Lyme arthritis in Western Europe: a multicentre retrospective study.

To characterize Lyme arthritis, with a focus on management, and outcome. Observational retrospective multicentre study in Western France, of all consecutive cases of Lyme arthritis, documented by Borrelia burgdorferi IgG on ELISA serological testing, confirmed by Western blot, with or without positive Borrelia PCR in synovial fluid, with no alternative diagnosis. We enrolled 52 patients (29 males), with a mean age of 43 ± 19.4 years. Most patients had monoarthritis (n = 43, 82.7%), involving the knee (n = 51, 98.1%), with a median delay between symptoms onset and Lyme arthritis diagnosis of 5 months (interquartile range, 1.5-8). Synovial fluid analysis yielded median white cell count of 16,000/mm3 (9230-40,500), and positive PCR in 16 cases (39%), for B. burgdorferi sensu stricto (n = 5), B. garinii (n = 5), B. afzelii (n = 3), and undetermined (n = 3). All patients received antibiotics, for a median duration of 28 days (21-30), with doxycycline (n = 44, 84.6%), ceftriaxone (n = 6, 11.5%), or amoxicillin (n = 2). Twelve patients (23.1%) also received intra-articular injection of glucocorticoids as first-line treatment. Of 47 patients with follow-up, 35 (74.5%) had complete resolution of Lyme arthritis. Lyme arthritis in Western Europe may be due to B. burgdorferi ss, B. afzelii, or B. garinii. Clinical presentation is similar to Lyme arthritis in North America (i.e. chronic knee monoarthritis), with low sensitivity of synovial fluid PCR (39%).

Surveillance and Molecular Identification of Borrelia Species in Ticks Collected at U.S. Army Garrison Humphreys, Republic of Korea, 2018-2019.

Tick-borne pathogens are contributing factors for the increased incidence of vector-borne diseases throughout the world, including Lyme borreliosis, one of the most prevalent spirochetes belonging to the Borrelia burgdorferi sensu lato group. The present study focused on the detection of Borrelia species from hard ticks collected at U.S. Army Garrison Humphreys, Republic of Korea (ROK), using molecular and genotypic analyses. Tick-borne disease surveillance was conducted from January to December, 2018-2019. A total of 24,281 ticks (2 genera and 5 species) were collected from road-killed Korean Water deer (KWD) and by tick drag. Haemaphysalis longicornis (92.0%) was the most commonly collected species, followed by Haemaphysalis flava (4.9%), Ixodes nipponensis (3.1%), Haemaphysalis phasiana (0.07%), and Haemaphysalis japonica (<0.01%). The ospA gene sequences of Borrelia afzelii were detected in 12/529 pools of I. nipponensis. Three and one pools were positive for B. afzelii and Borrelia miyamotoi, respectively, using the 16s rRNA gene. None of the pools of Haemaphysalis ticks collected from KWD or by tick drag were positive for Borrelia species. I. nipponensis was collected throughout the year from KWD and from February to November by tick drag, suggesting that they were active throughout the year, and expanding the risk period for acquiring Lyme borreliosis and Borrelia relapsing fever in the ROK. This study assessed disease risk factors associated with the prevalence of Lyme disease in ticks collected from KWD and by tick drag using molecular analysis. These results provide an understanding and awareness into the prevalence and molecular characteristics of Borrelia species in the ROK.

Association between statin use and clinical course, microbiologic characteristics, and long-term outcome of early Lyme borreliosis. A post hoc analysis of prospective clinical trials of adult patients with erythema migrans.

