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Background: Changing geographical and seasonal activity patterns of ticks may increase the risk of tick infestation and tick-borne pathogen (TBP) transmission for both humans and animals. Methods: To estimate TBP exposure of dogs and cats, 3000 female I. ricinus from these hosts were investigated for Anaplasma phagocytophilum and Borrelia species. Results: qPCR inhibition, which was observed for ticks of all engorgement stages but not questing ticks, was eliminated at a template volume of 2 µl. In ticks from dogs, A. phagocytophilum and Borrelia spp. prevalence amounted to 19.0% (285/1500) and 28.5% (427/1500), respectively, while ticks from cats showed significantly higher values of 30.9% (464/1500) and 55.1% (827/1500). Accordingly, the coinfection rate with both A. phagocytophilum and Borrelia spp. was significantly higher in ticks from cats (17.5%, 262/1500) than dogs (6.9%, 104/1500). Borrelia prevalence significantly decreased with increasing engorgement duration in ticks from both host species, whereas A. phagocytophilum prevalence decreased only in ticks from dogs. While A. phagocytophilum copy numbers in positive ticks did not change significantly over the time of engorgement, those of Borrelia decreased initially in dog ticks. In ticks from cats, copy numbers of neither A. phagocytophilum nor Borrelia spp. were affected by engorgement. Borrelia species differentiation was successful in 29.1% (365/1254) of qPCR-positive ticks. The most frequently detected species in ticks from dogs were B. afzelii (39.3% of successfully differentiated infections; 70/178), B. miyamotoi (16.3%; 29/178), and B. valaisiana (15.7%; 28/178), while B. afzelii (40.1%; 91/227), B. spielmanii (21.6%; 49/227), and B. miyamotoi (14.1%; 32/227) occurred most frequently in ticks from cats. Conclusions: The differences in pathogen prevalence and Borrelia species distribution between ticks collected from dogs and cats may result from differences in habitat overlap with TBP reservoir hosts. The declining prevalence of A. phagocytophilum with increasing engorgement duration, without a decrease in copy numbers, could indicate transmission to dogs over the time of attachment. The fact that this was not observed in ticks from cats may indicate less efficient transmission. In conclusion, the high prevalence of A. phagocytophilum and Borrelia spp. in ticks collected from dogs and cats underlines the need for effective acaricide tick control to protect both animals and humans from associated health risks. [ABSTRACT FROM AUTHOR]

People may be exposed to questing Ixodes ricinus ticks in urban settings, e.g. residential gardens. Little is known about the garden characteristics that support a tick population. To determine which features in and around residential gardens support or limit the occurrence and abundance of questing I. ricinus ticks, we sampled them in residential gardens in the Braunschweig region that differed in various intrinsic and extrinsic parameters. We recorded the number of questing nymphal and adult ticks on transects, and by using mixed-effects generalized linear regression models, we related their occurrence and abundance to garden characteristics, meteorological covariates, and landscape features in the vicinity. We detected questing I.ricinus ticks in about 90% of the 103 surveyed gardens. Our occurrence model (marginal R2 = 0.31) predicted the highest probability of questing ticks on transects with hedges or groundcover in gardens, which are located in neighborhoods with large proportions of forest. The abundance of questing ticks was similarly influenced. We conclude that I.ricinus ticks are frequent in residential gardens in Northern Germany and likely associated with intrinsic garden characteristics on a small scale, such as hedges, as well as extrinsic factors on a local scale, such as the proportion of nearby woodland. [ABSTRACT FROM AUTHOR]

Objectives: The purpose of the current study was to analyse the risks of Lyme borreliosis (LB) among 1,070 forestry workers, the influence of responsible behaviour (use of repellents, skin self-inspection) on Borrelia screening result status, and the occurrence of immediate and mid-term symptoms after tick bites and LB positive serological screening test., Methods: The questionnaire was conducted as well as blood tests for LB disease by one-stage serological screening procedure using ELISA for specific B. burgdorferi IgM and IgG antibodies (EuroImmun AG company, Germany)., Results: While 39.6% of foresters were LB positive among bitten foresters, as many as 27.0% were LB positive among those, who did not recall any tick attacks at all. Individuals with known history of tick bites had significantly higher odds (1.770×) of being LB positive (p < 0.05), while the use of repellents or skin self-inspection after visiting woods had no influence on LB results. The odds of skin discolouration after tick bites was significantly lower (0.682×) in case of LB positive test compared to LB negative test (p < 0.05), which can be explained by the fact that foresters could be unaware about erythema migrans appearance and timing, considering tick bite and developed later rash as completely separate events. Moreover, 69.1% of the bitten foresters with LB positive result developed no secondary symptoms (excluding those related to the skin), and the most frequent clinical symptoms were arthralgia (24.9%), followed by myalgia (7.6%), headache (5.7%), and damage to facial nerve (2.7%), which are non-specific and can be present in other illnesses., Conclusion: Therefore, the recommendations proposed would be the regular laboratory testing for LB of sensitive and at-risk population, who visits endemic woody areas, irrespective of all other factors involved.

Rickettsiae of the spotted fever group (SFG) are zoonotic tick-borne pathogens. Small mammals are important hosts for the immature life stages of two of the most common tick species in Europe, Ixodes ricinus and Dermacentor reticulatus. These hosts and vectors can be found in diverse habitats with different vegetation types like grasslands and forests. To investigate the influence of environmental and individual factors on Rickettsia prevalence, this study aimed to analyse the prevalence of SFG rickettsiae in ticks and small mammals in different small-scale habitats in central Germany for the first time. Small mammals of ten species and ticks of two species were collected from grasslands and forests in the Hainich-Dün region, central Germany. After species identification, DNA samples from 1098 ticks and ear snips of 1167 small mammals were screened for Rickettsia DNA by qPCR targeting the gltA gene. Positive samples were retested by conventional PCR targeting the ompB gene and sequencing. Rickettsia DNA was detected in eight out of ten small mammal species. Small mammal hosts from forests (14.0%) were significantly more often infected than those from grasslands (4.4%) (p < 0.001). The highest prevalence was found in the mostly forest-inhabiting genus Apodemus (14.8%) and the lowest in Microtus (6.6%), which inhabits grasslands. The prevalence was higher in D. reticulatus (46.3%) than in the I. ricinus complex (8.6%). Adult ticks were more often infected than nymphs (p = 0.0199). All sequenced rickettsiae in I. ricinus complex ticks were R. helvetica, and the ones in D. reticulatus were R. raoultii. Unlike adults, questing nymphs have had only one blood meal, which explains the higher prevalence in I. ricinus adults. Interestingly, habitat type did influence infection probability in small mammals, but did not in ticks. A possible explanation may be the high prevalence in Apodemus flavicollis and A. sylvaticus which were more abundant in the forest. [ABSTRACT FROM AUTHOR]

The newly discovered group of Jingmenviruses has been shown to infect a wide range of hosts and has been associated with febrile illness in humans. During a survey for Jingmenviruses in ticks from Lower Saxony, Germany, Alongshan virus (ALSV) was identified in Ixodes spp. ticks. Additional virus screenings revealed the presence of ALSV in the bodies and saliva of ticks collected at several locations in Lower Saxony. Vector competence studies that included Ixodes ricinus and Dermacentor reticulatus validated the replication of ALSV within those tick species. In vitro feeding experiments with ALSV-injected Ixodes ricinus demonstrated effective viral transmission during blood feeding. To evaluate the potential viral transmission during a natural blood meal, sera from wild game and domestic animals were investigated. One serum sample from a red deer was found to be positive for ALSV RNA, while serological screenings in game and domestic animals revealed the presence of ALSV-specific antibodies at different locations in Lower Saxony. Overall, those results demonstrate the broad distribution of ALSV in ticks in Lower Saxony and hypothesize frequent exposure to animals based on serological investigations. Hence, its potential risk to human and animal health requires further investigation. [ABSTRACT FROM AUTHOR]

Background: Hyalomma marginatum and H. rufipes are two-host tick species, which are mainly distributed in southern Europe, Africa to central Asia but may also be found in Central and Northern Europe through introduction by migratory birds., Methods: Ticks were collected while feeding or crawling on animals and humans, or from the environment, in different regions in Germany, between 2019 and 2021 in a citizen science study and from 2022 to 2023 in the wake of this study., Results: From 2019 to 2023, a total of 212 Hyalomma adult ticks were detected in Germany. This included 132 H. marginatum and 43 H. rufipes ticks sent to research institutions and 37 photographic records that were only identified to genus level. The number of detected ticks varied over the years, with the highest number of 119 specimens recorded in 2019, followed by 57 in 2020. Most of the specimens were collected from horses, while some were collected from other animals, humans or found crawling on human clothes or other objects inside or outside houses. The screening of 175 specimens for Crimean-Congo hemorrhagic fever virus and of 132 specimens for Babesia/Theileria spp. by PCR gave negative results, while human-pathogenic Rickettsia were detected in 44% (77/175) of the total samples. Subsequent amplicon sequencing and phylogenetic analysis of representative samples determined the species of 41 Rickettsia aeschlimannii and one R. slovaca sequences., Conclusions: Analysis of climatic factors indicated a significantly higher probability of Hyalomma occurrence at locations with higher average spring temperature during the years 2019 and 2020 compared to randomly generated pseudo-absence locations. Dry and hot conditions probably facilitated Hyalomma nymphs' survival and molting into adults during these years., (© 2024. The Author(s).)

