The European hedgehog (Erinaceus europaeus) – A suitable reservoir for variants of Anaplasma phagocytophilum?

The European hedgehog (Erinaceus europaeus) is a common insectivore in most parts of Europe and is frequently infested by the ticks Ixodes ricinus and I. hexagonus. I. ricinus ticks have been found infected with Anaplasma phagocytophilum, an obligate intracellular bacterium, but little is known about the potential of the hedgehog as a reservoir host. In this study, the infection with A. phagocytophilum and the genetic variants involved were investigated in a captive hedgehog population which was kept in a fenced, natural grass and bush garden habitat, and also in its ticks. Additionally hedgehogs from hedgehog caretaking stations were investigated. EDTA blood and ticks were collected from the captive hedgehog population once a month from March to October 2007 and in March and April 2008. All 3 developmental stages of I. ricinus and I. hexagonus occurred on the hedgehogs. After DNA extraction, the samples were screened for A. phagocytophilum with a real-time PCR, and selected samples were further investigated with a nested PCR targeting the partial 16S rRNA gene, followed by sequencing. One hundred thirty-six out of 220 hedgehog blood samples (61.8%) from altogether 48 individuals, 413 out of 563 I. ricinus samples and 90 out of 338 I. hexagonus samples were PCR-positive. Thirty-two hedgehogs were positive more than once, most frequently twice or 3 times, but also up to 9 times. Sequencing of the partial 16S rRNA gene resulted in 6 variants, but one variant (‘A’) was the most frequent which appeared in 93.8% of the positive hedgehogs. This variant (equaling Frankonia II, GenBank AF136712) has recently been reported from human, equine, and canine granulocytic anaplasmosis cases and thus, its specific association with hedgehogs is an important finding in the epidemiology of A. phagocytophilum in Europe.The high infection rate of both hedgehogs and ticks with A. phagocytophilum and the simultaneous infestation with 2 tick species of all developmental stages suggest that the hedgehog may be a suitable reservoir for at least some variants of A. phagocytophilum.     

Exposure of dogs and cats to Borrelia miyamotoi infected Ixodes ricinus ticks in urban areas of the city of Poznań, west-central Poland

Borrelia miyamotoi is an emerging human pathogen that causes a relapsing fever-like disease named B. miyamotoi disease. The bacterium belongs to the relapsing fever borreliae, and similar to spirochetes of the Borrelia burgdorferi sensu lato group, it is transmitted only by hard ticks of the Ixodes ricinus complex. To date, B. miyamotoi has not been demonstrated to cause illness in dogs or cats, and is poorly documented in veterinary medicine. The aim of this study was to determine the B. miyamotoi presence in (i) host-seeking ticks and (ii) engorged Ixodes sp. ticks collected from dogs and cats during their inspection in veterinary clinics of the city of Poznań, west-central Poland. Host-seeking ticks were sampled in dog walking areas localized in urban forested recreational sites of the city. In this study, 1,059 host-seeking and 837 engorged I. ricinus ticks collected from 680 tick-infested animals (567 dogs and 113 cats) were screened. Additionally, 31 I. hexagonus ticks (one larva, 13 nymphs, and 17 females) were collected from three cats; one larva and one nymph were collected from two dogs; and one dog was infested with a single Dermacentor reticulatus female.Borrelia DNA was identified by the amplification and sequencing of the V4 hypervariable region of the 16S rRNA gene and flaB gene fragments. DNA of B. miyamotoi was detected in 22 (2.1%) of the host-seeking ticks (in all developmental tick stages and in all study areas). In addition, the engorged I. ricinus ticks exhibited a similar B. miyamotoi presence (1.8%). Fifteen I. ricinus ticks collected from animals tested positive for the presence of B. miyamotoi DNA, and the DNA of B. miyamotoi was observed in three (9.1%; one female and two nymphs) I. hexagonus ticks. The single D. reticulatus female collected from a dog tested PCR-negative for the bacterium. The results of this study demonstrated the establishment and broad presence of the bacterium in tick populations from different urban ecosystems of the city of Poznań. The lack of difference in the mean infection presence of animal-derived and host-seeking I. ricinus ticks suggests that the systematic surveillance of pets may be useful for the evaluation of human exposure to B. miyamotoi infected ticks in urban areas. Additional studies are required to further elucidate the role of domestic and wild carnivores in the epidemiology of B. miyamotoi, which remains unknown.     

