Biofilms – Revised September 2015

Biofilms: A Hideout for Borrelia burgdorferi.

The following article came from the research conducted (and later published at ) by Eva Sapi, PhD and her research team.  Dr Sapi wrote this article years earlier in Lyme Times Magazine in Summer 2007 (requires a paid subscription) CALDA is now located at

The University of New Haven established a Lyme Disease Research group in 2004. To date, over forty graduate students have received training in Lyme disease related research. One of our recent projects studies the different formations of Borrelia burgdorferi, the Lyme disease bacteria. With coauthor Dr. Alan MacDonald, we recently suggested that Borrelia burgdorferi is capable of forming an organized structure called biofilm. Our proposal is based on recently published and several unpublished images of Borrelia. 

What is biofilm?

“Biofilm is a self-made protective environment for microbial populations in which they adhere to each other or to living or inert surfaces”. In the biofilm, single or multiple types of organisms can surround themselves with a complex matrix, better known as “slime”. The main purpose of the biofilm structure is to allow microbes to survive various environmental stresses, including the presence of attacking immune cells like phagocytes, or antibacterial agents. While conventional antibiotic therapy is usually effective against free-floating bacteria, it is frequently ineffective once pathogens have formed biofilms, because biofilm colonies can be up to 1,000-times more resistant to antibiotics.

Furthermore, even if a biofilm-related infection appears to respond to antibiotics, it could relapse weeks or even months later and turn into a very difficult to treat chronic infection. The National Institutes of Health estimate that nearly 80 percent of chronic microbial infections in the human body are due to biofilms, such as chronic lung infection in cystic fibrosis patients, catheter infections, chronic urinary and middle ear infections, gingivitis, sinusitis and even fatal endocarditis.

What do we know about biofilm? Biofilms start as just a few microorganisms adhering to each other or to a surface, and then begin to communicate3-4. This communication will initiate a change in gene expression and cells start to produce an exopolysaccharide, which will become the protective matrix 3.The colonies then can develop into complex, three dimensional structures housing millions of individual microbes. Like cities, they have towers, columns, bridges and channels for the flow of nutrients. A mature biofilm is usually composed of three layers: an inside film layer that binds the biofilm to the surface; another film made up of colonies of single or multiple species of bacterial and/or fungal organisms; and the surface film from which free-floating microorganisms can be released as individual cells that can colonizing other places.

So what are the mechanisms by which bacteria can evade the therapeutic interventions in biofilm? The first studies suggested that the bacteria deep within the biofilm live in an environment where diffusion of antibiotics might be difficult. There are major differences in the chemical composition of the biofilm such as low pH and anaerobic condition etc, which can either inactivate the antibiotics or render bacteria inactive so the antibiotics cannot kill them.

However, recent publications suggested that the main reason for antibiotics resistance is the changes in the gene expression profile of microbial cells harboring in biofilms. For example, researchers identified mutant bacteria that are capable of forming biofilm but are not resistant to antibiotics. The differential expression of a large number of genes is known to occur in the initial steps of biofilm formation, such as the up-regulation of exopolysaccharide synthesis.

So how about the immune system? Why cant they recognize and destroy the biofilm? Multiple studies demonstrate that phagocytes can be found attached to biofilm but they are not able to eliminate it. To answer this very puzzling question German scientists used marine bacteria as a model and studied the defensive mechanism against their environmental enemy, which is a phagocyte (called amoebae). They identified that this marine bacterial biofilm can release a paralyzing agent that deactivates and even kills the amoeba. So clearly biofilm is not just a defensive fortress, it can also fight back.

So how about chronic Lyme disease? Can it be explained by a biofilm formation of Borrelia burgdorferi? If yes, the possibility that Borrelia burgdorferi is capable of forming a biofilm can change the way we think about Lyme disease, especially in patients where it seems to be a persistent disease, despite long term antibiotic treatment. The elucidation of the molecular mechanisms responsible for the switch from free-living growth to a biofilm phenotype, with the development of antibiotic resistance, should provide novel therapeutic targets in chronic Lyme disease.

Despite the potential importance of this hypothesis, to date there has been no studies attempted to determine whether Borrelia burgdorferi is indeed capable of biofilm formation and whether such a formation results in increased antibiotic resistance. Borrelia burgdorferi sensitivity to antimicrobial agents has traditionally been studied in the free-living state. Conclusions drawn from many of these studies, therefore, need to be re-validated.

