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Background: Vietnam and its region are regarded as an ixodid tick biodiversity hotspot for at least two genera: Haemaphysalis and Dermacentor. To contribute to our knowledge on the tick fauna of this country, ticks from these two genera as well as an Ixodes species were analyzed morphologically and their molecular-phylogenetic relationships were examined in taxonomic and geographical contexts., Methods: For this study, seven Haemaphysalis sp. ticks were removed from dogs and collected from the vegetation. These showed morphological differences from congeneric species known to occur in Vietnam. In addition, three Ixodes sp. ticks were collected from pygmy slow lorises (Xanthonycticebus pygmaeus), and a Dermacentor female had been previously collected from the vegetation. After DNA extraction, these were molecularly or phylogenetically analyzed based on the cytochrome c oxidase subunit I (cox1) and 16S rRNA genes., Results: The three species were morphologically identified as (i) Ixodes granulatus, which had nearly or exactly 100% sequence identities to conspecific ticks reported from large (approximately 2000 km) geographical distances but was more different (having lower, only 94.2% cox1 and 96.7% 16S rRNA sequence identity) from samples collected within 1000 km of Vietnam in Southern China and Malaysia, respectively; (ii) Haemaphysalis bispinosa, which showed 100% sequence identity to samples reported within both narrow and broad geographical ranges; and (iii) a new species, Dermacentor pseudotamokensis Hornok sp. nov., described here morphologically and shown to be phylogenetically a sister species to Dermacentor tamokensis., Conclusions: Haemaphysalis bispinosa shows genetic homogeneity in the whole of South and Southeast Asia, probably owing to its frequent association with domestic ruminants and dogs (i.e. frequently transported hosts). However, I. granulatus, the Asian rodent tick, has a mixed geographical pattern of haplotypes, probably because it may associate with either synanthropic or wild-living rodents as primary hosts. This tick species is recorded here, for the first time to our knowledge, as parasitizing lorises in Vietnam and its region. Based on phylogenetic analyses, D. pseudotamokensis Hornok sp. nov., recognized and described here for the first time, was almost certainly misidentified previously as Dermacentor steini, drawing attention to the need to barcode all Dermacentor spp. in Southern Asia., Competing Interests: Declarations. Ethics approval and consent to participate: Tick samples were collected from all animals during regular veterinary care, therefore no ethical permission was needed. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Research into various bacterial pathogens that can be transmitted between different animals and may have zoonotic potential has led to the discovery of different strains of Bartonella sp. in bats and their associated ectoparasites. Despite their enormous species diversity, only a few studies have focussed on the detection of bacterial pathogens in insectivorous bats of boreal forests and their associated Macronyssidae and Spinturnicidae mites. We collected and molecularly analysed mite samples from forest-dwelling bat species distributed all along the boreal belt of the Palearctic, from Central Europe to Far East. Ectoparasitic mites were pooled for DNA extraction and DNA amplification polymerase chain reaction (PCRs) were conducted to detect the presence of various bacterial (Anaplasmataceae, Bartonella sp., Rickettsia sp., Mycoplasma sp.) and protozoal (Hepatozoon sp.) pathogens. Bartonella sp. DNA was detected in four different mite species (Macronyssidae: Steatonyssus periblepharus and Spinturnicidae: Spinturnix acuminata, Sp. myoti and Sp. mystacinus), with different prevalences of the targeted gene (gltA, 16-23S ribosomal RNA intergenic spacer and ftsZ). Larger pools (>5 samples pooled) were more likely to harbour Bartonella sp. DNA, than smaller ones. In addition, cave-dwelling bat hosts and host generalist mite species are more associated with Bartonella spp. presence. Spinturnicidae mites may transmit several distinct Bartonella strains, which cluster phylogenetically close to Bartonella species known to cause diseases in humans and livestock. Mites with ubiquitous presence may facilitate the long-term maintenance (and even local recurrence) of Bartonella-infestations inside local bat populations, thus acting as continuous reservoirs for Bartonella spp in bats., (© 2024 The Author(s). Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.)

Babesia and Theileria species (Apicomplexa: Piroplasmida) are tick-borne protozoan parasites that can cause mild to severe infection in humans, wildlife, livestock and companion animals. To date, reports on the molecular study of piroplasms from wild living small mammals and their ticks are still limited, especially in Asia. This study encompassed an extensive survey involving 907 liver samples and 145 ixodid ticks from 16 different species of small mammals (Rodentia, Lagomorpha, Eulipotyphla). These were collected in 13 cities and counties in northern China. DNA extracts from these samples were screened for the presence of piroplasm 18S rRNA gene. Samples that tested positive were further evaluated for other genetic markers of piroplasms, including the cox1 gene and the ITS1-5.8S rDNA-ITS2 region. Several piroplasm species were identified, including Babesia sp. tavsan2, Babesia occultans, Theileria sp. Xinjiang, Theileria equi, and Theileria sp. Kalecik. Among these, Theileria sp. Xinjiang was shown to be the most prevalent. Importantly, Babesia sp. tavsan2 was identified in the tick Rhipicephalus sanguineus from the Yarkand hare and Theileria sp. Kalecik in Hyalomma asiaticum from the long-eared hedgehog, in line with the detection of these pathogens in tissue samples of the relevant hosts. This study further disclosed the presence of DNA from B. occultans and T. equi, typically found in cattle and horses respectively, with an additional discovery in small mammals. Moreover, Theileria sp. Kalecik, which was first detected in small-sized mammals, and Babesia sp. tavsan2, were both reported for the first time in China., Competing Interests: Declaration of Competing Interest The authors have no relevant financial or non-financial interests to disclose., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Two species of Southeast Asian pangolins (the Chinese pangolin, Manis pentadactyla and the Malayan or Sunda pangolin, Manis javanica) are critically endangered species. Therefore, knowledge on their parasitic infections is very important, especially considering ticks that can transmit which pathogens. In this study, 32 pangolin ticks (Amblyomma javanense), that were collected in Vietnam and Laos, were analyzed with molecular methods for the presence of tick-borne pathogens. Two members of the family Anaplasmataceae were shown to be present in 14 pangolin ticks, i.e., Candidatus Anaplasma pangolinii and an Ehrlichia sp. In three ticks, a single Rickettsia genotype was also detected, and in seven ticks four 18S rRNA sequence variants of a Babesia sp. Most importantly, a novel protozoan agent, tentatively called here Trypanosoma sp. "PAT14" was detected in one A. javanense nymph. These results imply the first molecular finding of any species of Anaplasma, Ehrlichia and Babesia in pangolin ticks from Vietnam and Laos. On the other hand, detection of a new tick-associated Trypanosoma sp. in A. javanense from Southeast Asia is not only important from a taxonomic point of view, but it is also the first finding of any trypanosomes in the genus Amblyomma in Eurasia and adds pangolins to the potential placental mammalian hosts of any trypanosomes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Potentially zoonotic pathogens have been previously detected in bat-associated ticks, but their role in disease transmission and their frequency of feeding on non-bat hosts is poorly known., Methodology/principal Findings: We used molecular blood meal analysis to reveal feeding patterns of the bat-associated tick species Ixodes ariadnae, I. simplex, and I. vespertilionis collected from cave and mine walls in Central and Southeastern Europe. Vertebrate DNA, predominantly from bats, was detected in 43.5% of the samples (70 of 161 ticks) but in these ticks we also detected the DNA of non-chiropteran hosts, such as dog, Canis lupus familiaris, wild boar, Sus scrofa, and horse, Equus caballus, suggesting that bat-associated ticks may exhibit a much broader host range than previously thought, including domestic and wild mammals. Furthermore, we detected the zoonotic bacteria Neoehrlichia mikurensis in bat ticks for the first time, and other bacteria, such as Bartonella and Wolbachia., Conclusions/significance: In the light of these findings, the role of bat ticks as disease vectors should be urgently re-evaluated in more diverse host systems, as they may contribute to pathogen transmission between bats and non-chiropteran hosts., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Szentiványi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Historically, for more than one and a half centuries, only one so-called "long-legged bat tick" species, i.e., Ixodesvespertilionis Koch was known to science. However, during the past decade, it was recognized on a molecular basis that long-legged ixodid ticks associated with bats may represent at least six species. Of these, until recently, five have been morphologically described. In this study, Ixodes ticks were collected from two Myotis species in southeastern Asia, Vietnam. Based on the morphological and molecular characteristics of the female, nymph and larva, Ixodeslanigeri Hornok, sp. nov. is described here. The male is unknown. Like other members of the Ixodesariadnae complex, I.lanigeri Hornok apparently shows a preference for vesper bats as its typical hosts. In this context, host-association and geographical separation may explain the evolutionary divergence of I.lanigeri Hornok from its closest relative occurring on Murinahilgendorfi Peters in East Asia, Japan, because no Myotis or Murina spp. have overlapping distribution between Vietnam and the main islands of Japan. On the other hand, supposing that (similarly to I.ariadnae ) I.lanigeri Hornok probably occurs on other myotine bats and knowing that several Myotis species indigenous in Vietnam have a broad geographical range in southern and southeastern Asia, the new tick species most likely has a widespread distribution in this area., Competing Interests: The authors have declared that no competing interests exist., (Sándor Hornok, Jenő Kontschán, Ai Takano, Yasuhiro Gotoh, Alexandre Hassanin, Vuong Tan Tu.)

