A ground-breaking study on Biofilms of Borrelia has today been published in Plos One by a team based at New Haven including Dr Alan Macdonald and Dr Eva Sapi.
The full paper and accompanying images can be seen at the following link:
Biofilms, by definition, equate to chronic infection and provide a much better explanation for the GFP-Labelled amorphous clumps of Borrelia recently found by IDSA guidelines author Linda Bockenstedt in a study of antibiotic-treated mice, than her own claim that they are just the debris of "dead" Borrelia.
From the new paper:
"In this paper, we provide substantial evidence that features of Borrelia burgdorferi aggregate development correspond to well-established characteristic features of biofilm formation. The progression of biofilm formation by Borrelia burgdorferi closely resembles that of other biofilms described in literature [20].
This starts by either creating floating biofilms or by adherence of individual cells to biotic or abiotic surfaces, followed by a significant expansion of the aggregates into a three-dimensional structure. Critically, the colonies develop a protective EPS layer. The EPS matrix largely consists of alginate, the primary polymeric compound in the EPS of other well-studied bacterial biofilms [20]....
"Among the order Spirochaetales, Treponema denticola and Leptospira spp have already been reported to able to participate in biofilms...
" Using AFM, we were able to show the sequential steps of the initial adhesion event and the different morphological changes over a 2-week time period with unprecedented resolution. We have also shown evidence of potential EPS secretion in early aggregates.
Our AFM images of Borrelia burgdorferi aggregates demonstrated channel-like structures, which have been reported to be observed in other biofilms [37], including Leptospira spp biofilms [36]. The previous biofilm studies suggested that those channels provide oxygen and nutrient diffusion to embedded cells as well as waste removal [37].......
"In this study, we have also provided evidence that the surface of the aggregates, but not the surface of the surrounding individual spirochetes, contains significant amounts of eDNA and calcium.
eDNA is now widely accepted to be one of the major components of the biofilm matrix of many bacteria and has been shown to perform multifaceted functions in bacterial biofilms, such as maintaining architectural integrity and enhancing resistance to environmental stressors [31]�[33].
" A recent study showed that negatively charged extracellular DNA can chelate cations, including magnesium and calcium, and induce the expression of genes involved in the modification of cell surface components and genes involved in antimicrobial resistance in Pseudomonas aeruginosa [32]. In our future studies, we will investigate whether the previously observed increase in resistance of Borrelia aggregates to environmental stressors is due to similar interactions of the cationic environment and eDNA component of EPS.
The next very important question to be addressed is whether Borrelia burgdorferi biofilm exists in vivo and potential function of those structures might serve other than shelter individual spirochetes from environmental stressors. Borrelia aggregates were recently demonstrated in the midguts of naturally-infected nymphs during their blood meal and it was suggested that aggregates might have a role in successful transmission of Borrelia [16]. It was reported that aggregating spirochetes have undergone complex morphological and developmental changes especially in the initial adherence phase, during which non-motile spirochetes advance as networks toward the basolateral surface of the gut epithelium...
"In summary, our study is the first to characterize the biofilm formation by Borrelia species in vitro..."
Elena Cook
-------------------- Justice will be ours. Posts: 786 | From UK | Registered: Oct 2007
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desertwind
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posted
Great info....Someone else posting a link for the research article w/ pictures.
I believe this is a big missing piece. I just started on Serrapeptase for biofilms. Expereinced some symptoms that I have not had for about 8 years. I do believe the Serrapeptase is helping to dissolve the biofilms.
Posts: 1671 | From Tick Infested New Jersey | Registered: Apr 2010
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Haley
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posted
Can someone translate this into a simple summary for my lyme brain? I didn't read the entire article. I know that it says - the bacteria is creating biofilm.
How does this apply to us. Should we stay away from calcium? I think I read that it is composed of calcium. I just ordered a calcium powder product.
