Re: Good summary about biofilms

[Guest guest]

HI, !Thanks for caring! I can understand that you feel hesitating about doing something that is unsure if it helps and might be toxic. I really hope it is not that bad with the toxicity, cause it really helps my 8-year old. THe lesions/blisters (they are really small) are located only at the buttocks.(at least so far). She has no other symptoms, thank god! My little one (almost 2 years old now) is doing quite well she gets occasional blisters too, but not much. SHe has infections on and off, but I guess that is due to that she is at day-care with a lot of other kids when I work. It seems like it´s the same for the other small kids there. You are such a kind soul , I wish you the

best!CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Saturday, September 10, 2011 3:14 PMSubject: Re: Good summary about biofilms

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.

> > > > > > > >

[Guest guest]

I've just got around to watching this. What an amazing lady.On 9 September 2011 10:39, <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 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.”[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.”

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/seeding”, a biofilm colony differentiates to form an outer “wall” of stationary bacteria, while the inner region of the biofilm “liquefies”, 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…”[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.”[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.”

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.”

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.”

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.”[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.”[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.”[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.”

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.”

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.”

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.”[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.”

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,”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.”

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.”

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.”

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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[Guest guest]

Here's another short clip about biofilm. This is going to be huge in medicine in the next 10 years! All sorts of diseases caused by biofilms, not just Lyme Disease. I'm just now starting to understand the complexity of what we are looking at with LD. Since there are different strains of Lyme Disease, some simpler in structure than others, some LD without co-infections, we never know what we are getting or what we ended up with in our biofilm. Krys, maybe your mum will not end up with biofilm if she does not get Lyme Disease from the mites. I sure hope not. Others with bird mites have not gotten biofilm. I do not believe Zoe ended up with Lyme or biofilm.http://www.youtube.com/watch?v=lpI4WCM_9pM & NR=1From: "Krys Brennand" <krys109uk@...>bird mites Sent: Monday, September 12, 2011 7:37:38 AMSubject: Re: Good summary about biofilms

I've just got around to watching this. What an amazing lady.On 9 September 2011 10:39, <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 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,†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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Hi CeciliaAnd you too Cecilia. You have a wonderful heart! I sure wonder what these lesions/blisters are too. I might do the bleach at some point too. I got my liver/kidney testing done today so as soon as results come back from that I'll start some new drugs and see where I go from there. I got a very strange bite 4 days ago on my stomach. It blistered and formed a red ring around it. I had an identical bite last year on my leg. The blister got to be the size of a chick pea, with a red ring outside of it. I had something similar several years ago on my groin area - I can't imagine what caused that. Is that typical of a mite bite if any of you know.Hugs to all, From: "Cecilia Borg" <ceciliaborg@...>bird mites Sent: Sunday, September 11, 2011 10:03:32 PMSubject: Re: Good summary about biofilms

HI, !Thanks for caring! I can understand that you feel hesitating about doing something that is unsure if it helps and might be toxic. I really hope it is not that bad with the toxicity, cause it really helps my 8-year old. THe lesions/blisters (they are really small) are located only at the buttocks.(at least so far). She has no other symptoms, thank god! My little one (almost 2 years old now) is doing quite well she gets occasional blisters too, but not much. SHe has infections on and off, but I guess that is due to that she is at day-care with a lot of other kids when I work. It seems like it´s the same for the other small kids there. You are such a kind soul , I wish you the

best!CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Saturday, September 10, 2011 3:14 PMSubject: Re: Good summary about biofilms

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.

> > > > > > > >

[Guest guest]

Hi, linda!Thanks for your reply! Sounds strange with the bite with the ring around it. Mine (the mite-bites) never got rings around them, so I don´t know if that could be mite-bites or something else. Guess someone else here knows.Take care!CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Monday, September 12, 2011 8:57

PMSubject: Re: Good summary about biofilms

Hi CeciliaAnd you too Cecilia. You have a wonderful heart! I sure wonder what these lesions/blisters are too. I might do the bleach at some point too. I got my liver/kidney testing done today so as soon as results come back from that I'll start some new drugs and see where I go from there. I got a very strange bite 4 days ago on my stomach. It blistered and formed a red ring around it. I had an identical bite last year on my leg. The blister got to be the size of a chick pea, with a red ring outside of it. I had something similar several years ago on my groin area - I can't imagine what caused that. Is that typical of a mite bite if any of you know.Hugs to all, From:

"Cecilia Borg" <ceciliaborg@...>bird mites Sent: Sunday, September 11, 2011 10:03:32 PMSubject: Re: Good summary about biofilms

HI, !Thanks for caring! I can understand that you feel hesitating about doing something that is unsure if it helps and might be toxic. I really hope it is not that bad with the toxicity, cause it really helps my 8-year old. THe lesions/blisters (they are really small) are located only at the buttocks.(at least so far). She has no other symptoms, thank god! My little one (almost 2 years old now) is doing quite well she gets occasional blisters too, but not much. SHe has infections on and off, but I guess that is due to that she is at day-care with a lot of other kids when I work. It seems like it´s the same for the other small kids there. You are such a kind soul , I wish you the

best!CeciliaFrom: "Goldstein@..." <Goldstein@...>bird mites Sent: Saturday, September 10, 2011 3:14 PMSubject: Re: Good summary about biofilms

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.

> > > > > > > >

[Guest guest]

LINDA, please go a see a doctor straight away!! I have just been reading that

ticks cause bites with a red ring around after a couple of days. It says when

the ring is there, you should urgently go and see a doctor. Have you got

flu-like symptoms and/or stomach/guts problems as well ?

> >>>

> >>> 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.

> >>>

> >>>

> >>>

> >>>

> >>>

> >>>

> >>>

> >>>

[Guest guest]

Maybe you got reinfected with Lyme.On Sep 14, 2011, at 1:54 PM, Aggi wrote: LINDA, please go a see a doctor straight away!! I have just been reading that ticks cause bites with a red ring around after a couple of days. It says when the ring is there, you should urgently go and see a doctor. Have you got flu-like symptoms and/or stomach/guts problems as well ? > >>> > >>> 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. > >>> > >>> > >>> > >>> > >>> > >>> > >>> > >>>

[Guest guest]

Thanks Aggi. I will do that. I had gone to the lab on Monday and my liver enzymes and kidney function are not doing too well. Liver enzymes are up from taking so many medications... which happens with taking lots of meds. Affects kidneys too, so I don't know what to do... plus I already have Lyme Disease. My situation on medical is that I have two doctors, one doctor that I see for Lyme Disease and co-infections and the other doctor for everything else. I'll call my LLD tomorrow and see what she thinks. I had the same thing happen last year... I have all these antibiotics at home ready to be used, but after my liver and kidney function tests I'm conflicted about whether I should start an antibiotic right now. I don't want to end up with renal failure. But I'll talk to my LLD and see what she says. Thank you for your concern and for mentioning it. Maybe I should start cat's claw anyway (Samento) for the Lyme. The arthritic symptoms of Lyme have been terrible in the last few months.From: "Aggi" <aggi_assmann@...>bird mites Sent: Wednesday, September 14, 2011 11:54:41 AMSubject: Re: Good summary about biofilms

LINDA, please go a see a doctor straight away!! I have just been reading that ticks cause bites with a red ring around after a couple of days. It says when the ring is there, you should urgently go and see a doctor. Have you got flu-like symptoms and/or stomach/guts problems as well ?

> >>>

> >>> 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.

> >>>

> >>>

> >>>

> >>>

> >>>

> >>>

> >>>

> >>>