Archive for the ‘Borrelia Miyamotoi (Relapsing Fever Group)’ Category

2018 Review of Previous Pathogen Transmission Time Studies in Deer Ticks

https://www.ncbi.nlm.nih.gov/pubmed/29398603

2018 Mar;9(3):535-542. doi: 10.1016/j.ttbdis.2018.01.002. Epub 2018 Jan 31.

Pathogen transmission in relation to duration of attachment by Ixodes scapularis ticks.

Abstract

The blacklegged tick, Ixodes scapularis, is the primary vector to humans in the eastern United States of the deer tick virus lineage of Powassan virus (Powassan virus disease); the protozoan parasite Babesia microti (babesiosis); and multiple bacterial disease agents including Anaplasma phagocytophilum (anaplasmosis), Borrelia burgdorferi and Borrelia mayonii (Lyme disease), Borrelia miyamotoi (relapsing fever-like illness, named Borrelia miyamotoi disease), and Ehrlichia muris eauclairensis (a minor causative agent of ehrlichiosis).

With the notable exception of Powassan virus, which can be transmitted within minutes after attachment by an infected tick, there is no doubt that the risk of transmission of other I. scapularis-borne pathogens, including Lyme disease spirochetes, increases with the length of time (number of days) infected ticks are allowed to remain attached. This review summarizes data from experimental transmission studies to reinforce the important disease-prevention message that regular (at least daily) tick checks and prompt tick removal has strong potential to reduce the risk of transmission of I. scapularis-borne bacterial and parasitic pathogens from infected attached ticks.

The most likely scenario for human exposure to an I. scapularis-borne pathogen is the bite by a single infected tick. However, recent reviews have failed to make a clear distinction between data based on transmission studies where experimental hosts were fed upon by a single versus multiple infected ticks. A summary of data from experimental studies on transmission of Lyme disease spirochetes (Bo. burgdorferi and Bo. mayonii) by I. scapularis nymphs indicates that the probability of transmission resulting in host infection, at time points from 24 to 72 h after nymphal attachment, is higher when multiple infected ticks feed together as compared to feeding by a single infected tick.

In the specific context of risk for human infection, the most relevant experimental studies therefore are those where the probability of pathogen transmission at a given point in time after attachment was determined using a single infected tick. The minimum duration of attachment by single infected I. scapularis nymphs required for transmission to result in host infection is poorly defined for most pathogens, but experimental studies have shown that Powassan virus can be transmitted within 15 min of tick attachment and both A. phagocytophilum and Bo. miyamotoi within the first 24 h of attachment. There is no experimental evidence for transmission of Lyme disease spirochetes by single infected I. scapularis nymphs to result in host infection when ticks are attached for only 24 h (despite exposure of nearly 90 experimental rodent hosts across multiple studies) but the probability of transmission resulting in host infection appears to increase to approximately 10% by 48 h and reach 70% by 72 h for Bo. burgdorferi. Caveats to the results from experimental transmission studies, including specific circumstances (such as re-attachment of previously partially fed infected ticks) that may lead to more rapid transmission are discussed.

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**Comment**

There are a number of problematic issues with this study:

  1. This is a review of previous studies.  There is nothing NEW here.  
  2. It’s important to note that ticks typically carry more than just borrelia and transmission times have not taken this fact into account: https://madisonarealymesupportgroup.com/2017/05/01/co-infection-of-ticks-the-rule-rather-than-the-exception/ and https://www.lymedisease.org/lyme-basics/co-infections/about-co-infections/  Infection with more than one pathogen is associated with more severe illness.https://madisonarealymesupportgroup.com/2018/10/30/study-shows-lyme-msids-patients-infected-with-many-pathogens-and-explains-why-we-are-so-sick/  For the first time, Garg et al. show a 85% probability for multiple infections including not only tick-borne pathogens but also opportunistic microbes such as EBV and other viruses.  This is a BIG DEAL.  Finally, a study showing what we face as patients in the real world.  They also never take into account nematodes (worms), mycoplasma, tularemia, and/or Bartonella.  These are infections many if not most patients have to contend with.  Some have been bioweaponized.
  3. They assume that the most likely scenario is for a person to be bitten by one tick.  Assuming makes an ass out of u and me.  When you take into account the latest information on the Asian tick, you quickly realize the probability of coming into contact with hundreds if not thousands of ticks at one time:  https://madisonarealymesupportgroup.com/2018/09/12/three-surprising-things-i-learned-about-asian-longhorned-ticks-the-tick-guy-tom-mather/  While human infection has yet to be found in the U.S., this tick is responsible for plenty of misery in Asia:  https://madisonarealymesupportgroup.com/2018/06/12/first-longhorned-tick-confirmed-in-arkansas/  It spreads SFTS (sever fever with thrombocytopenia syndrome), “an emerging hemorrhagic fever,” but the potential impact of this tick on tickborne illness is not yet known. In other parts of the world, it has been associated with several tickborne diseases, such as spotted fever rickettsioses, Anaplasma, Ehrlichia, and Borrelia, the causative agent of Lyme Disease.
  4. While they discuss the probability of multiple tick attachment, they never discuss the issue of partially fed ticks, where spirochetes would be in the salivary glands – leading to quicker transmission: http://iai.asm.org/content/61/6/2396.full.pdf  Ticks can spontaneously detach – and the authors of this study found that they did so 15% of the time in mice.  They also state that about a tenth of questing nymphs appear distended with partially fed sub-adult ticks being common.
  5. While the current review states, “There is no experimental evidence for transmission of Lyme disease spirochetes by single infected I. scapularis nymphs to result in host infection when ticks are attached for only 24 h (despite exposure of nearly 90 experimental rodent hosts across multiple studies), this study shows transmission can occur in under 16 hours:  https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4278789/
  6. https://madisonarealymesupportgroup.com/2017/04/14/transmission-time-for-lymemsids-infection/  Within this video, microbiologist Holly Ahern discusses the numerous problems with animal Bb transmission studies.  Transmission Time:  Only one study done on Mice. At 24 hours every tick had transmitted borrelia to the mice; however, animal studies have proven that transmission can occur in under 16 hours and it occurs frequently in under 24 hours.  No human studies have been done and https://www.dovepress.com/lyme-borreliosis-a-review-of-data-on-transmission-time-after-tick-atta-peer-reviewed-article-IJGM  no studies have determined the minimum time it takes for transmission.  And, never forget the case of the little girl who couldn’t walk or talk after a tick bite attachment of 4-6 hours:  https://madisonarealymesupportgroup.com/2016/12/07/igenex-presentation/
  7. They continue to blame Lyme/MSIDS on the black legged tick as the sole perp when experience and studies show there’s more potential transmitters at play:  https://madisonarealymesupportgroup.com/2018/11/07/are-mosquitoes-transmitting-lyme-disease/https://madisonarealymesupportgroup.com/2016/07/23/german-study-finds-borrelia-in-mosquitos/https://madisonarealymesupportgroup.com/2019/01/17/remember-deer-keds-study-shows-bartonella-causing-deer-ked-dermatitis-in-humans/
Please, quit doing reviews of previous data and do something new using better laboratory techniques!  We don’t need MORE of the same thing.

Relapsing Fever Found at Popular Recreation Site in CA Ticks

https://www.ncbi.nlm.nih.gov/pubmed/30517684

2018 Dec 4. doi: 10.1093/jme/tjy213. [Epub ahead of print]

Borrelia parkeri in Ornithodoros parkeri (Ixodida: Argasidae) Collected Using Compact Dry Ice Traps in Madera County, California.

Abstract

Tick-borne relapsing fever (TBRF) is a potentially serious vector-borne disease endemic to the western United States. Vector surveillance is compromised by the nidicolous life history of the three Ornithodoros species that transmit TBRF to people in this region. Large-scale stationary trapping methods were developed to survey a wide geographical range of Ornithodoros spp. which are known to vector relapsing fever Borrelia spp. in California. Ninety-six Ornithodoros parkeri were collected from four locations in the foothills of Fresno and Madera Counties. Two of these O. parkeri nymphs were PCR positive for Borrelia parkeri, and their collection at a popular recreation site increases the public health concern.

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More on Relapsing Fever:  https://igenex.com/ticktalk/2018/01/11/borreliosis-relapsing-fever-disease/

Relapsing Fever Spirochete Uniquely Adapted to Highly Oxidative Salivary Glands of Soft-bodied Tick

https://www.ncbi.nlm.nih.gov/pubmed/30489694

2018 Nov 29:e12987. doi: 10.1111/cmi.12987. [Epub ahead of print]

The relapsing fever spirochete Borrelia turicatae persists in the highly oxidative environment of its soft-bodied tick vector.