[ 10-27-2012, 12:42 PM: Message edited by: Haley ]
Posts: 2232 | From USA | Registered: Aug 2009
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Razzle
Frequent Contributor (1K+ posts)
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posted
I think mineral avoidance would be bad because the bugs would then pull calcium out of your bones/teeth instead. So I say supplement and let the bugs have whatever, but at least you'll save your own body structure...
Just my humble opinion...
-------------------- -Razzle Lyme IgM IGeneX Pos. 18+++, 23-25+, 30++, 31+, 34++, 39 IND, 83-93 IND; IgG IGeneX Neg. 30+, 39 IND; Mayo/CDC Pos. IgM 23+, 39+; IgG Mayo/CDC Neg. band 41+; Bart. (clinical dx; Fry Labs neg. for all coinfections), sx >30 yrs. Posts: 4167 | From WA | Registered: Feb 2011
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sparkle7
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posted
Here's a 10 minute excerpt of a longer 70 minute talk...
I think you have to buy the longer version. It's also from 2010. The new study 8 legs posted is very recent.
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lymeboy
Unregistered
posted
Does Tindamax have activity against biofilms?
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posted
Haley, I will try to do such a summary and post it in next few days.
Calcium is an essential mineral in the body, necessary for nerve function, muscle contraction strong bones and teeth etc. and you should not eliminate it from your diet.
Everyone's need for calcium differs depending on highly individual factors such as age, sex, your blood levels, your illness etc.. It's best to discuss supplementation issues with your LLMD - there is no "one-size fits all" policy.
Elena
quote:Originally posted by Haley: Can someone translate this into a simple summary for my lyme brain? I didn't read the entire article. I know that it says - the bacteria is creating biofilm.
How does this apple to us. Should we stay away from calcium? I think I read that it is composed of calcium. I just ordered a calcium powder product.
-------------------- Justice will be ours. Posts: 786 | From UK | Registered: Oct 2007
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Without an in vivo study that confirms the existence of these so called biofilms reverting back into infectious spirochetes,it is just a test tube theory that has not been proven yet.
-------------------- nn Posts: 103 | From Northen Neck Va | Registered: Jun 2012
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The authors of the Biofilms article spoke about the need for in vivo studies.
Why do you quote studies by people like IDSA guidelines author Linda Bockenstedt? Do you feel we should have confidence in her science?
Do you have confidence in the IDSA guidelines?
Elena Cook
[QUOTE]Originally posted by nnecker: [QB] Show me where biofilms are reverting back into dividing infectious spirochetes in vivo after antibiotic treatment?
h
-------------------- Justice will be ours. Posts: 786 | From UK | Registered: Oct 2007
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poppy
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Member # 5355
posted
Wasting your time, Elena. People who read this forum frequently know that necker is a troll.
Posts: 2888 | From USA | Registered: Mar 2004
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posted
Only two of these studies are by Linda Bockenstedt,but they have the same results as the other three.
Why wouldn't I have any confidence in her science?Do you have any in vivo evidence that contradicts it?
As far as the IDSA guidelines go,yea they might have to be tweeked here and there a little bit,but I sure don't have any confidence in the ILADS guidelines nor does the mainstream medical community,or the British Heath Protection Agency.
As far as Sapi providing in vivo evidence to support her hypothesis,good luck with that.How many times over do you have to run the same study?I highly doubt that all of a sudden you are going to start seeing cysts and biofilms reverting back into infectious spirochetes.
-------------------- nn Posts: 103 | From Northen Neck Va | Registered: Jun 2012
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Lymedin2010
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Member # 34322
posted
Better such studies, as with (Eva Sapi) & putting forth postulations, rather than the idiocy of having had treatment for a month & being cured.
The idiocy that does no tests nor cares to investigate it further, only to proclaim it is a post syndrome.
The medical communities, big pharma, & the gov have all run amuck. The people who have been directly or indirectly affected by this are the best advocates and appear to be more trustworthy, as we have learned thus far.
