Candidatus Neoehrlichia Mikurensis Research Articles

Molecular detection of pathogens from ticks collected from dogs and cats at veterinary clinics in Finland

BackgroundTicks carry microbes, some of which are pathogenic for humans and animals. To assess this One Health challenge, 342 ticks were collected from pet dogs and cats at 10 veterinary clinics in Finland as part of the European project “Protect Our Future Too”.MethodsThe tick species were identified, and ticks were screened with quantitative PCR (qPCR) for tick-borne pathogens, including Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Ehrlichia canis, Anaplasma spp., Candidatus Neoehrlichia mikurensis, tick-borne encephalitis virus (TBEV), and Babesia spp. For comparison, a subset of tick DNA (20 qPCR-positive samples) was analysed with 16S next-generation sequencing (NGS).ResultsMost ticks were Ixodes ricinus (289, 84.5%), followed by Ixodes persulcatus (51, 14.9%). One hybrid tick (I. ricinus/I. persulcatus, 0.3%) and one Rhipicephalus sanguineus tick (0.3%) were identified. We found one or more of the analysed pathogens in 17% (59/342) of the ticks. The most prevalent pathogen was B. burgdorferi s.l. (36, 10.5%), followed by Anaplasma phagocytophilum (12, 3.5%), B. miyamotoi (5, 1.5%), Babesia venatorum (4, 1.2%), and TBEV (1, 0.3%). Candidatus Neoehrlichia mikurensis DNA was amplified from three (0.9%) ticks. Ehrlichia canis was not detected. In the 16S NGS, six samples produced enough reads for the analysis. In these six samples, we confirmed all the positive qPCR findings of Borrelia spp. and Ca. N. mikurensis.ConclusionsThe high prevalence of pathogenic microorganisms in the ticks of this study emphasizes the importance of awareness of ticks and tick-borne diseases and prevention. Furthermore, the results show that veterinary surveillance can facilitate early detection of tick-borne pathogens and new tick species and draw attention to possible co-infections that should be considered both in symptomatic humans and animals after tick bites.Graphical

Circulation of four species of Anaplasmataceae bacteria in ticks in Harbin, northeastern China

Ticks play an important role in the evolution and transmission of Anaplasmataceae bacteria which are agents of emerging infectious diseases. In this study, a total of 1286 adult ticks belonging to five species were collected from cattle, goats, horses and vegetation in Harbin area, Heilongjiang province, northeastern China. The tick-borne Anaplasmataceae bacteria were identified by amplifying and sequencing the 16S rRNA (rrs) and heat shock protein-60 encoding (groEL) genes. The results showed that Ixodes persulcatus was dominant (38.8%, 499/1283) among the five tick species, and Anaplasmataceae bacteria were detected in all tick species with an overall prevalence of 7.4%. Four species of Anaplasmataceae bacteria (Anaplasma phagocytophilum, Anaplasma ovis, Anaplasma bovis, and “Candidatus Neoehrlichia mikurensis”), which are pathogenic to humans and/or animals, were identified from tick samples by phylogenetic analyzes of the rrs and groEL gene sequences. Interestingly, the cluster 1 strains were first identified in Asian, and a novel cluster was also detected in this study. These data revealed the genetic diversity of Anaplasmataceae bacteria circulating in ticks in Harbin area, highlighting the need to investigate these tick-borne pathogens and their risks to human and animal health.

Candidatus Neoehrlichia mikurensis and Hepatozoon sp. in voles (Microtus spp.): occurrence and evidence for vertical transmission

Candidatus Neoehrlichia mikurensis (CNM) and Hepatozoon spp. are important vector-borne parasites of humans and animals. CNM is a relatively recently discovered pathogen of humans. Hepatozoon are parasites of reptiles, amphibians and mammals, commonly found in rodents and carnivores worldwide. The present study aimed to determine the prevalence of CNM and Hepatozoon spp. in three species of Microtus and to assess the occurrence of vertical transmission in naturally-infected voles. Molecular techniques were used to detect pathogen DNA in blood and tissue samples of captured voles and their offspring. The prevalence of CNM in the vole community ranged 24–47% depending on Microtus species. The DNA of CNM was detected in 21% of pups from three litters of six infected Microtus dams (two Microtus arvalis and one M. oeconomus) and in 3/45 embryos (6.6%) from two litters of eight CNM-infected pregnant females. We detected Hepatozoon infection in 14% of M. arvalis and 9% of M. oeconomus voles. Hepatozoon sp. DNA was detected in 48.7% of pups from seven litters (6 M. arvalis and 1 M. oeconomus) and in two embryos (14.3%) obtained from one M. arvalis litter. The high prevalence of CNM infections in the Microtus spp. community may be a result of a relatively high rate of vertical transmission among naturally infected voles. Vertical transmission was also demonstrated for Hepatozoon sp. in M. arvalis and M. oeconomus. Our study underlines the significance of alternative routes of transmission of important vector-borne pathogens.

