Saturday, 22 October 2016


Ceftriaxone Pulse Dosing Fails to Eradicate Biofilm-like Microcolony B. burgdorferi Persisters Which Are Sterilized by Daptomycin/Doxycycline/Cefuroxime Drug Combination without Pulse Dosing

Jie Feng1Shuo Zhang1Wanliang Shi1 and Ying Zhang1*
  • 1Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, USA
Although the majority of Lyme disease patients can be cured, at least 10-20% of the patients continue to suffer from persisting symptoms such as fatigue, muscular and joint pain, and neurologic impairment after standard 2-4 week antibiotic treatment. While the causes for this post-treatment Lyme disease symptoms are unclear, one possibility is due to B. burgdorferi persisters that are not effectively killed by current antibiotics such as doxycycline or amoxicillin used to treat Lyme disease. A previous study showed that four rounds of ceftriaxone pulse dosing treatment eradicated B. burgdorferi persisters in vitro using a relatively young late log phase culture (5 day old). In this study, we investigated if ceftriaxone pulse dosing could also eradicate B. burgdorferi persisters in older stationary phase cultures (10 day old) enriched with more resistant microcolony form of persisters. We found that ceftriaxone pulse dosing could only eradicate planktonic log phase B. burgdorferi spirochetal forms and round body forms but not more resistant aggregated biofilm-like microcolony persisters enriched in stationary phase cultures. Moreover, we found that not all drugs are suitable for pulse dosing, with bactericidal drugs ceftriaxone and cefuroxime being more appropriate for pulse dosing than bacteriostatic drug doxycycline and persister drug daptomycin. We also showed that drug combination pulse dosing treatment was more effective than single drug pulse dosing. Importantly, we demonstrate that pulse dosing treatment impaired the activity of the persister drug daptomycin and its drug combination against B. burgdorferi persisters and that the most effective way to kill the more resistant biofilm-like microcolonies is the daptomycin/doxycycline/ceftriaxone triple drug combination without pulse dosing. Our findings indicate pulse dosing may not always work as a general principle but rather depends on the specific drugs used, with cidal drugs being more appropriate for pulse dosing than static or persister drugs, and that drug combination approach with persister drugs is more effective at killing the more resistant microcolony form of persisters than pulse dosing. These observations may have implications for more effective treatment of Lyme disease. Future studies are required to validate these findings in animal models of B. burgdorferi persistence.

Tuesday, 18 October 2016


An excellent documentary shown on IrishTV recently on Lyme disease

Published on Aug 31, 2016
Nicci St.George Smith met James and Brian at the 2016 Irish Lyme Disease Conference organised by Ann Maher in conjunction with Tick Talk Ireland

Here they tell her about their journey toward completing their documentary LIVING WITH LYME

Further information from Tick Talk Ireland

Saturday, 15 October 2016


The Internet has been a buzz recently with the news that Willy Burgdorfer found patients with Lyme disease their blood showed a strong reaction for the Swiss Agent ( Rickettsia Helvetica)

The ‘Swiss Agent’: Long-forgotten research unearths new mystery about Lyme disease

'At a government lab in Montana, Willy Burgdorfer typed a letter to a colleague, reporting that blood from Lyme patients showed “very strong reactions” on a test for an obscure, tick-borne bacterium. He called it the “Swiss Agent.”
Now STAT has obtained those documents, including some discovered in boxes of Burgdorfer’s personal papers found in his garage after his death in 2014. The papers — including letters to collaborators, lab records, and blood test results — indicate that the Swiss Agent was infecting people in Connecticut and Long Island in the late 1970s.'
'On the top of a stack of documents in his garage was a mysterious note, penned boldly in red ink in the scientist’s unmistakable handwriting. “I wondered why somebody didn’t do something,” it said. “Then I realized that I am somebody.”'
The STAT article can be read at this link:-
'The STAT article reports that both Jorge Benach and Allen Steere now say it is time to take a closer look at Rickettsia helvetica’s role in Lyme disease. Benach says the research “should be done” because public health concerns warrant a closer look.
For patients, looking into pathogenic factors related to persistence in tick-borne disease is long overdue. The failure to note the existence of the Swiss agent along with Borrelia burgdorferi is bacteria in the initial publication about Lyme disease may have set back progress in understanding the pathogenicity of Lyme disease decades.' From Lyme Policy Wonk :-

An interesting angle is that from Lyme MD

In 2011 this was published:-

First detection of spotted fever group rickettsiae in Ixodes ricinus and Dermacentor reticulatus ticks in the UK. 