Introduction: Whales, dolphins, and porpoises are susceptible to infections by protozoan and metazoan parasites., Methods: In this study, tissue samples, as well as flatworms and roundworms, were collected from a common bottlenose dolphin ( Tursiops truncatus ), three short-beaked common dolphins ( Delphinus delphis ), two striped dolphins ( Stenella coeruleoalba ), a harbor porpoise ( Phocoena phocoena ), a long-finned pilot whale ( Globicephala melas ), and a fin whale ( Balaenoptera physalus ). These samples were molecularly analyzed., Results: In one D. delphis, Toxoplasma gondii was detected in multiple organs, including the cerebellum. The cysts of the tapeworms Clistobothrium delphini and Clistobothrium grimaldii were identified in G. melas . Flukes collected from D. delphis belong to Brachycladium atlanticum , while those removed from S. coeruleoalba probably represent a new species. Four species of lungworms were also identified: Halocercus delphini in S. coeruleoalba , Halocercus sp. in T. truncatus , Stenurus globicephalae in G. melas , and a potentially new Pharurus sp. in P. phocoena ., Conclusion: These findings show, to the best of our knowledge, for the first time, the presence of T. gondii DNA in D. delphis . The cerebellum of the animal was Toxoplasma -infected, which might be relevant to inadvertent stranding. In this study, new genetic markers were sequenced for several helminth parasites of marine mammals, possibly including undescribed species., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Specht, Keve, Fernández-Maldonado, Cerezo Caro, Takács, Kontschán and Hornok.)

Equine piroplasmosis (EP) is an important tick-borne disease of equids, caused by Theileria equi, Theileria haneyi, and Babesia caballi. Nonetheless, there has been a scarcity of systematic reports on EP parasites in donkeys in Kyrgyzstan, Central Asia. In this study, piroplasms were screened in 1900 blood samples from imported donkeys from the Osh Oblast (southwestern Kyrgyzstan) by targeting partial 18S ribosomal RNA using the polymerase chain reaction (PCR). Through molecular and phylogenetic analyses, all positive samples were sequenced to identify the species and genotypes. The results indicated the presence of both B. caballi and T. equi, with prevalence rates of 8.4% (160/1900) and 12.2% (232/1900), respectively. By amplifying part of the Erythrocyte Merozoite Antigen 1 (EMA-1) and Rhoptry-Associated Protein (RAP-1) genes, B. caballi genotype B and T. equi genotype A were identified. To the best of our knowledge, this is the first report on piroplasm infection among donkeys from Kyrgyzstan.

Background: Southeast Asia is regarded as a hotspot for the diversity of ixodid ticks. In this geographical region, Vietnam extends through both temperate and tropical climate zones and therefore has a broad range of tick habitats. However, molecular-phylogenetic studies on ixodid tick species have not been reported from this country., Methods: In this study, 1788 ixodid ticks were collected from cattle, buffalos and a dog at 10 locations in three provinces of northern Vietnam. Tick species were identified morphologically, and representative specimens were molecularly analyzed based on the cytochrome c oxidase subunit I (cox1) and 16S rRNA genes. Fifty-nine tick species that are indigenous in Vietnam were also reviewed in the context of their typical hosts in the region., Results: Most ticks removed from cattle and buffalos were identified as Rhipicephalus microplus, including all developmental stages. Larvae and nymphs were found between January and July but adults until December. Further species identified from cattle were Rhipicephalus linnaei, Rhipicephalus haemaphysaloides, Amblyomma integrum and Haemaphysalis cornigera. Interestingly, the latter three species were represented only by adults, collected in one province: Son La. The dog was infested with nymphs and adults of R. linnaei in July. Phylogenetically, R. microplus from Vietnam belonged to clade A of this species, and R. haemaphysaloides clustered separately from ticks identified under this name in China, Taiwan and Pakistan. Amblyomma integrum from Vietnam belonged to the phylogenetic group of haplotypes of an Amblyomma sp. reported from Myanmar. The separate clustering of H. cornigera from Haemaphysalis shimoga received moderate support., Conclusions: Three tick species (R. linnaei, A. integrum and H. cornigera) are reported here for the first time in Vietnam, thus increasing the number of indigenous tick species to 62. Clade A of R. microplus and at least R. linnaei from the group of Rhipicephalus sanguineus sensu lato occur in the country. There is multiple phylogenetic evidence that different species might exist among the ticks that are reported under the name R. haemaphysaloides in South and East Asia. This is the first report of A. integrum in Southeastern Asia., (© 2024. The Author(s).)