Bacteria and protozoa with pathogenic potential in Ixodes ricinus ticks in Viennese recreational areas.

Ixodes ricinus is the most relevant vector for tick-borne diseases in Austria and responsible for the transmission of Borrelia burgdorferi sensu lato (s. l.), which causes Lyme borreliosis in humans; however, also other bacteria and protozoa can be found in ticks and have the potential of infecting people and animals. In this study we collected ticks in popular recreational areas in the city of Vienna in the years 2019 and 2020 and analyzed them for the presence of such putative pathogenic microorganisms. By using reverse line blot (RLB) hybridization we detected DNA of B. burgdorferi s. l., Rickettsia spp., Babesia spp., Candidatus Neoehrlichia mikurensis (CNM) and Anaplasma phagocytophilum. Moreover, we also screened them for the relapsing fever spirochete Borrelia miyamotoi employing real-time PCR. The most frequently detected pathogens were B. burgdorferi s. l. in 28.6% of the ticks in 2019 and 21.3% of the ticks in 2020. The genus Rickettsia was detected in 13.8% of the ticks from 2019 and only in 4.6% from 2020. Babesia spp. were detected in 5.7% in 2019 and 4.2% in 2020. Furthermore, we detected CNM in 4.0% (2019) and 5.6% (2020), A. phagocytophilum in 0.5% (2019) and 1.3% (2020) and finally B. miyamotoi in 3.3% (2019) and 1.7% (2020). Collectively, we show that various microorganisms are prevalent in ticks collected in Vienna and identify hotspots for B. miyamotoi, which we have detected for the first time in the city.

Neoehrlichiosis: A Case Study of the Tick-Borne Pathogen That Can Cause Thromboembolic Events.

Candidatus Neoehrlichia mikurensis is a gram-negative bacterium carried and spread by Ixodes ricinus ticks often found in Europe and Asia. It causes a disease process called neoehrlichiosis, which can result in vasculitis and thromboembolic events. This pathogen does not grow in typical culture medium, and most laboratories do not carry the specific polymerase chain reaction (PCR) test needed to detect neoehrlichiosis. If a patient presents to an emergency department complaining of a recent tick bite and symptoms of a deep vein thrombosis or pulmonary embolism, consider that these symptoms may be related. Treat the tick bite with doxycycline for 3 weeks and manage the thromboembolic event according to standard treatment of care.

Novel Protozoans in Austria Revealed through the Use of Dogs as Sentinels for Ticks and Tick-Borne Pathogens.

We previously isolated and cultivated the novel Rickettsia raoultii strain Jongejan. This prompted us to ask whether this strain is unique or more widely present in Austria. To assess this issue, we retrospectively screened ticks collected from dogs in 2008. Of these collected ticks, we randomly selected 75 (47 females and 28 males) Dermacentor reticulatus, 44 (21 females, 7 males, and 16 nymphs) Haemaphysalis concinna, and 55 (52 females and 3 males) ticks of the Ixodes ricinus complex. Subsequently, these ticks were individually screened for the presence of tick-borne pathogens using the reverse line blot hybridization assay. In our current study, we detected DNA from the following microbes in D. reticulatus: Anaplasma phagocytophilum, Borrelia lusitaniae, Borrelia spielmanii, Borrelia valaisiana, and R. raoultii, all of which were R. raoultii strain Jongejan. In H. concinna, we found DNA of a Babesia sp., Rickettsia helvetica, and an organism closely related to Theileria capreoli. Lastly, I. ricinus was positive for Anaplasma phagocytophilum, Borrelia afzelii, Borrelia burgdorferi sensu stricto, Borrelia garinii/Borrelia bavariensis, B. lusitaniae, B. spielmanii, B. valaisiana, Candidatus Neoehrlichia mikurensis, Rickettsia helvetica, Rickettsia monacensis, and Theileria (Babesia) microti DNA. The detection of DNA of the Babesia sp. and an organism closely related to Theileria capreoli, both found in H. concinna ticks, is novel for Austria.

