Biofilm Basics by Bob Henke of The Post Star

One of the most pervasive life forms is called biofilm. A biofilm is most easily understood by thinking of it as simply a slimy colony of bacteria. It turns out very few bacteria are actually free-living; more than 99 percent live in biofilms.

The nature of these biofilms is exceptionally variable. Some are actually moveable, with the whole group oozing about seeking good habitat. Others fasten to some host or strata and remain there, at times virtually impossible to remove. These little bacteria cities may be composed of a single species of bacteria or there may be several different types living together.

Each bacteria in these aggregations exudes a slippery, sugary substance that forms a case over them all, protecting them from attack by other unicellular animals as well as ensuring that any food produced by the group remains with the group. In some of the larger biofilms, the species involved may have complementary metabolisms, that is one species uses as food the materials excreted by others and vice versa.

Biofilms are found everywhere from on sheer rock faces at the top of mountains to plastic surgery implants. The cause of most childhood inner ear infections is a biofilm and biofilm infections ranging from surgical wound infections to heart valve infections kill more than 40,000 Americans annually, more than die from cancer.

The reason they are so troublesome is because most drugs cannot penetrate the biofilm’s tough exterior. The reason for this and the reason biofilms are so intrinsically dangerous is their ability to communicate and thereby keep all the colony residents working toward a single result.

The nature of this communication is at once fascinating and a critical research need. In simplest form, the communication is chemically based. The individual bacteria pump out streams of chemicals which are varied, depending on the message. Other bacteria receive, make note and take appropriate action before duplicating the chemical sonnet and sending it along. In this way they can change direction, change the permeability of the film to let in nutrients or let out waste products or perform a host of other activities.

One of the key chemical phrases researchers are investigating is called a quorum sensor. This is, simply put, how a bacterium knows it is not alone.

The quorum sensing process works like this: A bacteria routinely produces what is called an autoinducer chemical. When the bacteria in turn senses a great concentration of autoinducer chemical returning to it, meaning there are a lot of other bacteria nearby, the gene for biofilm production is turned on and all members of the cohort begin producing the impermeable covering. Inside an animal, this makes the colony invulnerable to attack from, for example, antibiotics given to cure the sinus infection the biofilm is causing.

If researchers can discover the exact chemical signature of this autoinducer, it may be possible to convince the bacteria they are all alone causing them to abandon the biofilm and travel about, making treatment of the malady much easier and straightforward.

This esoteric-seeming research could ultimately save millions of lives. I wish they could also find something that would make me quit communicating long enough to save mine.

Bob Henke may be contacted by mail c/o The Post-Star, email at bobhenke@capital.net, on Twitter at @BobHenke, or on Facebook.

Dr. Dao Nguyen, now at McGill University, trained in the University of Washington lab of Dr. Pradeep Singh, a lung specialist who studies bacterial biofilm infections.

“A chief cause of the resistance of biofilms is that bacteria on the outside of the clusters have the first shot at the nutrients that diffuse in,” said Pradeep Singh, associate professor of medicine and microbiology at the University of Washington.

“This produces starvation of the bacteria inside clusters, and severe resistance to (their) killing,” added the senior study author, the journal Science reports.

“Bacteria become starved when they exhaust nutrient supplies in the (infected) body, or if they live clustered together in groups known as biofilms,” said study co-author Dao Nguyen, assistant professor of medicine at Montreal’s McGill University.

Preventing pathogenic bacteria from sensing nutrient starvation may present a new therapeutic approach to increasing antibiotic efficacy and preventing drug resistance, researchers claim. A team led by McGill University investigators has found that blocking an active mechanism used by bacteria to respond to starvation by slowing their growth significantly reduces the natural tolerance to antibiotics that infectious organisms develop when nutrient supplies become low.

