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.     

Dynamics of tick infestations in foxes in Thuringia,Germany

This study aimed to provide up-to-date information on the dynamics of tick infestations on foxes in Thuringia, as the most recent information available was published in 1997. Fox carcasses that had been sent to the Thuringian State Authority for Food Safety and Consumer Protection (Thüringer Landesamt für Lebensmittelsicherheit und Verbraucherschutz – TLLV), between January 1st and December 31st, 2009, were examined for the presence of ticks. All ticks collected were stored at ?20 °C before being identified and classified according to their developmental stage and sex. Out of a total of 1286 foxes examined, 989 (76.9%) were infested with ticks. A total of 13,227 ticks were collected from the foxes. The stage most frequently found was the larva (48.1%), followed by the adult (34.1%), and the nymphal stage (17.8%). Regarding the adult stage, Ixodes ricinus was the most frequent tick species detected (82.2%), followed by I. canisuga (10.8%) and I. hexagonus (6.7%). Dermacentor reticulatus ticks were very rare (0.3%). With regard to nymphs, I. canisuga and I. hexagonus were the most frequent tick species found, and this was also assumed for the larval stage. The results indicate the occurrence of tick infestations in foxes throughout the year, mainly by I. ricinus, I. canisuga, and I. hexagonus, with seasonal variations. Foxes were infested by I. ricinus ticks significantly more frequently from April to September. This applied to all tick developmental stages, but especially to adults. In contrast to I. ricinus, the infestation of foxes with I. canisuga and I. hexagonus was significantly higher from January to March and from October to December, especially with the immature developmental stages.     

Ixodid tick species found in northern Sweden – Data from a frontier area

Environmental and climatic changes in northern Europe have shaped a geographical area in which new tick species may become established and introduce new tick-borne pathogens. In recent decades, ticks have expanded their latitudinal and altitudinal range limits in northern Sweden. In this study, ticks were collected in 2018 and 2019 in northern Sweden from different hosts, mainly from dogs, cats and humans. The ticks in 2018 (n = 2141, collected from 65 municipalities in 11 provinces) were identified as Ixodes ricinus (n = 2108, 98.5%), Ixodes persulcatus (n = 18, 0.8%), Ixodes trianguliceps (n = 14, 0.7%) and Hyalomma marginatum (n = 1, 0.05%). The ticks collected in 2019 (n = 519, across a smaller area than in 2018, i.e. Sweden's four northernmost provinces) were identified as I. ricinus (n = 242, 46.6%) and I. persulcatus (n = 277, 53.4%). Among those collected in 2019, the majority of I. ricinus (n = 111, 45.9%) were submitted from the province of Västerbotten, while most I. persulcatus (n = 259, 93.5%) were collected in the province of Norrbotten. This study provides updated figures on the geographical distribution of two Ixodes species in northern Sweden. The results confirmed I. ricinus to be the dominant species and that I. persulcatus has enlarged its distributional area compared with previous reports. Updated knowledge of tick distribution is fundamental for the creation of risk maps and will allow relevant advice to be provided to the general public, suggesting measures to prevent tick bites and consequently tick-borne diseases.     

Cercopithifilaria spp. in ticks of companion animals from Asia: new putative hosts and vectors

Cercopithifilaria bainae, Cercopithifilaria grassi, and Cercopithifilaria sp. II sensu Otranto et al., 2013 tick borne filarioids are typically found in dogs. Among them, Cercopithifilaria bainae has a worldwide distribution according to the occurrence of its tick vector, Rhipicephalus sanguineus sensu lato (s.l.). Nevertheless, in Asian countries, despite the wide presence of this tick species, data on Cercopithifilaria spp. are scant. Therefore, this study aimed to assess the occurrence of these dermal filarioids in ixodid ticks collected on dogs and cats from Asian countries, providing a better epidemiological picture on their distribution in this continent. Ticks (n = 687) of the species Rhipicephalus sanguineus s. l. (n = 667), Rhipicephalus haemaphysaloides (n = 8), Haemaphysalis longicornis (n = 7), Haemaphysalis campanulata (n = 1), Haemaphysalis wellingtoni (n = 2), Haemaphysalis hystricis (n = 1), and Ixodes sp. (n = 1) were collected on dogs and cats under the frame of previous studies in China, India, Indonesia, Malaysia, Singapore, Taiwan, Thailand, the Philippines and Vietnam. Tick samples were molecularly screened for Cercopithifilaria spp. by conventional PCR and real-time PCR using two pair of primers targeting partial sequences of cytochrome c oxidase 1 gene. Overall, Cercophitifilaria spp. DNA was detected in 9.5% (n = 65/687) of the tick specimens tested, with C. bainae being the most prevalent species (8.9%), followed by C. grassii (0.6%). Most Cercophitifilaria spp. positive ticks were collected on dogs (92.3%; 60/65); whereas ticks collected on cats represented 7.7% of the positive specimens. In addition, Cercopithifilaria spp. were mostly detected in R. sanguineus s.l. ticks (96.9%; 63/65), followed by Rhipicephalus haemaphysaloides (3.1%; 2/65). Data herein presented demonstrate the occurrence of dermal tick borne filarioids of the genus Cercopithifilaria in several Asian countries, with C. bainae being the most prevalent species. We also report for the first time the molecular detection of C. bainae in R. sanguineus s.l. ticks collected on cats, as well as in R. haemaphysaloides ticks, suggesting that the biological cycle of this filarioid species may involve other intermediate and definitive hosts than R. sanguineus s.l. and dogs. However, confirmatory studies on the role of other tick species and domestic cats on the biology of C. bainae are advocated.     