Anaplasmosis in Poland – analysis of 120 patients

The aim of our study was to clarify the clinical picture of anaplasmosis through analysis of the symptoms and clinical signs presented by infected patients in a cohort of tick-bitten individuals. The study included 1375 patients with suspicion of tick-borne disease. Finally, 120 patients (8.7%) were diagnosed with anaplasmosis (HGA). Blood samples were examined by PCR for Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis, Borrelia burgdorferi sensu lato, Babesia spp., and Bartonella spp.. Based on analysis of 120 patients with HGA we concluded that anaplasmosis is not as rare in Europe, as it is thought to be and often appears as a co-infection with other tick-borne pathogens. The co-infection rate of patients with A. phagocytophilum infection in tick endemic areas is high.Co-infection of A. phagocytophilum with B. burgdorferi s.l. or tick-borne encephalitis virus may influence symptom frequency. PCR together with medical history, clinical picture and basic laboratory tests is a sufficient method for the diagnosis of anaplasmosis. Doxycycline is an effective drug leading to complete recovery.

Infections with Tickborne Pathogens after Tick Bite, Austria, 2015-2018.

Abstract The aim of this prospective study was to assess the risk for tickborne infections after a tick bite. A total of 489 persons bitten by 1,295 ticks were assessed for occurrence of infections with Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia spp., Babesia spp., Candidatus Neoehrlichia mikurensis, and relapsing fever borreliae. B. burgdorferi s.l. infection was found in 25 (5.1%) participants, of whom 15 had erythema migrans. Eleven (2.3%) participants were positive by PCR for Candidatus N. mikurensis. One asymptomatic participant infected with B. miyamotoi was identified. Full engorgement of the tick (odds ratio 9.52) and confirmation of B. burgdorferi s.l. in the tick by PCR (odds ratio 4.39) increased the risk for infection. Rickettsia helvetica was highly abundant in ticks but not pathogenic to humans. Knowledge about the outcome of tick bites is crucial because infections with emerging pathogens might be underestimated because of limited laboratory facilities.

A brief review: the prevalence of tick-borne pathogens in urban and suburban areas

Ticks are widely distributed blood-sucking ectoparasites and vectors for numerous zoonotic pathogens that cause infectious diseases in humans and animals. The increase in the incidence of tick-borne diseases (TBD) is partially associated with climatic changes, such as shorter and warmer winters, prolonged growing seasons, and also with increasing urbanisation. In recent decades, a rising number of established populations of medically important ticks have been reported in urban and suburban areas such as city parks or suburban forests over many regions in Europe. The transformation of natural ecosystems into urban areas becomes actual significant problem because it could affect the circulation of tick-borne pathogens and increase the risk of infection for humans and domestic animals. Tick-borne pathogens, including Borrelia burgdorferi s. l., Rickettsia spp., Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis, and Babesia spp., have been detected in urban tick populations in Europe. Such places as parks, leisure-time areas, green spaces, and gardens become endemic zones of tick-borne pathogens. This review describes the investigations on the prevalence of tick-borne pathogens in urbanised areas conducted in Europe during the last fifteen years (2005–2020).

Landscape epidemiology of neglected tick-borne pathogens in central Europe.

Studies of tick-borne diseases (TBDs) in Europe focus on pathogens with principal medical importance (e.g. Lyme disease and tick-borne encephalitis), but we have limited epidemiological information on the neglected pathogens, such as the members of the genera Anaplasma, Rickettsia, Babesia and Candidatus Neoehrlichia mikurensis. Here, we integrated an extensive field sampling, laboratory analysis and GIS models to provide first publicly available information on pathogen diversity, prevalence and infection risk for four overlooked zoonotic TBDs in the Czech Republic. In addition, we assessed the effect of landscape variables on the abundance of questing ticks at different spatial scales and examined whether pathogen prevalence increased with tick density. Our data from 13,340 ticks collected in 142 municipalities showed that A.phagocytophilum (MIR=3.5%) and Ca. Neoehrlichia mikurensis (MIR=4.0%) pose geographically uneven risks with localized hotspots, while Rickettsia (MIR=4.9%) and Babesia (MIR=1.1%) had relatively homogeneous spatial distribution. Landscape variables had significant effect on tick abundance up to the scale of 1km around the sampling sites. Questing ticks responded positively to landscape diversity and configuration, especially to forest patch density that strongly correlates with the amount of woodland-grassland ecotones. For all four pathogens, we found higher prevalence in places with higher densities of ticks, confirming the hypothesis that tick abundance amplifies the risk of TB infection. Our findings highlight the importance of landscape parameters for tick vectors, likely due to their effect on small vertebrates as reservoir hosts. Future studies should explicitly investigate the combined effect of landscape parameters and the composition and population dynamics of hosts on the host-vector-pathogen system.