Characterization of ferritin 2 for the control of tick infestations

Ixodes ricinus is one the most abundant tick species in Europe and these ticks transmit pathogens causing human and animal diseases. The cattle ticks, Rhipicephalus (Boophilus) spp., affect cattle production in tropical and subtropical regions of the world. Development of vaccines directed against tick proteins may reduce tick infestations and the transmission of tick-borne pathogens. However, a limiting step in tick vaccine development has been the identification of tick protective antigens. Herein, the tick iron metabolism pathway was targeted in an effort to identify new tick protective antigens. Recombinant I. ricinus (IrFER2) and Rhipicephalus microplus (RmFER2) ferritin 2 proteins were expressed in Escherichia coli and used to immunize rabbits and cattle, respectively. Vaccination with IrFER2 reduced I. ricinus tick numbers, weight and fertility in rabbits with an overall vaccine efficacy (E) of 98%. Control of cattle tick, R. microplus and Rhipicephalus annulatus infestations was obtained in vaccinated cattle with overall E of 64% and 72%, respectively. Notably, the efficacy of the RmFER2 vaccine was similar to that obtained with Bm86 against R. microplus. These collective results demonstrated the feasibility of using ferritin 2 to develop vaccines for the control of tick infestations.     

Winter activity of questing ticks (Ixodes ricinus and Dermacentor reticulatus) in Germany − Evidence from quasi-natural tick plots,field studies and a tick submission study

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.     

Investigation on haplotypes of ixodid ticks and retrospective finding of Borrelia miyamotoi in bank vole (Myodes glareolus) in Switzerland

The current status of tick species, important tick-borne bacteria and protozoan parasites is well-documented in Switzerland. However, reports on the genetic diversity and geographical relationships of tick species in this country appear to be in part lacking or outdated. Thus, the aim of this study was to collect ticks from various host species in southern Switzerland, to compare them in a geographical context and to screen in these samples rare tick-borne pathogens hitherto not reported or having low prevalence in Switzerland.In 2019–2020 altogether 177 ixodid ticks were collected from the vegetation, as well as from humans (n = 17), dogs (n = 23), cats (n = 41), red deer (n = 8), a European rabbit and a European hedgehog at 25 locations in three cantons of south Switzerland. Tick species were identified morphologically, followed by DNA extraction and comparison of mitochondrial haplotypes with molecular-phylogenetic methods. Tick DNA extracts, as well as sixty-two rodent liver or spleen tissue DNA extracts (representing six species) available from 2005 to 2006 were screened for trypanosomes, Occidentia massiliensis and Borrelia miyamotoi.Morphologically, three tick species were identified: Ixodes ricinus (n = 170), Rhipicephalus sanguineus sensu lato (n = 6) and I. hexagonus (n = 1). In contrast to companion animals (dogs, cats) immature ticks (larvae and nymphs) predominated on humans, which was a highly significant association (P < 0.0001). Molecular comparison of the cytochrome c oxidase subunit I (cox1) gene with GenBank data established the species as R. sanguineus sensu stricto and confirmed I. hexagonus, both showing 99.8–100% sequence identity to conspecific ticks from northern Italy. Seventy-nine specimens morphologically identified as I. ricinus revealed high 16S rRNA gene haplotype diversity and represented two phylogenetic groups. Two I. ricinus haplotypes from Switzerland belonged to the same haplogroup with I. inopinatus from Spain, Germany and Austria as well as with I. ricinus reported from a broad geographical range of Europe (including Italy, the Netherlands, Poland, Latvia and Sweden). All 141 tick DNA extracts (from five R. sanguineus s.l., 135 I. ricinus and one I. hexagonus) and 62 rodent tissue DNA extracts were negative for trypanosomes and O. massiliensis. However, B. miyamotoi was identified in a bank vole (Myodes glareolus) and three ticks by sequencing.From Switzerland, this is the first report of tick haplotypes that are phylogenetically closely related to I. inopinatus. However, based on their morphology, both specimens are considered as I. ricinus. These results highlight the importance that the identification of I. inopinatus should be based on coherent morphologic and molecular properties. This is also the first report of rodent-borne B. miyamotoi in Switzerland. Taking into account the year of collection (2005), in a chronological order this might be the first indication of B. miyamotoi in any rodent species in Europe.     