Abstract

The relapsing fever spirochete Borrelia turicatae possesses a complex life cycle in its soft-bodied tick vector, Ornithodoros turicata. Spirochetes enter the tick midgut during a bloodmeal, and during the following weeks spirochetes disseminate throughout O. turicata. A population persists in the salivary glands allowing for rapid transmission to mammalian hosts during tick feeding. Little is known about the physiological environment within the salivary glands acini in which B. turicatae persists. In this study, we examined the salivary gland transcriptome of O. turicata ticks and detected the expression of fifty-seven genes involved in oxidant metabolism or antioxidant defenses. We confirmed the expression of five of the most highly expressed genes including glutathione peroxidase (gpx), thioredoxin peroxidase (tpx), manganese superoxide dismutase (sod-1), copper-zinc superoxide dismutase (sod-2), and catalase (cat) by reverse-transcriptase droplet digital PCR (RT-ddPCR). We also found distinct differences in the expression of these genes when comparing the salivary glands and midguts of unfed O. turicata ticks.

Our results indicate that the salivary glands of unfed O. turicata nymphs are a highly oxidative environment where reactive oxygen species (ROS) predominate, while midgut tissues comprise a primarily nitrosative environment where nitric oxide synthase is highly expressed. Additionally, B. turicatae was found to be hyperresistant to ROS compared to the Lyme disease spirochete B. burgdorferi, suggesting that it is uniquely adapted to the highly oxidative environment of O. turicata salivary gland acini.

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**Comment**

Much can be learned about Borrelia turicatae by reading this case study:  https://wwwnc.cdc.gov/eid/article/23/5/16-2069_article

We learn:

  • Ornithodoros turicata soft bodied ticks, are endemic to Texas and Florida
  • They are found in caves and ground squirrel or prairie dog burrows  https://madisonarealymesupportgroup.com/2018/04/23/tick-borne-relapsing-fever-found-in-austin-texas-caves/
  • Once infected, they remain infected for the rest of their lives, which can be up to ten years.
  • Attachment is painless
  • They are rapid night feeders (5-60min)
  • Due to their rapid feeding they are rarely found or leave lesions
  • Patient in study suffered with headache, nausea, & pain behind knees
  • Had numerous lesions which resolved after 6 days (without treatment)
  • Developed persistent fever
  • Developed thrombocytopenia (low platelets)
  • Developed elevated Erythrocyte sedimentation rate & C-reactive protein
  • Improved rapidly with doxycycline
  • Platelet count normalized within 2 weeks
  • Asymptomatic soldiers with similar exposure were treated prophylactically
  • TBRF is a neglected and probably underdiagnosed disease
  • Published cases in Texas have been supported by serology for the TBRF group, exposure location, and tick collections, but the authors state successful identification of B. turicatae in a human has not been reported
  • Military training groups in Israel have declared certain caves off limits because of heavy tick presence https://madisonarealymesupportgroup.com/2017/10/27/israeli-kids-get-lyme-disease-from-ticks-in-caves/ and have prophylactically administered doxycycline to those suspected to have been exposed
  • Asymptomatic patients given doxy don’t have a Jarisch-Herxheimer reaction but those with active illness do
Another study demonstrating the wily and adaptable nature of spirochetes.

 

 

 

 

Tick-borne Relapsing Fever as a Potential Veterinary Medical Problem

https://www.ncbi.nlm.nih.gov/m/pubmed/29943903/

Tick-borne relapsing fever as a potential veterinary medical problem.

Elelu N. Vet Med Sci. 2018.

Abstract

Tick-borne relapsing fever (TBRF) caused by the bacteria Borrelia, is poorly documented in veterinary medicine. Given the widespread presence of the soft tick vectors – Ornithodoros and the recently discovered hard tick vectors, as well as their close association with animal hosts, it is highly likely that infection occurs, but is rarely reported to be of veterinary importance. Sporadic reports of canine infection, some being fatal through to probable cause of abortion in horses have been published. Some of these pathogens exist in regions where there are limited diagnostic facilities, hence, they are likely to be missed and their impact on productivity may be unquantified. Here we review available literatures on cases of TBRF in domestic and wild animals in order to show their potential veterinary medical impact. Future efforts using field and laboratory surveys are needed to determine pathogenesis, vector competence and distribution in animals, their impact on animal health and productivity as well as to prevent further spill to the human population, where it is already a public health problem in some parts of the world.

 

 

Five Genera of Pathogens Found in Ticks On Russian Dogs

https://www.ncbi.nlm.nih.gov/m/pubmed/30428925/

Dog survey in Russian veterinary hospitals: tick identification and molecular detection of tick-borne pathogens.

Livanova NN, et al. Parasit Vectors. 2018.

