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Sapi - Antibiotic susceptibility different forms of Bb

Posted: Fri 6 May 2011 15:36
by Martian

Sapi et al. finally managed to publish some of their findings in a respectable peer-reviewed journal the open access journal Dovepress! I think we will see their other findings here published as well.

The study is an evaluation of in-vitro antibiotic susceptibility of amongst others the "biofilm-like" colonies of Borrelia burgdorferi. Note that the term "biofilm-like" is used, instead of simply "biofilm".

Also note that one of the co-authors is Stricker.

Full-text: [PDF-alert]
Evaluation of in-vitro antibiotic susceptibility of different morphological forms of Borrelia burgdorferi

Original Research
(635) Article views

Authors: Sapi E, Kaur N, Anyanwu S, Luecke DF, Datar A, Patel S, Rossi M, Stricker RB
Published Date May 2011 , Volume 2011:4 Pages 97 - 113 DOI 10.2147/IDR.S19201

Eva Sapi1, Navroop Kaur1, Samuel Anyanwu1, David F Luecke1, Akshita Datar1, Seema Patel1, Michael Rossi1, Raphael B Stricker2
1Lyme Disease Research Group, Department of Biology and Environmental Sciences, University of New Haven, New Haven, CT, USA; 2International Lyme and Associated Diseases Society, Bethesda, MD, USA

Background: Lyme disease is a tick-borne illness caused by the spirochete Borrelia burgdorferi. Although antibiotic therapy is usually effective early in the disease, relapse may occur when administration of antibiotics is discontinued. Studies have suggested that resistance and recurrence of Lyme disease might be due to formation of different morphological forms of B. burgdorferi, namely round bodies (cysts) and biofilm-like colonies. Better understanding of the effect of antibiotics on all morphological forms of B. burgdorferi is therefore crucial to provide effective therapy for Lyme disease.

Methods: Three morphological forms of B. burgdorferi (spirochetes, round bodies, and biofilm-like colonies) were generated using novel culture methods. Minimum inhibitory concentration and minimum bactericidal concentration of five antimicrobial agents (doxycycline, amoxicillin, tigecycline, metronidazole, and tinidazole) against spirochetal forms of B. burgdorferi were evaluated using the standard published microdilution technique. The susceptibility of spirochetal and round body forms to the antibiotics was then tested using fluorescent microscopy (BacLight™ viability staining) and dark field microscopy (direct cell counting), and these results were compared with the microdilution technique. Qualitative and quantitative effects of the antibiotics against biofilm-like colonies were assessed using fluorescent microscopy and dark field microscopy, respectively.

Results: Doxycycline reduced spirochetal structures ~90% but increased the number of round body forms about twofold. Amoxicillin reduced spirochetal forms by ~85%–90% and round body forms by ~68%, while treatment with metronidazole led to reduction of spirochetal structures by ~90% and round body forms by ~80%. Tigecycline and tinidazole treatment reduced both spirochetal and round body forms by ~80%–90%. When quantitative effects on biofilm-like colonies were evaluated, the five antibiotics reduced formation of these colonies by only 30%–55%. In terms of qualitative effects, only tinidazole reduced viable organisms by ~90%. Following treatment with the other antibiotics, viable organisms were detected in 70%–85% of the biofilm-like colonies.

Conclusion: Antibiotics have varying effects on the different morphological forms of B. burgdorferi. Persistence of viable organisms in round body forms and biofilm-like colonies may explain treatment failure and persistent symptoms following antibiotic therapy of Lyme disease.

Keywords: Lyme disease, spirochetes, cysts, round bodies, biofilms

Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Fri 6 May 2011 16:17
by Claudia
Are there any bacteria that won't form a genuine biofilm, or at the very least a "biofilm-like" colony, when allowed to flourish in the unnatural conditions of a petri dish or on the surfaces of a glass test tube?

Shouldn't researchers be looking at whether or not Bb, a motile bacteria, is even capable of forming a complex biofilm in vivo first? Why hasn't this been observed in the bodies of research mice or dogs? (Finding some Bb spirochetes in the plaque of Alzheimer's patient brains isn't even close to the same thing).

Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Fri 6 May 2011 16:32
by Martian
BTW: one of the authors, David Luecke, will be at the following presentations program:

“Breakthroughs in Lyme Disease Research”
Join us to hear our news --- the latest findings of the
Lyme Disease Research Group at the University of New Haven
Saturday May 21, 1PM ~ 8PM
University of New Haven ~ Dodds Theatre
300 Boston Post Road, West Haven, CT 06516
~ Free and Open to the Public ~


Graduate Thesis Research Projects:

David Luecke – “Unraveling Unique Features of Borrelia burgdorferi Biofilm”

Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Fri 6 May 2011 17:05
by rlstanley

What is a 'biofilm-like' colony?