The investigators work is reported in Science in a paper titled “Active Starvation Responses Mediate Antibiotic Tolerance in Biofilms and Nutrient-Limited Bacteria.” Pradeep K. Singh, Ph.D., Dao Nguyen, Ph.D., and colleagues

Abstract:

Bacteria become highly tolerant to antibiotics when nutrients are limited. The inactivity of antibiotic targets caused by starvation-induced growth arrest is thought to be a key mechanism producing tolerance. Here we show that the antibiotic tolerance of nutrient-limited and biofilm Pseudomonas aeruginosa is mediated by active responses to starvation, rather than by the passive effects of growth arrest. The protective mechanism is controlled by the starvation-signaling stringent response (SR), and our experiments link SR-mediated tolerance to reduced levels of oxidant stress in bacterial cells. Furthermore, inactivating this protective mechanism sensitized biofilms by several orders of magnitude to four different classes of antibiotics and markedly enhanced the efficacy of antibiotic treatment in experimental infections.

You can read all of my biofilm posts here

Previous treatments: multiple rounds of antibiotics without eradication of the H. pylori bacteria.

A pre-test for H. pylori stool antigen was performed on February 07, 2011 and was found to be positive. After  44 days on my H. pylori protocol, a follow-up stool antigen test was performed. The results came back negative for H. pylori.

Current Status: GI symptoms are improving and I expect that with the addition of  a few nutritional supplements to support her specific needs, combined with certain dietary restrictions and additions, this patient will be symptom free in short-order.

Stress Made Me Solid and Less Human

When we are under acute or chronic episodes of physical or emotional stress, our body protects itself by shifting the relative balance of our nervous system, to  sympathetic dominance (self-control), thereby rapidly releasing specific stress hormones such as cortisol, adrenaline aka epinephrine and noradrenaline aka norepinephrine.   The long-term effects of the continual release of these hormones is not good at all and will eventually lead to significant degenerative changes within the body.

The stress hormone norepinephrine affects parts of the brain where attention and responding actions are controlled.   Along with epinephrine, norepinephrine also underlies the so-called fight-or-flight response.  During this stress response, heart rate increases, glucose is triggered to be released from energy stores, and blood flow is increased to skeletal muscle.  At the same time blood and energy is drawn away from the gastrointestinal tract and other internal organs.

1.  Norepinephrine is synthesized from dopamine by utilizing the enzyme dopamine β-hydroxylase.

2.  The gene for dopamine β-hydroxylase has shown some association linkages with the gene that controls the ABO blood types.(1)

3.  Norepinephrine and epinephrine possess a synergistic relationship with AI-3, an autoinducer* and may even substitute itself for the AI-3 auto-inducer, resulting in biofilm growth.

4.  The common denominator between type O blood and dopamine appears to be via the null allele (A null allele is a mutant copy of a gene  that completely lacks that gene’s normal function).  In this case the null allele is the type O blood allele in the human A, B and O blood type system – A, B and AB blood don’t possess it.

A hypothesis  could then be made, based on the above data, that type O blood individuals who are highly stressed (over-activated adrenal glands and sympathetic nervous system dominant) may possess a predisposition to infections or overgrowth of yeasts, bacterias and biofilm in the GI tract, since both epinephrine and norepinephrine are present throughout the gastrointestinal tract, and are involved in the stress response.  This may be especially relevant for those type O individuals who possess the ‘Hunter’ epigenotype.(2)

*Bacteria communicate via signaling molecules called auto-inducers, a type of bacteria pheromone.  These autoinducers can initiate or interfere with Quorum Sensing.  One of the two series of auto-inducer molecules are the Auto-Inducers AI-1, AI-2 and AI-3.

These autoinducers are one of the very few biologically active family of molecules that contain the element boron.  Some evidence indicates that grapefruit juice and its furocoumarins inhibits autoinducer signaling and biofilm formation in bacteria.  The most abundant source of furocoumarins in our diet would be grapefruit juice.  The average levels of furocoumarins were lower in the juice from red grapefruit than the white variety, with the highest level of  this component found in the meat of the grapefruit.

(1) AF Wilson, RC Elston, R M Siervogel, and LD Tran. Linkage of a gene regulating dopamine-beta-hydroxylase activity and the ABO blood group locus. Am J Hum Genet. 1988 January; 42(1): 160-166.

How Bacteria “Talk” An eight minute video explaining, in simple English, how bacteria communicate through Quorum Sensing (QS)

Bonnie Bassler is a Howard Hughes Medical Institute Investigator and professor in the Department of Molecular Biology at Princeton University

Bonnie Bassler responded to viewer questions and comments about “talking” bacteria.