Detection of Leishmania infantum,Babesia canis,and rickettsiae in ticks removed from dogs living in Italy

The aims of this study were to determine natural infections by Anaplasma phagocytophilum/Anaplasma platys, Bartonella henselae, Ehrlichia canis, Leishmania infantum, Rickettsia spp., Babesia spp., and Hepatozoon spp. by molecular methods in ticks (n = 91) removed from dogs with clinical signs and laboratory abnormalities compatible with tick-borne diseases (n = 22) living in Italy and to assess the distribution and species of ticks encountered. Ticks from dogs living in southern Italy were all identified as Rhipicephalus sanguineus (n = 25), ticks from central Italy included Rh. sanguineus (n = 8) and Ixodes ricinus (n = 9), ticks from northern Italy included Rh. sanguineus (n = 45), Dermacentor marginatus (n = 3), and one I. ricinus. Leishmania infantum, Rickettsia spp., and Babesia canis were the only pathogens detected in 7 (8%), 4 (4%), and 2 (2%) out of 91 ticks, respectively. L. infantum was detected in I. ricinus from central Italy and in Rh. sanguineus from northern and central Italy. Rickettsia conorii and Ri. massiliae were detected in Rh. sanguineus ticks from central and southern Italy (Sicily), respectively. Bab. canis was detected in D. marginatus ticks from northern Italy.     

Ticks,fleas and rodent-hosts analyzed for the presence of Borrelia miyamotoi in Slovakia: the first record of Borrelia miyamotoi in a Haemaphysalis inermis tick

In Slovakia, little knowledge is available on the occurrence, hosts and vectors of Borrelia miyamotoi of the relapsing fever group. In the current study, 2160 questing and rodent-attached ticks of six species (Ixodes ricinus, Ixodes trianguliceps, Dermacentor marginatus, Dermacentor reticulatus, Haemaphysalis concinna and Haemaphysalis inermis), 279 fleas belonging to 9 species (Ctenophthalmus agyrtes, Ctenophthalmus solutus, Ctenophthalmus assimilis, Megabothris turbidus, Amalareus penicilliger, Hystrichopsylla orientalis, Ctenophthalmus uncinatus, Doratopsylla dasycnema and Nosopsyllus fasciatus) and skin biopsies from 245 small mammals belonging to eight species (Apodemus agrarius, Apodemus flavicollis, Apodemus uralensis, Myodes glareolus, Crocidura leucodon, Micromys minutus, Microtus arvalis, Microtus subterraneus) were screened for the presence of B. miyamotoi DNA. The overall prevalence of B. miyamotoi found in questing and rodent-attached ticks was 1.8% (23 positive/1260 examined) and 3.4% (31 positive/900 examined), respectively. Borrelia miyamotoi was detected in questing I. ricinus, rodent-attached I. ricinus and H. inermis ticks, and in one male of the common vole (M. arvalis) in different habitats (mainly rural) in eastern Slovakia. However, B. miyamotoi was not found in any of the tested fleas. Our findings indicate that rural habitats with different species of tick vectors and hosts are appropriate for the occurrence of B. miyamotoi.     

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.     

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.     

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.     