There is mention to the need of in vivo studies. Again better than some under the rug swept explanation and the thoughless and robotically mindless explanations of those who in reality could care less.
Posts: 2094 | From NY | Registered: Oct 2011
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posted
Ok, have Dr Sapi infect the mice(with ticks not a needle)give the mice antibiotics(proper doses,proper amount of time,properly monitor blood levels,and no immunosuppessors)and let's see what happens.
I will bet the same thing happens to what always happens.They will not find any dividing infectious spirochetes.
-------------------- nn Posts: 103 | From Northen Neck Va | Registered: Jun 2012
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Summer3
Frequent Contributor (1K+ posts)
Member # 35286
posted
nnecker,
I'm interested to why you continuously try to dispute ILADS doctors and their guidelines along with quoting IDSA supported studies as if they are the only "facts" in this highly controversial issue. If you do not agree with ILADS, fine, that's your choice to not be treated under those guidelines.
For MANY people, IDSA guidelines do not work. They continue to progress with symptoms of Lyme and co-infections. It is their choice to seek alternative treatments.
There are no tests that prove that Lyme can or has been eradicated from the human body.
In my situation, I had absolutely NO health conditions prior to Lyme. I had never needed to go to a doctor for anything. I had a 105 degree fever and rash after a tick bite. I'm in a Lyme endemic area and have had 5-6 ticks on me per year for my entire life.
I was given a week of low-dose amoxicillin by the first doctor. A month later I was able to get a month of low-dose doxy. When I continued to get worse, my choice based on the IDSA guidelines was simple: symptomatically treat all of the symptoms, look for other completely unrelated causes, or do nothing. Obviously I'm not going to do nothing while I'm having seizures, paralysis of my limbs, severe joint pain, etc. I DID allow mainstream doctors to look for other conditions. They have found nothing other than positive tests for Lyme, co-infections and a very low CD57 and lymphocyte count.
Sometimes you have to use common sense. If you have a known condition that did not respond to one doctor's opinion of adequate treatment, find another treatment that works. I highly doubt that if you or one of your family members were not responsive to the IDSA treatment guidelines and you had a known case of Lyme, that you would continue to listen to the IDSA's position and assume that all of the symptoms are due to aches and pains of daily living.
poppy
Frequent Contributor (1K+ posts)
Member # 5355
posted
Give up people. This is a troll, probably one that has appeared here in past years under other names. I don't know why this person is allowed to continue to harass lyme patients. Previously, good longtime contributers have been banned for lesser offenses.
Posts: 2888 | From USA | Registered: Mar 2004
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If I did, I would have the privilege of meeting people with astounding intellectual abilities. Like for example, yourself.
After all, you are a man (or a woman) who was able to go, in only four months, from a state where he/she knew nothing at all about Lyme ("WOW,this conversation is unbelievable.I pray to God I don't have to go through this.I feel so sorry for you all and hope you get better." NNecker on Lymenet 28Jun 2012) - to a state where he/she now has a full handle on the Lyme controversy, and can quote fluently from the scientific literature, and of course, firmly believes in the IDSA guidelines.
Furthermore, even though you live in Virginia, you seem to know all about the HPA in my country, who incidentally now operate their Lyme lab out of Porton Down, our premier biological warfare centre.
Before you demand that Dr Sapi and her colleagues undertake experiments that you, the anonymous person who only got introduced to the world of Lyme a few months ago, why not say who you are, your medical or scientific qualifications to comment on the issue. I invite you to put your contribution on the comments board of PLOS itself.
Unfortunately, you will not be allowed to call yourself "NN" there, as they only accept comments from those who identify themselves fully. Also, if it should turn out that your location is not in that little town in Virginia, but somewhere else - say Crofton, Maryland or somewhere else - your comment will be disallowed.
Aint bureaucracy a pain in the nnnnnnneck? Elena
quote:Originally posted by nnecker: Ok, have Dr Sapi infect the mice(with ticks not a needle)give the mice antibiotics(proper doses,proper amount of time,properly monitor blood levels,and no immunosuppessors)and let's see what happens.