Q: Is it possible that even when you make a drug to keep the bacteria from “talking” that the bacteria will work their way around the drug that’s stopping them from communicating, just like they’ve worked around current antibiotics?
Alexandra, 7th grade, Manchester, New Hampshire

A: Hi Alexandra,

Absolutely. We know that bacteria will evolve mechanisms of resistance to anti-quorum-sensing therapies. The hope is that because anti-quorum-sensing strategies do not kill bacteria, only keep them from communicating, this is a less harsh treatment and a less stringent selection for resistance. Because of that, the hope is that resistance will develop more slowly than it does to traditional antibiotics. Thus anti-quorum-sensing therapies may have a “longer shelf life” than traditional antibiotics. Of course, we won’t know if that is indeed the case until we make the drug and monitor its effectiveness.

Q: Do bacteria use quorum sensing for a variety of purposes, e.g., fish form schools for protection, lions form prides to hunt? Also, do the same types of bacteria from different colonies communicate differently than the same types of bacteria from different colonies, i.e., do genetically related bacteria recognize each other aside from their being the same species?
Paula, Washington, D.C.

A: Dear Paula,

Quorum sensing is used to control hundreds of different group processes. In the cases we understand best, the processes controlled are ineffective when carried out by individual bacteria acting alone but become effective when carried out in synchrony by the group. Virulence is a good example of a quorum-sensing-controlled behavior. A few bacteria can’t make us sick. Even if they release toxins or other harmful agents, we are HUGE compared to them, so each bacterium’s measly few molecules of a toxin can’t harm us. But, if the bacteria wait, count themselves with quorum-sensing molecules, and then all of the bacteria release their toxins simultaneously, they can overcome an enormous host. There are all kinds of examples of quorum-sensing-controlled group behaviors: bioluminescence, virulence, exchange of DNA, biofilm formation, symbiosis….

Bacteria have multiple chemical languages. We think each bacterial species has a molecule that is unique and represents its own species-specific language. Each unique molecule allows a particular species to know and communicate with its relatives (i.e., have a private conversation). We know there is a universal molecule, a sort of bacterial trade language that bacteria use to converse between species (i.e., a Bacterial Esperanto). There is mounting evidence that many additional molecules remain to be discovered, such as species non-specific molecules that say “who” the neighbor is. This field is only about 10 years old. So far we have only managed to identify a few molecules. We know that bacteria interpret a rich and complex chemical world, and we are working hard to define the complexity of the lexicon.

Q: Can you provide some thoughts on individuals with knee or hip implants and the biofilm some unfortunately develop and if there is hope for probiotics in this issue?
[first name not given] Raunaque, LaCrosse, Wisconsin

A: Dear Mr. or Ms. Raunaque,

Biofilms are communities of bacteria attached to surfaces (for example, as described in the NOVA piece, your teeth in the morning). Biofilms are a cause of constant concern for people with medical implants, heart valves, etc., because these medical devices provide a niche for bacteria to adhere to and thus cause infection. We know that quorum sensing is required for biofilm formation: If the bacteria can’t talk, they can’t make these large, interacting, adherent communities. The goal of many researchers’ studies is to find methods to interfere with biofilm formation, and, of course, a key focus of study is on interrupting quorum sensing. One strategy is to make molecules that antagonize the natural quorum-sensing molecules to impede biofilm formation. Another strategy is the one you suggest, to develop probiotic quorum-sensing therapies that enhance the conversation among our commensal bacteria at the expense of the biofilm formers. Both these avenues are currently being explored.

microparticles "microbubbles" used for detection of bacterial biofilm

Certain types of bacteria, such as: Borrelia burgdorferi, Escherichia coli, Candida albicans, Clostridium difficile, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, and Vibrio cholerae, can join forces to form protective communities called biofilms.  These thin layers of bacteria, which grow on the surfaces of medical implants or directly on tissue in the body, can be difficult to treat because they are more resistant to drugs than the bacteria on their own.  Currently there is no established way to image biofilms in or out of the body.