Abstract

BACKGROUND: Species of Canidae in Russia can be infested with up to 24 different tick species; however, the frequency of different tick species infesting domestic dogs across Russia is not known. In addition, tick-borne disease risks for domestic dogs in Russia are not well quantified. The goal of this study was to conduct a nationwide survey of ticks collected from infested dogs admitted to veterinary clinics in Russian cities and to identify pathogens found in these ticks.

METHODS: Ticks feeding on dogs admitted to 32 veterinary clinics in 27 major cities across Russia were preserved in ethanol and submitted to a central facility for examination. After identification, each tick was evaluated for infection with known tick-borne pathogens using PCR.

RESULTS: There were 990 individual ticks collected from 636 dogs. All collected ticks belonged to the Ixodidae (hard ticks) and represented 11 species of four genera, Dermacentor, Ixodes, Rhipicephalus and Haemaphysalis. Four most common tick species were D. reticulatus, followed by I. persulcatus, I. ricinus and R. sanguineus. Ixodes persulcatus ticks were found to be infected with 10 different pathogens, and ticks of this species were more frequently infected than either D. reticulatus or I. ricinus. Ixodes persulcatus females were also more frequently co-infected with two or more pathogens than any other tick.

Pathogenic species of five genera were detected in ticks:

  • Anaplasma centrale, A. phagocytophilum & A. marginale (Anaplasma)
  • Babesia canis, B. microti, B. venatorum, B. divergens, B. crassa & B. vogeli (Babesia)
  • Borrelia miyamotoi, B. afzelii and B. garinii (Borrelia)
  • Ehrlichia muris, E. canis and E. ruminantu (Ehrlichia)
  • Theileria cervi (Theileria – a parasitic protozoan)
Anaplasma marginale, E. canis, B. crassa, B. vogeli and T. cervi were detected in I. persulcatus, and Babesia canis in D. marginatum, for the first time in Russia.

CONCLUSIONS: Multiple ticks from four genera and 11 species of the family Ixodidae were collected from domestic dogs across Russia. These ticks commonly carry pathogens and act as disease vectors. Ixodes persulcatus ticks present the greatest risk for transmission of multiple arthropod-borne pathogens.

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**Comment**

It’s getting harder and harder for The Cabal to hide the polymicrobial nature of Lyme/MSIDS.  The data just keeps pouring in:  https://madisonarealymesupportgroup.com/2018/10/30/study-shows-lyme-msids-patients-infected-with-many-pathogens-and-explains-why-we-are-so-sick/

For the first time, Garg et al. show a 85% probability for multiple infections including not only tick-borne pathogens but also opportunistic microbes such as EBV and other viruses.

I’m thankful they included Bartonella as that one is often omitted but definitely a player. I’m also thankful for the mention of viruses as they too are in the mix. The mention of the persister form must be recognized as well as many out there deny its existence.

Key Quote: “Our findings recognize that microbial infections in patients suffering from TBDs do not follow the one microbe, one disease Germ Theory as 65% of the TBD patients produce immune responses to various microbes.”

But there is another important point.

According to this review, 83% of all commercial tests focus only on Lyme (borrelia), despite the fact we are infected with more than one microbe.

And those tests miss half of all cases:  

https://madisonarealymesupportgroup.com/2018/09/12/lyme-testing-problems-solutions/  ...with the C6 Elisa its around 50% sensitive (in the context of the two tiered testing system on its own it has a sensitivity of 75%) because it misses about half of true positive cases….The Western Blot also has many problems with sensitivity at all stages but especially within the first month and again later on the more chronic it becomes.If you take the terrible sensitivity of both tests in the two tiered system you will start to see how testing positive consecutively on both is very unlikely, mathematically improbable and biologically almost impossible unless you are in the HLA autoimmune group which is comparatively rare.

https://madisonarealymesupportgroup.com/2018/01/16/2-tier-lyme-testing-missed-85-7-of-patients-milford-hospital/  Dr. Sin Lee identifying faulty serology tests for Lyme disease in 85.7% of the walk-in patients in the Emergency Room of Milford Hospital.

Please note that all the studies showing the polymicrobial nature of tick borne illness  are foreign.

The Cabal has everyone in the U.S. in a head-lock.

Will the real researchers please stand up and be counted?

 

 

 

 

 

 

 

 

 

Highlights from LDA/Columbia Lyme Disease Conference

https://globallymealliance.org/highlights-from-lda-columbia-lyme-disease-conference/

lyme disease conference

October 31, 2018

Highlights from LDA/Columbia Lyme Disease Conference

Highlights from LDA/Columbia 19th Annual Scientific Conference Lyme & Tick-Borne Diseases: Turning the Corner Through Research

Global Lyme Alliance (GLA) recently attended a conference jointly organized by Lyme Disease Association, Inc. and Columbia University. The meeting featured speakers from around the country who gave updates about diverse subjects relevant to Lyme and tick-borne diseases.