I will bet the same thing happens to what always happens.They will not find any dividing infectious spirochetes.
-------------------- Justice will be ours. Posts: 786 | From UK | Registered: Oct 2007
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posted
Your right, I did not know any thing about Lyme at first.I pulled a few ticks off of myself and got two bulls eye rashes and got scared and came to this site for help.
At first I took the advice that was given and took mass amounts of Doxy until I was puking my guts out all over the place.
After pulling my head out of the toilet one day I said this is stupid and said to myself let me start reading all I can about this disease.So I read and read and read all I could about it and now you know my opinion about it.
Since I have stopped taking the Doxy five months ago(about six weeks worth and against the advice of many here)I feel great.
But that's all academic,I think new people who come here should see both sides of the issue,and then they can decide for themselves what is the best path to take when it comes to treatment,don't you.
I post scientific evidence that supports my position just like you.I don't call you a troll for posting your study,I just don't agree with it and I show you why.
Don't you think new people should see all sides of this issue?If you are comfortable with your position about Lyme,why should you care if I post mine?If you do,then I think that is saying something about your position.
-------------------- nn Posts: 103 | From Northen Neck Va | Registered: Jun 2012
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Summer3
Frequent Contributor (1K+ posts)
Member # 35286
posted
Nnecker, there is a huge difference between people with an acute infection who can take a few weeks of doxy and then feel "great" and those with late stage infections who need more extensive treatment. Lyme has a wide range of manifestations.
I find that most people's first line of defense is to be treated via the mainstream IDSA guideline (that is if they can even get tested since the criteria is so stringent). Only after that fails, do they seek out support groups and alternative treatments. Therefore, the majority of people here and on other Lyme boards are not acute cases. They are the cases that IDSA treatment guidelines have failed. Likely, IDSA data and recommendations are not going to apply to them since they don't even recognize treatment failures as anything other than aches and pains of daily living.
The reason why I and some others here are getting so annoyed is not because we lack confidence in "our" position. It's because you are quoting data that has in some way contributed to a lot of Lyme patients (including myself) persisting to late stage infections which are VERY difficult to treat.
I am not in the fortunate position that you are. I don't feel "great" after a few weeks of doxy. I went from a case that would could have been easily treated to one that is causing serious neurological problems along with organ damage. The reason it got to this point is because of doctors basing diagnosis and treatment on IDSA guidelines and the "studies" that support them, many of which are outdated.
I don't think anyone on here wants to take high doses of antibiotics or herbs for years and years just to be able to function and prevent progression. However, we don't have a lot of alternatives, and there is significant evidence that antibiotics can help late stage Lyme cases. I know I for one cannot spend the rest of my life progressing and attributing seizures, paralysis and heart and liver involvement to "untreatable post-Lyme syndrome."
It would be similar to if you went to an oncologist with textbook symptoms of lymphoma. Suppose that doctor and others within his professional organization denied accurate testing and misdiagnosed you for 10 years until your Lymphoma reached stage 4 and your symptoms became debilitating. Would you then go back and quote that same doctor's studies and trust his judgement?
posted
Summer show me a long term patient study that shows that the majority of people who take 4 weeks of Doxy a couple of weeks after they have been bitten go chronic.Go ahead show me.
Those are mostly the people that I am talking to who may not be members but come here to learn about Lyme.
How do I know what happened to you and others here are not the exception.Does what happened to you happen to 1 in 10,1 in 50.Again show me scientific study that says it is not.
When I first came on here,I was being Pmed that I should take cystbusters and all this herbal crap or I was going to be in big trouble if I did not take it.Well I didn't and how am I doing.
This is what I try to warn people about.
-------------------- nn Posts: 103 | From Northen Neck Va | Registered: Jun 2012
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Summer3
Frequent Contributor (1K+ posts)
Member # 35286
posted
Exactly nnecker, but you were an early case which is much different than a late case. People should not experiment with antibiotics and herbs on their own and take massive doses in my opinion. They should work with a Lyme-literate doctor. One that will be willing to come up with a treatment if doxy doesn't work.