Ticks on the Channel Islands and implications for public health

The Channel Islands are British Crown dependencies located in the English Channel to the west of the Normandy coast in northern France. Whilst there have been studies investigating tick occurrence and distribution in different habitats on the mainland of the UK and in France, the Channel Islands have been relatively understudied. As such, little is known about whether the sheep tick, Ixodes ricinus, is present, and whether there is a potential risk of Lyme borreliosis on the Channel Islands. To ascertain the presence of I. ricinus on the three largest islands in the archipelago: Jersey, Guernsey and Alderney, surveys of ticks questing in the vegetation and ticks feeding on hosts were undertaken during April and May 2016. Across all three islands, the highest numbers of ticks were found in woodland habitats. Ixodes ricinus was the predominant questing tick species found on Jersey, and Ixodes ventalloi the most common questing tick species on Alderney and Guernsey, with little or no evidence of questing I. ricinus on either island. During field studies on small mammals, I. ricinus was the predominant tick species feeding on Jersey bank voles (Myodes glareolus caesarius), with Ixodes hexagonus the most common species infesting hedgehogs on Guernsey. We propose that the greater diversity of small mammals on Jersey may be important in supporting immature stages of I. ricinus, in contrast to Guernsey and Alderney. Morphological identification of tick species was confirmed by PCR sequencing based on amplification of the cytochrome c oxidase subunit one (cox1) gene (COI DNA barcoding). To date, there have been few records of human tick bites in the Channel Islands, suggesting that the current risk from tick-borne disease may be low, but continued reporting of any human tick bites, along with reporting of cases of Lyme borreliosis will be important for continued assessment of the impact of tick-borne diseases in the Channel Islands.     

Distribution of ticks in Bosnia and Herzegovina

Ticks are one of the important groups of parasites, whose populations have increased in Europe in the last 20 years and, as vectors of many emerging diseases, are a major threat to human and animal health. Bosnia and Herzegovina has both climate and environmental conditions favorable for ticks. Given this risk, the aim of this study was to identify tick species currently present in this country and to compare the findings with previous studies, in order to update information on their spatial distribution and other parameters of occurrence.From March 2017 to April 2020, ticks were collected in 42 sampling areas (municipalities) throughout the country. Prespecified identification keys were used for identifying the ticks to the species level.A total of 6193 tick specimens from the Ixodidae family were collected from animal hosts or from vegetation. Ixodes ricinus was the most dominant tick species found. However, our results indicate a decrease in the relative contribution of this tick species to the overall abundance of tick populations. Dermacentor marginatus, which almost doubled in abundance compared to earlier studies, is the second most common tick species. We further confirmed the presence of Dermacentor reticulatus, Rhipicephalus bursa, Rhipicephalus sanguineus sensu lato, Hyalomma marginatum and Haemaphysalis punctata. Ixodes hexagonus, Ixodes canisuga and Haemaphysalis concinna, which were previously reported at low abundance were not found amongst our specimens.     

Plasticity in the timing of detachment of an Eurasian-African songbird tick,Ixodes frontalis

In non-permanent parasites, host detachment should take place in an environment that ensures the continuation of their life cycle. Timing of detachment - in combination with the host's space use - affects dispersal and transmission success of the parasites and of the pathogens they vector. Before reaching the adult reproductive stage, ticks need to go through multiple immature developmental stages (larva and nymph), each feeding on host blood. In between the feeding bouts, they often remain in the off-host environment for considerable periods of time. With this study, we aimed to obtain more insight in Ixodes frontalis’ off-host habitat use by comparing its detachment pattern in different life stages with that of two habitat-specialized ticks also found on birds: the endophilic tree-hole tick (Ixodes arboricola) and the exophilic sheep tick (Ixodes ricinus), the latter living in humid understory vegetation of forests. For this, we artificially infested hole-roosting (great tits, Parus major) and open-roosting (blackbirds, Turdus merula) birds with ticks under laboratory conditions, and recorded whether detachment occurred during the day or the night. We hypothesize that nocturnal detachment improves off-host mating opportunities and host localization, whereas diurnal detachment optimizes tick dispersal. Ixodes frontalis nymphs detached during the night, especially when feeding on blackbirds. This behaviour was very similar to that of I. arboricola (larva and nymph) feeding on great tits. In contrast, I. frontalis larvae detached during the day, especially when feeding on great tits, which resembles that of I. ricinus’ feeding behaviour (larva and nymph). Ixodes frontalis left the host within seven days, immediately after completion of the blood meal. This is similar to both developmental stages of I. ricinus but contrasts with the very long (up to 20 days) feeding duration in I. arboricola. Thus I. frontalis shows strong plasticity, switching from dispersal-centered (larvae) to host-centered (nymphs) detachment behaviour. Findings are discussed with regard to the ticks’ habitat use, dispersal, life history and host specificity.     

