Hello to the forum,

Six years ago,a a Symposium at the University of New Haven I delivered a lecture on biofilms of Borrelia burgdorferi (Bb). The PDF File of the PPT and its Video have been posted on the Web and are freely downloadable.

Executive summary:

Spiral forms of borrelia burgdorferi (motile, corkscrew shaped) constitute the Planktonic forms of Bb.

In a biofilm community, the Borrelia undergo specialization, as in all biofilm communities. Their Extracellular matrix investment is derived from sloughed Outer Surface Membrane (glycolipids) DNA (extracellular DNA eDNA — like pseudomonas species ECMs), Microvesicles (blebs) and alginate-like-material with some calcium deposits. The specialized biofilm members include small, dot-like granular forms of Bb, Cystic and CWD Bb forms and the traditional spiral forms.

Our research leader is Eva Sapi, PhD, Research Scholar and Associate Professor of Biology and Environmental Science, University of New Haven, West Haven, Conn. Dr Sapi was the FIRST to suggest that Borrelia might exist as biofilm communities.

In the Image gallery which will accompany the Multiauthored PLOS ONE paper on In Vitro Biofilms of Bb, are many exceptional images of Biofilm communities of Bb including the very first use of Atomic Force Microscopy (AFM) to image spirochetal biofilms.

Our Research was buttressed by previous accepted manuscripts describing biofilm formation in Leptopsires (Institut Pasteur) and Biofilm formation by Oral Treponemes (Forsythe Dental Research Center, Harvard Univ).

Floating biofilms, as noted for the leptospira species by the Team at the Institut Pasteur, was a finding in some leptospira genera. Fixed (attached) Biofilms were also described for some genera of Leptospires. Oral Treponemes, to my knowledge do not form FLOATING biofilm communities.

Borrelia burgdorferi does indeed Form Floating Biofilm communities and Fixed sessile Biofilm communities.

We immediately began looking for models of IN VIVO Borrelia biofilms.

The tick midgut seems to be a very attractive model for IN VIVO Biofilms of Bb. Scanning EMicrographs of the tick midgut have demonstrated “carpets” of attached Bb punctuated by Cystic forms of Bb within the Carpet. It was our hope, and it remains our hope that the same rigorous evidence used to justify In Vitro Bb biofilms, will pay dividends in the study of the carpets of Bb in the tick midgut.

The Model of Human gastric mucosa covered by biofilms of Helicobacter pylori (fixed bioflm type communities) is especially attractive. Just recall that these Helicobater pylori microbes survive in an acid environment of pH=4.0.

We were delighted to receive a case study of human Bacterial Endocarditis always a biofilm infection) due to Borrelia burgdorferi from Europe in the peer reviewed literature, as a case report. PCR evidence of Borrelia burgdorferi DNA was produced as etiologic proof of Borreliosis for study of the Heart Valve vegetation’s attempts to stain individual borrelia forms (granular,spiral,cystic,or Cell wall deficient) were not undertaken.

I personally have long believed that the Plaques which accumulate incrementally as the severity of disease in Alzheimer’s Disease from Braak Stage I (Hippocampus limited) to Braak Stage VI ( generalized alzheimer plaque formation throughout the brain) — That these Plaques are Biofilms. Research by me is now underway to attempt to prove that Alzheimer’s Plaques (small, medium sized, and large diameter) are biofilms of borrelia burgdorferi species .

Dr Linda Bockenstedt of Yale School of Medicine, has recently published research demonstrating in a mouse model that chronic infection with laboratory introduced GFP labeled Borrelia burgodrferi produces arthritis ( Lyme arthritis) in the Tarsal joint of the Mice. She photographed and discusses in her paper a curious finding in the deep skin overlying the arthritic joints in her mice.

She describes large Amorphous “globs” of Borrelia in the Mouse skin comprised of huge numbers of Borrelia in mostly non-spiral form (including granular forms) which show a rounded profile at the interface of “glob” with Dermal tissue. The Number of organisms in the Bockenstedt Dermal “amorphous globs” is too numerous to count. She insists that ALL of the borrelial forms in her mouse Dermal globs are All DEAD. I come to a different conclusion. It is my analysis that the “amorphous Dermal globs” are Biofilm communities of Borrelia burgdorferi. Some of the organisms may be dead, as she asserts, but a considerable number of possible Viable borrelia [or viable-but-Non-cultivatible VBNC’s ] are actually the true interpretation of the Bockenstedt Dermal Amorphous Globs.

Indeed, one hallmark of biofilm communities is that the population number of organisms within the community often reaches numbers and densities which are not noted in traditional laboratory Bacteriology cultures on solid media.

I solicit comments from this forum on the ideas presented, with critiques and suggestions for improvement or for improved experimental design gratefully received by me.

Sincerely,

Alan B. MacDonald MD