Background: In Europe, canine babesiosis is most frequently caused by Babesia canis and Babesia vogeli, and occasionally by Babesia gibsoni.. In Germany, B. canis is recognized as endemic. The aims of this study were to assess how often Babesia spp. infections were diagnosed in a commercial laboratory in samples from dogs from Germany, and to evaluate potential risk factors for infection., Methods: The database of the LABOKLIN laboratory was screened for Babesia spp.-positive polymerase chain reaction (PCR) tests for dogs for the period January 2007-December 2020. Sequencing was performed for positive tests from 2018 and 2019. Binary logistic regression analysis was performed to determine the effects of sex, season, and year of testing. Questionnaires were sent to the submitting veterinarians to obtain information on travel abroad, tick infestation, and ectoparasite prophylaxis of the respective dogs. Fisher's exact test was used to calculate statistical significance and P < 0.05 was considered statistically significant., Results: In total, 659 out of 20,914 dogs (3.2%) tested positive for Babesia spp. by PCR. Of 172 sequenced samples, B. canis was identified in 156, B. vogeli in nine, B. gibsoni in five, and B. vulpes in two. Season had a statistically significant impact on test results when summer/winter (1.6% tested positive) was compared to spring/autumn (4.7%), with peaks in April (5.2%) and October (7.4%) [P < 0.001, odds ratio (OR) = 3.16]. Sex (male 3.5%, female 2.8%; P = 0.012, OR = 1.49) and age (< 7 years old 4.0%, ≥ 7 years old 2.3%; P < 0.001, OR = 1.76) of the tested dogs also had a statistically significant effect. A statistically significant impact was demonstrated for observed tick attachment (P < 0.001, OR = 7.62) and lack of ectoparasite prophylaxis (P = 0.001, OR = 3.03). The frequency of positive Babesia spp. tests did not significantly differ between the 659 dogs that had never left Germany and the 1506 dogs with known stays abroad (P = 0.088)., Conclusions: The possibility of canine infection with B. canis needs to be especially taken into consideration in spring and autumn in Germany as the activity of the tick Dermacentor reticulatus, a potential vector for canine babesiosis, is highest in these seasons. Travel and importation of dogs are considered major factors associated with canine babesiosis in Germany. However, autochthonous Babesia spp. infections also occur in a considerable number of dogs in Germany., (© 2023. The Author(s).)

Background: Ixodes spp. are vectors of zoonotic pathogens. All three active life stages (larvae, nymphs, adults) need to feed on a host in order to develop. Usually ticks parasitize attached to the external surface of their hosts' skin. Interestingly, in some cases ticks can also be found in the subcutaneous tissue in a variety of hosts, such as red foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) and dogs. Methods: The visceral side of 126 red fox-furs from Germany was examined visually searching for ticks. The localization of ticks was recorded and assigned to ten specific body parts. Morphological identification of ticks was performed according to standardized taxonomic protocols. Ticks which could not be further identified were examined genetically via conventional PCR targeting the 16S rRNA and cox1 gene. Hematoxylin and eosin (H&E) staining was used for histopathological examination. Results: In 111 out of 126 (88.1%) examined coats, at least one tick was found in the subcutaneous tissue. A total of 1203 ticks were removed from the subcutaneous tissue. Well-preserved ticks could be identified based on morphological criteria, but most ticks were in a progressed state of decomposition. Here, morphological species identification was not successful. Also, PCR methods did not lead to a successful species identification. The following species and development stages were found by morphological identification: Ixodes ricinus (female, n = 289; male, n = 8; nymph, n = 1), I. hexagonus (female, n = 2), I. canisuga (female, n = 1). Male I. ricinus were found individually or copulating in pairs with females. Subcutaneous ticks were localized at three predominant affected body parts: ears, axillar and inguinal region. Histological examination of subcutaneous ticks revealed a granulomatous panniculitis. Conclusions: To the authors' knowledge, this is the first finding of highly prevalent subcutaneous ticks in red foxes from Germany. Subcutaneous location of ticks seems to be very common in red foxes and the rule rather than the exception. Deep embedment of longirostra and long feeding times of females seem to put the subcutaneous location in favor. Most foxes were infested in the inguinal area, where the skin is thin and less hairy. [ABSTRACT FROM AUTHOR]

Purpose: Despite being vaccine-preventable, tick-borne encephalitis (TBE) continues to cause considerable morbidity in Germany. Limited insight into potentially debilitating consequences of TBE may partially underly low (~ 20%) TBE vaccine uptake. We aimed to systematically assess TBE sequelae and other consequences., Methods: Routinely notified TBE patients from 2018 to 2020 from Southern Germany were invited to telephone interviews acutely and again after 18 months. Duration of acute symptoms was prospectively assessed. Recovery was defined as score 0 on the modified RANKIN scale. Determinants of time to recovery were analysed with cox regression, adjusted for covariates identified using directed acyclic graphs, yielding hazard ratios (HR) and 95% confidence intervals (CI)., Results: Of 558 cases, 523 (93.7%) completed follow-up. Full recovery was reported by 67.3% (children: 94.9%, adults: 63.8%). Sequelae included fatigue (17.0%), weakness (13.4%), concentration deficit (13.0%), and impaired balance (12.0%). Compared with 18-39-year-olds, recovery rates were 44% lower in ≥ 50-year-olds (HR: 0.56, 95%CI 0.42-0.75) and 79% higher in children (HR: 1.79, 95%CI 1.25-2.56). The recovery rate was 64% lower after severe TBE (compared to mild; HR: 0.36, 95%CI 0.25-0.52) and 22% lower with comorbidities (HR: 0.78, 95%CI 0.62-0.99). Substantial health-care use was reported (90.1% hospitalisation, 39.8% rehabilitation). Of employed cases, 88.4% required sick leave; 10.3% planned/reported premature retirement due to sequelae., Conclusion: Half the adult and 5% of paediatric patients reported persisting sequelae after 18 months. Improved prevention could alleviate both individual (morbidity) and societal TBE burden (health-care costs, productivity losses). Insights into sequelae can help guide at-risk populations towards tick-avoidant strategies and encourage TBE vaccination., (© 2023. The Author(s).)

Background: Ixodid ticks are important vectors for zoonotic pathogens, with Ixodes ricinus being the most important in Europe. Rodents are hosts of immature life stages of I. ricinus ticks and are considered main reservoirs for tick-borne pathogens, e.g. Borrelia burgdorferi. The aim of this study was to analyse the prevalence as well as genospecies and sequence type (ST) diversity of Borrelia burgdorferi sensu lato in ticks and small mammals from central Germany and to elaborate on the influence of environmental and/or individual host and vector factors on Borrelia prevalence. Methods: After species identification, 1167 small mammal skin samples and 1094 ticks from vegetation were screened by B. burgdorferi sensu lato real-time polymerase chain reaction, and positive samples were characterized by multilocus sequence typing. Generalized linear (mixed) models were used to estimate how seasonality, small mammal species/tick life stage and habitat affect individual infection status. Results: In total, 10 small mammal species and three tick species, Ixodes ricinus, Ixodes inopinatus (both considered members of the I. ricinus complex) and Dermacentor reticulatus, were investigated. Borrelia DNA was detected in eight host species, i.e. the striped field mouse (Apodemus agrarius), the yellow-necked field mouse (Apodemus flavicollis), the wood mouse (Apodemus sylvaticus), the water vole (Arvicola amphibius), the bank vole (Clethrionomys glareolus), the field vole (Microtus agrestis), the common vole (Microtus arvalis), and the common shrew (Sorex araneus). Two species were Borrelia negative, the greater white-toothed shrew (Crocidura russula) and the pygmy shrew (Sorex minutus). The average prevalence was 6.2%, with two genospecies detected, Borrelia afzelii and Borrelia garinii, and at least three STs that had not been previously reported in small mammals. Borrelia prevalence in small mammals did not differ between seasons. Six genospecies of Borrelia—Borrelia afzelii, Borrelia valaisiana, Borrelia garinii, Borrelia lusitaniae, Borrelia spielmanii, and Borrelia burgdorferi sensu stricto—and 25 STs of Borrelia, of which 12 have not been previously described at all and five have not been previously reported in Germany, were detected in 13% of I. ricinus complex ticks. Prevalence was highest in adult females (25.3%) and lowest in nymphs (11.4%). Prevalence was significantly higher in ticks from grassland (16.8%) compared to forests (11.4%). Conclusions: The high level of small mammal diversity in this region of Germany seems to be reflected in a wide variety of genospecies and STs of B. burgdorferi. [ABSTRACT FROM AUTHOR]

Lyme borreliosis (LB) is caused by the transmission of Borrelia burgdorferi s.l. from ticks to humans. Climate affects tick abundance, and climate change is projected to promote shifts in abundance in Europe, potentially increasing human exposure. We analyzed serum samples collected between the years 2014-2019 from German National Cohort (NAKO) participants at four study sites (Augsburg, Berlin, Hanover, Münster) for immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies using an enzyme-linked immunosorbent assay (ELISA) and line blot immunoassay as confirmatory test for positive and equivocal ELISA samples. We reported crude and weighted seropositivity proportions for local estimates. We used mixed model analysis to investigate associated factors, such as age, sex, migration background, or animal contacts. We determined the serostatus of 14,207 participants. The weighted seropositivity proportions were 3.4% (IgG) and 0.4% (IgM) in Augsburg, 4.1% (IgG) and 0.6% (IgM) in northern Berlin, 3.0% (IgG) and 0.9% (IgM) in Hanover, and 2.7% (IgG) and 0.6% (IgM) in Münster. We found higher odds for IgG seropositivity with advancing age (p < 0.001), among males compared to females (p < 0.001) and reduced odds among participants with migration background compared to those without (p = 0.001). We did not find evidence for an association between serostatus and depression, children within the household, or animal contact, respectively. We found low seropositivity proportions and indications of differences across the study locations, although between-group comparisons did not yield significant results. Comparisons to earlier research are subject to important limitations; however, our results indicate no major increases in seropositivity over time. Nevertheless, monitoring of seropositivity remains critical in light of potential climate-related Borrelia exposure., (© 2023. The Author(s).)