Pavlos Anastasiadis and colleagues at the University of Hawaii at Manoa have developed a method to watch and measure growing biofilms with ultrasound.  The researchers used contrast agents, microparticles, more accurately, microbubbles, that are normally injected into the body to improve the quality of ultrasound images.  They modified the surface of bubbles in the agents to stick to two kinds of infectious bacteria that form biofilms (Staphylococcus aureus and Pseudomonas aeruginosa).   Acoustic pulses of ultrasound cause the bubbles to “ring” like a bell, revealing their location and the attached biofilm.

The research was done on isolated biofilms.  The next step will be to test it in living tissue.  Anastasiadis hopes to develop the technique to diagnose infective endocarditis, a disease in which bacterial biofilms form on the inner walls of damaged heart valves.

“Targeted ultrasound contrast agents for the imaging of biofilm infections” by Pavlos Anastasiadis  – Abstract: http://asa.aip.org/web2/asa/abstracts/search.may09/asa323.html

The discovery that bacteria are able to communicate with each other changed our general perception of many single, simple organisms inhabiting our world. Instead of language, bacteria use signaling molecules which are released into the environment. As well as releasing the signaling molecules, bacteria are also able to measure the number (concentration) of the molecules within a population. Nowadays we use the term ‘Quorum Sensing’ (QS) to describe the phenomenon whereby the accumulation of signaling molecules enable a single cell to sense the number of bacteria (cell density). In the natural environment, there are many different bacteria living together which use various classes of signaling molecules. As they employ different languages they cannot necessarily talk to all other bacteria. Today, several quorum sensing systems are intensively studied in various organisms such as marine bacteria and several pathogenic bacteria.

Quorum Sensing & Biofilm Formation

Why do bacteria talk to each other?

(QS) enables bacteria to co-ordinate their behavior. As environmental conditions often change rapidly, bacteria need to respond quickly in order to survive. These responses include adaptation to availability of nutrients, defense against other microorganisms (biofilm formation) which may compete for the same nutrients and the avoidance of toxic compounds (biofilm formation) potentially dangerous for the bacteria. It is very important for pathogenic bacteria during infection of a host (e.g. humans, other animals or plants) to co-ordinate their virulence in order to escape the immune response of the host in order to be able to establish a successful infection. The University of Nottingham Quorum Sensing Research Group

From Dr. Ettinger’s Biofilm Protocol for Lyme and Gut Pathogens: Pathogenic bacteria known to reside in biofilms include: Borrelia burgdorferi, Escherichia coli, Candida albicans, Clostridium difficile, Clostridium perfringens, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, and Vibrio cholerae. The number of human diseases shown to be associated with biofilms is expanding and includes chronic bacterial prostatitis, chronic rhinosinusitis, cystic fibrosis pneumonia, infective endocarditis, periodontitis, recurrent otitis media, and virtually all device and implant related infections. Strong evidence is also beginning to emerge for an etiologic role of pathogenic mucosal biofilms in gastrointestinal diseases, such as Irritable Bowel Disorders: Crohn’s disease and ulcerative colitis.

Biofilm Formation

A Brief history of “biofilm”
Center for Biofilm Engineering
Montana State University

Microbial communities attached to surfaces (biofilms) were observed long before people had the tools to study them in detail. In 1684 Antony van Leeuwenhoek remarked on the vast accumulation of microorganisms in dental plaque in a report to the Royal Society of London: “The number of these animalcules in the scurf of a man’s teeth are so many that I believe they exceed the number of men in a kingdom.”

The study of microbes took an important turn in the mid-1800s, when Robert Koch developed methods to create a solid nutrient medium in order to grow and isolate pure cultures of microorganisms. This development led to huge advances in medicine, agriculture, and industry. However, these advances were based on such a simplistic concept of microbial life that many ‘solutions’ generated by these techniques are now being reversed. Microorganisms have proved to be much more complex and less tractable than we ever imagined.