Tick-borne Pathogens Detected in the Blood of Immunosuppressed Norwegian Patients Living in a Tick-endemic Area.

The knowledge regarding the occurrence and the clinical implications of tick-borne infections in immunosuppressed patients living in tick-endemic areas is limited. Adult patients with autoimmune conditions requiring immunosuppressive treatment such as infliximab and rituximab were invited to participate in the study when they attended the hospital for treatment and/or control of the disease. Whole-blood samples were analyzed by real-time polymerase chain reaction for Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Anaplasma phagocytophilum, Rickettsia spp., Candidatus Neoehrlichia mikurensis, and Babesia spp. The occurrence of tick-borne pathogens in the blood of patients (n = 163) with autoimmune conditions requiring immunosuppressive treatment was evaluated. Pathogen DNA was detected in 8.6% (14/163) of the patients. The predominant pathogen was Ca. Neoehrlichia mikurensis (12/14), which was carried in the blood of infected patients for 10-59 days until treatment with doxycycline. B. burgdorferi s.l. and Rickettsia spp. were detected in 1 patient each. The B. burgdorferi-infected patient presented with fever, whereas the remaining patients were judged to have subclinical infections. B. miyamotoi, A. phagocytophilum, and Babesia spp. were not detected in any patient. Patients treated with biologicals and living in a tick-endemic area seem to have a high risk of contracting Ca. Neoehrlichia mikurensis infection, which, if left untreated, could result in thromboembolic complications.

Monitoring of ticks and tick-borne pathogens through a nationwide research station network in Finland

In 2015 a long-term, nationwide tick and tick-borne pathogen (TBP) monitoring project was started by the Finnish Tick Project and the Finnish Research Station network (RESTAT), with the goal of producing temporally and geographically extensive data regarding exophilic ticks in Finland. In the current study, we present results from the first four years of this collaboration.Ticks were collected by cloth dragging from 11 research stations across Finland in May–September 2015–2018 (2012–2018 in Seili). Collected ticks were screened for twelve different pathogens by qPCR: Borrelia afzelii, Borrelia garinii, Borrelia valaisiana, Borrelia burgdorferi sensu stricto, Borrelia miyamotoi, Babesia spp., Anaplasma phagocytophilum, Rickettsia spp., Candidatus Neoehrlichia mikurensis, Francisella tularensis, Bartonella spp. and tick-borne encephalitis virus (TBEV).Altogether 15 067 Ixodes ricinus and 46 Ixodes persulcatus were collected during 68 km of dragging. Field collections revealed different seasonal activity patterns for the two species. The activity of I. persulcatus adults (only one nymph detected) was unimodal, with activity only in May–July, whereas Ixodes ricinus was active from May to September, with activity peaks in September (nymphs) or July–August (adults). Overall, tick densities were higher during the latter years of the study. Borrelia burgdorferi sensu lato were the most common pathogens detected, with 48.9 ± 8.4% (95% Cl) of adults and 25.3 ± 4.4% of nymphs carrying the bacteria. No samples positive for F. tularensis, Bartonella or TBEV were detected.This collaboration project involving the extensive Finnish Research Station network has ensured enduring and spatially extensive, long-term tick data collection to the foreseeable future.