Birds are long-known as important disseminators of ixodid ticks, in which context mostly their latitudinal, south-to-north migration is considered. However, several bird species that occur in the eastern part of the northern Palaearctic are known to migrate westward. In this study, a female tick collected from the sedge warbler, Acrocephalus schoenobaenus , in Lithuania was identified morphologically and analyzed with molecular-phylogenetic methods. In addition, literature data were reviewed on ixodid tick species known to be associated with birds that have recorded east-to-west migratory route in the Palaearctic. The tick collected from A. schoenobaenus was morphologically identified as Ixodes apronophorus . Two mitochondrial genetic markers for this specimen showed 100% identity with a conspecific tick reported previously in Western Siberia, Russia. Based on literature data, as many as 82 bird species from 11 orders were found to have records of ringing in the easternmost part of the northern Palaearctic and recaptures in Europe. Of these bird species, 31 ixodid tick species were reported in the Euro-Siberian region. Nearly all passeriform bird species with east-to-west migration were reported to carry ticks, whereas no reports of tick infestation were documented from the majority of wetland-associated bird species, mostly from the orders Anseriformes and Charadriiformes. The first European sequences of bona fide I. apronophorus revealed genetic connectedness with conspecific ticks reported from Siberia. Since the principal hosts of this tick species are rodents which do not migrate large distances, the most likely explanation for genetic similarity in this direction is dispersal of this tick species via migratory birds. Given the high number of tick species that are known to associate with bird species migrating in westward direction, this appears to be an important means of the gene flow between geographically distant tick populations in the northern Palaearctic., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Background: Trichomonosis is a common infection in small animals, mostly manifesting in gastrointestinal symptoms such as diarrhea. Although oral trichomonads are also known, the species found colonizing the large intestine are more frequently detected protozoa., Methods: In the present study, four wildcats, 94 domestic cats, and 25 dogs, originating from 18 different locations in Hungary, were investigated for the presence of oral and large intestinal trichomonads based on the 18S rRNA gene and ITS2., Results: All oral swabs were negative by polymerase chain reaction (PCR). However, Tritrichomonas foetus was detected in a high proportion among tested domestic cats (13.8%) and dogs (16%), and Pentatrichomonas hominis only in two domestic cats. In addition, a novel Tritrichomonas genotype was identified in one cat, probably representing a new species that was shown to be phylogenetically most closely related to Tritrichomonas casperi described recently from mice. All positive dogs and half of the positive cats showed symptoms, and among cats, the most frequent breed was the Ragdoll., Conclusions: With molecular methods, this study evaluated the prevalence of oral and intestinal trichomonads in clinical samples of dogs and cats from Hungary, providing the first evidence of T. foetus in dogs of this region. In contrast to literature data, P. hominis was more prevalent in cats than in dogs. Finally, a hitherto unknown large intestinal Tritrichomonas species (closely related to T. casperi) was shown to be present in a cat, raising two possibilities. First, this novel genotype might have been a rodent-associated pseudoparasite in the relevant cat. Otherwise, the cat was actually infected, thus suggesting the role of a predator-prey link in the evolution of this trichomonad., (© 2024. The Author(s).)

Background: Multiple species of the genera Cytauxzoon and Hepatozoon can infect wild felines, but the diversity of these and other apicomplexan parasites in Eurasian lynx is scarcely known. The aim of this study was to detect Cytauxzoon and Hepatozoon species with molecular methods in Eurasian lynxes and their ticks in northwestern China., Methods: DNA was extracted from the heart, liver, spleen, lung, and kidney samples of three Eurasian lynxes as well as from their five ixodid ticks. These DNA samples were screened with polymerase chain reactions (PCRs) for Cytauxzoon with the partial cytochrome b gene (CytB), cytochrome c oxidase subunit I gene (COI), and small subunit ribosomal RNA gene (18S rRNA), and Hepatozoon with three different fragments of small subunit ribosomal RNA gene (18S rRNA). PCR products were sequenced, aligned, and phylogenetically analyzed., Results: One adult female of Eurasian lynx (#1, adult female) was co-infected with Cytauxzoon manul and Hepatozoon felis genotype I, while an adult male lynx (#2) was infected with C. manul. Interestingly, H. felis genotype I was both detected in a male cub (#3) and two out of five infesting Hyalomma asiaticum ticks., Conclusions: For the first time, Cytauxzoon manul is reported here from Eurasian lynx. In addition, H. felis has not been known to occur in this host species in China and Central Asia. Thus, the findings of this study extend our knowledge on the geographical distribution and host range of these haemoprotozoan parasites. Moreover, this is also the first evidence of C. manul and H. felis co-infection in Eurasian lynx., (© 2024. The Author(s).)

Background: Louse flies (Diptera, Hippoboscidae) are important blood-sucking parasites of birds and mammals with a worldwide distribution. The aim of our study was to collect louse flies from birds across multiple sites in Hungary and evaluate the effects of avian traits on louse fly-host relationships., Methods: Between 2015 and 2022, 237 louse flies were collected from birds at multiple locations in Hungary. The louse flies were identified to species level by morphological and molecular methods. Louse fly species and their seasonal dynamics were analyzed., Results: Six louse fly species were identified: Ornithomya avicularia, Ornithomya fringillina, Ornithomya biloba, Ornithomya chloropus, Ornithoica turdi and Ornithoctona laticornis. Results of statistical analyses indicated that habitat, migration habits and the feeding places of birds have significant effects on their possible role as hosts of O. avicularia, O. fringillina and O. turdi. Analysis of the temporal distribution of avian louse flies showed different seasonal patterns according to species. Phylogenetic analyses highlighted that O. turdi clustered separately from other members of the subfamily Ornithomyinae which thus did not form a monophyletic group., Conclusions: This study presents one of the longest continuous collections of ornithophilic louse fly species in Europe so far. Avian traits were shown to influence louse-fly infestation. To our best knowledge, this is the first report on O. laticornis in Europe. The ability of this African louse fly species to survive in Europe, as demonstrated in the present study, may be an indication of its future establishment. Our findings, in accordance with previous reports, also indicated that the subfamily Ornithomyinae should be taxonomically revised., (© 2024. The Author(s).)

Hard ticks (Acari: Ixodidae) are the economically and ecologically most important blood-sucking arthropod vectors that can transmit disease agents under temperate climate. In this group, the highest number of species (currently nearing 270) belongs to the genus Ixodes . For this review, more than 400 papers related to this genus in the context of Russia were checked for data on the host records, locations of collection, as well as ecology of assigned tick species. This monograph compensates for the lack of a similarly comprehensive English-language overview of Ixodes species in the region of Russia for nearly half century, and also makes a large set of data easily available for international readers, which is especially important if the original source is difficult to access from outside this country. In addition, the data from a significant number of papers on this topic available only in the Russian language are made accessible through this work., Competing Interests: The authors have declared that no competing interests exist., (Denis Fedorov, Sándor Hornok.)

Background: In the family Trypanosomatidae, the genus Trypanosoma contains protozoan parasites that infect a diverse range of hosts, including humans, domestic animals, and wildlife. Wild rodents, as natural reservoir hosts of various pathogens, play an important role in the evolution and emergence of Trypanosomatidae. To date, no reports are available on the trypanosomatid infection of pikas (Lagomorpha: Ochotonidae)., Methods: In this study, Mongolian pikas and their fleas were sampled at the China-Mongolia border, northwestern China. The samples were analyzed with polymerase chain reaction (PCR) and sequencing for the presence of Trypanosomatidae on the basis of both the 18S ribosomal RNA (18S rRNA) gene and the glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene. The morphology of trypomastigotes was also observed in peripheral blood smears by microscopy., Results: Molecular and phylogenetic analyses revealed a new genotype of the Trypanosoma lewisi clade that was found both in pika blood and flea samples. This genotype, which probably represents a new species, was provisionally designated as "Trypanosoma sp. pika". In addition, a novel genotype belonging to the genus Blechomonas of Trypanosomatidae was detected in fleas. On the basis of its molecular and phylogenetic properties, this genotype was named Blechomonas luni-like, because it was shown to be the closest related to B. luni compared with other flea-associated trypanosomatids., Conclusions: To the best of our knowledge, this is the first study to report any trypanosomatid species in Mongolian pikas and their fleas. Further studies are needed to investigate the epidemiology of these protozoan parasites, as well as to evaluate their pathogenicity for humans or domestic animals., (© 2024. The Author(s).)