Nobody is debating that doxy can treat early cases of pure acute Lyme. I'm sure it does resolve Lyme for certain uncomplicated early cases. It cannot however work for late cases and co-infections such as babesia and bartonella which many have.
I'm not sure how you would like a study to be conducted that allows patients to progress to late stage symptoms. That obviously would be unethical and is not a realistic proposition.
Since you do a lot of research, I'm sure you are aware of the animal studies showing persistence of Bb in dogs, mice and monkeys following antibiotic treatment. I'm sure you are also aware of the study showing that doxycycline at 200mg can increase the cystic form of Lyme at a rate of 200%.
What happened to someone like me may very well be an exception. However, the odds of being diagnosed quickly, at least in my area of the Northeast are slim. I had the rash, a known tick bite and flu-like symptoms. I couldn't get anyone to do a test until several months and doctor visits later. When I could get a test, they only did the ELISA and not a WB. Many months later I finally got a WB and it ended up being CDC positive. So in many cases the problem lies in the inability and unwillingness of doctors to clinically diagnose Lyme.
I can clearly with 100% positivity state that low dose doxy monotherapy did not work for me. I'm sure others can verify that as well. If it had worked, why would I continue to take abx or herbs? I am someone that has NEVER taken any medications for anything prior to Lyme.
I also think that it's a little bit early (based on your registration date) to definitively state that doxy cured you and you are fine. I hope that IS the case. If it isn't, I think that you will start to see that you'll need to look into other treatment options and the research, and patient testimonies that substantiate those options.
This is the last I will be posting regarding this debate as it's getting off the topic of the thread. Good luck nnecker and I hope that you continue to have substantial improvement from the treatment you chose.
Haley
Frequent Contributor (1K+ posts)
Member # 22008
posted
Necker - if you want to argue about this, start a new thread. I have not read all the posts, just the last few.
This is my theory - each person that has this illness has an entirely different illness, hence the new term MCIDS.
Each person is bitten by a different tick, in a different geographical area, that latched onto a different host (rat, dear, dog etc).
Some people like necker may get lucky and have one infection that can be treated quickly and efficiently, others may have 50 to 100 different pathogens that can't be treated with Doxycycline. There was a young girl recently that tested positive for the Bubonic plague. They found bug bites around her abdomen.
Think of the medicines we give to our horses and dogs - de-wormers. Why wouldn't these vectors pick up those types of pathogens. Large parasites don't get knocked out by your Immune system - they are too large.
That's why this illness is so complicated - because it manifests itself in so many different ways. Necker - just be happy that you were not one of the unsolvable mysteries.
Posts: 2232 | From USA | Registered: Aug 2009
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posted
Nobody is debating that Doxy can treat early cases of Lyme.What,are you kidding me?
As far as persistence in the early dog studies,once they monitored the blood levels correctly and increased the dosage a little they found no persistence.In one or two mice studies they found a couple of NON DIVIDING non infectious spirochetes.Up the antibiotic dosage a little bit and lets see what happens.
The monkey study has some issues,they found no persistence in short term infection period, only in the 4 to 6 month infection period.Some scientists are questioning the antibiotic blood levels and the way the monkeys were inoculated.
They were inoculated with a hypo-needle which they estimate is a 100 times the disease load of a tick.
So that's like a hundred ticks with Lyme biting a 25-30 pound child.Then you wait six months before you do any thing about it.
I would recommend more antibiotics then what the monkeys got.Even still they only got a couple of beat up spirochetes that they could not subculture.So what does that prove anyway biofilms cystform?More likely a dosage issue to me.
As far as SOME Dr's not treating early Lyme properly,I will agree with you on that, and I have always said that 2 weeks of Doxy is not enough.