BACKGROUND: Statins were shown to inhibit borrelial growth in vitro and promote clearance of spirochetes in a murine model of Lyme borreliosis (LB). We investigated the impact of statin use in patients with early LB. METHODS: In this post-hoc analysis, the association between statin use and clinical and microbiologic characteristics was investigated in 1520 adult patients with early LB manifesting as erythema migrans (EM), enrolled prospectively in several clinical trials between June 2006 and October 2019 at a single-center university hospital. Patients were assessed at enrollment and followed for 12 months. RESULTS: Statin users were older than patients not using statins, but statin use was not associated with Borrelia seropositivity rate, Borrelia skin culture positivity rate, or disease severity as assessed by erythema size or the presence of LB-associated symptoms. The time to resolution of EM was comparable in both groups. The odds for incomplete recovery decreased with time from enrollment, were higher in women, in patients with multiple EM, and in those reporting LB-associated symptoms at enrollment, but were unaffected by statin use. CONCLUSION: Statin use was not associated with clinical and microbiologic characteristics or long-term outcome in early LB.

A simple method to detect Borrelia burgdorferi sensu lato proteins in different sub-cellular compartments by immunofluorescence.

Spirochaetes constitute a unique phylum of bacteria, many of which cause severe clinical diseases. Borrelia burgdorferi sensu lato (B. burgdorferi s.l.)-the primary agent of Lyme borreliosis (LB)-is a quintessential member of this poorly understood phylum and the leading cause of tick-borne illness throughout most of the northern hemisphere. Despite its importance in human health, we lack a fundamental understanding of how B. burgdorferi s.l. is able to accomplish basic physiological tasks, such as DNA replication/segregation, and cell elongation or division. Recent advances in molecular tools to probe these essential cellular processes are great strides forward but require genetic manipulation. The latter is important since not all agents of LB are genetically tractable. Here, we describe a single method that is capable of fluorescently labeling B. burgdorferi s.l. proteins in different sub-cellular compartments. A comparative analysis of six different methods indicates that our optimized procedure outperforms all others and is the first to localize a cytoplasmic protein in B. burgdorferi s.l. by immunofluorescence. We contend that this strategy could be easily adapted to study the localization of any protein, in many Borrelia genospecies, information that will yield functional insights into the complex biology of this fascinating group of bacteria. In addition, it may provide new avenues of research in both in situ studies and in Lyme diagnostics.

Monitoring of pathogenic Borrelia burgdorferi sensu lato in the Almaty oblast, Kazakhstan.

Lyme borreliosis (LB) is one of the most common vector-borne diseases transmitted by ticks. It is caused by the Borrelia burgdorferi sensu lato (s.l.) genospecies. The Almaty oblast of Kazakhstan is considered endemic for LB. Nevertheless, there are significant gaps in the tick surveillance for LB agents in the region. We evaluated B. burgdorferi s.l. prevalence in 413 ixodid ticks collected from bitten people and the seroprevalence of antibodies to B. burgdorferi s.l. in 589 residents of the Almaty oblast, Kazakhstan. All samples were collected between 2018 and 2020. Borrelia burgdorferi s.l. DNA was detected in 24 % (9/38) of I. persulcatus ticks that attacked humans in the city of Almaty and the Talgar and Karasay districts. Multilocus sequence typing identified two novel B. afzelii sequence types in I. persulcatus. The seroprevalence of IgG antibodies against B. burgdorferi s.l. in the analyzed population was detected to be 5.8 %. The obtained results confirm active circulation of B. burgdorferi s.l. in the Almaty oblast of Kazakhstan and raise concern regarding LB preventive measures in the region.

The potential risk of exposure to Borrelia garinii, Anaplasma phagocytophilum and Babesia microti in the Wolinski National Park (north-western Poland).

Ixodes ricinus (Acari: Ixodida) is the main vector in Europe of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti. Wolinski National Park (WNP) is situated by the Baltic Sea and is frequently visited by tourists. The aim of the study was to determine the potential risk of exposure to tick borne infection with B. burgdorferi s.l., A. phagocytophilum and B. microti on the areas of WNP. In total, 394 I. ricinus were tested. The pathogens in ticks were detected by PCR, nested PCR, RFLP and sequencing. Altogether, pathogens were detected in 12.69% of the studied ticks. B. burgdorferi s.l., was shown in 0.25% of the studied I. ricinus, while A. phagocytophilum and B. microti were detected in 1.01% and 10.65% of studied ticks, respectively. Co-infection by A. phagocytophilum and B. microti was shown in only one I. ricinus nymph. Analysis of B. burgdorferi s.l., genospecies showed that 0.25% of the studied ticks were infected with Borrelia garinii. The obtained results show the potentially high human risk of exposure to tick-borne infection with B. microti, and the low potential risk of infection with B. garinii and A. phagocytophilum on the studied areas of WNP.

