Outer surface protein E (OspE) mediates Borrelia burgdorferi sensu stricto strain-specific complement evasion in the eastern fence lizard, Sceloporus undulatus.

In North America, Lyme disease is primarily caused by the spirochetal bacterium Borrelia burgdorferi sensu stricto (Bb), which is transmitted between multiple vertebrate hosts and ixodid ticks, and is a model commonly used to study host-pathogen interactions. While Bb is consistently observed in its mammalian and avian reservoirs, the bacterium is rarely isolated from North American reptiles. Two closely related lizard species, the eastern fence lizard (Sceloporus undulatus) and the western fence lizard (Sceloporus occidentalis), are examples of reptiles parasitized by Ixodes ticks. Vertebrates are known to generate complement as an innate defense mechanism, which can be activated before Bb disseminate to distal tissues. Complement from western fence lizards has proven lethal against one Bb strain, implying the role of complement in making those lizards unable to serve as hosts to Bb. However, Bb DNA is occasionally identified in distal tissues of field-collected eastern fence lizards, suggesting some Bb strains may overcome complement-mediated clearance in these lizards. These findings raise questions regarding the role of complement and its impact on Bb interactions with North American lizards. In this study, we found Bb seropositivity in a small population of wild-caught eastern fence lizards and observed Bb strain-specific survivability in lizard sera. We also found that a Bb outer surface protein, OspE, from Bb strains viable in sera, promotes lizard serum survivability and binds to a complement inhibitor, factor H, from eastern fence lizards. Our data thus identify bacterial and host determinants of eastern fence lizard complement evasion, providing insights into the role of complement influencing Bb interactions with North American lizards.

Extensive Diversity of RNA Viruses in Australian Ticks.

Understanding the microbiome of ticks in Australia is of considerable interest given the ongoing debate over whether Lyme disease and its causative agent, the bacterium Borrelia burgdorferisensu lato, are present in Australia. The diversity of bacteria infecting Australian ticks has been studied using both culture- and metagenomics-based techniques. However, little is known about the virome of Australian ticks, including whether this includes viruses with the potential to infect mammals. We used a meta-transcriptomics approach to reveal the diversity and evolution of viruses from Australian ticks collected from two locations on the central east coast of Australia, including metropolitan Sydney. From this we identified 19 novel RNA viruses belonging to 12 families, as well as 1 previously described RNA virus. The majority of these viruses were related to arthropod-associated viruses, suggesting that they do not utilize mammalian hosts. However, two novel viruses discovered in ticks feeding on bandicoot marsupials clustered closely within the mammal-associated hepacivirus and pestivirus groups (family Flaviviridae). Another bandicoot tick yielded a novel coltivirus (family Reoviridae), a group of largely tick-associated viruses containing the known human pathogen Colorado tick fever virus and its relative, Eyach virus. Importantly, our transcriptomic data provided no evidence for the presence of B. burgdorferisensu lato in any tick sample, providing further evidence against the presence of Lyme disease in Australia. In sum, this study reveals that Australian ticks harbor a diverse virome, including some viruses that merit additional screening in the context of emerging infectious disease.IMPORTANCE Each year a growing number of individuals along the east coast of Australia experience debilitating disease following tick bites. As there is no evidence for the presence of the causative agent of Lyme disease, Borrelia burgdorferisensu lato, in Australian ticks, the etiological basis of this disease syndrome remains controversial. To characterize the viruses associated with Australian ticks, particularly those that might be associated with mammalian infection, we performed unbiased RNA sequencing on 146 ticks collected across two locations along the coast of New South Wales, Australia. This revealed 19 novel RNA viruses from a diverse set of families. Notably, three of these viruses clustered with known mammalian viruses, including a novel coltivirus that was related to the human pathogen Colorado tick fever virus.

[Clinical presentation of Ixodes tick-borne borreliosis caused by Borrelia miyamotoi in the context of an immune response to the pathogen]. / Klinicheskie proiavleniia iksodovogo kleshchevogo borrelioza, vyzvannogo Borrelia miyamotoi, v kontekste immunnogo otveta na vozbuditel'.