Cystform and biofilm form, there is no in vivo evidence that proves that they are reverting back into infectious dividing spirochetes.So to me all this cystbuster and biofilm desolvers are a waste of money and potentially harmful.
-------------------- nn Posts: 103 | From Northen Neck Va | Registered: Jun 2012
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1.atherosclerosis � Biofilm may contribute to the development of atherosclerosis. Ott et al.'s work showed a diverse groups of bacterial �signatures� in atherosclerotic lesions of patients with coronary heart disease. In a commentary following Ott's paper, Katz and Shannon concluded that his work suggested that atherosclerotic plaques are composed of �functional biofilm.� The team noted that the characteristics of a �mature� arterial wall make it well-suited for biofilm formation and explains the inefficacy of antibiotics, such as macrolides or fluoroquinolones, in clinical trials. 2. chronic sinusitis � One study found that biofilms are present on the removed tissue of two-thirds of patients undergoing surgery for chronic inflammation of the sinuses."
and many other examples follow.
Looks like the ability to form a biofilm is a common way of bacteria surviving in vivo; and the main(obvious)thing about biofilms found in vivo is that they are chronicly infecting the patient.
I really look forward to your synopsis of the paper because patients need to be conversant with this and maybe we can try and educate our doctors. Especially those at primary care level, who seem to be so ignorant that Lyme is everywhere in the UK.
beths
Frequent Contributor (1K+ posts)
Member # 18864
posted
nnecker-perhaps you should be grateful you found this site. You took 6 weeks of doxy early on, and obviously were able to eradicate the infection.
If you hadn't found lymenet-would you have treated at all? I know you don't want to believe this but the advice you received may have saved you years of agony.
Most of us weren't treated early-giving the spirochetes time to disseminate.
Because you are ok, doesn't mean the rest of us are
Posts: 1276 | From maryland | Registered: Jan 2009
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posted
Hi all Here is my summary in lay terms of the paper. Best Elena Cook
Summary of Biofilm paper by Sapi et al at PLOS ONE Oct 2012
Biofilms are highly organised structures, created by microbes, which notoriously protect them from unfavourable conditions in their surroundings. Examples of the latter include an environment which is too acidic/alkaline; a change in temperature; and chemicals toxic to the microbe, such as antibiotics. Biofilms are known to be involved in serious chronic infection.
The researchers in this study set out to discover whether Borrelia burgdorferi (Bb), bacteria which cause Lyme disease, are capable of forming biofilms in the test tube.
They point out that Bb is well known to possess various strategies for survival and persistence in the infected host. For example, Bb has been proven to transform itself from its familiar spiral shape into other forms known as �Cystic�, �Granular� and �Cell-wall Deficient� varieties, which are adaptations to environmental conditions which might otherwise destroy it.
The authors note that organised collections, or �aggregates� of Bb containing not only spirals, but also Cystic and Granular forms have been seen both in the test tube and in living infected tissue. The formation of these aggregates dramatically impacts Bb�s ability to survive in unfriendly environments.
There are certain characteristics which are considered to be hallmarks, or �markers�, of biofilms. These include:
� Ongoing re-arrangement of structure and shape during development of the biofilm
� Ability to grow on a variety of living and non-living surfaces
� Secretion of EPS. [EPS stands for Extracellular Polymeric Substances. This is non-living material, distinct from the bacteria themselves, which envelopes the structure as a protective layer. Certain components of the EPS such as Alginate, Calcium and extracellular DNA are commonly seen in Biofilm EPS]
The researchers studied Bb cultures with a variety of microscopic techniques to determine which, if any, of the above Biofilm markers were present. They grew three sub-types of Bb in culture in polystyrene tubes (Bb 297, B31 and a variety of B31 genetically altered to glow fluorescently green for easier identification).
While initially they saw only individual spiral Borrelia, once the concentration had reached ten million bacteria per ml, aggregates began to appear.