Tijsse-Klasen EJameson LJFonville MLeach SSprong HMedlock JM.


A preliminary study was conducted to determine the presence of spotted fever rickettsiae in two species of British tick (Ixodes ricinus and Dermacentor reticulatus). The 16S rRNA gene of Rickettsia spp. was detected in 39/401 (9·7%) of ticks tested, including 22/338 (6·5%) I. ricinus and 17/63 (27%) D. reticulatus. Some positive I. ricinus samples showed 100% homology with Rickettsia helvetica (10/22), and most positive D. reticulatus showed 100% homology with R. raoultii (13/17). Five other Rickettsia spp. were detected exhibiting 96-99% homology. Ticks positive for rickettsiae were collected from various hosts and from vegetation from eight counties across Great Britain. The distribution of R. helvetica in various engorged and unfed stages of I. ricinus suggests that R. helvetica is widespread. R. raoultii was found in questing adult D. reticulatus in Wales and England. This is the first evidence of potentially pathogenic spotted fever rickettsiae in British ticks.

This study suggests the Swiss Agent ( Rickettsia Helvetica ) is widespread in the UK. More research is required to find out what role this infection is having on people sick in the UK following a tick bite. 

Monday, 12 September 2016


Testing for Lyme Disease is problematic for several reasons

'Scientifically, sensitivity (% of definite positive patients who test positive by the test in question) and specificity (% of well patients who test negative by the test in question) are measured against another test, such as CDC surveillance pos WB, which is how the patients are determined to be positive in the first place.  As ALL tests for Borrelia lack sensitivity to some extent this makes the sensitivity and specificity of the test in question highly suspect, or limited.  This, of course, means sensitivity and specificity are not really related to % of patients who ACTUALLY have the disease, but % who have been determined by a particular test of limited sensitivity to have the disease, a subset.  As patients who have been definitely determined to have Borreliosis by the CDC criteria for surveillance, and the CDC are measuring the sensitivity of a new test against that, of course the sensitivity of that new test will be inflated, a great advantage if you are the holder of the patent of that new test.  This would be especially so if the new test is measuring some parameter which is related in some way to the baseline test (such as both measuring antibodies to something).
In reality any of the various tests available if positive is a likely indication the patient has Borreliosis, but any one of them will have some false positives.  The false positive rate is not scientifically possible to accurately determine from evidence.  In Borreliosis the sceptics commonly cry "false positive" for any one test which shows positive, but they have no evidential basis on which to ground their claims.  It is certainly helpful to the patient's cause to have 2 separate tests positive, making a false positive far less likely.  In areas of medicine other than Lyme it is usually clinical suspicion plus one suggestive, not necessarily strong positive, test which would be taken as diagnostic evidence.
However, IF the judges (the CDC) are actively trying to protect the health insurance industry (which would appear to be the case, based on the evidence of their judgements), of course their current stance would make a lot of sense.  If they applied the same level of criteria for all other infectious diseases, only a small fraction of patients with infections of all kinds would be diagnosable or treatable.'

I read the above explanation recently and have permission to share, it explains very clearly one aspect of the problems we have with Lyme serology.

Prof Perronne discusses other aspects of Serolgy in this publication 

Calibration of serology

When Lyme serology was developed, no reliable method was available to be used as a gold standard for comparison. As most of the signs and symptoms are non-specific, no reliable clinical diagnostic score could be established. The low yield of culture and the difficulty involved in using the technique routinely were another major obstacle. A pragmatic cut-off level for the serologic tests had to be determined arbitrarily on blood donors (EUCALB, ; Assous, ). In the late seventies, when Lyme disease was first discovered, it was understandably thought to be a rare and regional phenomenon. Therefore, a low prevalence was set as experts were afraid the serologies would produce too many false positive diagnoses (EUCALB, ; Assous, ). Patients and control populations are ill-defined with a high variability in predictive positive and negative values from one test to another. Culture of B. burgdorferi or detection of its genome by polymerase chain reaction (PCR) may occasionally confirm the clinical diagnosis in seronegative patients, however none of these methods are sensitive enough to be considered reliable diagnostic methods, especially in routine practice (Schutzer et al., ; Nields and Kveton, ; Chmielewska-Badora et al.,; Brunner, ; Assous, ; Holl-Wieden et al., ; Aguero-Rosenfeld, ; Dietrich et al.,; Wallet et al., ). As a result, many patients suffering signs and symptoms compatible with Lyme disease, but whose test is negative, are falling by the wayside.