Changing climatic conditions and other anthropogenic influences have altered tick distribution, abundance and seasonal activity over the last decades. In Germany, the two most important tick species are Ixodes ricinus and Dermacentor reticulatus, the latter of which has expanded its range across the country during the past three decades. While I. ricinus was rarely found during the colder months in the past, D. reticulatus is known to be active at lower temperatures. To quantify tick appearance during winter, specimens were monitored in quasi-natural tick plots three times a week. Additionally, the questing activities of these two tick species were observed throughout the year at nine field collection sites that were regularly sampled by the flagging method from April 2020 to April 2022. Furthermore, tick winter activity in terms of host infestation was analysed as part of a nationwide submission study from March 2020 to October 2021, in which veterinarians sent in ticks mainly collected from dogs and cats. All three study approaches showed a year-round activity of I. ricinus and D. reticulatus in Germany. During the winter months (December to February), on average 1.1% of the inserted I. ricinus specimens were observed at the tops of rods in the tick plots. The average questing activity of I. ricinus amounted to 2 ticks/100 m² (range: 1-17) in the flagging study, and 32.4% (211/651) of ticks found infesting dogs and cats during winter 2020/21 were I. ricinus. On average 14.7-20.0% of the inserted D. reticulatus specimens were observed at the tops of rods in the tick plots, while the average winter questing activity in the field study amounted to 23 specimens/100 m² (range: 0-62), and 49.8% (324/651) of all ticks collected from dogs and cats during winter 2020/21 were D. reticulatus. Additionally, the hedgehog tick Ixodes hexagonus was found to infest dogs and cats quite frequently during the winter months, accounting for 13.2% (86/651) of the collected ticks. A generalized linear mixed model identified significant correlations of D. reticulatus winter activity in quasi-natural plots with climatic variables. The combined study approaches confirmed a complementary main activity pattern of I. ricinus and D. reticulatus with climate change-driven winter activity of both species. Milder winters and a decrease of snowfall, and consequently high winter activity of D. reticulatus, among other factors, may have contributed to the rapid spread of this tick species throughout the country. Therefore, an effective year-round tick control is strongly recommended to not only efficiently protect dogs and cats with outdoor access from ticks and tick-borne pathogens (TBPs), but also to limit the further geographical spread of ticks and TBPs to so far non-endemic regions. Further measures, including information of the public, are necessary to protect both, humans and animals, in a One Health approach., Competing Interests: Declaration of Competing Interest CS declares that she repeatedly has lectured for and acted as consultant for (veterinary) pharmaceutical and diagnostic companies and has previous and ongoing research collaborations with (veterinary) pharmaceutical and diagnostic companies. JP declares that she has been employed as a PhD student in the project, which was included in the financial support by Intervet Deutschland GmbH. HW is an employee of MSD Animal Health Innovation GmbH. OK is CEO of tick-radar GmbH and HD of IS Insect Services GmbH. MB is working as a freelancer for Global Health Press. Study conceptualization, investigation and data interpretation is completely independent of any companies´ opinions. The remaining authors declare that they have no conflict of interest., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Background: Tick-borne encephalitis (TBE) is the most important tick-borne viral disease in Eurasia and causes disease in humans and in a number of animals, among them dogs and horses. There is still no good correlation between tick numbers, weather conditions and human cases. There is the hypothesis that co-feeding due to simultaneous occurrence of larvae and nymphs may be a factor for the increased transmission of the virus in nature and for human disease. Based on long-term data from a natural TBEV focus, phylogenetic results and meteorological data we sought to challenge this hypothesis. Methods: Ticks from an identified TBE natural focus were sampled monthly from 04/2009 to 12/2018. Ticks were identified and pooled. Pools were tested by RT-qPCR. Positive pools were confirmed by virus isolation and/or sequencing of additional genes (E gene, NS2 gene). Temperature data such as the decadal (10-day) mean daily maximum air temperature (DMDMAT) were obtained from a nearby weather station and statistical correlations between tick occurrence and minimal infection rates (MIR) were calculated. Results: In the study period from 04/2009 to 12/2018 a total of 15,530 ticks (2,226 females, 2,268 males, 11,036 nymphs) were collected. The overall MIR in nymphs over the whole period was 77/15,530 (0.49%), ranging from 0.09% (2009) to 1.36% (2015). The overall MIR of female ticks was 0.76% (17/2,226 ticks), range 0.14% (2013) to 3.59% (2016). The overall MIR of males was 0.57% (13/2,268 ticks), range from 0.26% (2009) to 0.97% (2015). The number of nymphs was statistically associated with a later start of spring/vegetation period, indicated by the onset of forsythia flowering. Conclusion: There was no particular correlation between DMDMAT dynamics in spring and/or autumn and the MIR of nymphs or adult ticks detected. However, there was a positive correlation between the number of nymphs and the number of reported human TBE cases in the following months, but not in the following year. The hypothesis of the importance of co-feeding of larvae and nymphs for the maintenance of transmission cycle of TBEV in nature is not supported by our findings. [ABSTRACT FROM AUTHOR]

Background: Ixodes ricinus is the most common tick species in Europe and the main vector for Borrelia burgdorferi (sensu lato) and tick-borne encephalitis virus (TBEV). It is involved also in the transmission of Borrelia miyamotoi, a relapsing fever spirochete that causes health disorders in humans. Little is known regarding the circulation of Borrelia species and the natural foci of TBEV in north-eastern Germany. The goal of this study was to investigate the infection rates of Borrelia spp. and of TBEV in I. ricinus ticks from north-eastern Germany., Methods: Ticks were collected by flagging from 14 forest sites in Mecklenburg-Western Pomerania between April and October 2018. RNA and DNA extraction was performed from individual adult ticks and from pools of 2-10 nymphs. Real time reverse transcription PCR (RT-qPCR) targeted the 3' non-coding region of TBEV, while DNA of Borrelia spp. was tested by nested PCR for the amplification of 16S-23S intergenic spacer. Multilocus sequence typing (MLST) was performed on B. miyamotoi isolates., Results: In total, 2407 ticks were collected (239 females, 232 males and 1936 nymphs). Female and male I. ricinus ticks had identical infection rates (both 12.1%) for Borrelia spp., while nymphal pools showed a minimum infection rate (MIR) of 3.3%. Sequencing revealed four Borrelia species: B. afzelii, B. garinii, B. valaisiana and B. miyamotoi. Borrelia afzelii had the highest prevalence in adult ticks (5.5%) and nymphs (MIR of 1.8%). Borrelia miyamotoi was identified in 3.0% of adults and registered the MIR of 0.8% in nymphs. Borrelia valaisiana was confirmed in 2.5% adult ticks and nymphs had the MIR of 0.7%, while B. garinii was present in 1.1% of adults and showed a MIR of 0.1% in nymphs. The MLST of B. miyamotoi isolates showed that they belong to sequence type 635. No tick sample was positive after RT-qPCR for TBEV RNA., Conclusions: The prevalence of B. miyamotoi in I. ricinus ticks registered similar levels to other reports from Europe suggesting that this agent might be well established in the local tick population. The detection of B. burgdorferi (s.l.) indicates a constant circulation in tick populations from this region.