Structural Re-arrangements
The authors describe, with accompanying photographs, changes in shape and structure that appeared over time. Using the technique of Darkfield Microscopy, they saw a semi-spherical shape flatten to become film-like, then further change into a rigid structure with defined �hills, valleys and cracks�.
Examination of the same cultures using another technique, DIC microscopy, clarified that the �film� was not uniform but had a diverse composition. FITC staining demonstrated that spiral-shaped Bb create the original structure, and can be found throughout it after a few days, but some days later most of the Bb are in the form of �Round Bodies� (cysts).
The team next used the revolutionary technique of Atomic Force Microscopy (AFM). This allowed them to visualise the structural changes during the development of the Bb aggregates on an unprecedented nano-scale [ie one billionth of a metre; the diameter of atoms]. [Modern electron microscopes can achieve nano-scale resolutions, but the necessary preparation of the specimen by freezing or chemical techniques kill the organisms, so that the development in time could not be seen, and/or may damage the structure. Using AFM eliminated these drawbacks.]
The team observed spiral �shaped Bb combining in pairs, which later assembled themselves into a web-like mesh. In the centre they saw spiral Borrelia folding in on themselves, and Cystic forms throughout. Individual meshes then went on to combine together to form a progressively larger network, with new features appearing such as arches and loops. These latter were of uniform size, indicating that a highly organised and deliberate activity was taking place.
As time went on, more re-arrangements to the structure occurred, with pits and protrusions appearing. The authors have included 3D images documenting these changes.
Ability to Grow on Varied Surfaces
The New Haven team attempted to grow aggregates of Bb on a wide range of substances, including materials that are non-living ( glass and plastic) and those derived from living tissue, such as rat collagen and Matrigel (a product derived from mouse tumour ). Bb was able to attach to, and form aggregates on, all these materials, even when its density was only 5000 cells per ml. The authors also found that Bb could form free-floating aggregates, and have included a video demonstrating this.
EPS
The team biochemically analysed the covering of the structure to see if it conformed with the known composition of Biofilms. Using a staining method known as the Spicer and Meyer technique, they found that the material was either alginate or an alginate-like compound.
Alginate is a substance found in nature in brown algae, but is also a well-known constitutent of EPS in Biofilms. Tests with antibodies that bind specifically to Alginate proved that it was this.
Presence of Calcium on the surface of the EPS is another marker of Biofilm. Alginate reacts with Calcium to form an insoluble compound, Calcium Alginate. Using a specific stain for Calcium (Alizarin Red-S), it was shown that the centre of the aggregates were positive for presence of Calcium.
Another hallmark of Biofilm formation is the occurrence of extracellular DNA (eDNA) on the surface. This plays several roles, including helping microbes to attach to surfaces, and enabling them to resist unfavourable conditions. Using the chemical stain DDAO, the team found that eDNA was indeed present on the surface of the aggregates.
The researchers point out that eDNA bearing a negative electric charge is known to be able to chelate certain minerals such as Magnesium and Calcium. This can play a role in bacterial resistance to antibiotics. They hope to explore this connection with regard to Bb in the future.
Borrelia is part of the bacterial classification of Spirochetes, and other spirochetes are known to make biofilms, some of which are involved in disease.
The team note that Treponema denticola, a spirochete found in biofilms along with other microbes, is involved in gum disease. This bacterium also produces biofilm in the test tube.
Another spirochete, Leptospira, forms free-floating biofilms. Certain leptospira cause Weil�s Disease, a serious and often fatal malady transmitted by contact with rat urine.
Atomic Force Microscopy allowed the researchers to see channels running through the Bb aggregate structure, similar to those known to occur in Biofilms made by Leptospira and other bacteria. These channels may function to supply nutrients and oxygen to the bacteria resident in the Biofilm, and carry away waste products, much like the circulatory system in a human.
The team discuss the possibility of Bb forming Biofilms in living organisms, and the question of whether these might serve other functions in addition to protecting the bacteria.