Possible causes of seronegativity

Several factors leading to seronegativity have been identified in confirmed cases of Lyme disease: (i) the arbitrary cut-off level of tests, (ii) the sequestration of antibodies in immune complexes, (iii) the wide variety of species and subspecies of Borrelia that co-exist in different parts of the world, and (iv) coinfections with other pathogens which may be responsible for some or all of the symptoms or which may alter the immune response (Schutzer et al., ; Brunner, ). The complex B. burgdorferi sensu lato includes (Table (Table1):1): B. burgdorferi sensu stricto (including genetic diversity), B. afzelii, B. garinii (several serotypes) and additional species isolated in different parts of the world (Rudenko et al., ; Ogden et al., ). Some of these species have been isolated in symptomatic patients (Varela et al., ; Lopes de Carvalho et al., ; Rudenko et al., ; Stanek and Reiter, ; Branda and Rosenberg, ; Clark et al., ; Lee et al.,; Margos et al., ). B. spielmanii may cause early skin disease (Stanek and Reiter, ). B. bavariensis, B. bisettii, B. valaisiana, B. americana, B. andersonii, B. lonestari and more recently B. kurtenbachii have been isolated from patients with Lyme-like diseases (Varela et al., ; Rudenko et al.,; Rizzoli et al., ; Stanek and Reiter, ; Clark et al., ). The pathogenic role of B. lusitaniae, isolated in a case of vasculitis, remains to be substantiated (Rudenko et al., ). Despite such diversity in strains, most of the commercially available tests still rely on the original 1982 Massachusetts B31 isolate of B. burgdorferi. No diagnostic tool is available for routine detection of B. miyamotoi (Branda and Rosenberg,; Lee et al., ). Coinfections with other microbes add to the complexity of these illnesses (Table (Table1).1). Among patients with early Lyme disease in the USA, 2–12% were found to also have human granulocytic anaplasmosis, and 2–40% babesiosis (Wormser et al., ). In Brazil, a Lyme-like syndrome, due to the tickAmblyomma, has been described and mobile non cultivable spirochetes could be visualized in patients' blood using a dark field microscope (Mantovani et al., ). A new tick-borne bacterial pathogen, CandidatusNeoehrlichia mikurensis, was reported in Switzerland (Fehr et al., ). An illustration of the limits of serology is the Scottish example: the sensitivity of the immunoblot was improved by using local Scottish strains of Borrelia (Mavin et al., ).

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.
To read Prof Perronne full paper

Saturday, 3 September 2016


Published on Aug 25, 2016
Green fluorescent Borrelia interacting with the walls of a skin postcapillary venule of a live mice after injection of bacteria into the blood stream. Blood vessel walls are stained with a red dye. Posted by lab of Tara Moriarty ( From: Norman et al 2008:

Fascinating research from the Moriatry Lab which has just published the following paper

Biomechanics of Borrelia burgdorferi Vascular Interactions

Rhodaba Ebady, Alexandra F. Niddam Anna E. Boczula Yae Ram Kim Nupur Gupta Tian Tian Tang Tanya Odisho

Hui Zhi Craig A. Simmons Jon T. Skare Tara J. Moriarty


  • Biomechanical characterization of bacterial-vascular interactions
  • Identified catch bond-mediated bacterial vascular adhesion mechanism
  • Conserved vascular interaction strategies in bacteria and host cells


Systemic dissemination of microbes is critical for progression of many infectious diseases and is associated with most mortality due to bacterial infection. The physical mechanisms mediating a key dissemination step, bacterial association with vascular endothelia in blood vessels, remain unknown. Here, we show that endothelial interactions of the Lyme disease spirochete Borrelia burgdorferi under physiological shear stress mechanistically resemble selectin-dependent leukocyte rolling etc... ................................................

These findings provide insight into the biomechanics of bacterial-vascular interactions and demonstrate that disseminating bacteria and circulating host immune cells share widely conserved mechanisms for interacting with endothelia under physiological shear stress.
To read the paper go to 
This paper is causing a great stir in the media rocketting the paper up to -
  • top 2% of all research papers of same age
  • top 4% of Cell Reports papers of same age
  • top 3% of all Cell Reports papers ever!!!!!
with a growing list of media interest  that can be read from

David Michael Conner  explains so well in the Huffington Post why this paper is being read so widely.
For more information and videos visit the Moriatry Lab website

Friday, 2 September 2016


Ticks found on 'one third' of dogs, researchers say

Tuesday, 9 August 2016


My early morning walk in 'Tick Land' prior to being interviewed on BBC Surrey about Tick awareness. I am well booted and covered and Meg wears her tick collar which so far has been successful.