BORRELIA BURGDORFERI - LYME DISEASE FORMS DRUG TOLERANT PERSISTER CELLS

Borrelia burgdorferi, the causative agent of Lyme disease, forms drug-tolerant persister cells.

1. Bijaya Sharma1, 
2. Autumn V. Brown1, 
3. Nicole E. Matluck1, 
4. Linden T. Hu2and 
5. Kim Lewis1*

+Author Affiliations

1. 1Department of Biology, Northeastern University, Boston, Massachusetts, USA
2. 2Department of Medicine, Division of Geographic Medicine and Infectious Diseases, Tufts Medical Center, Boston, Massachusetts, USA.

ABSTRACT

Borrelia burgdorferi is the causative agent of Lyme disease, which affects an estimated 300,000 people annually in the US. When treated early, the disease usually resolves, but left untreated, can result in symptoms such as arthritis and encephalopathy. Treatment of the late stage disease may require multiple courses of antibiotic therapy. Given that antibiotic resistance has not been observed for B. burgdorferi, the reason for the recalcitrance of late stage disease to antibiotics is unclear. In other chronic infections, the presence of drug-tolerant persisters has been linked to recalcitrance of the disease. In this study, we examined the ability of B. burgdorferi to form persisters. Killing of growing cultures of B. burgdorferiwith antibiotics used to treat the disease was distinctly biphasic, with a small subpopulation of surviving cells. Upon regrowth, these cells formed a new subpopulation of antibiotic-tolerant cells, indicating that these are persisters rather than resistant mutants. The level of persisters increased sharply as the culture transitioned from exponential to stationary phase. Combinations of antibiotics did not improve killing. Daptomycin, a membrane-active bactericidal antibiotic, killed stationary phase cells, but not persisters. Mitomycin C, an anti-cancer agent that forms adducts with DNA, killed persisters and eradicated both growing and stationary cultures of B. burgdorferi. Finally, we examined the ability of pulse-dosing an antibiotic to eliminate persisters. After addition of ceftriaxone, the antibiotic was washed away, surviving persisters were allowed to resuscitate, and antibiotic was added again. Four pulse-doses of ceftriaxone killed persisters, eradicating all live bacteria in the culture.

http://aac.asm.org/content/early/2015/05/20/AAC.00864-15.abstract

Earlier post with vimeo of Prof Lewis as well as CDC Webinar  

http://lookingatlyme.blogspot.co.uk/2014/06/cdc-webinar-on-lyme-disease-persistence.html

Good to see Dr Linden T Hu working with Prof Lewis - Dr. Linden Hu, Tufts University

Borrelia burgdorferi Persistence: Consensus and Controversy – where do we go from here? This was presented in CDC Webinar on persistence see link above.

Researchers’ discovery may explain difficulty in treating Lyme disease - 

“This is the first time, we think, that pulse-​​dosing has been pub­lished as a method for erad­i­cating the pop­u­la­tion of a pathogen with antibi­otics that don’t kill dor­mant cells,” Lewis said. “The trick to doing this is to allow the dor­mant cells to wake up.”

He added: “This gives you an idea that you could, in prin­ciple, estab­lish a sim­ilar reg­i­ment for treating patients for this and other chronic diseases.”

http://www.northeastern.edu/news/2015/06/researchers-discovery-may-explain-difficulty-in-treating-lyme-disease/

Other videos of Prof Lewis Principles of Antibiotic Discovery - Kim Lewis 

https://www.youtube.com/watch?v=89MdHzNHGZI&list=UUComKOHir2WrDuRZXP8DT-A

Uncultured Bacteria - Kim Lewis 

https://www.youtube.com/watch?v=-ojRvlwanSA

What lights my fire https://www.youtube.com/watch?v=GBZ9hyrMsoM

Prof Lewis featured in a recent BBC documentary on Panorama on his research into finding new antibiotics 

http://www.bbc.co.uk/news/health-32701896 

and earlier http://www.bbc.co.uk/news/health-30657486 

HEADACHES FOLLOWING TICK BITE - COULD IT BE BARTONELLA?

Published on Apr 2, 2015

Bartonella infection is a recently identified emerging infectious disease associate with both acute and chronic disorders in humans and animals from cat scratch disease and trench fever to symptoms easily misdiagnosed as an autoimmune disorder.

Video credit: NC Museum of Natural Sciences http://naturalsciences.org/

Healthy teen boy develops debilitating #headaches. Fortunately his mother is a veterinarian.