What are the criteria for a 'biofilm-like' colony? I couldn't find anything that defined this thing; no explanation. Could be a biofilm-like colony is simply an artifact (clump of junk caused by process or prep); I saw some photos of clumps of live-dead stained bacteria (so it says). Like to know how all this stuff gets defined, how artifacts get ruled out.

...we developed novel evaluation methods
involving optimal culture conditions for three different forms
of B. burgdorferi (spirochetes, round bodies, and biofilm-like
colonies) and improved bacterial viability determination
techniques. These techniques were used to test the effectiveness
of antibiotics commonly used for Lyme disease treatment
against the different forms of B. burgdorferi.
So, Dove Press peer review evaluated these novel techniques? Right.

Who are these people trying to convince with this kind of stuff? Including a bunch of authors and names in the acknowledgements smacks of trying to keep LymeLand from questioning the veracity of this stuff.

If indeed there is some sort of novel method here, get the thing properly & stringently peer reviewed. And while you are at it, get some independent lab(s) to replicate the work. .

Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Fri 6 May 2011 22:27
by Martian
I think the publication of the study might serve as a neat stepping stone for a next Dove Press publication from Team Sapi: "Evaluation of in-vitro Samento and Banderol herbal extracts susceptibility of different morphological forms of Borrelia burgdorferi", in order to show that this stuff kicks the heck out of Borrelia morphological forms and alleged Borrelia biofilms biofilm-like structures, and to suggest that herbal agents could provide an effective therapeutic approach for Lyme disease patients (as a substitute for antibiotics).

Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Mon 9 May 2011 10:48
by RitaA
Sorry -- I just noticed this was under the "Science" section -- not the "General" one where personal opinions are welcome.


Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Mon 9 May 2011 16:48
by rlstanley
NOTE: the following was written as a response to the post above that was deleted soon after I posted.

The main point, the leading point, that was made in the removed post was that the poster felt Sapi used Dove to rapidly publish results. And that is exactly what I respond to below. I explain why that reasoning has major problems and that rapid communication of results is done at symposia and meetings. Following that, publication in a good peer reviewed journal is expected.

In addition, that deleted post indicated that it was important to get results out rapidly because that could be used for treatment. I respond to that as well.

Next time, this person's entire post will be quoted.

There is no valid excuse for publishing in what amounts to vanity (pay to publish) 'scientific/medical' 'journals'. The exceedingly tiresome excuse, of course, is that people can have easy, open access rapidly. In fact, rapid review of novel techniques--or any paper--for that matter is suspect. And it doesn't take a rocket scientist to figure that one out.

See the thread ... f=7&t=3045 about Predatory Open-Access publications.

Historically--and this still holds-- 'rapid response' of one's findings is presented at symposia/meetings. HOWEVER, since stringent peer review of the work cannot be done in this venue--although valid societies DO have standards--it is simply the first step in letting everyone know about one's findings. The next step is to get published as well as possible.

What's the problem with that? The work is 'out there', rapidily communicated. Then get the paper published via valid and stringent peer review in a respectable journal. Yes, it takes time, and that time is essential for critical evaluation. I would think this would be something one would want especially if one's health is involved.

In addition, there ARE many open-access journals that do not fall into the pay-to-publish category. There are many good journals, period. So, pray tell, why not TRY for some respectable peer review that holds some water. Try. Start somewhere at the top, or even the middle for publication, not at the bottom.

The old 'but it's peer reviewed!' is the foundation of publishing poorly. This dodge is easy to see through unless one is entirely naive of the publishing processs. And ANYONE who uses that platitude casts even more doubt on the validity of their work.

Publishing very poorly generates ongoing doubt in the individuals and societies that matter in decisions of setting standards, funding and furthering scientific/medical knowledge. One only does one's field a disservice by publishing badly. It's a vicious circle, and LymeLand is caught in it.

It takes time, real time to get funding, do competent work and publish well. Taking shortcuts only works against you. Time's a wastin', LymeLand! Still waiting after 2 decades.

Putting papers into the hands of naive people who are told to give to doctors without critical evaluation does more harm than good, too. This backfires in many ways. Along with those breathless press releases.

The question of being tested or treated by anything that is based on rapid communication presents many concerns, including ethical ramifications. People continue to be damaged in LymeLand in an on-going basis because of this kind of stuff. And when they get damaged--and dumped by those who do the damage--all they can do is howl to the wind, or occassionally take legal action.