Ixodes tick-borne borreliosis caused by Borrelia miyamotoi (ITBB-BM) is a previously unknown infectious disease discovered in Russia. AIM: The present study continues the investigation of the clinical features of ITBB-BM in the context of an immune system-pathogen interaction. SUBJECTS AND METHODS: The study enrolled 117 patients with ITBB-BM and a comparison group of 71 patients with Lyme disease (LD) that is ITBB with erythema migrans. All the patients were treated at the New Hospital, Yekateringburg. More than 100 clinical, epidemiological and laboratory parameters were obtained from each patient's medical history and included in the general database. A subset of patients hospitalized in 2015 and 2016 underwent additional laboratory examinations. Namely, the levels of B. miyamotoi-specific IgM and IgG antibodies were measured by the protein microarray containing GlpQ protein and four variable major proteins (VMPs): Vlp15/16, Vlp18, Vsp1, and Vlp5. The blood concentration of Borrelia was estimated by quantitative real-time PCR. RESULTS: In contrast to LD, first of all (p<0.001) the following clinical features were typical for ITBB-BM: the absence of erythema migrans (in 95% of patients), fever (93%), fatigue (96%), headache (82%), chill (41%), nausea (28%), lymphopenia (56%), thrombocytopenia (46%), the abnormal levels of alanine aminotransferase (54%) and C-reactive protein (98%), proteinuria (61%). Given the set of these indicators, the course of ITBB-BM was more severe in approximately 70% of patients. At admission, only 13% and 38% of patients had antibodies to GlpQ and VMPs, respectively; at discharge, antibodies to GlpQ and VMPs were detected in 88% of patients. There was no statistically significant association of the antibody response with individual clinical manifestations and laboratory parameters of the disease. However, patients with more severe ITBB-BM produced less IgM antibodies to VMPs and GlpQ at the time of discharge. CONCLUSION: ITBB-BM is a moderate systemic disease accompanied by the production of specific antibodies in virtually all patients.

[EPIDEMIOLOGIC SITUATION BY NATURAL-FOCI INFECTIONS IN THE CRIMEA FEDERAL DISTRICT IN 2014-2015].

AIM: Analysis of epidemic manifestations of natural-foci infections (NFI), clarification of spectrum of their causative agents, determination of epizootic activity of natural foci in the Crimea Federal District (KFD). MATERIALS AND METHODS: Epizootologic examination of 10 administrative districts of KDF was carried out. 291 pools (2705 specimens) of ixodes ticks and 283 samples of organs of small mammals were studied by PCRmethod for the presence of DNA/RNA of causative agents of a number of NFI. RESULTS: Morbidity by NFI in KFD was registered by 6 nosologies: Lyme borreliosis, Marseilles fever, leptospirosis; tularemia, intestine yersiniosis and tick-borne viral encephalitis, wherein, transmissive infections made up 91.6%. Circulation of causative agents of Crimea hemorrhagic fever, Q fever, group of tick-borne spotted fever, Lyme borreliosis, human granulocytic anaplasmosis, human monocytic ehrlichiosis, hemorrhagic fever with renal syndrome, West Nile fever, tularemia and leptospirosis was established. CONCLUSION: Due to activity of natural foci of NFI further monitoring of epidemiologic and epizootologic manifestations of these infections in the Crimea, including using genetic methods of analysis, is necessary for ensuring sanitary-epidemiologic welfare of KFD population.

Detecting Lyme disease using antibody-functionalized single-walled carbon nanotube transistors.

We examined the potential of antibody-functionalized single-walled carbon nanotube (SWNT) field-effect transistors (FETs) to use as a fast and accurate sensor for a Lyme disease antigen. Biosensors were fabricated on oxidized silicon wafers using chemical vapor deposition grown carbon nanotubes that were functionalized using diazonium salts. Attachment of Borrelia burgdorferi (Lyme) flagellar antibodies to the nanotubes was verified by atomic force microscopy and electronic measurements. A reproducible shift in the turn-off voltage of the semiconducting SWNT FETs was seen upon incubation with B. burgdorferi flagellar antigen, indicative of the nanotube FET being locally gated by the residues of flagellar protein bound to the antibody. This sensor effectively detected antigen in buffer at concentrations as low as 1 ng/ml, and the response varied strongly over a concentration range coinciding with levels of clinical interest. Generalizable binding chemistry gives this biosensing platform the potential to be expanded to monitor other relevant antigens, enabling a multiple vector sensor for Lyme disease. The speed and sensitivity of this biosensor make it an ideal candidate for development as a medical diagnostic test.

[The use of electron microscopy and virus isolation in the diagnosis of aseptic neuroinfections]. / Vyuzití elektronové mikroskopie a izolace viru v diagnostice aseptických neuroinfekcí.