We have recently established an in vitro model to study biofilm formation of Borrelia burgdorferi and proposed to use this system to evaluate the antimicrobial sensitivity of Borrelia burgdorferi in biofilm. Our goal is to test several known antibiotic agents frequently used in the Lyme disease treatment, as well as several natural agents, for the ability to interfere with or destroy biofilm production. This project recently received a generous support grant from the Turn the Corner Foundation (now Lyme Research Alliance.)

As mentioned above, the formation of a biofilm begins with the attachment of free-floating microorganisms to each other or to a surface. In the case of Borrelia burgdorferi, our preliminary results show that Borrelia burgdorferi is capable of forming biofilm.

We have monitored several environmental conditions for the initiation of biofilm structure and found that that cell density is one of the most important factors. When Borrelia burgdorferi reach a certain density, they started to stick to each other and start to form an organized structure.

Our working hypothesis for this finding is that nutrient depletion is the main stress factor for the initiation of biofilm. It was previously demonstrated that organisms within biofilms could withstand nutrient deprivation better than free-floating counterparts. Furthermore, as we presented at the 2007 ILADS conference, we have seen similar changes after exposure of Borrelia burgdorferi to penicillin. In the penicillin treated samples, as early as 24 hours, we observed formation of a granular/cystic form covered by a biofilm-like substance.

In our next set of experiments we will test other stressors that can initiate Borrelia burgdorferi biofilm formation, including different temperatures, pH, oxidative radicals, heavy metals and of course several synthetic and natural antibacterial agents. Our final goal is to identify antibacterial agents that are effective in killing Borrelia burgdorferi without inducing biofilm, or even capable of destroying Borrelia burgdorferi in biofilm.

In summary, if we can demonstrate that biofilm structure of Borrelia burgdorferi renders them resistant to antibiotics, it could provide a logical explanation as to why extensive antibiotic treatment for patients with a tick-bite history could fail. The end result from our study could provide novel therapeutic approaches for Lyme literate physicians to explore for chronically ill patients.

Watch these fascinating videos that support and expound on Dr. MacDonald and Dr. Sapi’s article:

Dr. Bill Costerton – The “Father” of Biofims   –   and (not about Lyme specifically)  Doctor “Bill” Costerton has studied biofilms for more than 40 years. His biofilm research and prolific publishing through the decades covers many fields: medical microbiology, microbial ecology, industrial microbiology and bio-remediation. For many, he’s a hero for revealing a profound truism: bacteria (et al) infect two ways, in their planktonic or by biofilm state.His research led to many medical breakthroughs, confirming that biofilms cause myriad chronic conditions: middle ear infections, kidney stones, gum disease and unfortunately many more that affect millions of people. Yet, his work also led to beneficial treatments of sewerage, industrial waste and polluted soil.                 