As a former researcher, my colleagues and I sought out the highest standards and most critical evaluation of our work. We WANTED, we DESIRED to have our work validated by the most critical in our field. When the paper ran the peer review gauntlet and passed muster, you earned that hard fought for respect & knew you were taken seriously. You ADDED to the knowledge base, not detracted from it. Oh, and you were not fooling anyone, ESPECIALLY yourself. Otherwise the findings are best just stuck in a drawer and ignored by all (as some of ours was).

To put a twist on an old saying: Publish in haste, repent at leisure. Or make excuses.

edit 1 to note that my response was to a post that was later deleted
edit 2 is to note the basic content to what I reponded to


Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Mon 9 May 2011 19:04
by RitaA
Again, my apologies for muddying the waters with a personal perspective -- which I have removed.


Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Mon 9 May 2011 20:55
by Camp Other
I've recently written about this paper here: ... vitro.html


I'm wondering about Sapi's choice of publication as well, especially given her previous publications on cancer research and the peer review process involved with, say, Immunology or Endocrinology or Genetics... it seems unusual to go with DP when she's already published under these other titles.

Maybe the best thing anyone could do here is email her and ask her why she decided to publish with DP, and if a more detailed paper will be published in the near future?

Thinking off the cuff here... and this is wild speculation on my part, but what if Sapi is working on a patent for novel culture methods? It won't show up on Google patents (I've already been there; haven't seen anything thus far and it only shows processed patents not ones mid-process) now but it would require an NDA. This might be some way to split the difference so that people know what she's working on without doing full disclosure? I don't know.

As for the rest of it, in terms of content:

I'm asking a number of the same questions you've already brought up here (methods? statistical analysis? "bio-film like" definiton?)

In surveying research that is already out there in other journals, tigecycline definitely looks more effective and reaches an MIC and MBC in vitro earlier than doxycycline does, regardless of the round body issue - and more research would be needed on tinidazole to treat Bb because there just isn't that much so far.

Either way, this is in vitro. More in vivo tests are needed that examine the relevance of the round body issue - if there is any. Barthold doesn't think they're as important - he's already found live whole spirochetes post-treatment that could be transmitted by ticks to a new host.

The argument about their viability just shifts to another target at that point... Such as "Are they really viable and infectious in such small numbers?" and "What if they're highly attenuated in the new host and do not cause that host infection?"

I'm somewhat intrigued by how many passages it takes for Bb to lose its infectivity... I think I remember reading something about one study showing that during post-abx xenodiagnosis, Bb lost their plasmids or they were damaged during transfer to the new host (Bockenstedt?) but in Barthold's they were found intact... I need to go reread my sources on this.

Re: Sapi - Antibiotic susceptibility different forms of Bb

Posted: Mon 9 May 2011 21:05
by Camp Other
I take that back about the patent. I just needed to look at the UNH web site.

I think I need more coffee. :roll:

From: ... ofile.html

So this is 2008... it's in process.
"Two UNH professors have joined the patent race. This time, it is for a new test that will more rapidly and accurately detect Lyme Disease. One of the professors, Eva Sapi, an associate professor in the Department of Biology and Environmental Science, remembers when she visited her doctor with a sore throat. A nurse took a throat culture, receiving the results within moments. Sapi thought to herself, "Why can't we do this with Lyme Disease?'

Once she returned to her lab, she and cohorts in the UNH Lyme Disease Research Group, including Saion Sinha, an associate professor of Physics, began experimenting with available technology at the University. The group was successful in their experimentation and jumped at the opportunity to patent their work.

They applied for the patent in October, and are prepared to wait. "It is a very long process," Sinha says. The group documented their work carefully, recording each day's experiments in a lab notebook, a process that was witnessed and signed by another party to prove that there was no fabrication.

Secrecy is equally as important. "Nothing can be published, discussed or presented," Sinha said. The secrecy is necessary while the process of applying for the patent is in progress. It will take two to three years before the patent becomes official. Not only is the application process lengthy but it is also expensive - about $15,000 to $20,000, Sinha estimated. The University grants money to groups associated with UNH; the UNH Lyme Disease Research Group was lucky enough to acquire the backing for the project.

Until then, Eva Sapi and Saion Sinha will continue to wait and hope that their device - one that will help with the ongoing struggle to successfully treat all aspects of Lyme Disease - will carry their names as the successful inventors."
Could be it's related to this, or she filed for more than one patent?