Dr. Claudia Molins from the Centers for Disease Control and Prevention (CDC) discussed her work on finding metabolic signatures for Lyme disease. These are byproducts of metabolism that are specific to Lyme patients, and since they are unique to different diseases, identifying these signatures might be useful as a diagnosis of Lyme disease. Even more compelling, they may yield clues about which subsets of patients are unlikely to be cured by first-line antibiotic therapy.

Another speaker, Dr. William Robinson of Stanford University, spoke about his collaboration with Dr. John Aucott of Johns Hopkins University, who is funded by GLA. Together, they work on samples from the landmark SLICE study, which deciphers immunological data from patients in various stages of Lyme disease. Dr. Robinson’s talk focused on antibody production in Lyme disease. These can be both protective against the pathogen, as well as directed against the host, that may help to explain both the failure to protect against the microbe as well as to harm the patient, painting a complex picture of the immune response against Borrelia. He also discussed how these may correlate with disease stage.

A potential new therapeutic drug candidate for Lyme disease was presented by Dr. Kim Lewis, a GLA grant recipient. His group has rediscovered hygromycin A, a previously known drug that was never screened for activity against B. burgdorferi, which it kills without any effect on other bacteria. Thus, it may potentially be developed as a treatment for Lyme disease that will not harm the commensal human microbiome.

Dr. Utpal Pal, Mark Blackman, and Dr. Chrysoula Kitsou

 

 

 

 

 

 

 

 

From the University of Maryland, Dr. Utpal Pal, who is on the GLA scientific advisory board, the conference heard of his findings about BBA57, a Borrelia gene that is critical for the initial infectivity of the bacteria into a mammalian host, via various strategies to block the host immune system. Curiously, mutations in BBA57 only transiently stop bacterial infection, and bacteria that rebound from the initial delay are able to persist in the host, attesting to the difficulty of bacterial eradication.

Dr. Shannon Delaney from Columbia University spoke about patients who suffer from symptoms that are similar to, but subtly different from Lyme disease. Her study reveals that infection by Borrelia miyamotoi, related to B. burgdorferi, may be responsible for these illnesses, which are missed by standard Lyme disease testing.

Finally, Dr. Anne Louise Oaklander from Massachusetts General Hospital discussed small-fiber peripheral neuropathy, disorders of the nervous system that may result from hyperinflammation in peripheral sites, leading in some cases to chronic pain and other symptoms. Characterization and documentation of these disorders has begun, and greater understanding of the underlying mechanism may reveal whether they are linked to persisting symptoms attributed to tick-borne infections. The conference overall spread a wealth of knowledge and inspired new research ideas.

Understanding Lyme and tick-borne infections is enormously complex. That’s why it’s so important for the scientific community to collaborate. Only through collaboration can we find the answers that will allow us to treat and, ultimately, cure those suffering from tick-borne diseases.

 

 

 

Study Shows Lyme/MSIDS Patients Infected With Many Pathogens and Explains Why We Are So Sick

https://www.nature.com/articles/s41598-018-34393-9?fbclid=IwAR3k-zPy2rJu8OuFl3HHqJ0twLPJvQrxiIUALUs0T-BuuJ50_1VQVwcflIQ (Please see comment at end of article)

Evaluating polymicrobial immune responses in patients suffering from tick-borne diseases

Kunal Garg, Leena Meriläinen, Ole Franz, Heidi Pirttinen, Marco Quevedo-Diaz, Stephen Croucher & Leona Gilbert
Scientific Reportsvolume 8, Article number: 15932 (2018)   https://doi.org/10.1038/s41598-018-34393-9

Abstract
There is insufficient evidence to support screening of various tick-borne diseases (TBD) related microbes alongside Borrelia in patients suffering from TBD. To evaluate the involvement of multiple microbial immune responses in patients experiencing TBD we utilized enzyme-linked immunosorbent assay. Four hundred and thirty-two human serum samples organized into seven categories followed Centers for Disease Control and Prevention two-tier Lyme disease (LD) diagnosis guidelines and Infectious Disease Society of America guidelines for post-treatment Lyme disease syndrome. All patient categories were tested for their immunoglobulin M (IgM) and G (IgG) responses against 20 microbes associated with TBD. Our findings recognize that microbial infections in patients suffering from TBDs do not follow the one microbe, one disease Germ Theory as 65% of the TBD patients produce immune responses to various microbes. We have established a causal association between TBD patients and TBD associated co-infections and essential opportunistic microbes following Bradford Hill’s criteria. This study indicated an 85% probability that a randomly selected TBD patient will respond to Borrelia and other related TBD microbes rather than to Borrelia alone.