OBJECTIVE: In aseptic neuroinfections, the etiology is usually known in 50-70% of cases. The aim was to increase the rates using electron microscopy (EM) and virus isolation in cell cultures. MATERIAL AND METHODS: The prospective study included 34 patients with aseptic neuroinfections hospitalized at the Department of Infectious Diseases in Ostrava fromJuly to November 2012. EM examined cerebrospinal fluid of all patients and virus isolation in tissue cultures was performed in all cerebrospinal fluid samples. Cerebrospinal fluid was examined by polymerase chain reaction for enteroviruses in 30 patients and for herpes simplex virus 1 and 2 in 29 patients. Detection of antibodies against Borrelia burgdorferi and tick-borne encephalitis was performed in all 34 patients. RESULTS: Possible etiological agents were discovered in 31 out of 34 patients (91%), with one agent being found in 23 patients (68%) and two agents being detected in 8 patients (24%). EM revealed the agents in 26 patients and virus isolation was successful in 10 patients. EM was the only method to identify 10 agents. A group of 23 patients with a single agent detected included 14 patients with enteroviral meningitis, 4 patients with Lyme borreliosis and 4 patients with tick-borne encephalitis; EM detected an undefined virus in the last patient. An unusual group of 8 patients with two agents detected comprised 5 patients with enteroviruses and spirochetes, 2 patients with tick-borne encephalitis and undefined viruses and 1 patient with a spirochete and an undetermined virus. CONCLUSION: EM can aid in explaining the etiology of aseptic neuroinfections. However, the clinical interpretation of results remains problematic, such as detection of unknown viruses or two possible agents in 8 out of 34 patients.

Use of tick surveys and serosurveys to evaluate pet dogs as a sentinel species for emerging Lyme disease.

OBJECTIVE: To evaluate dogs as a sentinel species for emergence of Lyme disease in a region undergoing invasion by Ixodes scapularis. SAMPLE POPULATION: 353 serum samples and 78 ticks obtained from dogs brought to 18 veterinary clinics located in the lower peninsula of Michigan from July 15, 2005, through August 15, 2005. PROCEDURES: Serum samples were evaluated for specific antibodies against Borrelia burgdorferi by use of 3 serologic assays. Ticks from dogs were subjected to PCR assays for detection of pathogens. RESULTS: Of 353 serum samples from dogs in 18 counties in 2005, only 2 (0.6%) contained western blot analysis-confirmed antibodies against B burgdorferi. Ten of 13 dogs with I scapularis were from clinics within or immediately adjacent to the known tick invasion zone. Six of 18 I scapularis and 12 of 60 noncompetent vector ticks were infected with B burgdorferi. No ticks were infected with Anaplasma phagocytophilum, and 3 were infected with Babesia spp. CONCLUSIONS AND CLINICAL RELEVANCE: Serosurvey in dogs was found to be ineffective in tracking early invasion dynamics of I scapularis in this area. Tick chemoprophylaxis likely reduces serosurvey sensitivity in dogs. Ticks infected with B burgdorferi were more common and widely dispersed than seropositive dogs. In areas of low tick density, use of dogs as a source of ticks is preferable to serosurvey for surveillance of emerging Lyme disease. IMPACT FOR HUMAN MEDICINE: By retaining ticks from dogs for identification and pathogen testing, veterinarians can play an important role in early detection in areas with increasing risk of Lyme disease.

Risk maps for range expansion of the Lyme disease vector, Ixodes scapularis, in Canada now and with climate change.

BACKGROUND: Lyme disease is the commonest vector-borne zoonosis in the temperate world, and an emerging infectious disease in Canada due to expansion of the geographic range of the tick vector Ixodes scapularis. Studies suggest that climate change will accelerate Lyme disease emergence by enhancing climatic suitability for I. scapularis. Risk maps will help to meet the public health challenge of Lyme disease by allowing targeting of surveillance and intervention activities. RESULTS: A risk map for possible Lyme endemicity was created using a simple risk algorithm for occurrence of I. scapularis populations. The algorithm was calculated for each census sub-division in central and eastern Canada from interpolated output of a temperature-driven simulation model of I. scapularis populations and an index of tick immigration. The latter was calculated from estimates of tick dispersion distances by migratory birds and recent knowledge of the current geographic range of endemic I. scapularis populations. The index of tick immigration closely predicted passive surveillance data on I. scapularis occurrence, and the risk algorithm was a significant predictor of the occurrence of I. scapularis populations in a prospective field study. Risk maps for I. scapularis occurrence in Canada under future projected climate (in the 2020s, 2050s and 2080s) were produced using temperature output from the Canadian Coupled Global Climate Model 2 with greenhouse gas emission scenario enforcing 'A2' of the Intergovernmental Panel on Climate Change. CONCLUSION: We have prepared risk maps for the occurrence of I. scapularis in eastern and central Canada under current and future projected climate. Validation of the risk maps provides some confidence that they provide a useful first step in predicting the occurrence of I. scapularis populations, and directing public health objectives in minimizing risk from Lyme disease. Further field studies are needed, however, to continue validation and refinement of the risk maps.