Background: Previous studies on the tick infestation of birds in the Carpathian Basin focused on songbirds (Passeriformes). Thus, the primary aim of the present work was to extend the scope of previous studies, i.e. to include aquatic (water-associated) bird species in a similar context, especially considering that these birds are usually long-distance migrants., Methods: Between March 2021 and August 2023, 11,919 birds representing 126 species were checked for the presence of ticks. From 352 birds belonging to 40 species, 905 ixodid ticks were collected. Tick species were identified morphologically and/or molecularly., Results: Ticks from avian hosts belonged to seven species: Ixodes ricinus (n = 448), I. frontalis (n = 31), I. festai (n = 2), I. arboricola (n = 36), I. lividus (n = 4), Haemaphysalis concinna (n = 382) and Dermacentor reticulatus (n = 2). Nymphs of I. ricinus occurred with a single activity peak around March-May, whereas its larvae typically infested birds in May, June or July. By contrast, H. concinna usually had its activity maximum during the summer (nymphs in June-July, larvae later in July-August). Interestingly, two ornithophilic species, I. frontalis and I. arboricola, were most active around winter months (between October and April). A significantly lower ratio of aquatic birds was found tick-infested than songbirds. Several new tick-host associations were revealed, including I. ricinus from Greylag Goose (Anser anser) and D. reticulatus from Great Egret (Ardea alba) and Sedge Warbler (Acrocephalus schoenobaenus). Ticks were collected for the first time in Europe from two species of predatory birds as well as from Little Bittern (Ixobrychus minutus). Bird species typically inhabiting reedbeds were most frequently infested with H. concinna, and most ticks localized at their throat, as opposed to forest-dwelling avian hosts, on which I. ricinus predominated and ticks were more evenly distributed., Conclusions: In the evaluated region, aquatic birds appear to be less important in tick dispersal than songbirds. However, newly revealed tick-host associations in this category attest to their hitherto neglected contribution. The results suggest that the habitat type will have significant impact not only on the species composition but also on the feeding location of ticks on birds., (© 2024. The Author(s).)

Eurasian lynx ( Lynx lynx ) is widely distributed in various habitats in Asia and Europe, and it may harbor multiple pathogens. Currently, the information on protozoan infection in Eurasian lynx is scarce. In this study, we performed nested polymerase chain reaction (nPCR) analysis to detect intestinal protozoan infection in three dead Eurasian lynxes, in northwestern China. Three dead Eurasian lynxes, an adult female (#1), an adult male (#2), and a cub male (#3), were sampled in West Junggar Mountain, the northwestern region of Xinjiang Uyghur Autonomous Region. The intestine samples were analyzed using nPCR. We used primers targeting the cytochrome C oxidase subunit I gene ( COI ) for detection of Sarcocystis and Eimeria species and targeting the small subunit 18 S ribosomal RNA gene ( 18S rRNA ) for detection of Cystoisospora species. The nPCR-positive products were sequenced, aligned, and phylogenetically analyzed. Three intestinal protozoa, Sarcocystis albifronsi , Eimeria alpacae , and Cystoisospora felis , were found in three Eurasian lynxes. The intestine sample of Eurasian lynx #2 was detected with S . albifronsi and E . alpacae . In addition, C . felis was only found in the intestine sample of Eurasian lynx #3. To the best of our knowledge, S . albifronsi and E . alpacae were detected in Eurasian lynx for the first time. In addition, C . felis was firstly found in Eurasian lynx in China. These findings extend our knowledge of the geographical distribution and host range of intestinal protozoa., Competing Interests: The authors declare that they have no competing interests., (© 2024 The Authors.)

South Africa has six species of primates, three of which are bushbabies (family Galagidae). Very little information is available on their parasites due to the lack of longitudinal studies, although Rhipicephalus appendiculatus, Amblyomma hebraeum and Haemaphysalis elliptica were previously reported from the brown greater galago (Otolemur crassicaudatus) in South Africa. During 2014-2019, 83 O. crassicaudatus (70 live-trapped and 13 deceased animals) were checked for the presence of hard ticks, all from Limpopo Province, South Africa. Seventy-three of 83 (88 %) galagos were found to be tick-infested. Among ixodid genera, Haemaphysalis had the highest prevalence (46 % of the bushbabies), followed by Rhipicephalus (25 %) and Ixodes (18 %). In total, ten tick species were identified. Importantly, all infestations were monospecific. Ticks occurred on various body parts of bushbabies, thus no predilection site was noted. In conclusion, while previously only three ixodid species were known to infest bushbabies in South Africa, the present study showed that these animals can be parasitized by a much broader range of hard ticks., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Birds play a crucial role in disseminating ticks that carry pathogens of high veterinary-medical importance. The aim of this study was to analyze data of a long-term tick collection from birds at a single stop-over site in Central Europe, Hungary. Over eight years (2015-2022) 5833 ticks (ten species) were collected from 2395 tick-infested birds. The most abundant species were Ixodes ricinus (n = 3971) and Haemaphysalis concinna (n = 1706). Ixodes ricinus nymphs and larvae were the most frequently occurring on resident and short-distance migratory birds with forest habitat but Ha. concinna was the most abundant species on reed-associated, long-distance migrants. Haemaphysalis concinna occurred mostly on birds feeding above the ground level, while I. ricinus predominated on ground feeding birds. Infestation with I. ricinus nymphs always peaked in the first half of the year, in contrast to larvae which were more abundant on avian hosts in the autumn. At the same time, Ha. concinna larvae and nymphs had their peak numbers in the summer. This is the first long-term study on the tick infestation of birds in Central Europe. The study shows that, migration distance, habitat type, and typical feeding level of birds, as well as characteristics of tick life cycle are all key factors in the role of birds as tick disseminators. It was revealed that Savi's Warbler (Locustella luscinioides) is the most frequent hosts of Ha. concinna in Central Hungary., (© 2024. The Author(s).)

Bat flies are one of the most abundant ectoparasites of bats, showing remarkable morphological adaptations to the parasitic habit, while the relationship with their hosts is characterized by a high level of specificity. By collecting bat flies from live hosts, our intention was to elucidate the seasonal differences in bat fly occurrence and to describe factors regulating the level of incipient host specificity. Our results indicate that the prevalence and the intensity of infestation is increasing from spring to autumn for most host species, with significant differences among different fly species. Males showed higher infestation levels than females in autumn, suggesting a non-random host choice by flies, targeting the most active host sex. Bat-bat fly host specificity shows seasonal changes and host choice of bat flies are affected by the seasonal differences in hosts' behavior and ecology, the intensity of infestation and the species composition of the local host community. Nycteribiid bat flies showed lower host specificity in the swarming (boreal autumn) period, with higher prevalence recorded on non-primary hosts. Choosing a non-primary bat host may be an adaptive choice for bat flies in the host's mating period, thus increasing their dispersive ability in a high activity phase of their hosts., (© 2024. The Author(s).)