Control of infestations by Ixodes ricinus tick larvae in rabbits vaccinated with aquaporin recombinant antigens

BackgroundTick-borne diseases greatly impact human and animal health worldwide, and vaccines are an environmentally friendly alternative to acaricides for their control. Recent results have suggested that aquaporin (AQP) water channels have a key function during tick feeding and development, and constitute good candidate antigens for the control of tick infestations.MethodsHere we describe the effect of vaccination with the Ixodes ricinus AQP1 (IrAQP) and a tick AQP conserved region (CoAQP) on I. ricinus tick larval mortality, feeding and molting.ResultsWe demonstrated that vaccination with IrAQP and CoAQP had an efficacy of 32% and 80%, respectively on the control of I. ricinus larvae by considering the cumulative effect on reducing tick survival and molting.ConclusionsThe effect of the AQP vaccines on larval survival and molting is essential to reduce tick infestations, and extended previous results on the effect of R. microplus AQP1 on the control of cattle tick infestations. These results supports that AQP, and particularly CoAQP, might be a candidate protective antigen for the control of different tick species.     

Two-year monitoring of tick abundance and influencing factors in an urban area (city of Hanover,Germany)

Ticks may transmit a variety of human and animal pathogens. Prevalence of Borrelia spp., Rickettsia spp. and Anaplasma phagocytophilum in ticks has been monitored in the city of Hanover, Germany, since 2005. However, to determine the infection risk for humans and animals, not only pathogen prevalence, but also tick abundance and seasonality need to be taken into account. Therefore, the aim of this study was to investigate tick abundance at ten different collection sites in the city of Hanover, Germany. Collection of questing ticks was performed by the flagging method in the first and second half of each month during the tick season (April-October) in 2017 and 2018. At each 200 m² collection site, one of four 50 m² fields was sampled per visit on a rotational basis, resulting in 100 m² sampled per month. In addition, data on weather conditions, near-ground temperature, relative humidity and vegetation composition were noted at each collection event. In 2017, a total of 1770 ticks were collected, while 1866 ticks were collected in 2018. Ixodes ricinus was the most prevalent species (97.0 % of all ticks, 98.0 % of nymphs, 91.6 % of adults) followed by I. inopinatus (2.3 % of all ticks, 1.1 % of nymphs, 8.0 % of adults), I. frontalis (0.6 % of all ticks, 0.6 % of nymphs, 0.3 % of adults) and I. hexagonus (0.03 % of all ticks, 0.03 % of nymphs, 0.0 % of adults). Using generalized linear mixed modeling, density of I. ricinus and I. inopinatus in 2017 was significantly higher than in 2018. Regarding different landscape types, ticks were significantly more abundant in mixed forests than in parks, with more than 50 ticks/100 m² on average in both years. In urban parks, average tick density amounted to 15 ticks/100 m² in 2017 and 11 ticks/100 m² in 2018 and in broad-leaved forests average tick density was 13 and 18 ticks/100 m² in 2017 and 2018, respectively. Tick density showed a marked peak in June 2017 and in May 2018 at most sites, whereas a less pronounced peak was recognizable in September. Tick density varied considerably between collection sites. However, no statistically significant effect of (micro-)climatic variables, including near-ground temperature, relative humidity and saturation deficit, was found. Thus, further factors, such as the abundance of wildlife hosts, need to be considered in future studies to explain the differences between collection sites.     