Bb aggregates have been found in nymphal (juvenile) ticks feeding on blood, and these may play a role in transmission of disease. Bb in non-motile form (unable to move as individual bacteria) have nevertheless been shown to advance as �networks� inside the gut of the tick. They hope to re-examine Bb aggregates in feeding adult and nymphal ticks to see whether hallmarks of Biofilm are present there.
Taken together, the weight of all the evidence above proves that Bb forms Biofilms under test tube conditions. This is the first study to demonstrate this conclusively. The researchers now look forward to investigating the prospect of Bb Biofilm formation in the living infected host, and its possible role in chronic Lyme disease.
Just thought I'd let you know that the offending "nnecker" has now been removed from LymeNet by the moderators.
I don't think "nnecker" was just any old "troll" - I believe he was one of the leading Denialists.
It's very sad that - as if we do not have enough to contend with already - certain amoral scientists and doctors find it necessary to come on this forum, pretending to be patients, to create confusion and sow further harm.
It's no surprise that the person posing as "nnecker" chose to intervene so vehemently in this post. I was almost expecting him to make an appearance.
The biofilms paper is sending shivers down the spine of people like Nnecker and other leading Denialists, as it provides such tangible evidence for the existence of chronic Lyme.
The Denialists are continually moving the goalposts, as new evidence piles up validating chronic Lyme. With so many scientifically verified sightings of Borrelia occurring in the tissues of animals and humans who had received "adequate" antibiotics, it has become impossible for them to sustain the fiction that all this suffering is due to "somatisation" or other psychosomatic causes.
The concept of a purely auto-immune disease with no active infection was also being discredited.
They then hoped to convince the medical and lay public that the persisting Borrelia were weakened, half-dead shadows of their former selves, unable to generate disease in the body.
Because doubt has been cast on this, they then delivered their latest salvo in the shape of the recent Bockenstedt study, purporting to show that "persisters" are actually just the debris of Borrelia, long dead, but possible able to cause some limited inflammatory damage such as arthritis in some people.
Dr Bockenstedt, an IDSA guidelines author, refuses to allow other researchers the means to validate her results. She refuses to use a simple and readily available test which would have shown if her "debris" was dead or alive.
Because technology is advancing forward rapidly in so many biological fields, it is inevitable that new techniques will explode old lies. The use of Atomic Force Microscopy enabled Dr Sapi et al to visualise Borrelia biofilms in exceptional details. People like "nnecker" know this and, panic-stricken, they attempt to hold back progress by the kind of disgraceful, pathetic behaviour you have seen here.
Elena
-------------------- Justice will be ours. Posts: 786 | From UK | Registered: Oct 2007
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surprise
Frequent Contributor (1K+ posts)
Member # 34987
posted
Thank you Elena---
-------------------- Lyme positive PCR blood, and positive Bartonella henselae Igenex, 2011. low positive Fry biofilm test, 2012. Update 7/16- After extensive treatments, doing okay! Posts: 2518 | From USA | Registered: Nov 2011
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posted
I would encourage people on this forum to write a comment on the website of the publishers (PLoS) expressing your appreciation of this important study. You don't have to be a doctor or researcher to add your comment, which will appear online, but you do have to go through the free registration process.
If you go to the article at the link below, you can click on "Make a General Comment" in the blue box just to the right of the autnors' names.
The more positive comments it receives, the more it is likely to come to the attention of medical professionals, who might just be prepared to think outside the box, or rather, outside the censorship zone.
Elena
-------------------- Justice will be ours. Posts: 786 | From UK | Registered: Oct 2007
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posted
Thanks, Elena, for the article, your excellent summary of it, and also for your actions with "nnecker."
Funny how some people can be counted on to appear when articles like this are posted, isn't it? Voice sounds awfully familiar to those of us who have been around for a while...
-------------------- "Looks like freedom but it feels like death.. It's something in between, I guess"
Leonard Cohen, from the song "Closing Time" Posts: 822 | From California | Registered: Jan 2006
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