Earlier post on Bartonella http://lookingatlyme.blogspot.co.uk/2015/05/bartonella-emerging-infectious-disease.html

LYME DISEASE -YOU MAY NEVER BE RID OF IT

Lyme disease: fatigue, paralysis, meningitis ... You may never be rid of it
Posted on 12-04-2015 

English Translation by Google to highlight some salient points.

http://leplus.nouvelobs.com/contribution/1352040-maladie-de-lyme-fatigue-paralysie-meningite-on-ne-s-en-debarrasse-jamais.html
Christian Perronne, Head of Infectious Diseases at the University Hospital Raymond Poincare in Garches (Paris-Ouest-France).
Lyme disease is mainly transmitted through tick bites 

1st phase: a red patch around the bite
2nd phase, which can be insidious, the symptoms are multiple yet they often go unnoticed.

'In my consultations, I see sick people every day, some are in absolutely dismal states. Some have a 20-year gap in their lives, some have had to stop working, some have lost their spouse and their work, some even commit suicide.
The disease is not rare: the tests are bad'

'The only thing experts have managed to agree upon is what should be done in the primary phase: at least two weeks of antibiotics should be prescribed. But the problem is that in 80% of cases it is not done, doctors are simply not willing to give antibiotics for a tick bite.'

'In the secondary and tertiary phases, the American and French consensus recommends that patients be treated for three to four weeks. If the illness persists, a second class of antibiotics can be tried, but this will not heal not every patient.
After this period, it is “terra incognita”: everyone does as s/he pleases' 

'Medicine is not capable today of properly treating this disease. The reasoning being : why should money be spent on scientific research for an “imaginary or rare” disease? Nobody is going to care.
A one-year waiting period for my consultations
When the disease has become chronic, because of lack of early diagnosis and treatment, 80% of patients relapse, and as a result, doctors are even more incredulous. Some tell me that they have "never seen any Lyme". My opinion is that they simply have not recognized that one of their patients had Lyme! and they have been treating him/her for something else. I personally have a waiting period of one year for my consultations, it is utter madness ...
It is known that the bacteria keeps waking up from dormancy in cycles. Once you are infected, in many cases, you will never get rid of it. Some patients suffer from moderate forms but others are very ill, and their whole lives can become a living hell. Some patients, for example, can become bedridden after 10 years of illness.
In the US, a vicious and brutal war has been taking place for decades. It has been fueled by a handful of “experts” who refuse to recognize Lyme in its chronic form.'

This was an excellent article go to the link for the full article 

 http://leplus.nouvelobs.com/contribution/1352040-maladie-de-lyme-fatigue-paralysie-meningite-on-ne-s-en-debarrasse-jamais.html 

Lyme and associated tick-borne diseases: global challenges in the context of a public health threat  Christian Perronne

Conclusion and Perspectives

The numerous complexities of Lyme disease make it an extremely difficult illness to fully comprehend. It remains a diagnostic challenge even for the best informed of clinicians. The lack of a gold standard for diagnosis renders the management of patients difficult and seriously hinders our ability to produce accurate statistics, especially as very similar syndromes could be due to other species of Borrelia. In some patients suffering from syndromes of unclear origin, following tick bite, other microbial agents could also be playing a role. Lyme disease has now entered the political debate as shown by the amendment (Section 54.1-2963.2) voted in 2013 by the State of Virginia, USA, that compels physicians to inform their patients that the “current laboratory testing for Lyme disease can be problematic.” The fact that politicians are being called upon to rule on these matters should prompt scientists to regain control of the situation. Politicians should instead become aware of the necessity to fund research and facilitate the setting up of independent international working groups. Reliable testing is essential to investigate the many syndromes of unclear origin that may mimic many other medical disorders. Proper fundamental and clinical research is urgently needed as it would be the most cost effective way of ensuring that patients are accurately diagnosed and that the best therapeutic strategies are decided upon (Stricker and Johnson, 2014). Development of new diagnostic methods is badly needed. New PCR methods and new genomic techniques, such as high throughput sequencing, could prove promising in identifying the complex mix of microbial agents that are probably involved (Vayssier-Taussat et al., 2013; Lee et al., 2014). Next generation sequencing allowed the identification of various bacteria from Ixodes ricinus ticks in France:Anaplasma phagocytophilum, Bartonella henselae, B. grahamii, Borrelia afzelii, B. garinii, B. burgdorferi, B. miyamotoi, Candidatus Neoerlichia mikurensis, Ehrlichia canis, Rickettsia canadensis, R. felis, and R. helvetica (Vayssier-Taussat et al., 2013). These new techniques should be applied to human samples. Other variables, such as genetic, environmental, or auto-immune factors should also be studied. The name “Lyme disease” is too restrictive as it focuses and fuels the controversy. A new term should be agreed upon for these syndromes with possible infectious involvement, often following tick bites. Closer collaboration between epidemiologists, microbiologists, immunologists, geneticians, environmental scientists, veterinarians, entomologists, and clinicians is needed to identify the main agents that could be causing these occult infections and to determine strain pathogenicity. A new multidirectional approach is crucial in order to widen the field of research and to move forward.

http://journal.frontiersin.org/article/10.3389/fcimb.2014.00074/full

LYME DISEASE PROTEST LONDON 2015 - SCIENCE NOT BELIEF.