Background: Rodents play an important role in the life cycle of ixodid and argasid ticks, particularly as hosts of larvae and nymphs. The great gerbil (Rhombomys opimus), the preferred prey item of several carnivores (e.g. the red fox and marbled polecat), is the dominant rodent species in the Gurbantunggut Desert in northwestern China. The aim of this study was to investigate tick species associated with different hosts in the habitat of great gerbils, including wildlife and livestock., Methods: During 2018-2023, ticks were removed from 326 great gerbils, two red foxes (Vulpes vulpes), three marbled polecats (Vormela peregusna), 35 pastured sheep (Ovis aries), and one long-eared desert hedgehog (Hemiechinus auritus) in the Gurbantunggut Desert. Ticks were identified according to standard morphological keys. Then, they were further analyzed by molecular and phylogenic methods based on two mitochondrial markers, 16S rDNA and cytochrome c oxidase subunit I (COI) genes., Results: A total of 889 ticks were collected, representing five species. These included Hyalomma asiaticum (n = 425: 24 larvae, 79 nymphs and 322 adults), Rhipicephalus turanicus (n = 153: 2 nymphs and 151 adults), Haemaphysalis erinacei (n = 298: 4 larvae, 7 nymphs and 287 adults), Ixodes acuminatus (n = 7: 4 nymphs and 3 adults) and Ornithodoros tartakovskyi (6 adults). Based on COI sequences, molecular and phylogenetic analyses showed that (i) I. acuminatus from great gerbils and marbled polecats clustered with I. acuminatus reported from Europe; (ii) O. tartakovskyi found in northwestern China belonged to an independent clade; (iii) Hy. asiaticum, R. turanicus and Ha. erinacei had 100% sequence identities to conspecific ticks sampled previously in China., Conclusions: The great gerbil is an important host for the developmental stages of I. acuminatus, O. tartakovskyi, Ha. erinacei, Hy. asiaticum and R. turanicus, thus supporting the life cycle of several tick species which, as adults, parasitize predators (red fox and marble polecat) as well as pastured sheep and hedgehogs in the Gurbantunggut Desert. Ixodes acuminatus and O. tartakovskyi were found for the first time on great gerbil and marbled polecat, respectively., (© 2024. The Author(s).)

Considering the original nature of carnivorism among dogs and cats, nowadays these pets are increasingly fed BARF (Biologically Appropriate Raw Food/Bone And Raw Food) diet to improve their health conditions. However, this diet may also carry health risks, such as vitamin or mineral deficiencies, and infection with pathogens including parasites. In our study, fecal samples of 89 pet animals kept on BARF diet were subjected to coprological examination followed by molecular analyses. Six of them shed eggs of Dicrocoelium dendriticum. This result was confirmed with PCR and sequencing, and in one case, the DNA of Fasciola hepatica was also demonstrated. In addition, oocysts of Cystoisospora canis, a Cystoisospora ohioensis-like sp. and Eimeria stiedai, as well as sporocysts of a Sarcocystis sp. were also detected. All samples were negative for Neospora caninum and Toxoplasma gondii. In conclusion, no evidence was found for the infection of BARF-fed dogs and cats with parasites that are usually associated with this diet and considered as clinico-pathological risk factors for these pets themselves (e.g., N. caninum, T. gondii). However, fluke eggs (probably originating from ruminants) and oocysts of E. stiedai (from rabbit liver in the food) were demonstrated as pseudoparasites. These species are usually not considered among parasite-associated risks of BARF-feeding, implying that other animals living near BARF-fed pets are neglected in this context. However, where intermediate hosts of D. dendriticum occur in urban areas, BARF-feeding may indirectly affect later other dogs and cats. It was also shown here that BARF-feeding may contribute to the contamination of the environment with E. stiedai oocysts, thus increasing the risks of biliary coccidiosis in nearby living pet rabbits that would otherwise not have access to oocysts of E. stiedai., (© 2024. The Author(s).)

Four Ixodes species represent the subgenus Scaphixodes Schulze, 1941 in Europe, but none of them were reported to be compared in a molecular-phylogenetic context. This study compensates for this lack of data. A tick larva, morphologically identified as Ixodes (Scaphixodes) caledonicus Nuttall, 1910, was collected from an Alpine swift (Tachymarptis melba) during its nesting period in Transylvania, Romania. Following DNA extraction, PCR analyses and sequencing in part with newly designed primers, three genetic markers of this specimen were amplified and compared to GenBank data, and two were analyzed phylogenetically. Based on sequence comparisons of its mitochondrial cytochrome c oxidase subunit I (cox1) and nuclear 28S rRNA genes I. caledonicus appeared to be closely related to members of the subgenus Pholeoixodes. However, the topology of the concatenated cox1 and 16S rRNA gene phylogenetic tree clearly showed its clustering with Ixodes (Scaphixodes) philipi. In conclusion, I. caledonicus is part of the tick fauna of Romania and is expected to occur also in other countries of the Carpathian Basin where rocky cliffs are available for nesting of swifts and other birds. This is the first species of the subgenus Scaphixodes in Europe, for which the traditional (morphology-based) taxonomic assignment is confirmed by molecular-phylogenetic analyses., Competing Interests: Declaration of Competing Interest We declare we have no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Background: Among hard ticks (Acari: Ixodidae), the genus Ixodes comprises the highest number of species, which in turn are most numerous in the Afrotropical zoogeographic region. In South Africa extensive morphological studies have been performed on Ixodes species but only few reports included molecular analyses., Methods: In this study, 58 Ixodes spp. ticks, collected from ten mammalian and eight avian host species in South Africa, were molecularly and phylogenetically analyzed. In addition, a newly collected sample of the Palearctic Ixodes trianguliceps was included in the analyses., Results: Among the ticks from South Africa, 11 species were identified morphologically. The majority of ticks from mammals represented the Ixodes pilosus group with two species (n = 20), followed by ticks resembling Ixodes rubicundus (n = 18) and Ixodes alluaudi (n = 3). In addition, single specimens of Ixodes rhabdomysae, Ixodes ugandanus, Ixodes nairobiensis and Ixodes simplex were also found. Considering bird-infesting ticks, Ixodes theilerae (n = 7), Ixodes uriae (n = 4) and ticks most similar to Ixodes daveyi (provisionally named I. cf. daveyi, n = 2) were identified. Molecular analyses confirmed two species in the I. pilosus group and a new species (I. cf. rubicundus) closely related to I. rubicundus sensu stricto. Phylogenetic trees based on concatenated mitochondrial or mitochondrial and nuclear gene sequences indicated that the subgenus Afrixodes forms a monophyletic clade with bird-associated exophilic ticks (subgenus Trichotoixodes). Ixodes trianguliceps clustered separately whereas I. alluaudi with their morphologically assigned subgenus, Exopalpiger., Conclusions: Phylogenetic analyses shed new lights on the relationships of Ixodes subgenera when including multiple sequences from subgenus Afrixodes and African as well as Palearctic species of subgenera Trichotoixodes and Exopalpiger. Subgenera Afrixodes and bird-associated Trichotoixodes share common ancestry, suggesting that the latter might have also originated in Africa. Regarding the subgenus Exopalpiger, I. alluaudi is properly assigned as it clusters among different Australian Ixodes, whereas I. trianguliceps should be excluded., (© 2023. The Author(s).)