Acrodermatitis chronica atrophicans: clinical and microbiological characteristics of a cohort of 693 Slovenian patients.

BACKGROUND: Information on large groups of patients with acrodermatitis chronica atrophicans (ACA) is limited. METHODS: We assessed clinical and microbiological characteristics of patients with ACA diagnosed at a single medical centre and compared findings in periods 1991-2004 vs. 2005-2018. The cohort is representative of Slovenian ACA patients. RESULTS: We assessed 693 patients: 461 females and 232 males, with median age of 64 years. Median duration of ACA before diagnosis was 12 months. In all but 2 patients, the skin lesions were located on extremities, more often on the lower (70.0%) than the upper (45.2%), bilaterally in 42.4%. Reddish-blue discoloration, swelling, thinning and wrinkling of skin were present in 95.2%, 28.1%, 46.4% and 20.5% of patients, respectively. Overall, 64.4% of patients reported constitutional symptoms, 23.1% had local symptoms, and 20.8% had symptoms/signs of peripheral neuropathy. Nodules, arthritis, joint deformity, muscle atrophy and paresis were rare (<3%). Borreliae were isolated from 200/664 (30.1%) skin samples; 92.8% were Borrelia afzelii. B. garinii and B. burgdorferi s.s. were more often isolated from the skin of male patients (OR = 4.17) and from those with arthropathy (OR = 11.74). Patients included in the more recent period were older, complained less often of constitutional symptoms but more often of local symptoms, and more often had local swelling but less often skin atrophy and bilateral involvement, probably as a consequence of earlier diagnosis. CONCLUSIONS: ACA, typically caused by B. afzelii, usually affects older women. Clinical presentation depends on the duration of illness and probably on the Borrelia species causing the disease.

Lyme neuroborreliosis in Swedish children-PCR as a complementary diagnostic method for detection of Borrelia burgdorferi sensu lato in cerebrospinal fluid.

The aim of this study was to evaluate polymerase chain reaction (PCR) as a diagnostic method for the detection of Borrelia burgdorferi s.l. in CSF of Swedish children with LNB. This study was performed retrospectively on CSF and serum samples collected from children evaluated for LNB (n = 233) and controls with other specific neurological disorders (n = 59) in a Swedish Lyme endemic area. For anti-Borrelia antibody index, the IDEIA Lyme Neuroborreliosis kit (Oxoid) was used. Two in-house real-time PCR assays targeting the 16S rRNA gene were evaluated (TaqMan® and LUX™). Among patients classified as LNB cases (n = 102), five children (5%) were Borrelia PCR-positive in CSF with the TaqMan® assay. In the Non-LNB group (n = 131), one patient was Borrelia PCR positive with the TaqMan® assay. Among controls (n = 59), all CSF samples were PCR negative. When amplifying and sequencing ospA, we found B. garinii (n = 2), B. afzelii (n = 2), B. bavariensis (n = 1), and one untypable (n = 1). With the LUX™ technology, all CSF samples were PCR negative. The TaqMan® assay could detect only few cases (n = 6) of B. burgdorferi s.l. in CSF among children with LNB and the sensitivity was very low (5%). However, using larger CSF volumes and centrifugation of samples, the PCR technique could still be useful as a complementary diagnostic method when evaluating LNB. Furthermore, detection of spirochete DNA in clinical matrices, including CSF, is the method of choice for studying epidemiological aspects of LNB, a tick-borne emerging disease.