A paradigm shift is required in current healthcare policies to diagnose TBD so that patients can get tested and treated even for opportunistic infections.
Please see link for full article.  Snippets below:

Introduction
Tick-borne diseases (TBDs) have become a global public health challenge and will affect over 35% of the global population by 20501. The most common tick-borne bacteria are from the Borrelia burgdorferi sensu lato (s.l.) group. However, ticks can also transmit co-infections like Babesia spp.2, Bartonella spp.3, Brucella spp.4,5,6,7,8, Ehrlichia spp.9, Rickettsia spp.10,11, and tick-borne encephalitis virus12,13,14. In Europe and North America, 4–60% of patients with Lyme disease (LD) were co-infected with Babesia, Anaplasma, or Rickettsia11,15,16. Evidence from mouse and human studies indicate that pathogenesis by various tick-borne associated microbes15,16,17 may cause immune dysfunction and alter, enhance the severity, or suppress the course of infection due to the increased microbial burden18,19,20,21,22. As a consequence of extensive exposure to tick-borne infections15,16,17, patients may develop a weakened immune system22,23, and present evidence of opportunistic infections such as Chlamydia spp.24,25,26,27, Coxsackievirus28, Cytomegalovirus29, Epstein-Barr virus27,29, Human parvovirus B1924, and Mycoplasma spp.30,31. In addition to tick-borne co-infections and non-tick-borne opportunistic infections, pleomorphic Borrelia persistent forms may induce distinct immune responses in patients by having different antigenic properties compared to typical spirochetes32,33,34,35. Nonetheless, current LD diagnostic tools do not include Borrelia persistent forms, tick-borne co-infections, and non-tick-borne opportunistic infections.

The two-tier guidelines36,37,38 for diagnosing LD by the Centers for Disease Control and Prevention (CDC) have been challenged due to the omission of co-infections and non-tick-borne opportunistic infections crucial for comprehensive diagnosis and treatment39,40. Emerging diagnostic solutions have demonstrated the usefulness of multiplex assays to test for LD and tick-borne co-infections41,42. However, these new technologies do not address seroprevalence of non-tick-borne opportunistic infections in patients suffering from TBD and they are limited to certain co-infections41,42. Non-tick-borne opportunistic microbes can manifest an array of symptoms24,29 concerning the heart, kidney, musculoskeletal, and the central nervous system as seen in patients with Lyme related carditis43, nephritis44, arthritis45, and neuropathy46, respectively. Therefore, Chlamydia spp., Coxsackievirus, Cytomegalovirus, Epstein-Barr virus, Human parvovirus B19, Mycoplasma spp., and other non-tick-borne opportunistic microbes play an important role in the differential diagnosis of LD24,29. As the current knowledge regarding non-tick-borne opportunistic microbes is limited to their use in differential diagnosis of LD, it is unclear if LD patients can present both tick-borne co-infections and non-tick-borne opportunistic infections simultaneously.

For the first time, we evaluate the involvement of Borrelia spirochetes, Borrelia persistent forms, tick-borne co-infections, and non-tick-borne opportunistic microbes together in patients suffering from different stages of TBD. To highlight the need for multiplex TBD assays in clinical laboratories, we utilized the Bradford Hill’s causal inference criteria47 to elucidate the likelihood and plausibility of TBD patients responding to multiple microbes rather than one microbe. The goal of this study is to advocate screening for various TBD microbes including non-tick-borne opportunistic microbes to decrease the rate of misdiagnosed or undiagnosed48 cases thereby increasing the health-related quality of life for the patients39, and ultimately influencing new treatment protocol for TBDs.

Results
Positive IgM and IgG responses by CDC defined acute, CDC late, CDC negative, PTLDS immunocompromised, and unspecific patients to 20 microbes associated with TBD (Fig. 1) were utilized to evaluate polymicrobial infections (Figs 2–4). Patient categories included CDC acute (n = 43), CDC late (n = 43), CDC negative (n = 46), PTLDS (n = 31), immunocompromised (n = 61), unspecific (n = 31), and healthy (n = 177).

Polymicrobial infections are present at all stages of tick-borne diseases.