Infection of hard ticks in the Caspian Sea littoral of Iran with Lyme borreliosis and relapsing fever borreliae

The Caspian Sea littoral of Iran is home to various hard tick species, including Ixodes ricinus, the notorious vector of Lyme borreliosis (LB) in Eurasia. Here, in this area, we examined I. ricinus and other hard ticks, along with common rodents and small mammals for LB and relapsing fever (RF) borreliae infection. Ticks were collected from various mammalian hosts, including sheep, goats, cattle, camels, horses, dogs, donkeys, rodents, and hedgehogs. Rodents and small mammals were live-captured from different habitats. A real-time PCR for 16S rRNA sequence revealed borrelial DNA in 71 out of 501 (≈14 %) specimens belonging to I. ricinus and Rhipicephalus ticks. None of the rodents and small mammals showed borrelial infection in the viscera. PCR amplification and sequencing of a 600-bp sequence of the flaB identified Borrelia bavariensis, Borrelia garinii, Borrelia afzelii, and Borrelia valaisiana, and the RF Borrelia, B. miyamotoi in I. ricinus ticks. The RF-like Borrelia in Rhipicephalus ticks shared the highest identity (98.97 %) with an isolate infecting Haemaphysalis megaspinosa ticks in Japan. Our phylogeny and BLAST analysis suggest the range extension of the European I. ricinus-associated borreliae into the north of Iran.     

Control of Ixodes ricinus and Dermacentor reticulatus tick infestations in rabbits vaccinated with the Q38 Subolesin/Akirin chimera

Diseases transmitted by ticks greatly impact human and animal health and their control is important for the eradication of tick-borne diseases. Vaccination is an environmentally friendly alternative for tick control. Recent results have suggested that Subolesin/Akirin (SUB/AKR) are good candidate antigens for the control of arthropod vector infestations. Here, we describe the effect of vaccination with the Q38 chimera containing SUB/AKR conserved protective epitopes on Ixodes ricinus and Dermacentor reticulatus tick larval mortality, feeding and molting. We demonstrated that Q38 vaccination had an efficacy of 99.9% and 46.4% on the control of I. ricinus and D. reticulatus larvae by considering the cumulative effect on reducing tick survival and molting. The effect of the Q38 vaccine on larval feeding and molting is essential to reduce tick infestations and supports that Q38 might be a candidate universal antigen for the control of multiple tick species that can infest the same host.     

Isolation of Arsenophonus nasoniae from Ixodes ricinus ticks in Slovakia

The tick Ixodes ricinus is the most prevalent and widely distributed tick species in Central Europe, commonly found in woodlands, heaths, and forests and particularly abundant in the Alpine region. This tick readily bites humans and transmits a number of bacterial and viral pathogens. We collected 10 live nymphs of I. ricinus ticks from vegetation in the Rovinka forest, Slovakia, and isolated a strain of Arsenophonus nasoniae from one tick using the BME/CTVM2 cell line. A new isolate was then subcultured on axenic media (Columbia agar supplemented with 5% sheep blood). To the best of our knowledge, this bacterium was never previously isolated from hard ticks or identified in ticks in Europe. We amplified and sequenced the 16S rRNA, rpoB, and ftsY genes. Limited genetic characterization showed that the isolated strain is almost identical to a strain from the parasitic wasp Nasonia vitripennis. Electron microscopy revealed a typical morphology of a Gram-negative bacterium, without pili or flagellae. Its role in human and animal pathology remains to be evaluated.     

Seasonal activity and tick-borne pathogen infection rates of Ixodes ricinus ticks in Hungary

Ixodes ricinus is the most important tick species in Europe as it is most widely distributed and transmits the majority of tick-borne zoonotic pathogens. As limited data are available for Hungary, the aim of the present study was to investigate the seasonal timing of questing by I. ricinus and the infection rate of this tick species with all major tick-borne zoonotic pathogens. Monthly collections of I. ricinus were carried out over 3 consecutive years by dragging a blanket in 6 biotopes representing different areas of Hungary. Altogether, 1800 nymphs (300 per collection point) were screened as pooled samples (each of 5 specimens) by PCR-based methods for tick-borne pathogens. I. ricinus larvae, nymphs, and adults had bimodal activity patterns with a major peak in the spring. As newly moulted ticks of all stages are thought to emerge in the autumn of each year, it appears that most newly emerged ticks delayed their questing until the following spring. The minimum prevalence of Borrelia burgdorferi sensu lato was 2.5%. Borr. afzelii, Borr. burgdorferi sensu stricto, Borr. garinii, Borr. lusitaniae, and Borr. valaisiana were identified by hybridization. The minimum infection rate with spotted fever group rickettsiae was 1.9%. Rickettsia helvetica was identified in all biotopes. The minimum prevalence of Anaplasma phagocytophilum, Babesia divergens and Bab. microti was low (0.3–0.5%). Bartonella spp.-, Francisella tularensis-, and TBE virus-specific amplification products were not detected. Relative to the results of comparable studies carried out in the Carpathian Basin, the prevalence of tick-borne pathogens was low in Hungary. This might be attributed to the climatic difference between the lowland areas of Hungary and submountain areas of the surrounding countries involved in the studies.     

