tone and other issuesWhy patients with chronic disease are disaffected and how online social networks meet their needsSun-blocking culture among the ChineseSecond-guessing the consensus on vitamin DTravels, papers, and more… an updateThree days at the J. Craig Venter InstituteThe bacteria boom – implications of the Human Microbiome ProjectUnderstanding BiofilmsInterview with Dr. Randall Wolcott, bacterial biofilm wound specialistInsights into horizontal gene transfer: conversations with Dr. Gogarten and Dr. LakeVoices of reason in the vitamin D debateInterview with evolutionary biologist EwaldWhat can medical research learn from the open source software movement?Interview with Dr. Greg Blaney: MP physicianBacteria and cancer: an interview with Dr. Alan CantwellInterview with Nadya Markova: L-form ExpertGerald Domingue: Pioneer of Atypical BacteriaA History of Cell Wall Deficient Bacteria: A Selection of Researchers Who Have Worked with the L-formPatient InterviewsInterview with Gene – sarcoidosis, bladder cancerInterview with Bonnie B – lupus, Sjogren’s SyndromeInterview with Eastlund – diabetes, sarcoidosis,

irritable bowel syndromeInterview with Roy P. – sarcoidosis, rheumatoid arthritisInterview with de Jager: chronic fatigue syndrome, multiple chemical sensitivityInterview with Ken L. – Post Treatment Lyme Disease Syndrome (PTLDS)Interview with Doreen V. – autism, ADHD depression, severe anxiety, CFSInterview with Jane -Aoki: Neurosarcoidosis, systemic sarcoidosis; spasticity, myasthenia, CNS dysfunction, joint pain, pulmonary, splenic and cardiac involvement.Interview with Melinda Stiles – Lyme, Irritable bowel syndrome/colitis, Radiculitis (inflammation of the nerve roots)Interview with Freddie Ash – Sarcoidosis of the heart, coronary artery disease, atrial fibrillationInterview with P. Bear R.N. – Chronic Borreliosis (“Lymeâ€), MCS (multiple chemical sensitivities), Chronic Spinal Inflammation, Peripheral NeuropathyInterview with Sherry Cook – Sarcoidosis, Cat Scratch Fever, Restless Leg SyndromeInterview with Leesa Shanahan – Sarcoidosis (Heerfordt’s Syndrome), UveitisInterview with Mirek Wozga – sarcoidosisInterview with Albert – CFS, depression, food sensitivitiesInterview with Carole – Sarcoidosis, fibromyalgia,

CFSInterview with Shirley J. (Saj) – SarcoidosisInterview with Robyn – Lyme, myoclonusInterview with Sue Andorn – Lyme, BabesiaInterview with Ival Meyer – Arthritis, dyslexiaInterview with Guss Wilkinson – Sarcoidosis, psoriasis,

insomniaAbout Amy ProalAmy Proal graduated from town University in 2005 with a degree in biology. While at town, she wrote her senior thesis on Chronic Fatigue Syndrome and the Marshall Protocol.Amy has spoken at several international conferences and authored several peer-reviewed papers on the intersection of bacteria and chronic disease.If you have questions about the MP, please visitCureMyTh1.org where volunteer patient advocates will answer your questions. Another good resource is the MP Knowledge Base, which is scheduled to be completed within the next year.Categories Select Category aging (1) biofilms (3) cancer (2) cardiovascular disease (1) cognitive dysfunction (3) conferences and trainings (7) diet (4) familial aggregation (3) featured articles (18) history (3) horizontal

gene transfer (2) interview (doctor/researcher) (5) interview (patient) (21) L-form bacteria (6) marshall protocol (21) medical research (5) mental illness (2) microbiome (3) News Flash (38) obesity (2) personal (2) presentations (3) statins (1) Uncategorized (1) videos (4) vitamin d (12) RSS FeedsPostsCommentsCopyright © 2009 - Bacteriality — Exploring Chronic DiseaseThis site uses a modified version of Illacrimo Theme created by Design DiseaseFrom: Goldstein@...To: bird mites Sent: Friday, September 9, 2011 8:28:49 AMSubject: Re: Good summary about biofilms