London Lyme Disease Protest 13 May 2015 as part of a Worldwide Lyme Disease Protest during May.

What do we protest for :-

More Awareness
Better tests
Better Diagnosis
Better treatment

In the news BBC Scotland - 

European Space Agency backs work mapping Scottish tick hotspots

'ESA has awarded the mapping effort a grant of almost £180,000.'

'Roger Evans, of NHS Highland, said: "Unfortunately, health organisations have limited effective tools at their disposal to reduce the incidence of Lyme disease.

"Resident and visiting populations need an approach that will help them prevent getting Lyme disease and manage exposure to ticks."

He added: "We believe that by using the latest technology and what's commonly called citizen science we can create an interactive and accurate Lyme disease identification and risk management system."

'Organisers plan to protest outside the Department of Health in London where they will call for doctors to be made more aware of the latest research, diagnosis and treatment techniques.

They also want current UK tests for the disease to be re-examined.'

The Bristol University project is also mentioned - (details posted previously 

http://lookingatlyme.blogspot.co.uk/2015/04/the-big-tick-project-thank-you-chris.html   )

http://www.bbc.co.uk/news/uk-scotland-highlands-islands-32692519?

The National -
Lyme Disease: Space agency funds new app to track tick hotspots

'The European Space Agency (ESA) has given £180,000 of financial backing to a project run by NHS Highland, the University of the Highlands and Islands and Scotland’s Rural College to test the new LymeMap app.'

Also in this article Nicola Seal talks about her struggles to get treated for Lyme Disease here in the UK.
http://www.thenational.scot/news/lyme-disease-space-agency-funds-new-app-to-track-tick-hotspots.2922

Recently Aeon Ideas - Pam Weintraub posed the Question 

Why do doctors keep fighting about Lyme disease?

There are some excellent answers found -
http://ideas.aeon.co/questions/why-do-doctors-keep-fighting-about-lyme-disease

My viewpoint was Because of Beliefs -unfounded in science

‘I am a doctor I don’t believe you’ - to quote one doctor on route from Downing St to Dept of Health building passing the London Lyme Disease protest on Wednesday May 13 2015. She had been offered a leaflet based on science already published about Lyme disease, she refused and that was what she said.

Such tragic consequences when basic science is ignored in place of beliefs -

All doctors are trained that antibody tests can miss cases - it depends on the immune state of patients and antibodies used to test for ( maybe a different strain or species) so why suddenly would Lyme Disease be different?

Since antibiotics were first introduced in USA and after the war eventually manufactured in UK , it has been recognised that antibiotics rarely eradicate infection, they reduce the burden of infection with the hope that the immune system can deal with what infection remains. So as in other infections if the immune system does not deal with what remains, further courses of antibiotics are usually required. This is antibiotic persistence which lately is being lumped in with antibiotic resistance, quite a different process.

A spirochetal disease that is capable of penetrating every organ and tissue - well documented in the scientific literature, why then would it magically be cured after only a short course of antibiotics if the patient remained with the same symptoms after the antibiotics as they had before the antibiotic treatment - especially when further courses of antibiotics continue to improve health and quality of life? This is often observed as visual signs by doctors prepared to treat longer.

As science moves forward much of it supporting the patients’ lived experience those with entrenched 'beliefs’ dig in deeper and produce paper after paper using opinionated hubris, tests that in their patent applications they acknowledge as unreliable ( when better tests are available in a research setting) and treatment regimes already shown not to eradicate the bacteria by the numerous scientific papers on seronegativity and persistent infection. We have Dr Barthold who says in 100% of animals the infection persists after a course of antibiotics and 12 months later the level of infection circulating is the same as before the antibiotic treatment. We have Johns Hopkins researchers struggling to find antibiotics to eradicate all persister cells in vitro, and the tri therapy combination of antibiotics that managed to do this, two of them are not widely available for doctors to prescribe.