Diversity of tick species and associated pathogens on peri-urban wild boars – First report of the zoonotic Babesia cf. crassa from Hungary

Wild boars show increasing numbers and population densities throughout Europe, including Hungary. While their presence is appreciated as game animals, they are also responsible for significant agricultural damage, habitat degradation and water quality issues. In addition, wild boars may harbor ticks and can act as reservoirs of tick-borne pathogens, thus posing a risk of transmission towards humans and domestic animals. This latter aspect of their veterinary-medical and epidemiological significance has become especially important in recent years, because increasing numbers of wild boars are reported to enter urban areas. Despite of this, reports on tick infestations of wild boars are scarce in Europe.For this study, 333 ixodid ticks were collected from 51 wild boars at 32 peri-urban locations in 14 counties of Hungary, during 2005-2008 (older samples) and 2019-2020 (new samples). Five species of ticks were identified: Dermacentor reticulatus (n = 165), Ixodes ricinus (n = 90) and Haemaphysalis concinna (n = 29) in both sample groups, while H. inermis (n = 29) and D. marginatus (n = 20) were only found among the old samples. The seasonality of collected ticks corresponded to their known activities.After DNA extraction, ticks were screened for three groups of tick-borne pathogens. All samples were negative for brucellae, recently reported to be carried and transmitted transovarially by D. marginatus. Four D. reticulatus contained Babesia canis DNA, while in one H. concinna nymph the recently discovered zoonotic B. cf. crassa (reported in Slovenia within 80 km of our sampling site) was detected. In addition, Anaplasma phagocytophilum was identified in D. reticulatus (n = 1), H. concinna (n = 3) and in its known vector, I. ricinus (n = 15). Phylogenetically, three out of four A. phagocytophilum genotypes clustered with zoonotic ones.In conclusion, despite of the high prevalence of Brucella suis in wild boars in Hungary, no evidence was found in support of the epidemiological role of ticks in transmitting brucellae. On the other hand, wild boars might introduce B. canis-carrier D. reticulatus into urban areas, unlike birds (which are not known to carry this tick species in the country). Most importantly, tick-infested wild boars can contribute to the spread of a novel zoonotic Babesia sp. and of the zoonotic variants of A. phagocytophilum.     

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.     

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.     

Ticks on humans in an Atlantic rainforest preserved ecosystem in Brazil: Species,life stages,attachment sites,and temporal pattern of infestation

We herein describe exposure and tick attachment to researchers and field staff within the greatest preserved inland Atlantic rainforest, located in Brazil, over a non-consecutive period of five years. Overall, 433 ticks from five species were collected, Amblyomma coelebs (n = 281), Amblyomma brasiliense (n = 79), Amblyomma incisum (n = 43), Amblyomma ovale (n = 4), Haemaphysalis juxtakochi (n = 4). Clusters of larvae belonged to the genus Amblyomma (n = 21) and Haemaphysalis (n = 1). Both Haemaphysalis and Amblyomma species were observed crawling on humans but only ticks from the latter genus were found attached. Most recovered ticks (59%) were found attached and, whereas A. brasiliense was the main host-seeking tick species in the forest, A. coelebs nymphs were the ones that overwhelmingly attached to humans (70% of all attachments). At the same time only 0.4% of human bites were of A. ovale, the Rickettsia parkeri strain Atlantic rainforest vector tick in Brazil. Tick attachment sites were recorded for 132 ticks and lower/upper limbs and the trunk were more parasitized than the head/neck. Tick bites were recorded throughout the year; while larvae did not show an evident seasonal pattern, nymphs attached to humans mainly in autumn and winter and the few adult bites were recorded in spring. Our results highlighted A. coelebs nymphs as major human-biters and evidenced the necessity of studies that discern human tick bites that occur after exposure to host-seeking ticks from those that occur after tick displacement from hosts that carry ticks to human dwellings.     

On the way between Africa and Europe: Molecular taxonomy of ticks collected from birds in Malta