The first data update of the atlas of ticks in Germany published in 2021 is presented here. This atlas provides maps based on georeferenced tick locations of 21 species endemic in Germany as well as three tick species that are regularly imported to Germany. The data update includes the following numbers of newly georeferenced tick locations: 17 Argas reflexus, 79 Carios vespertilionis, 2 Dermacentor marginatus, 43 Dermacentor reticulatus, 4 Haemaphysalis concinna, 3 Haemaphysalis punctata, 3 Hyalomma rufipes, 3 Ixodes apronophorus, 9 Ixodes arboricola, 1 Ixodes ariadnae, 30 Ixodes canisuga, 3 Ixodes frontalis, 80 Ixodes hexagonus, 3 Ixodes lividus, 497 Ixodes ricinus/inopinatus, 1 Ixodes rugicollis, 17 Ixodes trianguliceps, 14 Ixodes vespertilionis, and 45 Rhipicephalus sanguineus sensu lato. Old and new tick findings were mapped, such as the northernmost occurrence of D. marginatus in Germany observed in 2021, but also the historical records from the first descriptions of I. apronophorus and I. arboricola, which were georeferenced here for the first time. The digital dataset of tick locations available for Germany is supplemented by 854 new tick locations. These records increase the number of tick species mapped in the federal states Bavaria, Brandenburg and Mecklenburg Western Pomerania by five each, those in Berlin and Schleswig-Holstein by four each, those in Hamburg by three, those in Baden-Wuerttemberg, Bremen, Lower Saxony, Northrhine-Westphalia, Rhineland Palatinate and Thuringia by two each, and those in Hesse, Saxony and Saxony-Anhalt by one each. Thus, the first data update of the tick atlas in Germany and the underlying digital dataset significantly improve our knowledge of the distribution of these tick species and helps to investigate the effects of climate change and habitat changes on them. [ABSTRACT FROM AUTHOR]

Ixodes ricinus is the most abundant tick species and an important vector of pathogens in Germany and in large parts of Europe. A few other ixodid tick species, e.g., Dermacentor reticulatus, may also be of eco-epidemiological relevance. As ticks are not only found in natural but also in suburban areas (parks, gardens), the present study investigated whether ticks occur on and near football grounds thus posing a potential risk to players and visitors. Thirty-two football grounds from all 16 German federal states were selected, mainly situated adjacent to a green area (forest, park). Ticks were collected by the conventional flagging method in spring 2018, and nymphs and adults were counted and morphologically determined. Altogether 807 nymphal and adult ticks were collected from 29 football grounds: 714 I. ricinus, 64 Ixodes inopinatus, 2 Ixodes frontalis, 24 Ixodes sp. ticks, and 3 D. reticulatus. Ixodes inopinatus was found in 13 out of 16 German states. Three ticks were even found on the turf of two football fields. It can be concluded that ticks occur quite frequently and sometimes in high abundance near football grounds situated close or adjacent to a forest or a park. [ABSTRACT FROM AUTHOR]

Background: The European hedgehog (Erinaceus europaeus) is known for high levels of ectoparasitism that not only represents a health risk for the animals themselves, but also for pet animals and humans as hedgehogs are frequently taken into human care. In the present study, patterns of ectoparasite infestation were assessed in hedgehogs taken into care at northern German animal rehabilitation centres. Methods: Ectoparasites (ticks, fleas and mites) of 498 hedgehogs were collected over a period of 3 years from July 2018 to May 2021. Species were identified based on morphological characteristics and also via amplification and sequencing of the partial cytochrome c oxidase subunit 2 (COX-2) gene for fleas of the family Ceratophyllidae. Seasonal changes in infestation patterns as well as correlations with animal age, body weight and health status were assessed using generalised linear models. Results: Infestation with ticks, fleas and mites occurred throughout the year. Overall, 86.5% (431/498) of the examined hedgehogs were infested with ticks, 91.4% (455/498) with fleas and 17.7% (88/498) with mites.Ixodes ricinus and Ixodes hexagonus/Ixodes canisuga were the most common tick species detected, with the additional occurrence of one Ixodes frontalis. Significant seasonal changes were observed for I. ricinus, but not for I. hexagonus/I. canisuga. Additionally, I. ricinus nymph prevalence declined significantly as of 2020, probably as a consequence of the climate change-related drought as of 2018. In hedgehogs with flea infestations, Archaeopsylla erinacei, Ceratophyllus sciurorum, Nosopsyllus fasciatus and Ctenocephalides felis were identified. In all cases of mite infestation, Caparinia tripilis was detected, in addition to specimens of the family Macronyssidae and free-living mites of the family Acaridae. Statistical analyses showed correlations regarding the factors month, year, body weight and age, but no correlation was evident regarding the health status of the animals. Conclusions: With a detected infestation rate of 98.6%, almost all of the examined hedgehogs were infested with at least one ectoparasite species. The seasonal activity patterns of the different ectoparasite species together with the complex annual cycle of hedgehogs lead to different seasonal patterns in ectoparasite prevalence and infestation intensities. Due to the risk of transmission of zoonotic pathogens as well as the possible negative impact on the host itself, hedgehogs should be treated against ectoparasites when taken into care facilities. [ABSTRACT FROM AUTHOR]

Southern Germany is known as tick-borne encephalitis (TBE) risk area; however, the north of the country is almost free of human TBE cases. Due to its location in the transition zone between TBE risk areas and areas with only sporadic cases, Saxony is of importance in the surveillance of TBE. Roe deer (Capreolus capreolus), showing high seroprevalence of TBE virus (TBEV) antibodies, are considered to be sentinels for TBE risk assessment. Thus, roe deer could be used as indicators helping to better understand the focality of the TBEV in nature and as a possible source to isolate TBEV. Therefore, the aims of this study were to examine roe deer coats for the presence of ticks to establish the tick burden and to detect the TBEV in attached ticks. One hundred thirty-four roe deer coats were provided by hunters from the Hunting Association in Saxony (August 2017–January 2019). The coats were frozen at − 80 °C and after de-freezing examined on both sides—inside and outside. Attached and nonattached ticks were collected, morphologically identified and tested using real-time RT-PCR for the presence of TBEV. In total, 1279 ticks were found on 48 coats. The predominant species was Ixodes ricinus (99.76%; n = 1276). Three remaining specimens were Ixodes spp. (0.16%, 1 female and 1 nymph) and Dermacentor reticulatus (0.08%, 1 male). The average infestation rate was 26.7 (SD = 69.5), with maximum of 439 ticks per animal. Females were the dominant life stage of ticks (n = 536; 42%), followed by nymphs (n = 397; n = 31.1%), males (n = 175; 13.7%), and larvae (n = 168; 13.2%). Only half of collected ticks were attached (n = 662; 51.8%). TBEV was detected only in one tick out of 1279 tested ticks. It was a female infesting a roe deer from Saxon Switzerland-East Ore Mountain. The results show that the method used in this study is not sufficient as a sentinel marker to predict TBEV spreading in nature. Although previous studies demonstrated the usefulness of serological testing of roe deer in order to trace TBE-endemic regions, using ticks attached to them to get virus isolates is not productive. [ABSTRACT FROM AUTHOR]

Background: Borreliosis is the most frequently transmitted tick-borne disease in Europe. It is difficult to estimate the incidence of tick bites and associated diseases in the German population due to the lack of an obligation to register across all 16 federal states of Germany., Objective: The aim of this study is to show that Google data can be used to generate general trends of infectious diseases on the basis of borreliosis and tick bites. In addition, the possibility of using Google AdWord data to estimate incidences of infectious diseases, where there is inconsistency in the obligation to notify authorities, is investigated with the perspective to facilitate public health studies., Methods: Google AdWords Keyword Planner was used to identify search terms related to ticks and borreliosis in Germany from January 2015 to December 2018. The search volume data from the identified search terms was assessed using Excel version 15.23. In addition, SPSS version 24.0 was used to calculate the correlation between search volumes, registered cases, and temperature., Results: A total of 1999 tick-related and 542 borreliosis-related search terms were identified, with a total of 209,679,640 Google searches in all 16 German federal states in the period under review. The analysis showed a high correlation between temperature and borreliosis (r=0.88), and temperature and tick bite (r=0.83), and a very high correlation between borreliosis and tick bite (r=0.94). Furthermore, a high to very high correlation between Google searches and registered cases in each federal state was observed (Brandenburg r=0.80, Mecklenburg-West Pomerania r= 0.77, Saxony r= 0.74, and Saxony-Anhalt r=0.90; all P<.001)., Conclusions: Our study provides insight into annual trends concerning interest in ticks and borreliosis that are relevant to the German population exemplary in the data of a large internet search engine. Public health studies collecting incidence data may benefit from the results indicating a significant correlation between internet search data and incidences of infectious diseases., (©Cora Scheerer, Melvin Rüth, Linda Tizek, Martin Köberle, Tilo Biedermann, Alexander Zink. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 16.10.2020.)

In Germany, the knowledge about ticks infesting bats is limited, and is restricted only to a few studies, most of them dating back decades. To further improve our knowledge on ticks parasitising bats, healthy and sick bats in central Germany were examined for ticks. In total 519 larvae and one nymph of Carios vespertilionis were collected from nine bat species: Eptesicus nilssonii, Eptesicus serotinus, Myotis daubentonii, Myotis myotis, Nyctalus leisleri, Pipistrellus nathusii, Pipistrellus pygmaeus, Pipistrellus pipistrellus, and Vespertilio murinus. Either the presence of C. vespertilionis was new for some areas or it was confirmed in some federal states in central Germany. The infestation rate was mostly low (n = 1–5 larvae/bat). However, in two cases a high number of ticks was observed. The highest infestation of 97 C. vespertilionis larvae was recorded on one Parti-coloured bat (V. murinus). [ABSTRACT FROM AUTHOR]

Background: Arthropod-borne diseases remain a major health-threat for humans and animals worldwide. To estimate the distribution of pathogenic agents and especially Bartonella spp., we conducted tick microbiome analysis and determination of the infection status of wild animals, pets and pet owners in the state of Hesse, Germany., Results: In total, 189 engorged ticks collected from 163 animals were tested. Selected ticks were analyzed by next generation sequencing (NGS) and confirmatory PCRs, blood specimens of 48 wild animals were analyzed by PCR to confirm pathogen presence and sera of 54 dogs, one cat and 11 dog owners were analyzed by serology. Bartonella spp. were detected in 9.5% of all ticks and in the blood of 17 roe deer. Further data reveal the presence of the human and animal pathogenic species of genera in the family Spirochaetaceae (including Borrelia miyamotoi and Borrelia garinii), Bartonella spp. (mainly Bartonella schoenbuchensis), Rickettsia helvetica, Francisella tularensis and Anaplasma phagocytophilum in ticks. Co-infections with species of several genera were detected in nine ticks. One dog and five dog owners were seropositive for anti-Bartonella henselae-antibodies and one dog had antibodies against Rickettsia conorii., Conclusions: This study provides a snapshot of pathogens circulating in ticks in central Germany. A broad range of tick-borne pathogens are present in ticks, and especially in wild animals, with possible implications for animal and human health. However, a low incidence of Bartonella spp., especially Bartonella henselae, was detected. The high number of various detected pathogens suggests that ticks might serve as an excellent sentinel to detect and monitor zoonotic human pathogens.