In a 1940 issue of the Journal of Bacteriology, authors H. Heukelekian and A. Heller wrote, “Surfaces enable bacteria to develop in substrates otherwise too dilute for growth. Development takes place either as bacterial slime or colonial growth attached to surfaces.” Claude ZoBell described many of the fundamental characteristics of attached microbial communities in the 1940s. In the late decades of the 20th century, numerous articles were written about microbial films or slime layers; German researchers sometimes used the term “Schmutzdecke.” As the unique properties of microbial communities vs planktonic microbes grew more apparent, it became helpful to use a special term to describe them. “Biofilm” was used colloquially among researchers for some years before it was considered a term acceptable for use in publication. The earliest use of “biofilm” in publication is in the Swedish journal Vatten: Harremoës, P. 1977. “Half-order reactions in biofilm and filter kinetics,” Vatten, 33 122-143. (If you know of an earlier publication with “biofilm” in it, please let us know; we would be happy to make a correction.)

Early biofilm researchers studied the implications of biofilms in waste-water filtration, biofouling of industrial equipment, and dental plaque (Leewenhoek would have been pleased). Since bacteria preferentially attach to surfaces, biofilms are virtually ubiquitous. Biofilm formation is also implicated in microbiologically influenced corrosion (MIC), product contamination, medical device-related infections, and chronic wounds. Biofilm can also be used for positive effects, especially in water pretreatment systems and contaminated soils.

In 1990, recognizing the significance of microbial activity, as well as the tremendous economic costs associated with microbial communities on surfaces, the US National Science Foundation founded the Center for Biofilm Engineering at Montana State University in Bozeman (though, interestingly, NSF would not initially accept the word “biofilm” in the Center’s name; instead the award funded the “Center for Interfacial Microbial Process Engineering”). Since that time, the field of biofilm research has exploded. New tools and techniques are continually pioneered to help understand the secrets of microbial community interactions. In addition to numerous research laboratories in the US, several groups study biofilms worldwide, including centers in Denmark, England, Germany, Australia, and Singapore.

What is a biofilm?

Most of you have never heard of the term “biofilm”, but you have certainly encountered “biofilm” on a routine basis. If you’ve ever been to the dentist and he’s scraped “plaque”, which causes tooth decay, off your teeth; that’s a type of bacterial biofilm. The “slim” that clogs your drains is also biofilm. The slippery coating on rocks, at the water’s edge of a stream or river, is just a  bacterial biofilm-coating. Pond-scum – a biofilm. If you’ve ever been diagnosed with Candida albicans; H. pylori; chronic sinus or prostate infection; or Lyme disease, chances are they’re living, hiding and replicating in a biofilm colony.

Iodine staining of biofilm plaque (upper right)

This is the best product for removing the bacterial biofilm that causes plaque – Biotene PBF Chewing Gum.

These microorganisms (biofilm colonies) are usually encased in an extracellular polysaccharide that they themselves synthesize, via the release of signaling molecules through quorum sensing (QS). This glue-like substance allows them to anchor to all kinds of surfaces – such as metals, plastics, soil particles, medical implant materials, and tissue. As long as sufficient moisture and nutrients are available, a bacterial biofilm can form just about anywhere. In your body that would be from your mouth, especially the teeth, through the stomach and GI tract, all the way down to the rectum. Biofilm in the environment can be found, most often, in ponds, streams, rivers, etc.  A biofilm can be formed by a single bacterial species, but more often than not, biofilms consist of many species of bacteria, as well as fungi/yeast, algae, protozoa, debris and corrosion products. Once anchored to a surface, biofilm microorganisms carry out a variety of detrimental or beneficial reactions, depending on the surrounding environmental or body conditions.

In the human body, biofilm colonies are the main reason that certain conditions take so long to get handled. In my opinion, if it were not for “biofilm”, conditions caused by the microorganisms – Candida albicans, Candida spp,  H. pylori, Lyme’s bacteria (Borrelia burgdorferi) and many others, would be far easier to diagnose and/or treat. It is crucial in any treatment protocol to first handle the biofilm.  By doing so, it will make a significant difference in the amount of time, money and effort spent on treating many, so called, stubborn condition – like the above.