Comparison of tick-borne pathogen prevalence in Ixodes ricinus ticks collected in urban areas of Europe

Tick-borne diseases are a major threat to human and animal health. An increasing number of natural habitats have been transformed into urban areas by human activity; hence, the number of reported tick bites in urban and suburban areas has risen. This retrospective analysis evaluated 53 scientific reports concerning infections of Ixodes ricinus ticks collected from urban and suburban areas of Europe between 1991 and 2017. The results indicate significant differences in many variables, including a higher number of Anaplasma phagocytophilum infections in Eastern Europe than in Western Europe. The opposite result was observed for Candidatus Neoehrlichia mikurensis infections. A comparison of climate zones revealed that Borrelia burgdorferi s.l. infections have the greatest median incidence rate in subtropical climate zones. No statistical significance was found when comparing other tick-borne pathogens (TBPs), such as Borrelia miyamotoi, Rickettsia spp., Babesia spp., Bartonella spp., Ehrlichia spp., Coxiella burnetii and Francisella tularensis. The analysis also showed significant differences in the overall prevalence of TBPs according to average temperatures and rainfall across Europe. This retrospective study contributes to the knowledge on the occurrence and prevalence of TBPs in urbanized areas of Europe and their dependence on the habitats and geographical distributions of ticks. Due to the increased risk of tick bites, it is of great importance to investigate infections in ticks from urban and suburban areas.

Long-term persistence of Anaplasma phagocytophilum and Ehrlichia muris in wild rodents

Wild animals are reservoir hosts for a number of tick-transmitted agents, and long-term persistence of the agents is a key factor for their effective transmission from animal hosts to ticks. To study the persistence of Anaplasmataceae in rodents, 59 adult Myodes spp. voles (M. rutilus, M. rufocanus, and M. glareolus) were captured in Omsk Province, Asian Russia, freed from all ectoparasites, and kept in individual cages. Their blood samples were regularly analyzed for the presence of DNA of Anaplasma phagocytophilum, Ehrlichia muris, and “Candidatus Neoehrlichia mikurensis”. Anaplasma phagocytophilum, E. muris, and mixed infections were found in 29 (49 %), 17 (29 %), and 14 (24 %) voles, respectively. DNA of “Candidatus N. mikurensis” was not identified. Long-term persistence of A. phagocytophilum and E. muris in naturally infected Myodes spp. was shown for the first time. Thus, 12 animals with A. phagocytophilum were found infected for more than three months and four of them carried the bacterium throughout 22–55 weeks of observation. Four voles were infected with E. muris for 26–40 weeks. Fragments of groESL operon were sequenced for genotyping A. phagocytophilum and E. muris. In voles with A. phagocytophilum, the determined sequences belonged to two phylogenetic clusters. Most sequences were identical or closely related to those found in small mammals and Ixodes trianguliceps ticks from Western Siberia and European countries; one sequence was previously identified in rodents and Ixodes persulcatus and Ixodes pavlovskyi ticks from Asian Russia. Sequences of groESL fragments from E. muris were close to those determined previously in small mammals and I. persulcatus ticks from Siberia. In addition, a new groESL sequence of E. muris was identified in one vole and the sequence substantially differed from all known corresponding E. muris sequences (≥ 14 mismatches).

Assessment of Anaplasma phagocytophilum presence in early Lyme borreliosis manifested by erythema migrans skin lesions

BackgroundTo investigate to what extent early Lyme borreliosis patients with erythema migrans are infected with Anaplasma phagocytophilum. MethodsThree hundred ten patients from Poland with erythema migrans were included in the study. One hundred and eighty-three patients (59%) agreed to have both skin biopsy and blood samples analysed for Borrelia burgdorferi, A. phagocytophilum and ‘Candidatus Neoehrlichia mikurensis’, with PCR. Positive samples were confirmed with sequencing. ResultsB. burgdorferi DNA was detected in 49.7% of the skin samples and in 1.1% of the blood samples. A. phagocytophilum DNA was found in 7.1% blood samples, and in 8.2% of the skin biopsies. In four patients, A. phagocytophilum DNA was detected only in blood; in one case A. phagocytophilum DNA was found simultaneously in blood and skin, and additionally in this patients’ blood Borrelia DNA was detected. In four skin samples B. burgdorferi DNA was detected simultaneously with A. phagocytophilum DNA, indicative of a co-infection. ConclusionsA. phagocytophilum may be present in early Lyme borreliosis characterized by erythema migrans and should always be considered as a differential diagnostic following a tick bite and considered in treatment schemes, as these differs (in early stage of Lyme borreliosis doxycycline, amoxicillin, cefuroxime axetil and azithromycin are recommended, while in anaplasmosis the most effective courses of treatment are doxycycline, rifampin and levofloxacin). Consequently, the role of A. phagocytophilum in erythema migrans should be further studied.