Small mammals serve as reservoir hosts for tick-borne pathogens, especially for those which are not transmitted transovarially in ticks - such as Babesia microti. Molecular investigations on the prevalence of B. microti in wild small mammals and on attached ticks from differently structured areas may provide information on the circulation of B. microti in different ecological niches. In 2012 and 2013, 622 rodents (396 Myodes glareolus, 178 Apodemus flavicollis, 36 Apodemus sylvaticus, 4 Apodemus agrarius, 7 Microtus arvalis, 1 Microtus agrestis) were captured from three differently structured habitats (urban, sylvatic, recultivated) in Germany. Attached ticks were collected from 449 small mammals (3250 Ixodes ricinus, 7 Ixodes trianguliceps, 133 Dermacentor reticulatus). A representative selection of a maximum of 5 ticks per developmental stage and species per 30 rodents of each species, location and year resulting in 965 ticks was further investigated. DNA was extracted from tick, blood and spleen samples, and tested by PCR for the partial 18S rRNA gene of B. microti with subsequent sequencing. The prevalence was significantly higher in rodents from the sylvatic site (4.6%) than in rodents captured at both other sites (-0.6%) (χ(2)=11.95; p=0.00125). Body and spleen weight of infected M. glareolus from the sylvatic site were significantly higher compared to those from non-infected individuals from that site (p=0.00288 and p=0.00017, respectively). Babesia microti DNA was detected in 3 out of 965 attached ticks (0.3%; 95%CI: 0-1) from all sites, but they derived exclusively from rodents captured at the sylvatic site. At the same site, I. ricinus nymphs (7.7%; 95%CI: 1-25.3) were significantly more often infected than I. ricinus larvae (0%; 95%CI: 0-1.3)(χ(2)=26.72; p<0.0001). The majority of positive rodents was also found at that site. I. trianguliceps occurred exclusively and the majority of M. glareolus at that site. Thus, it may be assumed that the circulation of B. microti is more efficient where this tick species and voles exist sympatrically than in areas with a predominant occurrence of Apodemus species., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Models can be applied to extrapolate consequences of climate change for complex ecological systems in the future. The acknowledged systems' behaviour at present is projected into the future considering climate projection data. Such an approach can be used to addresses the future activity and density of the castor bean tick Ixodes ricinus, the most widespread tick species in Europe. It is an important vector of pathogens causing Lyme borreliosis and tick-borne encephalitis. The population dynamics depend on several biotic and abiotic factors. Such complexity makes it difficult to predict the future dynamics and density of I. ricinus and associated health risk for humans. The objective of this study is to force ecological models with high-resolution climate projection data to extrapolate I. ricinus tick density and activity patterns into the future. We used climate projection data of temperature, precipitation, and relative humidity for the period 1971–2099 from 15 different climate models. Tick activity was investigated using a climate-driven cohort-based population model. We simulated the seasonal population dynamics using climate data between 1971 and 2099 and observed weather data since 1949 at a specific location in southern Germany. We evaluated derived quantities of local tick ecology, e.g. the maximum questing activity of the nymphal stage. Furthermore, we predicted spatial density changes by extrapolating a German-wide tick density model. We compared the tick density of the reference period (1971–2000) with the counter-factual densities under the near-term scenario (2012–2041), mid-term scenario (2050–2079) and long-term scenario (2070–2099). We found that the nymphal questing peak would shift towards early seasons of the year. Also, we found high spatial heterogeneity across Germany, with predicted hotspots of up to 2,000 nymphs per 100 m2 and coldspots with constant density. As our results suggest extreme changes in tick behaviour and density, we discuss why caution is needed when extrapolating climate data-driven models into the distant future when data on future climate drive the model projection. [ABSTRACT FROM AUTHOR]

An updated and increased compilation of georeferenced tick locations in Germany is presented here. This data collection extends the dataset published some years ago by another 1448 new tick locations, 900 locations of which were digitized from literature and 548 locations are published here for the first time. This means that a total of 3492 georeferenced tick locations is now available for Germany. The tick fauna of Germany includes two species of Argasidae in the genera Argas and Carios and 19 species of Ixodidae in the genera Dermacentor, Haemaphysalis, and Ixodes, altogether 21 tick species. In addition, three species of Ixodidae in the genera Hyalomma (each spring imported by migratory birds) and Rhipicephalus (occasionally imported by dogs returning from abroad with their owners) are included in the tick atlas. Of these, the georeferenced locations of 23 tick species are depicted in maps. The occurrence of the one remaining tick species, the recently described Ixodes inopinatus, is given at the level of the federal states. The most common and widespread tick species is Ixodes ricinus, with records in all 16 federal states. With the exception of Hamburg, Dermacentor reticulatus was also found in all federal states. The occurrence of the ixodid ticks Ixodes canisuga, Ixodes frontalis, Ixodes hexagonus and I. inopinatus were documented in at least 11 federal states each. The two mentioned argasid tick species were also documented in numerous federal states, the pigeon tick Argas reflexus in 11 and the bat tick Carios vespertilionis in seven federal states. The atlas of ticks in Germany and the underlying digital dataset in the supplement can be used to improve global tick maps or to study the effects of climate change and habitat alteration on the distribution of tick species. [ABSTRACT FROM AUTHOR]

Rickettsiales (Anaplasma phagocytophilum and Rickettsia spp.) are regarded as potentially emerging tick-borne pathogens and may change in abundance in response to global climate change. However, continuous monitoring on their prevalence in questing ticks is only available for the northern German city of Hanover. In the presented follow-up of this long-term study, 2100 questing ticks of the Ixodes ricinus/Ixodes inopinatus-complex collected from April to October 2020 at ten different recreation sites in Hanover were individually analysed for Rickettsia and A. phagocytophilum infection by quantitative real-time PCR. Together with previous results from years 2005, 2010 and 2015, the current study allows to assess potential changes in tick infection rates with Rickettsiales over a 15-year monitoring period. In 2020, 3.0% (63/2100) of ticks were infected with A. phagocytophilum, 36.0% (756/2100) with Rickettsia spp. and 1.2% (26/2100) with both pathogens. Regarding the different developmental tick stages, nymphs showed a significantly lower A. phagocytophilum prevalence of 0.5% (5/1050) than adult ticks (5.5% [58/1050]) as well as compared to females (5.4% [38/700]) and males (5.7% [20/350]). For Rickettsia spp., nymphs also showed a lower prevalence of 33.2% (349/1050) with a significant difference to adult ticks (38.8% [407/1050]) and female ticks (40.7% [285/700]), while males had a Rickettsia infection rate of 34.9% (122/350). Comparison with previous years indicated a stable A. phagocytophilum prevalence over the 15-year monitoring period. In contrast, fluctuating Rickettsia prevalences were observed, with a peak in 2015 in all developmental stages, but similar infection rates in 2005 and 2020. Therefore, epidemiological changes in response to climate change are not (yet) evident. Nevertheless, the long-term monitoring study will be continued in the future, as climatic impacts on tick and reservoir host populations may have a delayed effect on pathogen prevalence and, consequently, transmission to humans and domestic animals., (Copyright © 2022. Published by Elsevier GmbH.)