In Eurasia, the geographically most widespread ixodid tick species of the bat families Rhinolophidae Gray, Vespertilionidae Gray, and Miniopteridae Dobson were considered to belong to four species, Ixodesvespertilionis Koch, I.collaris Hornok, I.ariadnae Hornok, and I.simplex Neumann. Previous data attest that bat-associated tick species from Eastern Asia show remarkable genetic difference from the above four tick species, but in the absence of detailed morphological comparison these were regarded as conspecific. In this study we compensate for this lack of data on three bat-associated tick species, reporting their morphological comparison, as well as molecular and phylogenetic relationships. According to the results we describe the females of three tick species new to science, i.e., I.nipponrhinolophi Hornok & Takano, sp. nov. , I.fuliginosus Hornok & Takano, sp. nov. , and I.fujitai Hornok & Takano, sp. nov. In case of all three new tick species the cytochrome c oxidase subunit (coxI) gene showed remarkably high sequence differences from the species that they previously were thought to belong to, well exceeding the average limit delineating ixodid tick species. This, as well as observed morphological differences fully justify their taxonomical status as new species., Competing Interests: The authors have declared that no competing interests exist., (Ai Takano, Takeo Yamauchi, Mamoru Takahashi, Hiroshi Shimoda, Yasuhiro Gotoh, Junko Mizuno, Michio Natsume, Jenő Kontschán, Dávid Kováts, Vuong Tan Tu, Sándor Hornok.)

The global epidemiological significance of bats and their blood-sucking ectoparasites is increasingly recognized. However, relevant data are scarce from Pakistan where the Palearctic and Oriental zoogeographic regions meet. In this study, 200 bats belonging to five species were examined for the presence of ectoparasites in Pakistan. Bat flies were found only on Leschenault's fruit bat (Rousettus leschenaultii). The prevalence of infestation did not correlate with habitat type and host traits including age, reproductive status, and sex. All bat flies represented the same Eucampsipoda species which was shown to be morphologically different from all species of its genus with known south Asian distribution and belonged to a separate phylogenetic group. These results highlight the existence of a hitherto undescribed bat fly species in southern Asia, which is not shared by the fruit bat species (R. leschenaultii) and insectivorous ones (e.g., Rhinopoma microphyllum) thus probably playing a role only in intraspecific transmission of pathogens., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Among vector-borne protozoa Hepatozoon felis and Cytauxzoon europaeus are considered emerging species in felids in Europe. To investigate the presence of these two protozoa 127 domestic cats and 4 wildcats were screened by PCRs targeting the 18S rRNA gene of Hepatozoon spp. and piroplasms, as well as the cytb gene of Cytauxzoon spp. The samples were collected inside and outside a region of Hungary, where both protozoan groups are endemic in wildcats. Among domestic cats, one proved to be infected with H. felis . Furthermore, spleen samples of four wildcats were also examined, among which three tested positive for H. felis , and one had co-infection with C. europaeus . Importantly, H. felis from the co-infected wildcat belonged to genogroup II, similarly to H. felis from the positive domestic cat. Based on phylogenetic evidence, this genogroup probably represents a separate species from genogroup I of H. felis , which was hitherto reported from Mediterranean countries in Europe. The two other wildcats also harbored H. felis from genogroup I. Neither Hepatozoon nor Cytauxzoon infections were detected outside the recently discovered endemic region. In conclusion, this study demonstrates for the first time in Europe that H. felis from genogroup II may emerge in free-roaming domestic cats in regions where this protozoan parasite is endemic in wildcats.

Reports on adult Hyalomma ticks in certain regions of the Carpathian Basin date back to the 19th century. These ticks were thought to emerge from nymphs dropping from birds, then molting to adults. Although the role of migratory birds in carrying ticks of this genus is known from all parts of Europe, in most countries no contemporaneous multiregional surveillance of bird-associated ticks was reported which could allow the recognition of hotspots in this context. From 38 passeriform bird species 956 ixodid ticks were collected at seven locations in Hungary. Tick species were identified as Ixodes ricinus ( n = 598), Ixodes frontalis ( n = 18), Ixodes lividus ( n = 6), Haemaphysalis concinna ( n = 321), and D. reticulatus ( n = 1). All 12 Hyalomma sp. ticks (11 engorged nymphs and 1 unengorged larva) were identified as H. rufipes based on three mitochondrial markers. This species was only found in the Transdanubian region and along its southeastern border. The Common Blackbird and the European Robin were the two main hosts of I. ricinus and I. frontalis , whereas H. concinna was almost exclusively collected form long-distance migrants. The predominant hosts of H. rufipes were reed-associated bird species, the Sedge Warbler and the Bearded Reedling, both in their nesting period. This study provides ornithological explanation for the regional, century-long presence of adult Hyalomma ticks under continental climate in the Transdanubian Region of the Carpathian Basin. More importantly, the autochthonous occurrence of a H. rufipes population was revealed for the first time in Europe, based on the following observations: (1) the bird species infested with H. rufipes are not known to migrate during their nesting period; (2) one larva was not yet engorged; (3) the larva and the nymphs must have belonged to different local generations; and (4) all H. rufipes found in the relevant location were identical in their haplotypes based on three maternally inherited mitochondrial markers, probably reflecting founder effect. This study also demonstrated regional and temporal differences in tick species carried by birds., Competing Interests: Authors TC, AB, AH, AM, ÁN, BK, ET, JG, OK, DK, and ZK were employed by BirdLife Hungary. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Keve, Csörgő, Benke, Huber, Mórocz, Németh, Kalocsa, Tamás, Gyurácz, Kiss, Kováts, Sándor, Karcza and Hornok.)

Background: The subgenus Pholeoixodes contains Ixodes species typically associated with birds that nest in cavities or with carnivorous mammals that are burrow-dwelling. Among ticks infesting the latter, Ixodes rugicollis is regarded as the rarest species in the western Palearctic. Despite the unique morphology of this species, its identification (especially of subadult stages) is difficult, and molecular-phylogenetic data to offer other diagnostic methods and a better understanding of its taxonomy are not available., Methods: In this study, a female and a male of I. rugicollis were collected in Romania. The female was compared morphologically to another female of this species collected in France and to the lectotype of Ixodes cornutus (from Tajikistan), which has similar morphology and host association. Following DNA extraction, two mitochondrial (cytochrome c oxidase subunit I: cox1 and the 16S rRNA gene) and two nuclear genetic markers (18S and 28S rRNA genes) of I. rugicollis were amplified and analyzed in a phylogenetic context., Results: Females of I. rugicollis and I. cornutus differed in the shape of their palps, scutum and areae porosae and the size of peritremes, but they were similar in palpal setal length, dental formula and arrangement of anal setae. Measurements of two I. rugicollis females examined were not less different from each other than from I. cornutus. Phylogenetically, I. rugicollis clustered with other members of its subgenus. The topology of all trees showed the position of bat-associated tick species of the subgenus Eschatocephalus among Pholeoixodes species., Conclusions: For the first time to our knowledge, this study provides high-resolution digital pictures of male and female I. rugicollis as well as corresponding molecular data. Morphological comparison of this species with I. cornutus could not resolve uncertainties in the validity of the latter species, which can only be accomplished after collecting new specimens of I. cornutus and consequent molecular comparisons. This study includes the first comprehensive molecular-phylogenetic analysis of western Palearctic Pholeoixodes species based on both nuclear and mitochondrial genetic markers and including I. rugicollis. The results of these confirm the phylogenetic position of subgenus Eschatocephalus within Pholeoixodes, justifying the need to merge them to comply with the taxonomic criterion of monophyly., (© 2023. The Author(s).)