Microbes include Borrelia burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia burgdorferi sensu stricto persistent form, Borrelia afzelii persistent form, Borrelia garinii persistent form, Babesia microti, Bartonella henselae, Brucella abortus, Ehrlichia chaffeensis, Rickettsia akari, Tick-borne encephalitis virus (TBEV), Chlamydia pneumoniae, Chlamydia trachomatis, Coxsackievirus A16 (CVA16), Cytomegalovirus (CMV), Epstein-Barr virus (EBV), Mycoplasma pneumoniae, Mycoplasma fermentans, and Human parvovirus B19 (HB19V).

In Fig. 2A, 51% and 65% of patients had IgM and IgG responses to more than one microbe, whereas 9% and 16% of patients had IgM and IgG responses to only one microbe, respectively. Immune responses to Borrelia persistent forms (all three species) for IgM and IgG were 5–10% higher compared to Borrelia spirochetes in all three species (Fig. 2B). Interestingly, the probability that a randomly selected patient will respond to Borrelia persistent forms rather than the Borrelia spirochetes (Fig. S2) is 80% (d = 1.2) for IgM and 68% for IgG (d = 0.7). Figure 2A and B indicated that IgM and IgG responses by patients from different stages of TBDs are not limited to only Borrelia spirochetes.

In Fig. 3 sub-inlets, more than 50% of the patients reacted to only the individual Borrelia strains suggesting that Borrelia antigens are not cross-reactive. If patients were cross-reacting among antigens, a larger percentage of the patients would be seen with the combination of all three species (Fig. S2). These results provide evidence to suggest that the inclusion of different Borrelia species and their morphologies in current LD diagnostic tools will improve its efficiency.

Discussions
The study outcome indicated that polymicrobial infections existed at all stages of TBD with IgM and IgG responses to several microbes (Fig. 2). Results presented in this study propose that infections in patients suffering from TBDs do not obey the one microbe one disease Germ Theory. Based on these results and substantial literature11,15,16,17,27,49,50,51 on polymicrobial infections in TBD patients, we examined the probability of a causal relationship between TBD patients and polymicrobial infections following Hill’s nine criteria47.

An average effect size of d = 1.5 for IgM and IgG (Fig. 4A) responses is considered very large52. According to common language effect size statistics53, d = 1.5 indicates 85% probability that a randomly selected patient will respond to Borrelia and other TBD microbes rather than to only Borrelia. Reports from countries such as Australia27, Germany49, Netherlands11, Sweden50, the United Kingdom51, the USA15,16, and others indicate that 4% to 60% of patients suffer from LD and other microbes such as Babesia microti and human granulocytic anaplasmosis (HGA). However, previous findings11,15,16,27,49,50,51 are limited to co-infections (i.e., Babesia, Bartonella, Ehrlichia, or Rickettsia species) in patients experiencing a particular stage of LD (such as Erythema migrans). In contrast, a broader spectrum of persistent, co-infections, and opportunistic infections associated with diverse stages of TBD patients have been demonstrated in this study (Fig. 2). From a clinical standpoint, the likelihood for IgM and IgG immune responses by TBD patients to the Borrelia spirochetes versus the Borrelia persistent forms, and responses to just Borrelia versus Borrelia with many other TBD microbes has been quantified for the first time (Fig. S2).

Borrelia pathogenesis could predispose individuals to polymicrobial infections because it can suppress, subvert, or modulate the host’s immune system18,19,20,21,22 to create a niche for colonization by other microbes54. Evidence in animals55 and humans11,15,16,27,49,50,51 frequently indicate co-existence of Borrelia with other TBD associated infections. Interestingly, IgM and IgG immune levels by patients to multiple forms of Borrelia resulted in immune responses to 14 other TBD microbes (Fig. 4B). In contrast, patient responses to either form of Borrelia (spirochetes or persistent forms) resulted in reactions to an average of 8 other TBD microbes (Fig. 4B). Reaction to two forms of Borrelia reflected an increase in disease severity indicating biological gradient for causation as required by Hill’s criteria47.