Background: Ticks and tick-borne diseases play a major role in companion animal health. Additionally, the European tick fauna is changing, for instance due to the spread of Dermacentor reticulatus, displaying a higher likelihood of winter activity than Ixodes ricinus. Therefore, we investigated current tick infestations in dogs and cats in Germany and in parts of Austria and the seasonal infestation risk. Methods: Overall, 219 veterinary practices were invited to collect ticks from cats and dogs on a monthly basis. Ticks were morphologically identified and female I. ricinus specimens were measured to estimate attachment duration. Results: In total, 19,514 ticks, 17,789 (91.2%) from Germany and 1506 (7.7%) from Austria, were received between March 2020 and October 2021, with 10,287 specimens (52.7%) detached from dogs, 8005 from cats (41.0%) and 1222 from other species (6.3%). In Germany, the most common tick species collected from dogs were I. ricinus (78.0%) and D. reticulatus (18.8%), while cats mainly harboured I. ricinus (91.3%) and I. hexagonus (5.5%) and only few D. reticulatus (0.6%). In Austria, collected I. ricinus reached similar proportions in dogs (90.4%) and cats (95.3%), followed by D. reticulatus in both dogs (5.2%) and cats (1.5%), with I. hexagonus (0.9%) collected only marginally from cats. The average infestation intensity amounted to 1.62 ticks/dog and 1.88 ticks/cat. The single to multiple infestation ratio was 79.1% to 20.9% in dogs and 69.0% to 31.0% in cats, with cats being significantly more often multiple infested than dogs, while the proportion of mixed-species infestations was 2.0% for both dogs and cats. The average attachment duration of female I. ricinus specimens amounted to 78.76 h for dogs and 82.73 h for cats. Furthermore, year-round tick exposure was confirmed, with 108 D. reticulatus and 70 I. ricinus received on average per month during December 2020 to February 2021. Conclusions: The study shows a year-round tick infestation risk, with activity of both D. reticulatus and I. ricinus during winter, and confirms the widespread occurrence of D. reticulatus in Germany. Additionally, long average attachment durations and frequent multiple infestations underline the need for adequate year-round tick control, even during the winter months. [ABSTRACT FROM AUTHOR]

Background: As the vector-borne diseases tick-borne encephalitis (TBE) and Lyme borreliosis (LB) are common in Germany and transmitted by tick bites, the aim of this study was to assess differences in the number of tick bites in various professions and regions across southern Germany to evaluate the differences in tick-associated risk., Materials and Methods: The analysis is based on three cross-sectional studies that were conducted in 2016 and 2017 in two real-life settings and in one medical setting in Bavaria. All participants filled in a paper-based questionnaire about their history with tick bites. Only adult participants (≥ 18 years) were included in this study., Results: Overall, 3503 individuals (mean age 50.8 ± 15.2 years, median age 53.0 ± 12.2 years, 54.0% female) were included. Of these, 50% worked in an outdoor profession and 56% lived in environs. Around 70% of participants reported at least one previous tick bite. In comparison to indoor workers, forestry workers (OR = 2.50; 95% CI: 1.10-5.68) had the highest risk for a tick bite followed by farmers (OR = 1.22; 95% CI: 1.01-1.47). Furthermore, people living in rural areas (OR = 1.97, 95% CI:1.49-2.59) and environs (OR = 1.98, 95% CI: 1.54-2.55) were twice as likely to have a previous tick bite than people living in urban areas. In general, slightly more tick bites were reported by people living in eastern Bavaria., Conclusion: Rising numbers of TBE and LB indicate the need for further prevention strategies, which should focus on outdoor professions with a higher risk and people living in environs and rural areas., (© 2022. The Author(s).)

Tick-borne diseases are recognized as a growing public health concern and cause significant issues in humans and animals by serving as dispersal agents of ticks and their associated pathogens. Birds contribute to the spread of ticks and tick-borne diseases, with migratory birds playing a role in regional dispersal. Only a few studies have been carried out in Germany so far to investigate tick fauna on birds and the pathogens they harbour. In 2017 and 2018, we collected ticks from a total of 358 birds in the northern region of the Federal State of Hesse, Germany. Most ticks were Ixodes ricinus larvae and nymphs (95.4%). However, two ornithophilic species were also collected: Ixodes frontalis (larvae, nymphs and females) and Ixodes arboricola (larvae and nymphs). Ticks were tested individually or in pools, depending on the tick life stage and species for each bird host, in order to detect tick-borne encephalitis (TBE) virus and Rickettsia spp. All samples tested negative for TBE virus. Of 684 investigated tick pools, 162 tested positive for Rickettsia spp. by screening PCR. Of these, 117 carried Rickettsia helvetica detected by specific real-time PCR. The samples which were negative in the R. helvetica PCR were further investigated using five targets (gltA, 16S, ompA IV, ompB, 23S-5S intergenic spacer region) using multi locus sequence typing and phylogenetic analysis. The obtained sequences showed strong phylogenetic relationship to Candidatus Rickettsia vini, which is most closely related to R. japonica and R. heilongjiangensis, and occur in I. arboricola. Three more sequences were determined as R. helvetica in I. ricinus and I. frontalis. This is the first detection of Ca. R. vini in Germany and the first 23S-5S data published for Ca. R. vini, a useful target for Rickettsia species identification., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Background: Tick-borne encephalitis (TBE) is the most common viral CNS infection with incidences much higher than all other virus infections together in many risk areas of central and eastern Europe. The Odenwald Hill region (OWH) in southwestern Germany is classified as a TBE risk region and frequent case numbers but also more severe infections have been reported within the past decade. The objective of the present study was to survey the prevalence of tick-borne encephalitis virus (TBEV) in Ixodes ricinus and to associate TBEV genetic findings with TBE infections in the OWH. Methods: Ticks were collected by the flagging methods supported by a crowdsourcing project implementing the interested public as collectors to cover completely and collect randomly a 3532 km2 area of the OWH TBE risk region. Prevalence of TBEV in I. ricinus was analysed by reversed transcription quantitative real-time PCR. Phylogeographic analysis was performed to classify OWH TBEV isolates within a European network of known TBEV strains. Mutational sequence analysis including 3D modelling of envelope protein pE was performed and based on a clinical database, a spatial association of TBE case frequency and severity was undertaken. Results: Using the crowd sourcing approach we could analyse a total of 17,893 ticks. The prevalence of TBEV in I. ricinus in the OWH varied, depending on analysed districts from 0.12% to 0% (mean 0.04%). Calculated minimum infection rate (MIR) was one decimal power higher. All TBEV isolates belonged to the European subtype. Sequence analysis revealed a discontinuous segregation pattern of OWH isolates with two putative different lineages and a spatial association of two isolates with increased TBE case numbers as well as exceptional severe to fatal infection courses. Conclusions: TBEV prevalence within the OWH risk regions is comparatively low which is probably due to our methodological approach and may more likely reflect prevalence of natural TBEV foci. As for other European regions, TBEV genetics show a discontinuous phylogeny indicating among others an association with bird migration. Mutations within the pE gene are associated with more frequent, severe and fatal TBE infections in the OWH risk region. [ABSTRACT FROM AUTHOR]

Ectoparasitic arthropodes, like ticks, confer numerous pathogenic bacterial and viral germs upon human beings and/or animals. Since these vectors are poikilothermic, there is a strong dependence on environmental conditions. Climatic as well as microclimatic conditions play a major role. Against this background, it can be anticipated that climatic changes also influence the bionomics of ticks and of the pathogens conferred by them.

To determine Borrelia spp. (Spirochaetales: Spirochaetaceae) prevalence and species distribution in Northern Germany, Ixodes ticks were sampled from April to October in 2018 and 2019 by the flagging method at three locations each in five regions. Analysis by quantitative real‐time PCR of 3150 individual ticks revealed an overall prevalence of 30.6%, without significant differences between tick stages (31.7% positive adults, 28.6% positive nymphs). Significant differences were observed in seasonal infection rates, but not between regions, landscape types or sampling years. Analysis of co‐infections with Rickettsiales indicated a negative association between Borrelia and Anaplasma phagocytophilum infection. The most frequent Borrelia species differentiated by Reverse Line Blot were B. afzelii and B. garinii/B. bavariensis, followed by B. valaisiana, B. burgdorferi sensu stricto, B. spielmanii and B. lusitaniae. Furthermore, B. miyamotoi was identified in 12.9% of differentiable samples. No effect of region nor landscape type on species composition was found, but significant variations in the distribution at the different sampling sites within a region were observed. The detected monthly fluctuations in prevalence and the differences in intra‐regional Borrelia species distribution underline the importance of long‐term and multi‐location monitoring of Borrelia spp. in ticks as an essential part of public health assessment. [ABSTRACT FROM AUTHOR]

Background: Small mammals are important maintenance hosts of ectoparasites as well as reservoir hosts for many arthropod-borne pathogens. In Germany, only a few studies have investigated ectoparasite communities on small mammals in their natural habitats. The aim of this study was to assess the species diversity and parameters influencing the mean intensity and prevalence of macroscopically visible ectoparasites, such as fleas, predatory mites and ticks., Methods: A total of 779 small mammals and 3383 ticks were available from earlier investigations for the data analysis of the current study from three differently structured study sites. In addition, fleas and predatory mites were collected from the captured rodents and taxonomically identified. Regression analyses were conducted on the group (ticks/mites/fleas) and species levels using hurdle models for the abundance of ectoparasite groups and a negative binomial model for the abundance of species., Results: Nearly 90% of the small mammals analyzed were infested with ectoparasites, with an average of 7.3 specimens per host. Hosts were infested with up to six species of ectoparasites simultaneously. In total, 12 flea, 11 mite and three tick species were detected. Ticks were more prevalent than fleas or mites, with > 80% of the hosts in urban and forest areas hosting ticks and around 60% of hosts presenting fleas, and only 20-40% of hosts presenting mites. Polyparasitism had a statistically significant influence on the prevalence of the investigated tick, mite and flea species, with odds ratios of > 1.0. Trapping location, season and host characteristics had significant influences on some-but not all-of the investigated species., Conclusions: The diversity of flea species was unexpectedly high and higher than that reported in comparable studies, which can be explained by the differently structured habitats and regions examined in this study. Polyparasitism was a key influencing factor and had a positive effect on the prevalence and/or abundance of the predominant tick, flea and mite species occurring on small mammals. Season, trapping location, host species and sex of the host species also had an influence on the prevalence and mean intensity of certain, but not all, ectoparasite species.