Candidatus Neoehrlichia mikurensis is widespread in questing Ixodes ricinus ticks in the Czech Republic

Candidatus Neoehrlichia mikurensis, the causative agent of tick-borne “neoehrlichiosis” has recently been reported in humans, mammals and ticks in Europe. The aim of this study was to map the distribution of this bacterium in questing ticks in the Czech Republic. A total of 13,325 Ixodes ricinus including 445 larvae, 5270 nymphs and 7610 adults were collected from vegetation by flagging in 140 Czech towns and villages from every region of the Czech Republic. The ticks were pooled into 2665 groups of 5 individuals respecting life stage or sex and tested for the presence of Ca. Neoehrlichia mikurensis by conventional PCR targeting of the groEL gene. The bacterium was detected in 533/2665 pools and 125/140 areas screened, showing an overall estimated prevalence of 4.4 % in ticks of all life stages. Phylogenetic analysis revealed only small genetic diversity among the strains found. Two pools of questing larvae tested positive, suggesting transovarial transmission. According to this study, Ca. Neoehrlichia mikurensis is another tick-borne pathogen widespread in I. ricinus ticks in the Czech Republic.

Monitoring of vector-borne diseases in the west part of Ukraine

During recent years, infectious and invasive animal diseases caused by viruses, bacteria, and protozoa and transmitted by ticks have been a new problem in medical and veterinary practice. Many of these diseases are zoonoses and lead to the disability and mortality of humans and animals. Ixodes ricinus and Dermacentor reticulatus are species of ticks that are commonly attack animals and humans in Europe. These ticks are spread throughout the EU and are involved in the transmission of a large number of vector-borne diseases. To date, relatively limited data on the circulation of tick-borne diseases and their transmission in western Ukraine are presented. The purpose of this study was to monitor the types of ixodid ticks, as well as to identify pathogens in ticks collected from the environment in urban parks and from dogs and cats. A total of 215 ticks (22 Ixodes ricinus and 193 Dermacentor reticulatus) collected in two regions of Lviv and Ivano-Frankivsk were investigated. Pathogens, namely, Rikettsia spp., Candidatus Neoehrlichia mikurensis, Anaplasma phagocytophilum and Bartonella spp., were detected in 64.0% of the I. ricinus ticks. Overall, 18.2% of I. ricinus ticks were positive for Rikettsia spp., 54.5% for Candidatus Neoehrlichia mikurensis, 9.1% for A. phagocytophilum and Bartonella spp. confirmed in 4.5% of the studied ticks. Mixed infestations were found in 5 samples of I. ricinus DNA, representing 22.7%. In addition, 74.6% of D. reticulatus ticks were infested with these pathogens. Overall, 30.0% of D. reticulatus ticks were positive for Rikettsia spp., Candidatus Neoehrlichia mikurensis was confirmed in 54.9%, A. phagocytophilum in 1.6%, and Bartonella spp. in 6.2% of the studied ticks. Mixed infestations were found in 35 D. reticulatus DNA samples, representing 18.1%. The prevalence of Rikettsia spp. in two regions of Lviv and Ivano-Frankivsk, among the studied ticks was 28.8%, Bartonella spp. – 6.0%, A. phagocytophilum – 2.3%, Candidatus Neoehrlichia mikurensis – 54.9%. Our data indicate the presence of pathogens in urban populations of ticks of I. ricinus and D. reticulatus in west part of Ukraine. Monitoring of tick-borne diseases is an important tool in the prevention and control of infections transmitted to humans and animals.

Molecular detection of tick-borne pathogens in bovine blood and ticks from Khentii, Mongolia.