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled

things that pop up, then crust over and go away and new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Hi Cecilia,No, I'm not taking the bleach baths... have felt pretty weak lately, so until I feel stronger I'm not going to do them, but may (I have to think about the pros and cons at this point--the MMS baths did not seem to help this either). I'm glad the bleach baths have helped your little 8 year old. Does she have any other health issues going on? I wish she didn't have to go through this. She is almost entirely dependent on you to help figure some of this stuff out to help her. How is the little one doing?From: "Cecilia Borg" <ceciliaborg@...>bird mites Sent: Friday, September 9, 2011 11:08:26 PMSubject: Re: Good summary about biofilms

HI, !Thanks for the info. Then I know we don´t have biofilm in hair or on body (only plaque as Krys explained:)Are you doing the bleach baths? Does it help you?Take care, !CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Friday, September 9, 2011 5:28 PMSubject: Re: Good summary about biofilms

Hi Cecilia,Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice. Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled

things that pop up, then crust over and go away and new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on it. Maybe Aandraya knows of something too.From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?Cecilia

From: "Goldstein@..." <Goldstein@...>

bird mites Sent: Friday, September 9, 2011 6:35 AM

Subject: Re: Good summary about biofilms

Yes. True. L.From: "Aandraya Da Silva" <aandraya@...>

bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my

summer vacation (long)Reply-VitaminK

> > I didn't mean to write a novel but this turned out to be somewhat long. > These are my observations and results as of mid-September; we are not > finished yet so I will update this. We started Interfase enzymes at the

> beginning of July. We are at the maintenance level and I don't know yet how > long we will continue to need Interfase. I seem to have uncovered many more > infections than I anticipated and eliminating them requires persistence.

> > > > All of the existing "biofilm protocols" assume that biofilm is limited to > the intestines, which just couldn't possibly be correct. Biofilm must be > colonizing all parts of the children's bodies, including tissues, glands,

> organs, membranes, joints, and all of the cranial openings including > sinuses, nose, eyes, ears, and mouth. Researchers know that

biofilm causes > heart valve infections, middle ear infections (i.e. otitis media, which is > rampant in children with autism), prostate inflammation, and periodontal > disease, none of which are located in the intestines. The mucous membrane

> surfaces in the head are known to be prime sites for biofilm colonization, > which means that toxins are being produced in close proximity to the brain. > Thus we have to assume that biofilm is everywhere in the body and that we

> have to treat biofilm everywhere in the body. > > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi > secrete sticky slime in order to anchor themselves to a surface, allowing

> them to stay in one place and build a colony rather than be swept away by > moving fluid such as blood. A wide variety of different microorganisms > reside within a

biofilm colony. The biofilm colony secretes significant > quantities of metabolic wastes, much of which is acid and ammonia, and the > children's kidneys must excrete all this acid and ammonia Metabolic wastes

> generated by biofilm can place a huge burden on the kidneys and if the > kidneys are unable to keep up with the flow of microbial wastes then the > result will be high levels of circulating toxins.

> > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys > in excreting microbial acids faster. The baths and the electrolyte drink > help in maintaining pH at a normal level. Liquid phosphorus helps the

> kidneys get rid of acid. Vitamin K2 will activate proteins that pull > calcium out of the slime and cause it to disintegrate. So the Interfase > enzymes need to be used along with all the other components of the

Vitamin K > protocol. If the child is not supported nutritionally during slime removal, > the child will not be able to tolerate the die-off. > > > > My approach differs from other "biofilm protocols" in that I am assuming

> biofilm colonies are everywhere in the body, not just in the intestines, so > biofilm must be dissolved from the tissues and organs as well as the > intestines. I have been using Klaire Labs' Interfase enzymes to dissolve

> the slime and they really do seem to work. It's critically important to > recognize that as the slime dissolves live microbes are released into the > bloodstream, so the enzymes should be started at a low dose with plenty of

> antimicrobials to kill the released microbes. > > > > Microbes and parasites can be divided into three categories, each of which > needs to be

treated: > > > > Category I: Extracellular microbes, including bacteria, fungi, and other > microorganisms living in a biofilm community attached to, but outside of, > the host's cells. It appears that the biofilm structure can be dissolved

> using specialized enzymes and Vitamin K-activated proteins. However enzymes > do not kill microbes - herbal and perhaps prescription antimicrobials will > be needed for that job. > >

> > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa, > worms, etc., which are living in and protected by the slime. These will > start to emerge and cause symptoms as the biofilm dissolves.