Simple Summary: The bacterium Anaplasma phagocytophilum can cause granulocytic anaplasmosis in domestic animals, wildlife, and humans. The pathogen is transmitted by ticks and is predominantly endemic in Central and Northern Europe. We discuss the percentages of dogs that tested positive for A. phagocytophilum by direct detection of the pathogen (4.9%) and by detecting antibodies (23.3%). We found a statistically significant impact of seasonality and years of testing on PCR results as well as sex, age, years of testing and seasonality on antibody results. The dynamics of infections with A. phagocytophilum in dogs in Germany are consistent with peaks in vector activity. There is a notable increase in canine granulocytic anaplasmosis in Germany over the time of this study, which calls for increased attention and demonstrates its rising importance in clinical practice. Anaplasma phagocytophilum is an obligate intracellular bacterium that causes granulocytic anaplasmosis in domestic animals, wildlife, and humans and is primarily transmitted by ticks of the Ixodes persulcatus complex. This retrospective study aims to determine the percentages of dogs that tested positive for A. phagocytophilum in Germany. It included the results of direct (polymerase chain reaction [PCR]) and indirect (immunofluorescence antibody test [IFAT], antibody-enzyme-linked immunosorbent assay [ELISA]) detection methods performed in the laboratory LABOKLIN on canine samples provided by German veterinarians from 2008 to 2020. Out of a total of 27,368 dogs tested by PCR, 1332 (4.9%) tested positive, while 24,720 (27.4%) of the 90,376 dogs tested by IFAT/ELISA had positive serology. High rates of positive PCR results were observed in months with known peaks in vector activity, showing that the dynamics of A. phagocytophilum infections in dogs in Germany are consistent with vector activity. In dogs with a positive PCR result, peaks in serology could be observed four weeks after initial testing. Male and senior dogs had higher rates of positive serology. A possible impact of environmental factors such as changes in climate should be investigated further. Overall, the upward trend in positive test results over the years indicates that canine granulocytic anaplasmosis will continue to become increasingly important for veterinary medicine. [ABSTRACT FROM AUTHOR]

Since the publication of 'An annotated checklist of the ticks of Germany' in 2012, the list now includes two additional ixodid species. Ixodes acuminatus is endemic in several countries bordering Germany. Sufficient specimens have now been found in southwestern Germany to show that this species is also native there. The other species, Ixodes inopinatus, has its main area of distribution in the western Mediterranean, and it is not yet clear whether the two females and one male found in Rhineland-Palatinate were accidental introductions or are part of a viable population. [ABSTRACT FROM AUTHOR]

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., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Background: The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts., Methods: Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice), were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated., Results: 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected., Conclusion: Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures.

Tick-associated Rickettsiales are important pathogens with relevance for public and animal health; therefore, knowledge regarding their distribution is essential for risk assessment and disease prevention. To investigate the prevalence of Anaplasma phagocytophilum and Rickettsia spp. in northern Germany, Ixodes ticks were flagged monthly from April to October in 2018 and 2019 at three collection sites each in the regions of Bremen, Emsland, Hanover, Kassel and Uelzen. A total of 3150 ticks (1052 females, 1048 males and 1050 nymphs) were individually examined for rickettsial infections using probe-based quantitative real-time PCR. Overall prevalence of A. phagocytophilum was 6.4 % (202/3150; 6.7 % [71/1052] in females, 7.5 % [79/1048] in males and 5.0 % [52/1050] in nymphs). For Rickettsia spp., the overall prevalence was 29.6 % (931/3150; 33.4 % [351/1052] in females, 28.3 % [297/1048] in males and 27.0 % [283/1050] in nymphs). Rickettsia species identification by real-time pyrosequencing on a subset of 409 positive samples was successful in 407 cases (99.5 %). Rickettsia helvetica was the predominant species with a detection rate of 99.8 % (406/407). Additionally, Rickettsia monacensis was detected in one tick (0.2 %). Generalized linear mixed models showed significant regional as well as monthly differences regarding the prevalence of both pathogens. In addition, the prevalence of both pathogens was significantly higher in 2018 (A. phagocytophilum: 8.0 % [126/1575], Rickettsia spp.: 35.4 % [558/1575]) than in 2019 (A. phagocytophilum: 4.8 % [76/1575], Rickettsia spp.: 23.9 % [373/1575]). In contrast, no effect of landscape type on pathogen prevalence was found. As Rickettsia spp.-detection was based on the single-copy gene gltA, it was possible to calculate the individual pathogen load per tick, which was significantly higher in female ticks than in nymphs (mean values: 8.19 × 10 4 vs. 9.58 × 10 3 ). Regional, seasonal and biennial prevalence differences of tick-transmitted Rickettsiales show the necessity to investigate ticks from multiple locations, over several months and in more than one year to reliably assess the infection risk on a larger geographical scale., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Background: Anaplasma phagocytophilum is a Gram-negative obligate intracellular bacterium that replicates in neutrophil granulocytes. It is transmitted by ticks of the Ixodes ricinus complex and causes febrile illness called granulocytic anaplasmosis primarily in humans, horses, dogs, sheep, cattle and goats. In comparison, clinically apparent disease has been described rarely in cats especially compared to dogs and horses. It is currently unknown whether cats are less susceptible to A. phagocytophilum or whether granulocytic anaplasmosis might be underdiagnosed in cats. Methods: To address this question, we examined clinical signs and laboratory findings in seven A. phagocytophilum infected cats from Germany and Switzerland. We then genetically characterized feline A. phagocytophilum strains and compared them to those from other hosts showing clinically apparent disease. For this purpose, ankA-based, groEL-based and multilocus sequence typing (MLST) were applied. Furthermore, the concordance between these typing methods was assessed. Results: Fever, lethargy and anorexia were the most common clinical signs in cats suffering from granulocytic anaplasmosis. The most frequent laboratory finding was thrombocytopenia. All three typing methods consistently indicated that the A. phagocytophilum strains found infecting cats are the same as those that cause disease in humans, dogs and horses. In general, the three typing methods applied exhibited high concordance. Conclusions: The genetic characterization of the feline A. phagocytophilum strains indicates that strain divergence is not the explanation for the fact that granulocytic anaplasmosis is much less frequently diagnosed in cats than in dogs and horses. Otherwise, it may be possible that cats are less susceptible to the same strains than dogs and horse are. However, due to the unspecific clinical signs, it should be considered that granulocytic anaplasmosis may be under-diagnosed in cats. [ABSTRACT FROM AUTHOR]

The importance of established and emerging tick-borne pathogens in Central and Northern Europe is steadily increasing. In 2007, we collected Ixodes ricinus ticks feeding on birds (n = 211) and rodents (n = 273), as well as host-seeking stages (n = 196), in a habitat in central Germany. In order to find out more about their natural transmission cycles, the ticks were tested for the presence of Lyme disease borreliae, Anaplasma phagocytophilum, spotted fever group (SFG) rickettsiae, Francisella tularensis, and babesiae. Altogether, 20.1% of the 680 ticks examined carried at least one pathogen. Bird-feeding ticks were more frequently infected with Borrelia spp. (15.2%) and A. phagocytophilum (3.2%) than rodent-feeding ticks (2.6%; 1.1%) or questing ticks (5.1%; 0%). Babesia spp. showed higher prevalence rates in ticks parasitizing birds (13.2%) and host-seeking ticks (10.7%), whereas ticks from small mammals were less frequently infected (6.6%). SFG rickettsiae and F. tularensis were also found in ticks collected off birds (2.1%; 1.2%), rodents (1.8%; 1.5%), and vegetation (4.1%; 1.6%). Various combinations of coinfections occurred in 10.9% of all positive ticks, indicating interaction of transmission cycles. Our results suggest that birds not only are important reservoirs for several pathogens but also act as vehicles for infected ticks and might therefore play a key role in the dispersal of tick-borne diseases. [ABSTRACT FROM AUTHOR]

In Europe, Lyme borreliosis (LB) and tick-borne encephalitis (TBE) are the two vector-borne diseases with the largest impact on human health. Based on data on the density of host-seeking Ixodes ricinus ticks and pathogen prevalence and using a variety of environmental data, we have created an acarological risk model for a region where both diseases are endemic (Czech Republic-South Bohemia and Germany-Lower Bavaria, Upper Palatinate). The data on tick density were acquired by flagging 50 sampling sites three times in a single season. Prevalence of the causative agents of LB and TBE was determined. Data on environmental variables (e.g., altitude, vegetation cover, NDVI, land surface temperature) were obtained from various sources and processed using geographical information systems. Generalized linear models were used to estimate tick density, probability of tick infection, and density of infected ticks for the whole area. A significantly higher incidence of human TBE cases was recorded in South Bohemia compared to Bavarian regions, which correlated with a lower tick density in Bavaria. However, the differences in pathogen prevalence rates were not significant. The model outputs were made available to the public in the form of risk maps, indicating the distribution of tick-borne disease risk in space.