Related Posts: Biofilm Protocol, Quorum Sensing, Lactonase

Biofilm Research and Links/Resources

THE ROLE OF EXTRACELLULAR DNA IN MAINTENANCE OF BIOFILMS FORMED BY E. COLI, H. INFLUENZAE, K. PNEUMONIAE, P. AERUGINOSA, S. AUREUS, S. PYOGENES AND A. BAUMANNII George V. Tetz & Victor V. Tetz Dept. of Microbiology, Virology and Immunology; Saint-Petersburg State Pavlov Medical University, Russia Email: vtetzv@yahoo.com

It is known that bacteria within biofilms are much less susceptible to antibiotics particularly because of poor antimicrobial penetration through surface film that covers microbial community and inactivating role of extracellular matrix. Combined effects of DNase (Enzyme for digesting single and double-stranded DNA) and antibiotics on established biofilms of different unrelated bacteria were displayed. A Combination of antibiotics with DNase I resulted in significant decrease of established biofilm biomass compared to the reduction of biomass achieved when antibiotics or DNase I were used alone.

DETECTION OF HELICOBACTER PYLORI IN BIOFILMS BY USING REAL-TIME POLYMERASE CHAIN REACTION (PCR) Linke, S., Gebel, J., Büttgen, S., Exner, M. Institute for Hygiene and Public Health, University of Bonn

Our results confirmed a possible existence of H. pylori in drinking-water biofilms.

ANALYSIS AND IDENTIFICATION OF THE BIOFILM WOUND MICROFLORA IN HORSE WOUNDS Samantha J. Westgate1, Steven L Percival2*, Derek C. Knottenbelt1 and Christine A. Cochrane1 1University of Liverpool, Department of Veterinary Clinical Science, Division of Equine Studies, Leahurst, Neston, South Wirral, UK *2ConvaTec Wound Therapeutics, Deeside, Flintshire CH5 2NU, UK

Equine wound healing is notoriously problematic on the lower limb, specifically when biofilms are evident. Equine chronic wounds display similar characteristics to chronic wounds in humans thus these cases provide an effective model for human cases. Whether wounds are caused by trauma or surgery their high prevalence is of concern and treatment can be both challenging and costly. Biofilms are considered detrimental to normal healing in non-healing and infected chronic wounds because of their recalcitrant nature towards antimicrobial agents. Biofilms are also known to be resistant to the effects of the immune system. Because of this fact more research in the area of chronic wounds and biofilms is warranted.

Culturable analysis of the microflora revealed that the majority of bacteria isolated from the chronic wounds of horses were Staphylococcus spp, Pseudomonas spp, Micrococcus spp, Enterococcus spp, Corynebacterium spp, Streptococcus spp, Bacillus spp, Aerococcus spp and Clostridium spp. Further analysis of all isolates highlighted their biofilm forming potential and antibiotic resistance profiles. Biofilms were shown to be evident in a large percentage of the chronic wounds. In conclusion these studies provide evidence that biofilms exist in the chronic wounds of horse which may well provide an underlying reason as to why a large percentage of chronic wounds are recalcitrant to antimicrobial therapies, do not heal a timely manner and often become infected.

BACTERIAL BIOFILMS IN SURGICAL SPECIMENS OF PATIENTS WITH CHRONIC RHINOSINUSITIS (sinusitis).
Sanclement JA, Webster P, Thomas J, Ramadan HH. Department of Otolaryngology, West Virginia University, Morgantown, West Virginia 26506-9200, USA.

CONCLUSIONS: Biofilms were demonstrated to be present in 80% the 30 patients undergoing surgery for chronic rhinosinusitis (CRS); none of the (control) patients without CRS had any evidence of biofilms.

Note: According to Andrew Foreman, B.M.B.S., Ph.D., and colleagues from the University of Adelaide in Australia, the most common bacteria reeking havoc in those experiencing CRS is Staphylococcus aureus (S. aureus).

First a little background on biofilm:

Fig. 1: The biofilm life cycle. 1: individual cells populate the surface. 2: extracellular polymeric substance (EPS) is produced and attachment becomes irreversible. 3 & 4: biofilm architecture develops and matures. 5: single cells are released from the biofilm. Related Post – Biofilm Basics and Quorum Sensing and Biofilm

This is an excerpt from a Klaire Labs product monograph which is a basic primer on the topic (My additions are in RED) The National Institutes of Health estimates that 60% of all human infections and 80% of refractory infections (def. unresponsive to medical treatment) are attributable to biofilm colonies.  I have seen this, most commonly, in cases I’ve worked-up, where the pathogen is: Chlamydia pneumoniae, Pseudomonas aeruginosa, Helicobacter pylori, [Lyme disease - Borrelia burgdorferi] and Candida albicans.