To obtain new data on the species diversity, host associations and spatiotemporal occurrence of flies and blood-sucking lice of veterinary importance in Malta, ectoparasites were collected at cattle, sheep and goat, pig farms, as well as dog shelters, and in two places in the absence of domestic animals. The species were identified morphologically, but voucher specimens were also analyzed with molecular-phylogenetic methods following DNA extraction. Altogether 3,095 flies (Diptera: Muscidae, Calliphoridae) were collected at farms and kennels near domestic animals, as well as 37 blowflies (Calliphoridae) in rural and urban areas without animals nearby. Regarding Muscidae, the great majority of flies ( n = 3,084) were identified as the common housefly ( Musca domestica ). Eight flies represented the stable fly ( Stomoxys calcitrans ). Three blowflies associated with dogs and small ruminants belonged to Lucilia cuprina . By contrast, all 37 blowflies collected without domestic animals nearby, were identified as Lucilia sericata . In addition, 22 sucking lice were collected from goats, and all belonged to Linognathus africanus . Molecular identification of 28 flies and four lice confirmed the above species. Considering the sex ratio of M. domestica among samples collected randomly at cattle farms, females predominated in the whole study period, but the abundance of males increased significantly toward the autumn. Stomoxys calcitrans was associated with cattle and dogs, whereas L. cuprina was found near small ruminants and dogs. To our knowledge, this is the first study including the molecular analysis of flies and lice of veterinary-medical importance from Malta. The most important finding of this study is the first evidence for the autochthonous occurrence of L. cuprina in Malta. The exclusive presence of L. cuprina at animal-keeping facilities in rural areas and association of L. sericata with urban areas void of livestock might reflect similar habitat preference of these species in Malta to what was reported in South Africa. Based on the sucking-louse burden in the examined goat herds, the situation in Malta was similar to northern Africa where the exclusive presence of L. africanus was reported, unlike toward the north in the Mediterranean Basin where populations of this species are mixed with Linognathus stenopsis ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Hornok, Cini Bruno, Takács, Keve, Sándor and Kontschán.)

Trichomonas gallinae is a geographically widespread protozoan parasite of birds. In this study, oropharyngeal swab samples were collected in Hungary and Romania from 99 columbiform birds, including 76 feral pigeons ( Columba livia domestica : 42 kept for racing, 32 with urban and two with rural habitat), four common wood pigeons ( C. palumbus ), 16 ring doves ( Streptopelia risoria ) and three Eurasian collared doves ( S. decaocto ). These samples were analyzed for the presence of T. gallinae using molecular methods. Racing feral pigeons had significantly higher prevalence of T. gallinae infection than urban feral pigeons. The rate of PCR-positivity was the highest among wood pigeons and ring doves. Based on 18S rRNA gene, T. gallinae was the most heterogenous among racing feral pigeons sampled in a trading-breeding place. Clinical signs were associated with only one 18S rRNA gene subtype. The most divergent 18S rRNA gene subtype, Trichomonas sp. Hu-TG37 clustered with T. canistomae and T. tenax and represents probably a new species. To our knowledge, this is the first report on the genetic diversity of T. gallinae in the southeastern European region. The results suggest that most detected T. gallinae 18S rRNA gene subtypes are not host-specific and do not cause clinical signs. The highest number of 18S rRNA gene subtypes was demonstrated among racing feral pigeons. Significantly more captive than free-living columbiform birds had T. gallinae infection. These data highlight the importance of epizootic monitoring of the genetic diversity and presence of T. gallinae in trading-breeding places of pigeons and doves., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tuska-Szalay, Sipos, Takács, Kontschán, Sándor, Péter, Berta, Kerek, Jerzsele, Votýpka and Hornok.)

Reptiles are frequently kept as pet animals. They are considered as important reservoirs of protozoa with veterinary-medical significance. At a reptile farm in Ireland, fecal samples were collected from 98 captive reptiles, representing 43 species of three orders (Squamata, Testudines, and Crocodylia). After DNA extraction, all samples were screened by conventional PCRs, targeting the ribosomal small subunit (SSU) RNA and alpha-tubulin genes of trichomonads and SSU RNA gene of Acanthamoeba spp. One leopard gecko (Eublepharis macularius) was positive for a not yet reported species/genotype of the genus Monocercomonas, different from M. colubrorum. Various Acanthamoeba genotypes were detected in six reptilian species, i.e., Acanthamoeba genotype T11 in Eunectes notaeus and Heloderma suspectum/horridum; genotype T4 in Varanus exanthematicus, Chlamydosaurus kingii, and Macrochelys temminckii; and the genotype T13 in Iguana iguana. Some of these amoeba species might have clinicopathological significance in both humans and animals. Our findings highlight the importance to monitor pathogenic protozoa in pet as well as wildlife reptiles, as a source of possible infection for animals and humans living nearby., (© 2022. The Author(s).)

Polyctenidae bugs are rarely studied, hematophagous, and highly specialized ectoparasites of bats. There are only 32 described species worldwide, including six species in the Afrotropical region. Knowledge on these parasites is limited, and most studies are restricted to the New World polyctenid species. Here we report additional records of Adroctenes horvathi from Kenya and South Africa, as well as Hypoctenes faini from Rwanda. We present an updated list of published polyctenid records in the Afrotropical region indicating their host specificity and their geographical distribution. We report global infection patterns and sex ratio of polyctenids based on previously published data, including Old and New World species. Lastly, we demonstrate the first molecular phylogeny of Polyctenidae, showing their phylogenetic relationship with the closely related family Cimicidae., Competing Interests: The authors declare no competing interests., (© 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

This study was initiated to assess the seasonality and to investigate the morphology of questing ixodid ticks in an urban habitat in Central Europe, Hungary. A neglected part of a large cemetery, with sparse tree covering and dense lower vegetation, was sampled monthly from February 2019 to May 2021. All ticks were analyzed morphologically, and selected specimens by amplifying and sequencing two genetic markers. During the study 3818 ticks were collected, including Ixodes ricinus (n = 2772), Ixodes frontalis (n = 350) and Haemaphysalis concinna (n = 696). Ixodes ricinus adults and nymphs showed year-round activity, whereas H. concinna was not active during winter months and early spring. Most I. frontalis nymphs were collected in late winter and early spring, whereas the peak activity of larvae was during late autumn. Interestingly, during the spring, the peak activity of I. ricinus adults and nymphs was later (in May) when preceded by a warm winter in 2020. In contrast, the 2019 and 2021 spring activity peaks occurred in March and April after sharply rising temperatures in February. This shift in the peak activity of I. ricinus coincided with the initiation of questing activity of H. concinna. Three notably different morphotypes and four malformed specimens of I. ricinus were found. However, these were not significantly different in their mitochondrial haplotypes and phylogenetic clustering from typical specimens of this species. On the other hand, I. frontalis was represented by two remarkably different haplogroups, between which in the nymph stage there were no recognizable morphological differences, suggesting the status of these as cryptic species., (© 2022. The Author(s).)