> > > > Category III: Intracellular parasites, such as toxoplasma and the various > tick-borne diseases, living inside the host's cells. These need to be

> treated for long periods of time. It is probable that the slime prevents > medications from reaching the infected cells so removal of biofilm colonies > should improve the treatment of intracellular microbes.

> > > > ENZYMES > > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase > Regular and Interfase Plus on all my family members plus a few friends'

> children and the EDTA has not tested positive for anyone so far. Thus my > advice is to use regular Interfase because the kidneys are already stressed; > adding a chelating agent during the early stages of slime removal is too

> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that > they are absorbed into the bloodstream. > > >

> Interfase by

Klaire Labs: I use my pendulum to test my children and myself > every day (I drive them crazy, actually) and these doses are based on my > experience. Depending on the child's size, start with no more than 1/4 to 1

> capsule and stay at that dose for several days to observe. Increase the > dose slowly, over a 12-week period, to reach the maintenance dose. > Recommended amounts of Interfase: > >

> > Children up to 4 years old: Start with 1/4 capsule/day and work up to a > maintenance dose of 12 capsules/day. > > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a

> maintenance dose of 24 capsules/day. > > > > Children 10-15: Start with no more than 1 capsule/day and work up to a > maintenance dose of 36 capsules/day. > >

> > Children 16+ and

adults: Start with no more than 1 capsule/day and work up > to a maintenance dose of 48 capsules/day. > > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin

> K2. Nattokinase dissolves fibrin which helps hold together the slime > structure. Recommended ratio is one Nattokinase capsule per three Interfase > capsules. I use Natto-K from Enzymedica. >

> > > Protease Enzymes: These might be helpful in because they will break down > the proteins in dead organisms. Give 4-8 or more per day. Use with caution > if the child has a history of GI pain. I use ViraStop from Enzymedica.

> > > > The dying microbes will produce lots of acid! I can't emphasize this > enough! Continue to support the children with the Vitamin K protocol so > they can clear the acid from their bodies.

If it's at all possible, take > your child to a mineral hot springs for a few days because it's a great way > to alkalinize the body quickly. > > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!

> The liver will have to process large quantities of toxins from the > dissolving slime. Vitamin A activates the genes in the liver that run the > detoxification enzymes so the liver will be on overdrive and consuming large

> quantities of Vitamin A which is why it's so important to increase the dose. > VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's > current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps

> from Pure Encapsulations and add in enough of those per week so that the > total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was > before. >

> > > ANTI-MICROBIALS AND ANTI-FUNGALS > > > > Category I anti-microbials: Use a variety of anti-microbials to target as > many different microbes as possible. We used a lot of goldenseal, which

> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic > and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from > Mediherb, which contains cranberry and uva ursi and got rid of something in

> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to > get something in the sinuses. Oil of oregano and colloidal gold (from > WaterOz) were helpful. It's advisable to use a variety of herbs in order to

> hit as many different microbes as possible. > > > > Category II anti-parasitics: treatment depends on what turns up. In my > family we have roundworms,

which were diagnosed in my younger son six years > and were obviously not treated adequately. I am using Biltricide and Vermox > which are prescription - I don't think herbals are sufficient to kill off

> the really large parasites. My pendulum testing indicates that the > prescription medicines need to be taken for MUCH longer than the PDR > indicates. On the plus side though, the prescription medicines seem to be

> acting against some Category III intracellular parasites too. > > > > Category III anti-parasitics: It takes a LONG time to kill off > intra-cellular parasites, for the obvious reason that they are located

> inside the cells and are thus well protected. For information on long-term > herbal treatments I recommend the book "Healing Lyme" by Harrod > Buhner. My older son has been taking all of the main herbs (andrographis,

> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The > prescription antihelmintics (e.g. Vermox and Biltricide) also seem to be > killing off intracellular parasites. >

> > > SINUSES AND OTHER CRANIAL OPENINGS > > > > Out of curiosity I used my pendulum to test whether fungus was growing in my > eyes and the answer was "yes" which got me thinking about infections in the

> various mucous membranes of the head. Biofilm is well known to colonize the > sinuses which means that microbes are producing toxins in close proximity to > the brain. Just reducing the infection load in his cranial openings has

> been surprisingly helpful for my older son. Use the neti pot twice a day if > possible, adding twice as much salt as recommended (use 1/2 teaspoon salt > per 1 cup water). Then drop colloidal silver into the

eyes and ears; use a > colloidal silver inhaler to get silver into the nostrils; and have your > child gargle and swish with colloidal silver. Argentyn 23 is supposed to be > the best brand, and it is available in dropper bottles, inhaler bottles, and

> regular bottles. I am also using a product called Neti-Wash Plus by > Himalayan Institute, found at Whole Foods, which contains goldenseal and > zinc and is effective against microbes in the sinuses and eyes.

> > > > The longer the neti pot routine can be maintained the better the results. > My testing with the silver suggests the following guidelines: use it in the > eyes once/day for a week, in the ears once/day for about 5 days, gargling

> once/day for about 5 days, and in the inhaler on an ongoing basis. This > cycle may need to be repeated. Be prepared for a smoldering infection in >

the sinuses or ears to flare as it is being eliminated. Colloidal silver in > the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > > > > CHELATION >

> > > Here is my two cents on chelation: Just put it aside until, at minimum, the > maintenance dose of Interfase has been reached. Attempting to chelate > while simultaneously inducing heavy die-off is much too stressful for the

> kidneys, which are already struggling to eliminate the microbial wastes. > EDTA is known to break apart biofilm, and my experience is that DMPS does > the same thing. When biofilm is broken apart abruptly a huge load of live

> microbes is released into the bloodstream, which release large quantities of > acids in response which puts further stress on the kidneys. Chelation > should not take priority over dissolving biofilm

or killing off intestinal > parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the > kidneys are acidic. Renal pH will not stabilize until the microbes have > been substantially eliminated - only then should chelating agents be used.

> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will > displace arsenic and will bind to aluminum (which is then eliminated), just > due to the chemistry of the molecules. Thus the Vitamin K protocol causes

> some metals to be eliminated naturally. > > > > So now, here is what I have seen in my older son who is going on 13: I > started giving him the herb andrographis last February, and although it was

> slow to act it helped a lot in calming him down. I did not see much > initially after starting Interfase in July but over the summer he

stopped > repeating things, stopped pacing, seemed to become much more mature. He is > more affectionate and is definitely more social, and this year had by far > the smoothest start to a school year he has ever had. He's much more

> responsible about his homework and is remembering to hand it in. His > teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our

> cranial therapist reiterates. He is taking about half as much magnesium > which is significant as he has been very dependent on high doses of > magnesium since he was about 5. He needs less liquid phosphorus and less

> trace minerals. ATP requirement has increased a little and of course > Vitamin A requirement has increased significantly. He was taking a lot of > goldenseal about a month ago but

doesn't need it currently; however the > andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have > all remained the same for many months now. He made big strides this summer.

> > > > My younger son, NT, is getting essentially the same supplements although he > doesn't need andrographis. He is definitely nicer and much easier to get > along with.

> > > > > > > >

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Krys!

Yes, it is interesting and that may be why it is so hard to get rid of staph-infections...

KR

Cecilia

From: Krys Brennand <krys109uk@...>bird mites Sent: Friday, September 9, 2011 8:32 PMSubject: Re: Good summary about biofilms

We know carbohydrates feed bacteria in plaque, also, if my memory serves, I think carbs encourage some types of fungus which inhabit the body. This could be a link where biofilms in warm bloodied animals are concerned.

I found the theory, in the last section, about using low level antibiotics in "pulses" to kill the biofilm, particularly interesting. I'd like to be able to see the results of more studies on this method.

I was reading that Staph aureus ( & other Staph species), apparently, are quorum sensing & biofilm producing.