The Maltese Archipelago is situated in the middle of the Mediterranean Basin, between Europe and Africa, therefore representing an important stopover site for migratory birds between these two continents. Despite this, up-to-date information is not available on tick species associated with birds in Malta. Therefore, in this study, birds mist-netted for ringing by BirdLife Malta were examined for the presence of ticks between September, 2019 and May, 2021. Ticks were identified morphologically and molecularly, using three genetic markers.During the study period, 57 individuals of 22 bird species were found tick-infested, from which altogether 113 ixodid ticks were collected. The majority of developmental stages were nymphs, but 13 larvae and one female were also found. These ticks belonged to nine species: Ixodes cumulatimpunctatus (n=1), Ixodes ricinus (n=2), Ixodes acuminatus (n=2), Ixodes frontalis (n=5), Ixodes festai (n=1), one species of the Amblyomma marmoreum complex (n=8), Hyalomma rufipes (n=78), Hyalomma marginatum (n=7) and Hyalomma lusitanicum (n=1). Eight Hyalomma sp. ticks could only be identified on the genus level. Regarding seasonality, all Palearctic Ixodes species were carried by birds exclusively in the autumn (i.e., north to south), whereas H. rufipes (with predominantly Afrotropical distribution) was exclusively collected in the spring (i.e., carried south to north). Two tick species that occurred on birds in Malta, i.e., a species of the A. marmoreum complex and I. cumulatimpunctatus are only indigenous in the Afrotropical zoogeographic region. This is the first finding of the latter tick species in Europe, and four tick species were identified for the first time in Malta.In conclusion, the diversity of tick species regularly arriving in Europe from Africa is most likely higher than reflected by data obtained in Mediterranean countries of mainland Europe. Most notably, ticks of the genus Amblyomma appear to be underrepresented in previous datasets. Ticks of the subgenus Afrixodes (represented by I. cumulatimpunctatus) might also be imported into Europe by migratory birds.     

Diversity and geographic distribution of rickettsial agents identified in brown dog ticks from across the United States

Rhipicephalus sanguineus sensu lato, or brown dog ticks, transmit a variety of pathogens of veterinary and public health importance globally. Pathogens vectored by brown dog ticks and identified in the United States include Babesia vogeli, Ehrlichia canis, and several spotted fever group Rickettsia spp. (SFGR). Due to the challenge of collecting canine blood samples nationwide to screen for exposure to these pathogens, we took an indirect approach and tested brown dog ticks for molecular evidence of infection. Brown dog ticks (616 adults and 65 nymphs) collected from dogs and cats across the nation were tested by separate PCR assays detecting Babesia spp., E. canis, and SFGR. While no Babesia sp. was found, we identified rickettsial agents in 3.5% (24/681; 95% CI 2.4–5.2%) of the ticks. Pathogens and related organisms detected in ticks included E. canis (n = 1), Rickettsia amblyommatis (n = 3), Rickettsia massiliae (n = 11), Rickettsia monacensis (n = 3), Rickettsia montanensis (n = 5), and an undefined Rickettsia species (n = 1). These data demonstrate a wider geographic distribution of R. massiliae than previously known, and to the authors’ knowledge, reports R. monacensis in brown dog ticks for the first time. Due to the close association that brown dog ticks have with domestic dogs and humans, more research is needed to understand the full array of organisms, some of which are zoonotic, potentially transmitted by this widespread tick complex.     

The brown dog tick Rhipicephalus sanguineus sensu Roberts, 1965 across Australia: Morphological and molecular identification of R. sanguineus s.l. tropical lineage

The brown dog tick Rhipicephalus sanguineus (Latreille, 1806) is the most widely distributed tick species globally. Throughout the world there are at least two divergent lineages on dogs that are traditionally grouped into what was known as R. sanguineus. The species R. sanguineus was recently redescribed using a neotype reported from countries with a temperate climate. The second lineage distributed in countries with primarily tropical climates is currently designated R. sanguineus s.l. tropical lineage. Here, we present a comprehensive genetic evaluation of Australian brown dog ticks from across the continent that complements the morphological study of R. sanguineus sensu Roberts (1965). A total of 294 ticks were collected from dogs around Australia ― including New South Wales, Queensland, the Northern Territory and Western Australia ― for morphological identification. All ticks were morphologically identified as R. sanguineus sensu Roberts (1965). DNA was isolated from a single leg from morphologically characterised individuals from New South Wales (n = 14), Queensland (n = 18), Northern Territory (n = 7) and Western Australia (n = 13), together with ticks from Fiji (n = 1) and the Seychelles (n = 1) for comparison with Australian ticks. The study revealed three cox1 haplotypes clustered only with R. sanguineus s.l. tropical lineage’. An updated distribution of R. sanguineus sensu Roberts (1965) is compared to the 1965 distribution. In the Australian context, R. sanguineus s.l. has appeared in north-western New South Wales but remains absent from coastal New South Wales. Despite both temperate and tropical climates being present in Australia, only R. sanguineus s.l. tropical lineage was found. The evidence does not support the presence of the strictly defined brown dog tick, R. sanguineus by Nava et al. (2018) in Australia, because the examined ticks are genetically and morphologically distinct. We recommend using the term brown dog tick, R. sanguineus sensu Roberts (1965) for specimens from Australia.     