• The protection conferred upon microorganisms by biofilm allows them to achieve a high level of antibiotic resistance, stealth and invisibility.

• Biofilm not only provide a physical barrier to antimicrobial agents (pharmaceutical antibiotics) and host antibodies, but facilitate the exchange of antibiotic-resistant genetic material between organisms and may contain antibiotic-degrading (hydrolysing) enzymes such as b-lactamase, effectively neutralizing incoming antibiotic (b-lactam antibiotics) molecules.

• In fact, biofilm communities can be 1000 times more resistant to antibiotics than free-floating bacteria.

• The decreased growth rate of sessile microorganisms (def. Permanently attached to a substrate; not free to move about; “an attached oyster”) also reduces their antibiotic susceptibility as most antimicrobial agents require rapid cell growth in order to effectively kill or inhibit the microbes.  Biofilm thus render pathogenic microorganisms enormously difficult to eradicate, and can almost single-handedly contribute to localized or systemic inflammatory reactions and delayed wound healing.

• Depending on the type of biofilm, one or more species of pathogens may be found embedded in the extracellular polymeric substance (def. Composed primarily of polysaccharides and can either stay attached to the cell’s outer surface, or be secreted into its growth medium).  Bacterial extracellular polymeric substance (EPS) maybe a carrier of, or may have heavy metals embedded in them, thus the indication for chelation w/EDTA. EDTA, ethylenediaminetetraacetic acid, is a chelating agent used to lower one’s body burden of heavy metals).

Pathogenic bacterial known to reside in biofilms include, but are not limited to: Borrelia burgdorferi (Lyme bacteria), Escherichia coli, Candida albicans (yeast and fungal mutation), Clostridium difficile, Clostridium perfringens, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, and Vibrio cholerae. The number of human diseases shown to be associated with biofilms is ever expanding and includes: chronic bacterial prostatitis, chronic rhinosinusitis (chronic sinus infections), cystic fibrosis pneumonia, infective endocarditis, periodontitis, recurrent otitis media, and virtually all device and implant related infections.  Strong evidence is also beginning to emerge for an etiologic (causative) role of pathogenic mucosal biofilm in gastrointestinal diseases, such as Irritable Bowel Disorders (IBS): Crohn’s disease and ulcerative colitis.

S. aureus biofilm

Dr. Marcus Ettinger’s Biofilm Protocol – Only the eradication phase is presented here.  There is a pre,  post and toxin reduction step as well.  You can get help with any of these steps – HERE

A. Products (mandatory products in red). These are ONLY the basics. Additional nutraceuticals may be needed, based on each individuals unique situation.

1. Monolaurin [AKA Glyceryl laurate or glycerol monolaurate] (monolaurin information) or Lauricidin
2. Nutiva Extra-Virgin Coconut Oil (42-52% Medium Chain Fatty Acids [MCFA], lauric acid, by volume)
3. Nattokinase (a potent fibrinolytic enzyme) I like this better than Lumbrokinase.
4. InterFase Plus™ (broad-spectrum enzyme formula w/EDTA)
5. Serrapeptase (a potent fibrinolytic enzyme)
6. Vitamin C (ascorbic acid – Not buffered, as most of these contain metals)
7. NAC (N-Acetyl-Cysteine)
8. Lactoferrin (specifically Nutricillin by Ecological Formulas) Dr. Anju Usman of Illinois states, “Our bodies make proteins, transferrin and lactoferrin, which mop up iron and block the ability of biofilm to form,” she said. “But pathogenic bacteria secrete iron chelators to snatch up iron and thus compete with the transferrin and lactoferrin for what they need to survive.”

B. Avoid supplemental forms of: magnesium, iron and calcium during the biofilm protocol, as they may contribute to  biofilm formation or decrease the effectiveness of the biofilm protocol.