Things have moved on so very far since I was at college, in fact I think much of what I learnt back then is obsolete.

I wonder whether the description of the quorum sensing bacteria as using a "language" & being "intelligent" could be misleading. I suppose it depends how we define "intelligent". We're not speaking telepathy or any actual thought. I'd think of it more like a signal which is responded to.

All the best,

Krys

On 9 September 2011 12:58, <Goldstein@...> wrote:

This was a good article if you have time to read it in its entirety. A couple of things that I did not know that I learned:

1. After antibiotics (all) there are bacteria they have referred to in science as persisters. This causes the rebound of the infection to come again as in ear infections, or in Cecilia's daughter's or my case, staph infection on the skin.

2. Since bacteria are quorum sensing they form complex grouping called biofilm and each have jobs to do... some provide nutrients, some are defenders, etc. Much like a family or civilization. They have their own language; they are intelligent.

3. Many, many, many known diseases are caused by biofilms, including ear infections in children, and plaque on teeth that harm tooth and gums. The fact that the infections come back year after year is a signal that there are persisters and there is biofilm in the inner ear.

4. One of the best ways you can help your health is by being fastidious with brushing, flossing and/or water picking teeth. As Krys said, biofilm is what makes plaque and as the teeth and gums degenerate, the biofilm finds its way to the rest of the tissues of the body. My father-in-law had endocarditis at one point and the article said the more teeth that are pulled (he recently had 9 more teeth removed), the more likely this biofilm finds its way to the brain causing strokes, or to the heart, causing endocarditis.

5. Research is picking up on the study of biofilms. The big push in the 90's until today has been the study of genetics (and in my opinion very necessary); author says biofilms will be one of the next big areas of study in science.

6. Biofilm is made up of a complex city of workers, including fungi, viruses and bacteria.

7. Biofilm can hide from the immune system, but it is just as harmful or more so than bacteria free roaming in the body.

I am curious about what foods stimulate the growth of biofilm... raw garlic obviously is a big fighter of biofilm or at least bacteria, viruses and fungi. Maybe there is more information out about that... I'll look.

So much to be gleaned from this article.

From: "Aandraya" <aandraya@...>bird mites Sent: Friday, September 9, 2011 10:27:53 AM

Subject: Re: Good summary about biofilms

I had that film too big time. I know of a girl who scraped it off and sent it to a lab, they said it was mold- Aspergillus (I think Niger). That's when I sought out treatment with Itraconazole and Voriconazle which worked. I still have some kind of mold in my skin, don't know if it's the same as before, another species, or just a lot less of the same. Either way, treating my chronic infections including the antifungals is knocking it out.

Aandraya

On Sep 9, 2011, at 10:28 AM, Goldstein@... wrote:

Hi Cecilia,

Two things, first, you are right. I don't think it is good to use Hibiclens long term. When we finish these bottles, I think that will be the end of that. Normally have not used antibacterial soaps here... Dr. told us to use it. But, what do they know? Right? Sometimes they give incorrect or bad advice.

Secondly about biofilms. I can just speak from experience. At the beginning of this attack of Mites and Morgellons we had no biofilm on the skin. Later on we developed a waxy, greasy film that is hard to remove, primarily in the hair. Husband has it too. Before his shower he looks greasy and after using something like the Hibiclens it seems to remove the surface biofilm. I have the same, and we both have those weird fluid filled things that pop up, then crust over and go away and new ones appear. I've tried many things already for this and the Doxy got rid of the biofilm for a while, but it came back after we finished the Doxy. Biofilm is a collection of organisms, but we are concerned about the biofilm from Lyme since this biofilm seems to be created by Lyme organisms and other organisms together. I think there is some research being done on this. I'll see if I can locate anything on

it. Maybe Aandraya knows of something too.

From: "Krys Brennand" <krys109uk@...>bird mites Sent: Friday, September 9, 2011 4:34:30 AMSubject: Re: Good summary about biofilms

I don't know much about biofilms, but dental plaque is one well known biofilm which affects everyone.

On 9 September 2011 02:45, Cecilia Borg <ceciliaborg@...> wrote:

How do you know you have biofilms?