Dermacentor reticulatus is a vector of tick-borne encephalitis virus

Tick-borne encephalitis virus (TBEV; family Flaviviridae) is the most medically important tick-borne virus in Europe and Asia. Ixodes ricinus and I. persulcatus ticks are considered to be the main vector ticks of TBEV in nature due to their specific ecological associations with the vertebrate hosts. Nevertheless, recent TBEV prevalence studies in ticks suggest that Dermacentor reticulatus ticks might play a relevant role in the maintenance of TBEV in nature. The goal of this study was to evaluate the vector competency of D. reticulatus for TBEV through experimental tick infections and comparative in vivo transmission studies involving D. reticulatus and I. ricinus ticks.We observed that after a transcoxal micro-capillary inoculation, adult female D. reticulatus ticks efficiently replicated TBEV during the observed period of 21 days. The mean virus load reached up to 2.5 × 10⁵ gene copies and 6.4 × 10⁴ plaque forming units per tick. The infected D. reticulatus ticks were able to transmit the virus to mice. The course of infection in mice was comparable to the infection after a tick bite by I. ricinus while the virus spread and clearance was slightly faster. Moreover, D. reticulatus ticks were capable of tick-to-tick non-viraemic transmission of TBEV to the Haemaphysalis inermis nymphs during co-feeding on the same animal. The co-feeding transmission efficiency was overall slightly lower (up to 54 %) in comparison with I. ricinus (up to 94 %) and peaked 1 day later, at day 3.In conclusion, our study demonstrated that D. reticulatus is a biologically effective vector of TBEV. In line with the recent reports of its high TBEV prevalence in nature, our data indicate that in some endemic foci, D. reticulatus might be an underrecognized TBEV vector which contributes to the expansion of the TBEV endemic areas.     

Control of tick infestations in wild roe deer (Capreolus capreolus) vaccinated with the Q38 Subolesin/Akirin chimera

Ticks (Acari: Ixodidae) are considered to be the most important vectors of disease-causing pathogens in domestic and wild animals, and emerging and re-emerging tick-borne diseases (TBD) exert an enormous impact on them. Wild ungulates are hosts for a wide variety of tick species and tick-borne pathogens that affect human and animal health. Consequently, the control of tick infestations and tick-borne pathogen prevalence is essential in some regions. Acaricides and animal management or culling have been used for the control of tick infestations and TBD, but tick vaccines constitute the best alternative to reduce the impact of acaricides on tick resistance and the environment. Previous results of controlled vaccination trials have shown that the Q38 Subolesin/Akirin chimera containing conserved protective epitopes could be a candidate universal antigen to control multiple tick species infestations. Thus, vaccination trials are necessary to validate these results under field conditions. In this study, we characterized the effect of Q38 vaccine on a wild population of European roe deer (Capreolus capreolus) in the Andalusian roe deer Reference Station (Junta de Andalucía, Cádiz, Spain). In this location, roe deer suffer especially severe parasitic conditions in some periods and commercial pesticides and ixodicides that are authorized to control ticks without specificity are frequently applied in the field, posing a threat to the environment. Animals vaccinated over a three-year period showed an antibody response to the vaccine antigen and a reduction in tick infestations by multiple species including Hyalomma marginatum, H. lusitanicum, Rhipicephalus bursa and Ixodes ricinus previously identified in roe deer, when compared to untreated controls. These results suggest the efficacy of Q38 for the control of tick infestations in wildlife.     