Recent studies reported the detection of DNA from tick-borne pathogens (TBPs) of veterinary relevance such as Anaplasma marginale, Babesia bigemina, Babesia bovis and Theileria orientalis in bovine blood samples from Mongolia. These findings were unexpected, as the known tick vectors of these pathogens are not known to occur in Mongolia. We therefore conducted a study in May and June 2013 in six districts of Khentii province where DNA of the said TBPs was previously found. Ticks collected from the vegetation and rodents, as well as blood samples from cattle, were screened for the presence of TBPs by reverse line blot (RLB) hybridization. Tick larvae collected from rodents were pooled. A total of 310 adult ticks were collected from the vegetation, and 249 tick larvae were collected from 24 rodents. Adult ticks (n=2,318) and blood samples were collected from 481 heads of cattle. All adult ticks were identified as Dermacentor nuttalli. DNA from Rickettsia raoultii (252/310; 81.3%), an uncharacterized Anaplasma species preliminary named Anaplasma sp. Mongolia (26/310; 8.4%), Candidatus Midichloria sp. (18/310; 5.8%), Theileria equi (16/310; 5.2%), Babesia caballi (5/310; 1.6%), T. orientalis (1/310; 0.3%), Borrelia afzelii (1/310; 0.3%) and Candidatus Neoehrlichia mikurensis (1/310; 0.3%) was detected in ticks collected from the vegetation. DNA of R. raoultii (27/28; 96.4%) and Midichloria sp. (2/28; 7.1%) was detected in the pooled tick larvae. Anaplasma sp. Mongolia, a species related to Anaplasma ovis based on a multi-locus analysis, was also detected in 153/481 (31.8%) of the bovine blood samples. DNA of B. bovis, B. bigemina and A. marginale was not detected in the ticks or bovine blood samples from Khentii district.

Detection of Candidatus Neoehrlichia mikurensis in Norway up to the northern limit of Ixodes ricinus distribution using a novel real time PCR test targeting the groEL gene

BackgroundCandidatus Neoehrlichia mikurensis is an emerging tick-borne pathogen. It is widely distributed in Ixodes ricinus ticks in Europe, but knowledge of its distribution in Norway, where I. ricinus reaches its northern limit, is limited. In this study we have developed a real time PCR test for Ca. N. mikurensis and used it to investigate the distribution of Ca. N. mikurensis in Norway.ResultsReal time PCR targeting the groEL gene was developed and shown to be highly sensitive. It was used to detect Ca. N. mikurensis in 1651 I. ricinus nymphs and adults collected from twelve locations in Norway, from the eastern Oslo Fjord in the south to near the Arctic Circle in the north. The overall prevalence was 6.5% and varied locally between 0 and 16%. Prevalence in adults and nymphs was similar, suggesting that ticks acquire Ca. N. mikurensis predominantly during their first blood meal. In addition, 123 larvae were investigated; Ca. N. mikurensis was not found in larvae, suggesting that transovarial transmission is rare or absent. Sequence analysis suggests that a single variant dominates in Norway.ConclusionsCa. N. mikurensis is widespread and common in ticks in Norway and reaches up to their northern limit near the Arctic Circle. Ticks appear to acquire Ca. N. mikurensis during their first blood meal. No evidence for transovarial transmission was found.

Ixodes persulcatus/pavlovskyi natural hybrids in Siberia: Occurrence in sympatric areas and infection by a wide range of tick-transmitted agents

Ixodes persulcatus and Ixodes pavlovskyi ticks, two closely related species of the I. ricinus - I. persulcatus group, are widely distributed in the southern part of Western Siberia. Recently, the existence of natural hybrids of I. persulcatus and I. pavlovskyi ticks has been demonstrated. The aim of this study was to evaluate the abundance of I. persulcatus/pavlovskyi hybrids in several locations with different ratios of parental tick species and to investigate the prevalence and genetic variability of a wide range of infectious agents in these hybrids compared to the parental tick species. Natural hybrids of I. persulcatus and I. pavlovskyi ticks were identified in all examined locations in Altai and Novosibirsk, Western Siberia, Russia. The abundance of hybrids varied from 7% to 40% in different locations and was maximal in a location with similar proportions of I. persulcatus and I. pavlovskyi ticks. For the first time, it was shown that hybrids can be infected with the same agents as their parental tick species: tick-borne encephalitis and Kemerovo viruses, Borrelia afzelii, Borrelia bavariensis, Borrelia garinii, Borrelia miyamotoi, Rickettsia helvetica, Rickettsia raoultii, Rickettsia sibirica, “Candidatus Rickettsia tarasevichiae”, Anaplasma phagocytophilum, Ehrlichia muris, “Candidatus Neoehrlichia mikurensis”, and Babesia microti. The prevalence of most bacterial agents in hybrids was intermediate compared to their parental tick species. Most genetic variants of the identified agents have been previously found in the parental tick species. Wide distribution of I. persulcatus/pavlovskyi natural hybrids implies that I. persulcatus, I. pavlovskyi and their hybrids coexist in all I. persulcatus - I. pavlovskyi sympatric areas.