Cecilia

From: "Goldstein@..." <Goldstein@...>bird mites Sent: Friday, September 9, 2011 6:35

AMSubject: Re: Good summary about biofilms

Yes. True. L.

From: "Aandraya Da Silva" <aandraya@...>bird mites Sent: Thursday, September 8, 2011 9:04:37 PMSubject: Good summary about biofilms

Those of us with chronic infections have lots of biofilms in us as these microbes live in colonies all together.

Date: September 8, 2011 9:54:24 PM CDT

VitaminK

Subject: [VitaminK] Re: Slime and bug removal, or, what I did for my summer vacation (long)

Reply-VitaminK

>> I didn't mean to write a novel but this turned out to be somewhat long.> These are my observations and results as of mid-September; we are not> finished yet so I will update this. We started Interfase enzymes at the> beginning of July. We are at the maintenance level and I don't know yet how> long we will continue to need Interfase. I seem to have uncovered many more> infections than I anticipated and eliminating them requires persistence.> > > > All of the existing "biofilm protocols" assume that biofilm is limited to> the intestines, which just couldn't possibly be correct. Biofilm must be> colonizing all parts of the children's bodies, including tissues,

glands,> organs, membranes, joints, and all of the cranial openings including> sinuses, nose, eyes, ears, and mouth. Researchers know that biofilm causes> heart valve infections, middle ear infections (i.e. otitis media, which is> rampant in children with autism), prostate inflammation, and periodontal> disease, none of which are located in the intestines. The mucous membrane> surfaces in the head are known to be prime sites for biofilm colonization,> which means that toxins are being produced in close proximity to the brain.> Thus we have to assume that biofilm is everywhere in the body and that we> have to treat biofilm everywhere in the body.> > > > Biofilm is slime. There is nothing mysterious about it. Bacteria and fungi> secrete sticky slime in order to anchor themselves to a surface, allowing> them to stay in one place and build a colony

rather than be swept away by> moving fluid such as blood. A wide variety of different microorganisms> reside within a biofilm colony. The biofilm colony secretes significant> quantities of metabolic wastes, much of which is acid and ammonia, and the> children's kidneys must excrete all this acid and ammonia Metabolic wastes> generated by biofilm can place a huge burden on the kidneys and if the> kidneys are unable to keep up with the flow of microbial wastes then the> result will be high levels of circulating toxins. > > > > One of the main thrusts of the Vitamin K protocol is to assist the kidneys> in excreting microbial acids faster. The baths and the electrolyte drink> help in maintaining pH at a normal level. Liquid phosphorus helps the> kidneys get rid of acid. Vitamin K2 will activate proteins that pull> calcium out of the slime and cause it to

disintegrate. So the Interfase> enzymes need to be used along with all the other components of the Vitamin K> protocol. If the child is not supported nutritionally during slime removal,> the child will not be able to tolerate the die-off.> > > > My approach differs from other "biofilm protocols" in that I am assuming> biofilm colonies are everywhere in the body, not just in the intestines, so> biofilm must be dissolved from the tissues and organs as well as the> intestines. I have been using Klaire Labs' Interfase enzymes to dissolve> the slime and they really do seem to work. It's critically important to> recognize that as the slime dissolves live microbes are released into the> bloodstream, so the enzymes should be started at a low dose with plenty of> antimicrobials to kill the released microbes.> > > > Microbes and parasites

can be divided into three categories, each of which> needs to be treated:> > > > Category I: Extracellular microbes, including bacteria, fungi, and other> microorganisms living in a biofilm community attached to, but outside of,> the host's cells. It appears that the biofilm structure can be dissolved> using specialized enzymes and Vitamin K-activated proteins. However enzymes> do not kill microbes - herbal and perhaps prescription antimicrobials will> be needed for that job. > > > > Category II: Gastrointestinal parasites such as giardia, amoebas, protozoa,> worms, etc., which are living in and protected by the slime. These will> start to emerge and cause symptoms as the biofilm dissolves. > > > > Category III: Intracellular parasites, such as toxoplasma and the various> tick-borne diseases, living inside