Detection of pathogens in ixodid ticks collected from animals and vegetation in five regions of Ukraine.

The distribution and prevalence of zoonotic pathogens infecting ixodid ticks in Western Europe have been extensively examined. However, data on ticks and tick-borne pathogens in Eastern Europe, particularly Ukraine are scarce. The objective of the current study was, therefore, to investigate the prevalence of Anaplasma phagocytophilum, Anaplasmataceae, Rickettsia spp., Babesia spp., Bartonella spp., and Borrelia burgdorferi sensu lato in engorged and questing ixodid ticks collected from five administrative regions (oblasts) of Ukraine, namely Chernivtsi, Khmelnytskyi, Kyiv, Ternopil, and Vinnytsia. The ticks were collected from both wild and domestic animals and from vegetation. Of 524 ixodid ticks collected, 3, 99, and 422 ticks were identified as Ixodes hexagonus, Ixodes ricinus, and Dermacentor reticulatus, respectively. DNA samples individually extracted from 168 questing and 354 engorged adult ticks were subjected to pathogen-specific PCR analyses. The mean prevalence in I. ricinus and D. reticulatus were, respectively: 10 % (10/97) and 3 % (12/422) for A. phagocytophilum; 69 % (67/97) and 52 % (220/422) for members of the Anaplasmataceae family; 25 % (24/97) and 28 % (117/422) for Rickettsia spp.; 3 % (3/97) and 1 % (6/422) for Babesia spp.; and 9 % (9/97) and 5 % (20/422) for Bartonella spp. Overall, between the five cities, there was no significant difference in the prevalence of any of the pathogens for the respective ticks (p > 0.05). The prevalence of B. burgdorferi s. l. in the questing and engorged I. ricinus varied from 0 to 27 % and 14-44%, respectively, with no statistical significance identified between the five cities (p > 0.05). In addition to reporting the updated data for Kyiv and Ternopil, this study is the first to provide the prevalences of the tick-borne pathogens for Chernivtsi, Khmelnytskyi, and Vinnytsia. This investigation is also the first to detect Neoehrlichia mikurensis in ixodid ticks from Ukraine. These new data will be useful for medical and veterinary practitioners as well as public health officials when diagnosing infections and when implementing measures to combat tick-borne diseases in Ukraine.

Spatial variability in prevalence and genospecies distributions of Borrelia burgdorferi sensu lato from ixodid ticks collected in southern Germany.

Lyme borreliosis (LB) is the most common arthropod-borne disease in Europe and North America and is caused by members of the Borrelia burgdorferi sensu lato (Bbsl) species complex. These bacteria are transmitted by ixodid tick vectors and therefore human LB risk is influenced by the prevalence and distribution of Bbsl genospecies within tick vectors throughout the wild. These distributions can easily change over spatiotemporal scales and, to understand LB risk fully, up to date information on prevalence and distribution of Bbsl is required. The last survey of Bbsl in southern Germany, including parts of the Munich metropolitan area, was completed in 2006 and new data is needed. Ixodid ticks were collected in seven plots located in and around Munich, Germany, from March to July 2019 and were screened for Bbsl. Borrelia burgdorferi s. l. positive ticks (52 adults, 158 nymphs) were found in all plots and adults (0-61.5 % Bbsl positive/plot) and nymphs (17.4-59.5 % Bbsl positive/plot) did not differ significantly in their overall Bbsl prevalence. The number of Bbsl positive nymphs did vary significantly between plots but the number of positive adults did not. In total, six Bbsl genospecies were located with B. afzelii and B. garinii dominating. Additionally, the relapsing-fever species B. miyamotoi was found in two sampling plots. Our results highlight the variability in Bbsl prevalence and genospecies distribution over short geographic distances and aid in understanding LB risk in and around the Munich metropolitan area.

Development of a PCR for Borrelia burgdorferi sensu lato, targeted on the groEL gene.