Multiple microbial infections in TBD patients seem plausible because ticks can carry more than eight different microbes depending on tick species and geography56,57. Moreover, Qiu and colleagues reported the presence of at least 18 bacterial genera shared among three different tick species and up to 127 bacterial genera in Ixodes persulcatus58. Interestingly, research indicates Chlamydia-like organism in Ixodes ricinus ticks and human skin59 that may explain immune responses to Chlamydia spp., seen in this study (Fig. 2). Additionally, prevalence of TBD associated co-infections such as B. abortus, E. chaffeensis, and opportunistic microbes such as C. pneumoniae, C. trachomatis, Cytomegalovirus, Epstein-Barr virus, and M. pneumoniae have been recorded in the general population of Europe and the USA (Table S2). However, true incidence of these microbes is likely to be higher considering underreporting due to asymptomatic infections and differences in diagnostic practices and surveillance systems across Europe and in the USA. More importantly, clinical evidence for multiple microbes has been reported in humans11,15,16,27,49,50,51, and livestock55 to mention the least. Our findings regarding the presence of polymicrobial infections at all stages of TBD further supports the causal relationship between TBD patients and polymicrobial infections (Fig. 2). Various microbial infections in TBD patients have been linked to the reduced health-related quality of life (HRQoL) and increased disease severity39.

An association between multiple infections and TBD patients relates well to other diseases such as periodontal, and respiratory tract diseases. Oral cavities may contain viruses and 500 different bacterial species60. Our findings demonstrate that TBD patients may suffer from multiple bacterial and viral infections (Fig. 4). In respiratory tract diseases, influenza virus can stimulate immunosuppression and predispose patients to bacterial infections causing an increase in disease severity61. Likewise, Borrelia can induce immunosuppression that may predispose patients to other microbial infections causing an increase in disease severity.

Traditionally, positive IgM immune reaction implies an acute infection, and IgG response portrays a dissemination, persistent or memory immunity due to past infections. Depending on when TBD patients seek medical advice, the level of anti-Borrelia antibodies can greatly vary as an Erythema migrans (EM) develops and may present with IgM, IgG, collective IgM/IgG, or IgA62. This study recommends both IgM and IgG in diagnosing TBD (Figs 5 and S4–S6) as unconventional antibody profiles have been portrayed in TBD patients. Presence of long-term IgM and IgG antibodies have been reported in LD patients that were tested by the CDC two-tier system. In 2001, Kalish and colleagues reported anti-Borrelia IgM or IgG persistence in patients that suffered from LD 10–20 years ago63. Similarly, Hilton and co-workers recorded persistent anti-Borrelia IgM response in 97% of late LD patients that were considered cured following an antibiotic treatment64.

Similar events of persistent IgM and IgG antibody reactions were demonstrated in patients treated for Borrelia arthritis and acrodermatitis chronica atrophicans65, chronic cutaneous borreliosis66, and Lyme neuroborreliosis67. A clear phenomenon of immune dysfunction is occurring, which might account for the disparities in LD patient’s antibody profiles and persistence. Borrelia suppresses the immune system by inhibition of antigen-induced lymphocyte proliferation18, reducing Langerhans cells by downregulation of major histocompatibility complex class II molecules on these cells19, stimulating the production of interleukin-10 and anti-inflammatory immunosuppressive cytokine20, and causing disparity in regulation and secretion of cytokines21. Other studies have demonstrated low production or subversion of specific anti-Borrelia antibodies in patients with immune deficiency status22.

In the USA alone, the economic healthcare burden for patients suffering from LD and ongoing symptoms is estimated to be $1.3 billion per year69. Additionally, 83% of all TBD diagnostic tests performed by the commercial laboratories in the USA accounted for only LD70. Globally, the commercial laboratories’ ability to diagnose LD has increased by merely 4% (weighted mean for ELISA sensitivity 62.3%) in the last 20 years71. This study provides evidence regarding polymicrobial infections in patients suffering from different stages of TBDs. Literature analyses and results from this study followed Hill’s criteria indicating a causal association between TBD patients and polymicrobial infections. Also, the study outcomes indicate that patients may not adhere to traditional IgM and IgG responses.

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**Comment**

For the first time, Garg et al. show a 85% probability for multiple infections including not only tick-borne pathogens but also opportunistic microbes such as EBV and other viruses.

I’m thankful they included Bartonella as that one is often omitted but definitely a player.  I’m also thankful for the mention of viruses as they too are in the mix.  The mention of the persister form must be recognized as well as many out there deny its existence.

Key Quote:  Our findings recognize that microbial infections in patients suffering from TBDs do not follow the one microbe, one disease Germ Theory as 65% of the TBD patients produce immune responses to various microbes.”

But there is another important point.

According to this review, 83% of all commercial tests focus only on Lyme (borrelia), despite the fact we are infected with more than one microbe.  The review also states it takes 11 different visits to 11 different doctors, utilizing 11 different tests to be properly diagnosed.  https://www.news-medical.net/news/20181101/Tick-borne-disease-is-multiple-microbial-in-nature.aspx?

This is huge.  Please spread the word.