A practical protocol to prepare paraffin-embedded whole tick histology sections

Ticks are important ectoparasites that are capable of transmitting multiple classes of pathogens and are currently linked with many emerging tick-borne diseases worldwide. With increasing occurrences of tick-borne diseases in both humans and veterinary species, there is a continuous need to further our understanding of ticks and the pathogens they transmit. Whole tick histology provides a full scope of the tick internal anatomy, allowing researchers to examine multiple organs of interest in a single section. This is in contrast to other techniques that are more commonly utilized in tick-borne disease research, such as electron microscopy and light microscopy of individual organs. There is a lack of literature describing a practical technique to process whole tick histologic sections. Therefore, the current study aims to provide researchers with a workable protocol to prepare high quality paraffin-embedded whole tick histology sections. Amblyomma americanum adults were used as an example species for this study. After a series of pilot experiments using a combination of various fixatives, softening agents and processing techniques, we elected to compare two common fixatives, 10% neutral-buffered formalin (NBF) and Bouin's solution for whole ticks. Equal numbers of A. americanum unfed adults (n = 10/fixative) were processed identically and their whole tick histology coronal sections were individually scored. Higher scores were assigned to whole tick sections that contained more internal organs that are crucial for tick-borne disease research (e.g. salivary glands and midgut), high integrity of tissues and exoskeleton on the section, and good fixation and staining quality of the tissues. The mean total scores for Bouin's-fixed ticks were significantly higher compared to NBF-fixed ticks (p = 0.001). To further assess our preferred technique, we also demonstrated the feasibility of producing high quality whole tick sections for three other common tick species of medical importance (Rhipicephalus sanguineus, Ixodes scapularis, and Dermacentor variabilis) using Bouin's solution. While this technique may require further optimization for other tick species, we described a feasible protocol that uses commonly available tools, reagents and standard histologic equipment. This should allow any investigator to easily make adjustments to this protocol as needed based on their experimental goals.     

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.     

Some new findings on the endophilic vector tick Ixodes hexagonus in Germany

The authors of the present study received a number of hard ticks (Acari: Ixodidae) from different regions of Germany in 2018?2021. In most cases, the ticks were removed from pet dogs or cats. In one case, engorged and detached ticks were found in a house, and it was unclear how they had gained entrance. Morphological examination revealed that all the specimens (n = 18, larvae, nymphs, and adults) were hedgehog ticks, Ixodes hexagonus. Morphological identification for two larvae was confirmed by 16S rRNA gene amplification. These two larvae were also examined for tick-borne encephalitis virus RNA, but were found to be negative. In a previous, unreported study by one of the authors, which commenced in the late 1980s, it was observed that unfed larvae of I. hexagonus were extremely long-lived. They were still alive 5 years after the engorged female had detached from the host when maintained at a moderate temperature under controlled not too warm laboratory conditions (constant 15 °C, 91% relative humidity, and darkness).     

Anaplasma phagocytophilum in European bison (Bison bonasus) and their ticks from Lithuania and Poland

The increasing population of European bison (Bison bonasus) can contribute to the prevalence of zoonotic pathogens. The aim of the present study was to assess the presence of A. phagocytophilum infection in European bison tissues as well as ticks removed from European bison in Lithuania and Poland. A further objective of this work was to compare the detected A. phagocytophilum strains. A total of 85 tissue samples (spleen) of European bison and 560 ticks belonging to two species, Ixodes ricinus (n = 408) and Dermacentor reticulatus (n = 152) were tested. DNA of A. phagocytophilum was detected based on RT-PCR in 40% of the European bison samples, 8.8% of the I. ricinus and 5.9% of the D. reticulatus ticks. Analysis of the obtained partial 16S rRNA gene sequences of A. phagocytophilum revealed the presence of three variants with two polymorphic sites. Furthermore, phylogenetic analysis with partial msp4 gene sequences grouped A. phagocytophilum variants into three clusters. This study revealed that the groEL gene sequences of A. phagocytophilum from European bison and their ticks grouped into ecotype I and only one sequence from Lithuanian European bison belonged to ecotype II. The results of the present study indicated that European bison may play a role as a natural reservoir of A. phagocytophilum.