the host's cells. These need to be> treated for long periods of time. It is probable that the slime prevents> medications from reaching the infected cells so removal of biofilm colonies> should improve the treatment of intracellular microbes. > > > > ENZYMES> > > > I am not using Interfase Plus with EDTA. I have energy tested Interfase> Regular and Interfase Plus on all my family members plus a few friends'> children and the EDTA has not tested positive for anyone so far. Thus my> advice is to use regular Interfase because the kidneys are already stressed;> adding a chelating agent during the early stages of slime removal is too> hard on them. > > > > All of the following enzymes should be given on an empty stomach so that> they are absorbed into the bloodstream. > > > > Interfase

by Klaire Labs: I use my pendulum to test my children and myself> every day (I drive them crazy, actually) and these doses are based on my> experience. Depending on the child's size, start with no more than 1/4 to 1> capsule and stay at that dose for several days to observe. Increase the> dose slowly, over a 12-week period, to reach the maintenance dose.> Recommended amounts of Interfase:> > > > Children up to 4 years old: Start with 1/4 capsule/day and work up to a> maintenance dose of 12 capsules/day.> > > > Children 5-9: Start with no more than 1/2 capsule/day and work up to a> maintenance dose of 24 capsules/day.> > > > Children 10-15: Start with no more than 1 capsule/day and work up to a> maintenance dose of 36 capsules/day.> > > > Children 16+ and adults: Start with no more than 1

capsule/day and work up> to a maintenance dose of 48 capsules/day.> > > > Nattokinase: Nattokinase is produced by the same bacteria that make Vitamin> K2. Nattokinase dissolves fibrin which helps hold together the slime> structure. Recommended ratio is one Nattokinase capsule per three Interfase> capsules. I use Natto-K from Enzymedica.> > > > Protease Enzymes: These might be helpful in because they will break down> the proteins in dead organisms. Give 4-8 or more per day. Use with caution> if the child has a history of GI pain. I use ViraStop from Enzymedica.> > > > The dying microbes will produce lots of acid! I can't emphasize this> enough! Continue to support the children with the Vitamin K protocol so> they can clear the acid from their bodies. If it's at all possible, take> your child to a mineral hot

springs for a few days because it's a great way> to alkalinize the body quickly.> > > > VITAMIN A INTAKE MUST BE INCREASED DRAMATICALLY ONCE INTERFASE IS STARTED!> The liver will have to process large quantities of toxins from the> dissolving slime. Vitamin A activates the genes in the liver that run the> detoxification enzymes so the liver will be on overdrive and consuming large> quantities of Vitamin A which is why it's so important to increase the dose.> VITAMIN A REQUIREMENT WILL TRIPLE. However, don't change your child's> current dose of cod liver oil; instead, get a bottle of Vitamin A gelcaps> from Pure Encapsulations and add in enough of those per week so that the> total amount of Vitamin A (cod liver oil plus gelcaps) is triple what it was> before.> > > > ANTI-MICROBIALS AND ANTI-FUNGALS> > >

> Category I anti-microbials: Use a variety of anti-microbials to target as> many different microbes as possible. We used a lot of goldenseal, which> seems to be a good broad-spectrum anti-microbial. Carrot-juice-and-garlic> and pau d'Arco are potent anti-fungals. We also used Cranberry Complex from> Mediherb, which contains cranberry and uva ursi and got rid of something in> the kidneys, and Resveratrol Extra from Pure Encapsulations, which seemed to> get something in the sinuses. Oil of oregano and colloidal gold (from> WaterOz) were helpful. It's advisable to use a variety of herbs in order to> hit as many different microbes as possible.> > > > Category II anti-parasitics: treatment depends on what turns up. In my> family we have roundworms, which were diagnosed in my younger son six years> and were obviously not treated adequately. I am using