Borrelia burgdorferi sensu lato (s.l.) is the etiological agent of Lyme disease, transmitted by ticks of the genus Ixodes Latreille. Diagnosis of Lyme disease in humans is often difficult and a detailed knowledge of the circulation of B. burgdorferi s.l. in tick hosts is therefore fundamental to support clinical procedures. Here we developed a molecular approach for the detection of B. burgdorferi s.l. in North Italian Ixodes ricinus (Linnaeus). The method is based on the amplification of a fragment of the groEL gene, which encodes a heat-shock protein highly conserved among B. burgdorferi s.l. species. The tool was applied in both qualitative and Real-time PCR approaches testing ticks collected in a North Italian area. The obtained results suggest that this new molecular tool could represent a sensitive and specific method for epidemiological studies aimed at defining the distribution of B. burgdorferi s.l. in I. ricinus and, consequently, the exposure risk for humans.

Update on the presence of Ixodes ricinus at the western limit of its range and the prevalence of Borrelia burgdorferi sensu lato.

It is often suggested that due to climate and environmental policy changes, the risk from tick-borne disease is increasing, particularly at the geographical limits of the vector distribution. Our project aimed to determine whether this was true for the risk of Lyme borreliosis in Ireland which is the western-most limit of Ixodes ricinus, the European vector of Borrelia burgdorferi sensu lato. The availability of a historical data set of tick infection rates compiled in the 1990s represented a unique opportunity as it provided a baseline against which current data could be compared. Following construction of a spatial predictive model for the presence and absence of I. ricinus based on data from 491 GPS locations visited between 2016 and 2019, 1404 questing nymphs from 27 sites were screened for the presence of Borrelia spp. using a TaqMan PCR aimed at the 23S rRNA gene sequence. All positive ticks were further analysed by nested PCR amplification and sequence analysis of the 5 S-23 S intergenic spacer. The model indicated that areas with the highest probability of tick presence were mostly located along the western seaboard and the Shannon and Erne river catchments, coinciding with historical high incidence areas of bovine babesiosis, while the infection rate of questing nymphs with B. burgdorferi s.l. and the prevalence of the various genospecies have remained surprisingly stable over the last 3 decades. Clear communication of the potential disease risk arising from a tick bite is essential in order to allay undue concerns over tick-borne diseases among the general public.

Seroprevalence of Antibodies against Borrelia burgdorferi s. l. and Leptospira interrogans s. l. in Cats in district of Brno and its environs, the Czech Republic.

OBJECTIVES: The aim of this study is to evaluate the seroprevalence of antibodies of Borrelia burgdorferi sensu lato (Bbsl) and Leptospira interrogans sensu lato (Lisl) and their possible concurrence in domestic cats living in variable conditions in South Moravia in the district of Brno and its environs. Additional objectives were to discover possible differences in seroprevalence between groups of cats living in different living conditions, and to determine the spectrum of Leptospira serogroups in cats in the same places. MATERIAL AND METHODS: A total of 360 blood sera from domestic cats of 3 different sets were collected during the period 2013-2015. All samples were examined using ELISA for the detection of IgM and IgG antibodies against Bbsl, and the microscopic agglutination test (MAT) for the detection of antibodies against 8 serogroups of Lisl. RESULTS: The ELISA method determined 15.8%, 4.8% and 10.3% IgM anti-Borrelia antibodies in the patient group, shelter cats and street cats, respectively. IgG anti-Borrelia antibodies were found in 6.2%, 9.5%, 5.2%, respectively. Antibodies specific for 5 Leptospira serogroups were detected by the use of MAT in 8.8%, 9.5% and 10.3% of cats from the investigated groups. The total positivity of all examined cats for anti-Borrelia antibodies was 18.0% and for anti-Leptospira - 9.2%. CONCLUSIONS: Cats can be infected with both Bbsl and Lisl. The obtained results are exclusive to the city of Brno and its environs, and are comparable to the limited previous studies. There is a need for further studies of clinical signs of both infections and the possible transmission of Leptospira by ticks.