[Exposure to ticks and erythema chronicum migrans among borreliosis patients in Lower Silesia]. / Ekspozycja na uklucia przez kleszcze a wystepowanie rumienia wedrujacego u pacjentów z borelioza z lyme na terenie Dolnego Slaska.

The aim of the study was the analysis of Lyme borreliosis cases notified by Clinic for Infections Diseases in Wroclaw in connection with exposure to ticks in year 2002. In the analyzed group of patients tick bites were confirmed in 77.9% and the most common places of exposition were forest regions in Lower Silesia (Masyw Slezy, Wzgórza Twardógorskie, Kotlina Klodzka, Wzgórza Trzebnickie, Bory Dolnoslaskie) and in Wroclaw area. EM appeared in 55.8% cases. In cases of multiple ticks professionally exposured persons EM appeared only in 22.2%.

Problems in the study and prophylaxis of mixed infections transmitted by ixodid ticks.

The spread of mixed infections with natural focality transmitted by ixodid ticks is a normal phenomenon attributable to trends in the relationships of different pathogens in the vector organism and ecosystem as a whole. Any disease developing as a result of tick bite should be regarded as a potentially mixed infection. Clinically, tick-borne mixed infections proceed more severely than the corresponding diseases caused by a single agent. The residual course of the disease may sometimes be accounted for by the persistence of two or even several pathogens. This implies the necessity of a comprehensive approach to the study, diagnosis, treatment, management and prophylaxis of infections belonging to this group.

Pathogen-tick-host interactions: Borrelia burgdorferi and TBE virus.

Both tick-borne encephalitis (TBE) virus and Borrelia burgdorferi depend for their survival on a tick-borne transmission cycle involving diverse vertebrate hosts and ixodid (primarily Ixodes) tick species. Increasing evidence strongly suggests that a key factor in successful transmission is the ability of these pathogens to exploit the pharmacological properties of their vector tick's saliva. Proteins and other chemicals, secreted in tick saliva, control the vertebrate hosts' haemostatic, inflammatory and immune responses in order to facilitate blood-feeding. Such bioactive saliva molecules include immunoglobulin-binding proteins, histamine-binding proteins, natural killer cell and interferon regulators, and complement inhibitors. By unravelling the contents of tick saliva and understanding their interactions with host and pathogen, we may identify new strategies for disease control.

Serologic evidence for tick-borne pathogens other than Borrelia burgdorferi (TOBB) in Lyme borreliosis patients from midwestern Germany.

The seroprevalence of antibodies against the human granulocytic ehrlichiosis agent (HGE) and Babesia microti was retrospectively determined in 76 Lyme borreliosis patients and in 44 asymptomatic individuals with a positive borreliosis serology, in comparison to 100 healthy blood donors from the Rhein-Main area. Additionally, seroreactivity for tick-borne encephalitis virus (TBEV) was investigated. For antibody detection, commercially available immunofluorescence assays (MRL Diagnostics, USA) and a TBEV-ELISA (Immuno, Germany) were used. In the control group, the positivity rate for anti-Borrelia burgdorferi (IgG/IgM) and anti-Babesia microti-antibodies in the population of the Rhein-Main area (Midwestern Germany) may be estimated at 15% and 8%, respectively. Examination for both HGE and TBEV demonstrated seroreactivity (IgG) in 1% of tested individuals. Specific anti-HGE IgG and/or IgM antibodies were more often discovered in cases of early Borrelia infection (stage I: 13.6%, stage II: 18.4%) than in patients with stage III disease (0%) or in seropositive but asymptomatic patients (6.8%). Investigation for TBEV revealed seroreactivity for IgG in 13% of these cases. No TBEV-IgM was found. Interestingly, the prevalence of anti-HGE and anti-TBEV antibodies among Lyme borreliosis patients and seropositive patients without active Lyme disease symptoms was significantly higher than that in the control group of healthy blood donors (p < 0.05). Likewise, antibody titers reflecting a recent infection with Babesia microti could be demonstrated more often in patients with Lyme borreliosis stage I or II (p < 0.05). Analysis of 50 samples from patients with florid or recent syphilis infection revealed no crossreactivity between Babesia microti, HGE and Treponema pallidum. Our findings suggest that concomitant or serial infection due to TOBB may be common in tick exposed patients from the Rhein-Main area and in European countries in general. Hence, in addition to TBEV, human babesiosis and HGE should always be considered by European physicians in the differential diagnosis of acute febrile illness following a tick bite.