Biltricide and Vermox> which are prescription - I don't think herbals are sufficient to kill off> the really large parasites. My pendulum testing indicates that the> prescription medicines need to be taken for MUCH longer than the PDR> indicates. On the plus side though, the prescription medicines seem to be> acting against some Category III intracellular parasites too.> > > > Category III anti-parasitics: It takes a LONG time to kill off> intra-cellular parasites, for the obvious reason that they are located> inside the cells and are thus well protected. For information on long-term> herbal treatments I recommend the book "Healing Lyme" by Harrod> Buhner. My older son has been taking all of the main herbs (andrographis,> resveratrol, cat's claw), plus pau d'Arco, for 7 months now. The> prescription antihelmintics (e.g. Vermox and Biltricide)

also seem to be> killing off intracellular parasites. > > > > SINUSES AND OTHER CRANIAL OPENINGS> > > > Out of curiosity I used my pendulum to test whether fungus was growing in my> eyes and the answer was "yes" which got me thinking about infections in the> various mucous membranes of the head. Biofilm is well known to colonize the> sinuses which means that microbes are producing toxins in close proximity to> the brain. Just reducing the infection load in his cranial openings has> been surprisingly helpful for my older son. Use the neti pot twice a day if> possible, adding twice as much salt as recommended (use 1/2 teaspoon salt> per 1 cup water). Then drop colloidal silver into the eyes and ears; use a> colloidal silver inhaler to get silver into the nostrils; and have your> child gargle and swish with colloidal silver.

Argentyn 23 is supposed to be> the best brand, and it is available in dropper bottles, inhaler bottles, and> regular bottles. I am also using a product called Neti-Wash Plus by> Himalayan Institute, found at Whole Foods, which contains goldenseal and> zinc and is effective against microbes in the sinuses and eyes. > > > > The longer the neti pot routine can be maintained the better the results.> My testing with the silver suggests the following guidelines: use it in the> eyes once/day for a week, in the ears once/day for about 5 days, gargling> once/day for about 5 days, and in the inhaler on an ongoing basis. This> cycle may need to be repeated. Be prepared for a smoldering infection in> the sinuses or ears to flare as it is being eliminated. Colloidal silver in> the eyes feels like tapwater in the eyes - uncomfortable but not terrible. > >

> > CHELATION> > > > Here is my two cents on chelation: Just put it aside until, at minimum, the> maintenance dose of Interfase has been reached. Attempting to chelate> while simultaneously inducing heavy die-off is much too stressful for the> kidneys, which are already struggling to eliminate the microbial wastes.> EDTA is known to break apart biofilm, and my experience is that DMPS does> the same thing. When biofilm is broken apart abruptly a huge load of live> microbes is released into the bloodstream, which release large quantities of> acids in response which puts further stress on the kidneys. Chelation> should not take priority over dissolving biofilm or killing off intestinal> parasites. Moreover, DMPS and DMSA have been shown to be ineffective if the> kidneys are acidic. Renal pH will not stabilize until the microbes have> been

substantially eliminated - only then should chelating agents be used.> > > > The phosphate in the supplemental ATP, used in the Vitamin K protocol, will> displace arsenic and will bind to aluminum (which is then eliminated), just> due to the chemistry of the molecules. Thus the Vitamin K protocol causes> some metals to be eliminated naturally.> > > > So now, here is what I have seen in my older son who is going on 13: I> started giving him the herb andrographis last February, and although it was> slow to act it helped a lot in calming him down. I did not see much> initially after starting Interfase in July but over the summer he stopped> repeating things, stopped pacing, seemed to become much more mature. He is> more affectionate and is definitely more social, and this year had by far> the smoothest start to a school year he has ever

had. He's much more> responsible about his homework and is remembering to hand it in. His> teachers are obviously pleased with his classroom behavior. > > > > From the supplement perspective, he is definitely less acidic which our> cranial therapist reiterates. He is taking about half as much magnesium> which is significant as he has been very dependent on high doses of> magnesium since he was about 5. He needs less liquid phosphorus and less> trace minerals. ATP requirement has increased a little and of course> Vitamin A requirement has increased significantly. He was taking a lot of> goldenseal about a month ago but doesn't need it currently; however the> andrographis, the resveratrol, the cat's claw, and the pau d'Arco doses have> all remained the same for many months now. He made big strides this summer.> > > > My younger

son, NT, is getting essentially the same supplements although he> doesn't need andrographis. He is definitely nicer and much easier to get> along with.> > > > > > > >

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