Molecular characterization of tick-borne bacteria and protozoans in yaks (Bos grunniens), Tibetan sheep (Ovis aries) and Bactrian camels (Camelus bactrianus) in the Qinghai-Tibetan Plateau Area, China.

Due to the specific plateau climate, a variety of unique animals live in the Qinghai-Tibetan Plateau Area (QTPA) including yaks (Bos grunniens), Tibetan sheep (Ovis aries) and Bactrian camels (Camelus bactrianus). However, information on tick-borne diseases (TBDs) in the QTPA and on the molecular characteristics of tick-borne pathogens (TBPs) in the area is limited. Therefore, the aim of this study was to investigate Anaplasma spp., Babesia spp., Theileria spp., Borrelia burgdorferi sensu lato and Rickettsia spp. infecting yaks, Tibetan sheep and camels in this area. A total of 276 animals were screened. Overall, 84.5% (164/194) of yaks, 58% (23/40) of Tibetan sheep, and 38% (16/42) of camels tested positive for at least one pathogen. Theileria spp., Anaplasma ovis and spotted fever group (SFG) Rickettsia spp. were detected as TBPs in the current study with overall infection rates of 10.9% (30/276), 8.3% (23/276) and 62.9% (171/276), respectively. Further study revealed that 1.5% (3/194) of the yaks were infected with Theileria sp. OT3, 1.5% (3/194) with T. luwenshuni, 6.2% (12/194) with T. uilenbergi, 1.1% (2/194) with T. ovis and 82% (159/194) with SFG Rickettsia spp. It was also shown that 58% (23/40) of the Tibetan sheep were infected with A. ovis and 15% (6/40) with T. ovis. Among the camels, 10% (4/42) were infected with T. equi, while 29% (12/42) were positive for Rickettsia spp. Sequence analysis revealed that the Rickettsia spp. infecting yaks and camels were Rickettsia raoultii and Rickettsia slovaca. To the best of our knowledge, this study reports the first detection and characterization of these pathogens in yaks, Tibetan sheep and camels in the country, except for T. luwenshuni infections in yaks.

Longitudinal study of prevalence and spatio-temporal distribution of Borrelia burgdorferi sensu lato in ticks from three defined habitats in Latvia, 1999-2010.

Members of the Borrelia burgdorferi sensu lato (s.l.) species complex are known to cause human Lyme borreliosis. Because of longevity of some reservoir hosts and the Ixodes tick vectors' life cycle, long-term studies are required to better understand species and population dynamics of these bacteria in their natural habitats. Ticks were collected between 1999 and 2010 in three ecologically different habitats in Latvia. We used multilocus sequence typing utilizing eight chromosomally located housekeeping genes to obtain information about species and population fluctuations and/or stability of B. burgdorferi s.l. in these habitats. The average prevalence over all years was 18.9%. From initial high-infection prevalences of 25.5%, 33.1% and 31.8%, from 2002 onwards the infection rates steadily decreased to 7.3%. Borrelia afzelii and Borrelia garinii were the most commonly found genospecies but striking local differences were obvious. In one habitat, a significant shift from rodent-associated to bird-associated Borrelia species was noted whilst in the other habitats, Borrelia species composition was relatively stable over time. Sequence types (STs) showed a random spatial and temporal distribution. These results demonstrated that there are temporal regional changes and extrapolations from one habitat to the next are not possible.

Prevalence of antibodies against Anaplasma spp., Borrelia burgdorferi sensu lato, Babesia gibsoni, and Ehrlichia spp. in dogs in the Republic of Korea.