Background: Hyalomma marginatum and Hyalomma rufipes are two-host tick species, which are mainly distributed in southern Europe, Africa and middle-eastern Asia. They are well-known vectors of Crimean Congo hemorrhagic fever (CCHF) virus and other viruses as well as Rickettsia aeschlimannii. In recent years, these tick species have been found sporadically in Germany, but they do not belong to the autochthonous tick fauna in Germany., Methods: Ticks with unusual morphology were collected and sent from private persons or public health offices to involve institutions for morphological identification and further testing. All ticks identified as Hyalomma spp. were tested using molecular detection methods for CCHF virus, Rickettsia spp., Coxiella burnetii and Coxiella-like organisms, Babesia spp. and Theileria spp., Results: Thirty-five ticks with an unusual appearance or behaviour were reported to us during summer-autumn 2018. For 17 of them, the description or photos implied that they belong to the hard tick genus Hyalomma. The remaining 18 ticks were sent to us and were identified as adult Hyalomma marginatum (10 specimens) or adult Hyalomma rufipes (8 specimens). All ticks tested negative for CCHF virus, Coxiella burnetii, Coxiella-like organisms, Babesia spp. and Theileria spp. The screening for rickettsiae gave positive results in 9 specimens . The Rickettsia species in all cases was identified as R. aeschlimannii., Conclusions: These results show that exotic tick species imported into Germany were able to develop from the nymphal to the adult stage under appropriate weather conditions. Fifty percent of the ticks carried R. aeschlimannii, a human pathogen, while CCHF virus or other pathogens were not detected. Imported Hyalomma ticks may be the source of exotic diseases acquired in Germany.

Background: Tick-borne encephalitis (TBE) virus is transmitted to humans and animals through tick bites and is thought to circulate in very strictly defined natural environments called natural foci. The most common tick serving as a vector for the TBE virus in central Europe is Ixodes ricinus; it is rarely found in other tick species and in Dermacentor reticulatus it has, so far, only been reported in Poland., Methods: Between autumn 2016 and spring 2018 ticks were collected by the flagging method in a new TBE focus in the district of northern Saxony, Germany, outside the known risk areas as defined by the national Robert Koch Institute. Ticks were morphologically identified and tested in pools for the presence of TBE virus using a real-time RT-PCR. TBE virus from positive pools was isolated in A549 cells, and the E gene sequences were determined after conventional RT-PCR, followed by a phylogenetic comparison., Results: TBE virus was detected in 11 pools, 9 times in flagged adults D. reticulatus (n = 1534; MIR: 0.59%, CI: 0.29-11.3%) and only twice in I. ricinus nymphs (n = 349; MIR: 0.57%, CI: 0.02-2.2%). All other ticks, I. ricinus males (n = 33), females (n = 30) and larvae (n = 58), as well as 5 I. inopinatus (2 females, 3 males) and 14 Haemaphysalis concinna (3 females, 11 nymphs), tested negative for TBE virus. TBE virus was not detected in I. ricinus during the summer, when D. reticulatus is not active. Sequence comparison of the entire E gene of the isolated virus strains resembled each other with only 3 nucleotide differences. The most closely related viral sequences belong to TBE virus strains from Poland and Neustadt an der Waldnaab (county of Neustadt an der Waldnaab, Bavaria), approximately 200 km east and 200 km south-west of the new focus, respectively., Conclusions: TBE virus was found in northern Saxony, Germany, with similar MIRs in D. reticulatus and I. ricinus, indicating that D. reticulatus plays an equal role to I. ricinus in virus circulation when both tick species are sympatric.

In the recent years matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) has become a useful tool to characterize arthropod species and their different stages of development. It was reported for sand flies and mosquitoes at immature stages and also assumed for ticks that geographic location can have a subtle influence on MALDI-TOF mass spectra which allows the discrimination of animals with specific local variations of the MALDI-TOF MS phenotype. It is so far uncertain, however, if these mass-spectrometric differences are based on genetic variation or on spectral features which depend on environmental or temporal features. The aim of this study was to analyze the influence of the geographic location, environmental factors and the season of the year on the MALDI-TOF mass spectra of Ixodes (I.) ricinus nymphs and if spectral variation would allow to draw conclusions with respect to the tick's provenience or conditions that influence the tick life cycle. Application of multivariate statistical models on spectra of ticks collected in different seasons and different habitats and locations within Germany showed that the impact of the location seemed to be small while season and habitat seemed to have stronger impact on the MALDI-TOF mass spectra. Possibilities and limitations of MALDI-TOF mass spectra to draw conclusions on the tick life cycle are discussed., Competing Interests: The authors have declared that no competing interests exist.

Questing ticks were sampled monthly over a period of 11 months from February, 2011 to December, 2011 at 13 sites in southern Germany using the flagging method. The ticks were identified to species, gender, and stadium. Although both I. ricinus and D. reticulatus were sampled, this study concentrated on I. ricinus, since it was the most abundant tick to be found. Additional weather data (air and soil temperature, relative air humidity, precipitation, sunshine duration) were recorded on each sampling site and the local vegetation described. A total of 14, 394 ticks was collected (7,862 larvae, 5,568 nymphs, 964 adults) and their activity was recorded in order to determine the seasonal activity pattern over different periods of the year. In contrast to the widely accepted pattern of a bimodal seasonal activity in moderate areas with a dominant peak in spring and a minor peak in autumn, a unimodal activity pattern was found for all development stages on six of the 12 sampling sites. Tick abundance was compared to weather variables. Tick host-seeking activity was found to be significantly dependent on the temperature at ground level, precipitation, and sunshine duration as well as relative air humidity. Adult ticks showed a positive correlation with the duration of sunshine, whereas nymphs were mostly unaffected by this phenomenon., (© 2014 The Society for Vector Ecology.)

It is commonly assumed that Dermacentor reticulatus immature life stages are nidicolous and therefore cannot be collected from vegetation. However, in June and July of 2018 and 2019, a total of 47 questing D. reticulatus larvae and two nymphs were collected by the flagging method in two different sites close to the city of Leipzig, Germany. To confirm their role in the transmission of tick-borne pathogens, 45 larvae (pooled by 2 in 21 pools and 1 pool with three individuals) and one nymph were tested either by conventional or real-time PCR for the presence of Bartonella spp., Neoehrlichia mikurensis, Rickettsia spp., and Babesia spp. All samples tested negative for Bartonella spp., N. mikurensis, and Babesia spp.; while the minimal infection rate of larvae for Rickettsia spp. was 42%, and the one tested nymph was also positive. Sequencing partial ompB genes revealed the presence of Rickettsia raoultii in larvae and nymph. Further research needs to be done to determine under which circumstances immature D. reticulatus ticks are found outside the burrows of their hosts and can be collected from vegetation., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

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., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Lyme borreliosis caused by spirochaetes of the Borrelia burgdorferi sensu lato (s.l.) complex is the most common tick-borne disease in Europe. In addition, the relapsing-fever spirochaete Borrelia miyamotoi, which has been associated with febrile illness and meningoencephalitis in immunocompromised persons, is present in Europe. This study investigated Borrelia prevalence and species distribution in ticks removed from humans and sent as diagnostic material to the Institute for Parasitology, University of Veterinary Medicine Hannover, in 2013-2017. A probe-based real-time PCR was carried out and Borrelia-positive samples were subjected to species determination by reverse line blot (RLB), including a B. miyamotoi-specific probe. The overall Borrelia-infection rate as determined by real-time PCR was 20.02 % (510/2547, 95 % CI: 18.48-21.63 %), with annual prevalences ranging from 17.17 % (90/524, 95 % CI: 14.04-20.68 %) in 2014 to 24.12 % (96/398, 95 % CI: 19.99-28.63 %) in 2015. In total, 271/475 (57.1 %) positive samples available for RLB were successfully differentiated. Borrelia afzelii was detected in 30.53 % of cases (145/475, 95 % CI: 26.41-34.89), followed by B. garinii/B. bavariensis (13.26 % [63/475], 95 % CI: 10.34-16.65). Borrelia valaisiana occurred in 5.89 % (28/475, 95 % CI: 3.95-8.41), B. spielmanii in 4.63 % (22/475, 95 % CI: 2.93-6.93), B. burgdorferi sensu stricto (s.s.)/B. carolinensis in 2.32 % (11/475, 95 % CI: 1.16-4.11), B. lusitaniae in 0.63 % (3/475, 95 % CI: 0.13-1.83) and B. bisettiae in 0.42 % (2/475, 95 % CI: 0.05-1.51) of positive ticks. Borrelia kurtenbachii was not detected, while B. miyamotoi was identified in 7.37 % (35/475, 95 % CI: 5.19-10.10) of real-time PCR-positive samples. Sanger sequencing of B. garinii/B. bavariensis-positive ticks revealed that the majority were B. garinii-infections (50/52 successfully amplified samples), while only 2 ticks were infected with B. bavariensis. Furthermore, 6/12 B. burgdorferi s.s./B. carolinensis-positive samples could be differentiated; all of them were identified as B. burgdorferi sensu stricto. Thirty-nine ticks (8.21 %, 95 % CI: 5.90-11.05) were coinfected with two different species. Comparison of the species distribution between ticks removed from humans in 2015 and questing ticks collected in the same year and the same area revealed a significantly higher B. afzelii-prevalence in diagnostic tick samples than in questing ticks, confirming previous observations. The obtained data indicate that Borrelia prevalence fluctuated in the same range as observed in a previous study, analysing the period from 2006 to 2012. Detection of B. miyamotoi in 7.37 % of Borrelia-positive samples points to the fact that clinicians should be aware of this pathogen as a differential diagnosis in cases of febrile illness., (Copyright © 2020 Elsevier GmbH. All rights reserved.)