Watch: “Biofilm Discovery in Lyme Disease with Dr. Eva Sapi –

Dr. Alan MacDonald – educated at Columbia University and mentored by Jorge Benach, PhD (Stony Brook University). Dr. MacDonald, a pathologist, is considered the man who first discovered the role of biofilms in chronic Lyme disease with Sr. Eva Sapi. 
For the past 30 years Dr. Alan MacDonald has worked to revive the Model of Syphilis and draw attention to clinical and laboratory parallels between Treponema pallidum infection and Lyme Borreliosis. Dr. MacDonald hypothesized that Alzheimer’s disease might be the late neural borreliosis equivalent of General paresis of the insane. He further hypothesized that syphilitic Tabes Dorsalis might have a “spinal cord only” neurodegenerative equivalent in Borreliosis, namely Amyotrophic Lateral Sclerosis (Lou Gehrig’s Disease). He continues to do ground-breaking work in this field. DONATE to Dr. MacDonald NOW at
He has gone far in proving that syphilitic Temporal arteritis has a Borrelia equivalent in Temporal arteritis (previously of of unknown cause). In addition to the ongoing Alzheimer’s studies, which has occuppied much of his most recent research, Alan has made many new observations in pathobiology with respect to the many faces of Lyme disease. He was the first to publish evidence for a cystic form of Bb, granular forms of Bb, and cell wall deficient forms. Although officially retired now, Dr. MacDonald has started a research collaboration with Dr. Eva Sapi of University of New Haven.
 Watch now: “The Biology of Lyme Disease: An Expert’s Perspective”
Dr. Eva Sapi – Dr. Eva Sapi, professor of cellular and molecular biology and director of the Lyme Disease Research Group at the University of New Haven (UNH), continues to be awarded each year with various grants to continue this exciting research to help complete the final steps in this project that could lead to significant improvements in the diagnosis and treatment of Lyme disease.As seen, previous research by Dr. Sapi and UNH’s Lyme Disease Research Group has demonstrated that the bacteria that causes the disease, Borrelia burgdorferi, is capable of resisting antibiotic treatment by “hiding” in a self-made protective layer called biofilm. 
This discovery could explain why some people infected with Lyme disease continue to have symptoms even after treatment with traditional antibiotic therapy.According to a report previously released by Sapi and the UNH researchers, “While conventional antibiotic therapy is usually effective against free-floating bacteria, it is frequently ineffective once pathogens have formed biofilms, because biofilm colonies can be up to 1,000 times more resistant to antibiotics.”
This latest project is aimed at understanding the molecular mechanisms taking place during biofilm development, in order to be better able to prevent and destroy it. Without the protection of the biofilm, the hope is that diagnostic tests and antibiotic treatments for Lyme disease can be made much more effective.
This video is a 10 minute clip, part of a 70 minute interview with Dr. Sapi from the University of New Haven.
Her article in 2010 published in Townsend Letter caused a big stir, and gave hope to many for an alternative method of fighting Borrelia and destroying biofilms using Samento and Banderol.  The article has numerous slides to show her research.  Read at
She is credited with being the first researcher to demonstrate that Lyme spirochetes can actually create their own complex biofilm community to survive indefinitely within their hosts; both human and animal.
Watch “Biofilm Discovery in Lyme Disease”:
Dr. Vince Fischetti studies how gram-positive bacteria interact with human tissues and cause disease. His lab produces lysins that kill major gram-positive pathogens aE” Streptococcus pyogenes, Streptococcus pneumoniae, Staphylococcus aureus, Enterococcus faecalis and Bacillus anthracis. Using phage lytic enzymes as a tool, Dr. Fischettis lab has developed and patented methods of eradicating biofilms which contribute to chronic bacterial diseases –
Watch Dr. Costerton and Dr. Wolcott:
Dr. Randy Wolcott has been practicing medicine for almost thirty years and focusing on treating “unhealable” wounds the last twelve. His personal research at the Wound Care Center’s Research and Testing Laboratories and his collaboration with biofilm experts in the wound care field have earned him international recognition.
Recent: Dr. Randy Wolcott on Molecular Validation of Biofilm Infection:
Dr. Scot Dowd, PhD – work experience with the USDA, diagnostics, bioinformatics and wound care has led to some remarkable discoveries. This knowledge is practiced daily within PathoGenius, his new company, and seems to help patients with chronic bacterial infections get properly diagnosed and treated.
Dr. Ehrlich fields questions from a range of patient-centric topics: what has been learned collectively in 20 years of biofilm-specific research, the bacteria-biofilm life cycle, how patients with prosthetics could be at higher risk for biofilm-related health conditions and more.
Dr. Marty Ross – Biofilms are protective communities where various forms of the Lyme bacteria can hide from prescriptive antibiotics, herbal antimicrobials, and the immune system. In chronic Lyme and associated diseases they can cause treatment resistance and relapse once antibiotics or herbal anti-microbials are stopped.
In chronic Lyme and associated diseases, Banderol and Samento extract (homeopathy) have proven very effective for killing spirochetes and breaking down the biofilms ( See   Also,  Lumbrokinase breaks apart biofilms, improves blood flow to the tissues by decreasing sticky blood (hypercoagulation), decreases crampy muscle pain, and improves supplement or prescription medicine delivery deep into the tissues.
Future of Biofilm Research: Dr. Bonnie Bassler on “Quorum Sensing” Bacteria Communicating in a cell and/or in a Biofilm?


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4 Responses to Biofilms

  1. Lesli says:

    Hi Jenna –

    Do you have a direct link for this article? The provided link just goes to the main page. When dealing with trying to educate a reluctant Dr. you need original sources. 🙂


  2. Pingback: New Biofilm Busting Protocols For Chronic Lyme Disease | Jenna's Lyme Blog

  3. David Slade says:

    Hi Jenna!

    Great article on Lyme Disease, few touch on the subject of the biofilms which bacterial organisms are known to secrete. I would love for you to check out my website on Lyme Disease. I myself recovered from Lyme Disease and had a large exposure to deer ticks in my teenager years. I’ve was permanently disabled for 7 years and wheel chair bound. I healed myself through a nontoxic nutritional approach that involved herbal tea remedies and homeopathic countermeasures. I am now 100% symptom free and serve my community as a holistic health coach who helps others recovery from their chronic health problems and Lyme diseases. Take a look at my website when you get a chance I would love to get your thoughts on it. — I offer proprietary herbal teas which are the same blends that I used to rid myself of Lyme Disease –

    – David

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