C. Take a broad-spectrum probiotic and prebiotic.  I like the combination of Now Foods brand Probiotic-10 and their Probiotic Defense Powder (contains gluten).  These products will help to crowd out the bad bacteria, and also help disrupt and replace biofilm colonies along the mucus membrane.

D. Specific additions based on condition (not a complete list):

1. Candida albicans – SF722* (10-Undecenoic Acid  50 mg) Thorne Research. This is as close as you can get to a medication and still be a natural substance.  There are a few chat rooms blasting this product, based on who knows what – can’t make everyone happy.  I’ve used SF722 for over 15 years and it is amazing – never a problem!  *Do not take SF722 if you are allergic to fish. There are many other amazing products that can be added to complement the SF722. It’s really a matter of how many pills someone wants/doesn’t want to take per day or the severity of one’s condition, that will determine, if or which, additional products will be added. If the Candida albicans overgrowth is severe, has not responded to holistic methods or has mutated into its more virulent hyphal form/fungal infection (nails, underarms, groin or skin); Diflucan (fluconazole), a prescription medication, is my personal preference, but Nizarol (ketoconazol) can also be used. In Azole-resistant Candida albicans, lactoferrin must be added to either medication in order to increase their effectiveness. There is a certain B vitamin, mineral and amino acid that possesses synergistic qualities and I find them indispensable when taking Diflucan (fluconazole), Nizarol (ketoconazole) or for supporting candida die-off symptoms.
2. Chlamydia pneumonia, Klebsiella pneumoniae or Pseudomonas aeruginosa – Pneumotrophin PMG by Standard Process, Inc. How it works.  I use this because it helps direct the body’s attention and healing efforts to the lung, where it’s needed most. Apex Energetics, H-PLR is also a mandatory addition. I also like to use OOrganik-15™ and Pneuma-Zyme™ by Biotics Research with some of my patients who manifest asthma, a chronic cough and/or emphysema like symptoms.     
3. H. pylori – Complete write-up on another post.
4. Chronic bacterial prostatitis – Quercitin (600mg’s) and Bromelain (200mg’s) combination by Now Foods. Decreases inflammation and oxidant stress in the prostate while increasing local concentrations of beta-endorphins. Apex Energetics, H-PLR is also a mandatory addition.

E. Certain dietary restrictions and additions will need to be taken. These are determined on a case by case basis.

Important Note:

All dosages will be provided if you purchase some or all of your “biofilm protocol” products through my office. I truly do want to help all who are interested, but it’s finally gotten to the point where too many people want free advice and an increasing amount of my time, and then buy all of their products elsewhere. I am a firm believer in fair exchange and I feel I have done that by providing the information in this post.

I also offer tailor made protocols for your individual situation, please contact our office for product prices and distance patient information (714) 639-4360. 

 

 

 

Biofilm testing is also available through Fry Laboratories. Fry Laboratories, L.L.C. is an independent clinical diagnostic and research laboratory located in Scottsdale, Arizona. We are committed to understanding chronic diseases and contributing to their cure through advancements in diagnostics and basic science research with emphasis on chronic inflammatory diseases, vector-borne diseases, and their intersection. Our clinical diagnostic laboratory offers general and targeted immunology services in conjunction with standard and cutting edge infectious disease detection and identification technologies. Our signature services include microscopy for visual identification and quantification of a wide range of blood-borne pathogens, co-infection serology, biofilm detection, and genus wide molecular detection technology with sequencing for individualized species and/or strain identification. We participate in both CAP and API quality control programs and provide worldwide testing service.

Diseases of Interest: Chronic Fatigue Syndrome, Fibromyalgia, Gulf War Veterans Illness, Chronic Lyme Disease, ALS (Lou Gehrig’s Disease), Parkinson’s Disease, Multiple Sclerosis, Autism, Lupus, Ulcerative Colitis, Scleroderma, Rheumatoid Arthritis, Osteoarthritis, Crohn’s Disease.

Infections of Interest: Borrelia (Lyme), Babesia, Bartonella, Anaplasma, Ehrlichia, Q-Fever (Coxiella), Toxoplasma, Rickettsia, Plasmodium, XMRV

Important: This post is not a substitute for medical advise or treatment and is for informational purposes only. Please consult with a physician before starting any nutritional or biofilm protocol on your own.