There is a lack of comprehensive studies on the seroprevalence of tick-borne pathogens in the Republic of Korea. Therefore, the aim of this study was to investigate the seroprevalences of Anaplasma spp. (A. phagocytophilum/A. platys), Borrelia burgdorferi sensu lato, Babesia gibsoni, Ehrlichia spp. (E. canis/E. ewingii), and Ehrlichia chaffeensis in dogs across the Republic of Korea in 2017 and 2018. A total of 2215 serum samples collected from 938 companion dogs, 969 shelter dogs, and 308 military working dogs were examined using commercial enzyme-linked immunosorbent assay (ELISA) and indirect fluorescence immunoassay (IFA) kits. Data collected for each animal, including breed, sex, age, region, season, and dog type, were used for statistical analysis. The overall seroprevalence was highest for Anaplasma spp. (15.1 %), followed by Ehrlichia spp. (10.3 %), B. burgdorferi sensu lato (6.4 %), E. chaffeensis (2.3 %), and B. gibsoni (1.7 %). One hundred and sixty-one dogs had antibodies against two or three different pathogens. The most common combinations were Anaplasma spp. - Ehrlichia spp. (2.1 %), Anaplasma spp. - E. chaffeensis (1.4 %), and Anaplasma spp. - B. burgdorferi sensu lato (1.2 %). Season was significantly associated with the seroprevalences of B. burgdorferi sensu lato and Ehrlichia spp., with dogs presenting the highest percentage of positive results during summer. Anaplasma spp. and B. gibsoni were significantly more prevalent in the northern and southern regions, respectively. The seroprevalences of Anaplasma spp., B. burgdorferi sensu lato, and Ehrlichia spp. were significantly higher in military working dogs, while the seroprevalence of E. chaffeensis was higher in companion dogs. The current findings are important for future surveillance of canine tick-borne pathogens and designing appropriate approaches for the diagnosis and control of these pathogens in the Republic of Korea.

Turkey tick news: A molecular investigation into the presence of tick-borne pathogens in host-seeking ticks in Anatolia; Initial evidence of putative vectors and pathogens, and footsteps of a secretly rising vector tick, Haemaphysalis parva.

This Turkey-based study investigated the presence of various tick-borne microorganisms in a broad-range of host-seeking ticks (n = 1019) that exhibit both hunter and ambusher characteristics. All collected ticks were analyzed individually via PCR-sequencing, resulting in the identification of 18 different microorganisms: six Babesia spp., including one putative novel species (Ba. occultans, Ba. crassa, Ba. rossi, Babesia sp. tavsan1, Babesia sp. tavsan2, and Babesia sp. nov.); six SFG rickettsiae (Ri. aeschlimannii, Ri. s. mongolitimonae, Ri. slovaca, Ri. raoultii, Ri. monacensis, and Ri. hoogstraalii); two Borrelia burgdorferi sensu lato spp. (Bo. afzelii and Bo. lusitaniae); two unnamed Hepatozoon spp.; Theileria annulata; and Hemolivia mauritanica. This provided evidence for the natural transstadial survival of these tick-borne microorganisms in adult ticks (in addition a nymph) of Turkey. Surprisingly, this study determined the presence of five different microorganisms (Ba. crassa, Ba. rossi, Babesia sp. Ucbas, Hepatozoon sp., and Ri. hoogstraalii) in host-seeking Haemaphysalis parva adults, for which poor data exist on its vectorial competence. Therefore, this study provides important data indicating the potential vectorial capacity of Ha. parva. This study also revealed the presence of the close ecological and evolutionary relationships between two important vector ticks, Hyalomma marginatum and Hy. aegyptium and determined genetic variations (distinct phylogenetic divergences inside the main clades) in some pathogenic SFG rickettsiae that are found in these ticks. Additionally, the presence of two Babesia species described very recently in hares with unknown vectors, namely Babesia sp. tavsan1 and Babesia sp. tavsan2, were detected for the first time in ticks. Finally, two unnamed Hepatozoon spp. were detected in Haemaphysalis ticks and their phylogenetic positions were demonstrated. Consequently, this study provides important data on the diversity of tick-borne microorganisms in host-seeking ticks and on potentially novel microorganisms (Babesia and Hepatozoon species) and their possible vectors (Ha. parva, Ha. sulcata, Hy. aegyptium, Hy. marginatum, and Rh. turanicus).