Tick and flea infestations in captive Tapirus terrestris and Tapirus kabomani (Perissodactyla: Tapiridae) in the Brazilian Amazon

The present study aimed to identify tick and flea species infesting captive tapirs (Tapirus terrestris and Tapirus kabomani) in the Brazilian Amazon. Ectoparasites were collected from tapirs chemically restrained in Zoo-botanical Parks, breeding facilities, conservationists and, environmental compensation areas in the states of Amapá, Amazonas, and Pará. After collection, the tick and flea specimens were placed in plastic pots containing isopropanol and identified according to dichotomous keys. Ectoparasite infestations were observed in 55% (18/33) tapirs, of which 61% (11/18) were single infestations with ticks or fleas, and 39% (7/18) were mixed infestations with different species of ticks and/or fleas. In total, 227 ticks (15 larvae, 107 nymphs, 38 females and 67 males) and 14 fleas (eight females and six males) were collected, identifying four tick species (Amblyomma cajennense sensu stricto , Amblyomma naponense, Amblyomma oblongoguttatum, and Amblyomma pacae) and three flea species and/or subspecies (Ctenocephalides felis felis, Rhopalopsyllus australis australis, and Tunga penetrans). In conclusion, infestations with Ixodidae ticks and Rhopalopsyllidae, Pulicidae and Tungidae fleas were found in captive tapirs in the Brazilian Amazon, and A. cajennense s.s. was the most frequent species. Additionally, the present study reports new associations between A. pacae and R. a. australis with T. terrestris and between A. cajennense s.s. with T. kabomani.     

Prevalence of Rickettsia spp., Anaplasma phagocytophilum,and Coxiella burnetii in adult Ixodes ricinus ticks from 29 study areas in central and southern Sweden

A total of 887 adult Ixodes ricinus ticks (469 females and 418 males) from 29 different localities in Sweden were screened for Rickettsia, Anaplasma, and Coxiella DNA using PCR and then subjected to gene sequencing. Rickettsial DNA was detected in 9.5–9.6% of the ticks. Most of the positive ticks were infected with Rickettsia helvetica. One tick harbored another spotted fever rickettsia, closely related to or identical with R. sibirica not previously found in I. ricinus nor in Sweden. Six of the ticks (0.7%) were infected with an Anaplasma sp., presumably A. phagocytophilum. Coxiella burnetii DNA was not detected in any of the ticks. The detection of R. helvetica and A. phagocytophilum in several of the localities sampled suggests that these potentially human-pathogenic agents are common in Sweden.     

The distribution limit of the common tick,Ixodes ricinus,and some associated pathogens in north-western Europe

In north-western Europe, the common tick, Ixodes ricinus, is widely established, its distribution appears to be increasing and the spread of tick-borne diseases is of increasing concern. The project ‘Flått i Nord’ (Ticks in northern Norway) commenced in spring 2009 with the intention of studying the tick’s distribution and that of its pathogens in northern Norway. Several methods were used: cloth-dragging, collecting from trapped small mammals, and collecting from pets. Since 2010, the occurrence of ticks in the region of northern Norway was determined directly by cloth-dragging 167 times in 109 separate locations between the latitudes of 64 °N and 70 °N (included seven locations in the northern part of Trøndelag County). The northernmost location of a permanent I. ricinus population was found to be Nordøyvågen (66.2204 °N, 12.59 °E) on the Island of Dønna. In a sample of 518 nymphal and adult ticks, the Borrelia prevalence collected close to this distribution limit varied but was low (1–15 %) compared with the locations in Trøndelag, south of the study area (15–27 %). Five specimens (1 %) were positive for Rickettsia helvetica. The length of the vegetation growing season (GSL) can be used as an approximate index for the presence of established populations of I. ricinus. The present study suggests that the threshold GSL for tick establishment is about 170 days, because the median GSL from 1991 to 2015 was 174–184 days at sites with permanent tick populations, showing a clear increase compared with the period 1961–1990. This apparent manifestation of climate change could explain the northward extension of the range of I. ricinus.     

The Ixodes ricinus tick as a vector of Borrelia burgdorferi in Slovenia

In many parts of Slovenia, the vegetation, climate and animals form an ideal natural environment for the persistence of different tick species. In this study, Ixodes ricinus ticks, the prevalent tick species in our country, are identified as the vectors of Borrelia burgdorferi in Slovenia. Ticks were collected by flagging between May and October 1990. All the ticks collected from different parts of Slovenia were I. ricinus and each one was examined individually. The ticks were tested to determine the presence of B. burgdorferi by indirect immunofluorescent assay. Among the 496 ticks examined for borreliae there were 85 adults and 411 nymphs. Twenty (23.5%) adults and 18 (4.4%) nymphs were found positive for Borreliae. This study discusses the importance of I. ricinus ticks in the role of Lyme borreliosis in Slovenia.