So as long as doctors ignore science and rely on opinions the patients are abused and neglected.

http://ideas.aeon.co/viewpoints/joanne-drayson-on-why-do-doctors-keep-fighting-about-lyme-disease?recently_published=true

LETS GET LYME DISEASE TESTING RIGHT

Three new Lyme Disease tests on the horizon

DNA Sequence Testing

DNA sequencing-based tests for Lyme disease and Lyme disease-like Borrelioses DNA sequencing, namely the Sanger sequencing technology, is the commonly used “gold standard” for validation of the results of all other nucleic acid-based clinical tests. Ötzi the Iceman who died 5,300 years ago had Lyme disease which was diagnosed in 2012 by DNA sequencing of the genomic DNA remnants of Borrelia burgdorferi preserved in the cadaver. We are using the same approach to diagnose  Lyme disease and Lyme disease-like borrelioses when the pathogens are still in the blood, in the joint fluid or in the cerebrospinal fluid of a patient. This technology is especially useful in testing for Borrelia miyamotoi infections because its “diagnosis currently relies on the use of tests to detect DNA of the organism”, as stated in a CDC guideline [3].  To read more visit http://www.dnalymetest.com/dnasequencetesting.html 'Researchers at Columbia University are beginning a study of a new test for early Lyme disease. The study is examining SpiroFind, a test developed in the Netherlands.'  "The nice thing about this test — if it works as we anticipate it will — is that you will be able to identify active infections in patients in early disease," said Dr. Brian Fallon, director of the Lyme and Tick-Borne Diseases Research Center at Columbia University. "Right now, the tests are only 30-50 percent sensitive in early disease." This was published by the Poughkeepsie Journal last year. http://www.poughkeepsiejournal.com/story/news/health/lyme-disease/2014/04/02/new-lyme-test-gets-study/7231729/  and  http://www.poughkeepsiejournal.com/article/20140403/NEWS01/304030023/New-Lyme-test-gets-study Nanotrap® Lyme Antigen Test (Nanotrap® LA Test) A GAME CHANGING TOOL FOR LYME DISEASE DIAGNOSIS      There is a critical need for a fast, reliable, and accurate Lyme disease test The Nanotrap® Lyme Antigen (LA) Test is a novel, high sensitivity, DIRECT test for Lyme that will provide valuable information to assist a physician in diagnosing and treating Lyme disease at most stages of infection. Unlike other clinical Lyme disease tests that are INDIRECT, the Nanotrap® LA Test is a non-invasive test that provides highly sensitive and specific direct detection of the Lyme antigen, Borrelia burgdorferi (Bb), in a patient’s urine sample. The Nanotrap® LA Test can provide valuable information, at anytime in the infection cycle, about the presence of the Bb antigen when used at the time of initial infection, during and after a course of treatment, and in some cases of recurrent disease. http://www.ceresnano.com/#!nanotrap-lyme-test/c64d "The winds," says Dr. Richard Horowitz, medical director of the Hudson Valley Healing Arts Center, "are starting to change." Horowitz believes that in the coming years, research will reveal that a vast number of other conditions commonly viewed as being distinct — such as arthritis or Chronic Fatigue Syndrome — are linked to Lyme. Dr. Kenneth Liegner, a Pawling-based internist, said treatments will also depend on better tests. The above is an interesting article in the Poughkeepsie Journal link to the full article  http://www.poughkeepsiejournal.com/story/news/health/lyme-disease/2015/05/03/lyme-disease-fact-book/25735589/  Dr Brian Fallon Key note speaker at a Public Health England meeting in March 2015 said that these are exciting times, regarding research into Lyme Disease. He said that there were one or two promising tests being developed.  http://lookingatlyme.blogspot.co.uk/2015/03/dr-fallon-presenting-at-lyme-disease.html The consequences of finding a reliable test for Lyme Disease could have a huge impact on the management of treatment for Lyme disease patients throughout the World.

BARTONELLA - AN EMERGING INFECTIOUS DISEASE

Dr. Edward Breitschwerdt on Bartonellosis - NorVect 2014

Dr. Breitschwerdt talked about Bartonella at the NorVect conference 2014. This is an excerpt of his presentation: Bartonellosis: A One Health Approach to An Emerging Infectious Disease To see all the presentations from both conference days, go to http://norvect.no/conference/conference-2014/movie-access/ 

The full presentation is well worth watching with fascinating insights and research that is being done on Bartonella an infection known about for 100 years but very difficult to culture and test for, commonly referred to as cat scratch disease and considered a mild and self limiting disease, Dr Breitschwerdt is finding that is not always the case.
Details of the Norvect Conference 2014 and 2015  http://norvect.no/conference/

In his presentation Dr Breitschwerdt  talks about the difficulties he had in funding his research hence his involvement now with Galaxy Diagnostics 

http://www.galaxydx.com/web/

You can follow some of the interesting developments through Galaxy Diagnostics Facebook page 

https://www.facebook.com/galaxydx?fref=ts

I posted previously about Bartonella 

http://lookingatlyme.blogspot.co.uk/2012/06/bartonella-another-part-of-puzzle-of.html

Links to research papers by Dr Breitschwerdt click here