Circulation of Tick-Borne Spirochetes in Tick and Small Mammal Communities in Santa Barbara County, California, USA.

A diversity of Borrelia burgdorferi sensu lato (Johnson, Schmid, Hyde, Steigerwalt & Brenner) (Spirochaetales: Spirochaetaceae) genomospecies, including the Lyme disease agent, Borrelia burgdorferi sensu stricto (s.s.), have been identified in the western United States. However, enzootic transmission of B. burgdorferi s.l. in small mammals and ticks is poorly characterized throughout much of the region. Here we report prevalence of B. burgdorferi s.l. in small mammal and tick communities in the understudied region of southern California. We found B. burgdorferi s.l. in 1.5% of Ixodes species ticks and 3.6% of small mammals. Infection was uncommon (~0.3%) in Ixodes pacificus Cooley and Kohls (Acari: Ixodidae), the primary vector of the Lyme disease agent to humans in western North America, but a diversity of spirochetes-including Borrelia bissettiae, Borrelia californiensis, Borrelia americana, and B. burgdorferi s.s.-were identified circulating in Ixodes species ticks and their small mammal hosts. Infection with B. burgdorferi s.l. is more common in coastal habitats, where a greater diversity of Ixodes species ticks are found feeding on small mammal hosts (four species when compared with only I. pacificus in other sampled habitats). This provides some preliminary evidence that in southern California, wetter coastal areas might be more favorable for enzootic transmission than hotter and drier climates. Infection patterns confirm that human transmission risk of B. burgdorferi s.s. is low in this region. However, given evidence for local maintenance of B. burgdorferi s.l., more studies of enzootic transmission may be warranted, particularly in understudied regions where the tick vector of B. burgdorferi s.s. occurs.

Prevalence of tick-borne encephalitis virus and Borrelia burgdorferi sensu lato in Ixodes ricinus ticks in Lower Bavaria and Upper Palatinate, Germany.

Lyme borreliosis and tick-borne encephalitis (TBE) are the most common tick-borne diseases in Germany. We collected Ixodes ricinus ticks from 16 high-risk and four low-risk sites distributed in Lower Bavaria and Upper Palatinate based on the number of human TBE cases recorded at the Robert Koch Institute from 2001 to 2009. A total of 8805 questing ticks (8203 nymphs, 602 adults) were collected in 2010 and examined in pools for the presence of tick-borne encephalitis virus (TBEV) using real-time RT-PCR. Overall TBEV prevalence evaluated as the minimum infection rate (MIR) was 0.26 % (23 positive pools/8805 ticks in 1029 pools). TBEV was detected at seven of the 16 high-risk sites, where MIR ranged from 0.16 to 2.86 %. A total of 3969 ticks were examined by PCR for infection with Borrelia burgdorferi sensu lato (s.l.) targeting the 5 S-23 S rRNA intergenic spacer (IGS) region. IGS nucleotide sequences were used to determine genospecies. Selected positive Borrelia samples were subjected to PCR and sequencing targeting the OspA gene, providing 46 sequences for molecular phylogenetic analysis. Of the 3969 questing ticks, 506 (12.7 %) were positive for B. burgdorferi s.l. Seven B. burgdorferi s.l. genospecies were identified: B. afzelii (41.3 %), B. garinii (19 %), B. valaisiana (13.8 %), B. burgdorferi sensu stricto (11.1 %), B. spielmanii (0.4 %), B. lusitaniae (0.2 %), and Candidatus B. finlandensis (0.6 %). Mixed infections were identified in 13.6 % of the ticks. The rate of infection in questing ticks varied among sites from 5.6 % (72 examined, four positive) to 29.5 % (88 examined, 26 positive). B. burgdorferi s.l. occurred at all 20 sites, whereas TBEV was detected only at the high-risk sites where more human TBE cases were reported compared to low-risk sites.