Purpose: To date, ten validated Arthrostoma species were reported. Here, a new hookworm species was found from Asian badger (Meles leucurus)., Methods: Nineteen hookworms (9 males and 10 females) were collected from the small intestine of two Asian badgers in Xinjiang Uygur Autonomous Region, northwestern China. The hookworms were morphologically examined according to key taxonomic characters, such as anterior extremity direction, structures of oral opening (cutting plates or teeth), vulva location, buccal capsule anatomy (integrated or formed by articulating plates), the length of spicule and gubernaculum, number of plates of buccal capsule, and presence or absence of vulvar papillae., Results: The hookworm species from Asian badger, here named as Arthrostoma leucurus sp. n., was different from the previously described ten Arthrostoma species. The phylogenetic tree based on the cox1 gene showed that Arthrostoma leucurus sp. n. formed a separate clade, as a sister group to Ancylostoma and Uncinaria species., Conclusion: Arthrostoma leucurus sp. n., the eleven validated Arthrostoma species, was identified from Asian badger in China., (© 2022. The Author(s).)

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., (Copyright © 2022. Published by Elsevier GmbH.)

Anaplasma phagocytophilum is the causative agent of granulocytic anaplasmosis in humans, dogs, cats, horses and tick-borne fever in ruminants. In Europe, its main vector is the tick species Ixodes ricinus. In this study, spleen and liver samples, as well as ticks from 18 wild-living mammals (belonging to seven species) were analysed for the presence of A. phagocytophilum with molecular methods. The zoonotic ecotype-I of A. phagocytophilum was identified in a European wildcat (Felis silvestris) and its tick, a European pine marten (Martes martes) and a Eurasian red squirrel (Sciurus vulgaris). All PCR-positive samples were collected in 2019 and originated in the same geographic area. These results indicate that taxonomically diverse mammalian species can maintain the local enzootic cycle of the same genotype of A. phagocytophilum. To the best of our knowledge, this is the first report of the zoonotic variant of A. phagocytophilum in the wildcat and in the European pine marten in a broad geographical context, as well as in the red squirrel in Hungary. Since all these host species are well known for their urban and peri-urban presence, the results of this study verify their role in the synanthropic enzootic cycle of granulocytic anaplasmosis and tick-borne fever.

Hard ticks (Acari: Ixodidae) are considered the most important transmitters of pathogens in the temperate zone that covers most of Europe. In the era of climate change tick-borne diseases are predicted to undergo geographical range expansion toward the north through regions that are connected to southern areas of the continent by bird migration. This alone would justify the importance of synthesized knowledge on the association of tick species with avian hosts, yet birds also represent the most taxonomically and ecologically diverse part of urban vertebrate fauna. Birds frequently occur in gardens and near animal keeping facilities, thus playing a significant role in the dispersal of ticks and tick-borne pathogens in synanthropic environments. The primary aim of this review is to provide a comprehensive reference source (baseline data) for future studies, particularly in the context of discovering new tick-host associations after comparison with already published data. The records on the ixodid tick infestations of birds were assessed from nearly 200 papers published since 1952. In this period, 37 hard tick species were reported from 16 orders of avian hosts in Europe. Here we compile a list of these tick species, followed by the English and Latin name of all reported infested bird species, as well as the tick developmental stage and country of origin whenever this information was available. These data allowed a first-hand analysis of general trends regarding how and at which developmental stage of ticks tend to infest avian hosts. Five tick species that were frequently reported from birds and show a broad geographical distribution in the Western Palearctic ( Ixodes arboricola, I. frontalis, I. ricinus, Haemaphysalis concinna and Hyalomma marginatum ) were also selected for statistical comparisons. Differences were demonstrated between these tick species regarding their association with bird species that typically feed from the ground and those that rarely occur at the soil level. The ecology of these five bird-infesting tick species is also illustrated here according to avian orders, taking into account the ecology (habitat type) and activity (circadian rhythm and feeding level) of most bird species that represent a certain order., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Keve, Sándor and Hornok.)

European beaver (Castor fiber L. 1758) is the biggest rodent species living in Europe. Beavers are semi-aquatic animals; they are defecating directly into the water; thus, they have an important role in spreading parasites related to water (e.g., protozoa and flukes). The first specimens of this once extinct rodent species in Hungary turned up in Szigetköz (upper flow of the Hungarian Danube) in 1991 dispersed from Austria. The reintroduction to Hungary started in 1996, and the population slowly increased in number up to around 4000 individuals, but the knowledge about their parasites is lacking. This is the first report on the metazoan parasites of beavers in Hungary and their molecular taxonomy. In the 5-year study, 47 beavers were trapped in four locations and euthanized with permission. Three different metazoan parasites were collected: larvae and adults of Platypsyllus castoris beetles, nymphs and adults of Schizocarpus sp. mites and eggs and adults of Stichorchis subtriquetrus flukes. From these three parasite species, molecular taxonomic studies were also carried out. The low number of metazoan parasites species detected in Hungarian beavers compared to other European countries (e.g., Poland) might be attributed to host population bottleneck effect during reintroduction. As parasites represent a significant component of the biodiversity and ecosystem, the conservation efforts should focus not only on host species but also on their parasites., (© 2022. The Author(s).)

Background: Among live wild mammals adapted to urban and peri-urban habitats in Europe, members of the families Felidae, Mustelidae and Sciuridae deserve special attention as pathogen reservoirs because all of these families include members that are kept as pets. We report here the results of our study on two important groups of tick-borne protozoan parasites in ticks and tissues of wild cats, mustelids and red squirrels., Methods: DNA was extracted from the tissues of carnivores (wild cats, mustelids; n = 16) and red squirrels (n = 4), as well as from ixodid ticks (n = 89) collected from these hosts. These DNA extracts were screened for piroplasms and Hepatozoon spp. using conventional PCR analysis and sequencing. In addition, 53 pooled samples of 259 questing Haemaphysalis concinna ticks were evaluated for the presence of Hepatozoon DNA, followed by phylogenetic analyses., Results: One wild cat was found to be coinfected with Cytauxzoon europaeus and a new genotype of Hepatozoon felis, and two additional wild cats were infected with H. felis from a different phylogenetic group. In mustelids, Hepatozoon martis and two further Hepatozoon genotypes were detected. The latter clustered separately, close to others reported from eastern Asia. In addition, Hepatozoon sciuri was detected in red squirrels. Morphologic and molecular analyses verified eight tick species. One wild cat was infected with a H. felis genotype that was significantly different from that in Ixodes ricinus females infesting this cat. Only three pools of questing H. concinna nymphs tested positive for Hepatozoon, one of which contained H. martis., Conclusions: This study provides the first evidence of the occurrence of any Cytauxzoon species and of three Hepatozoon species in Hungary. In addition to H. martis, two further mustelid-associated Hepatozoon genotypes were detected, one of which was new in terms of phylogenetic and broader geographical contexts. This may be the first indication that H. felis genotypes from both of its phylogenetic groups occur in Europe. This also appears to be the first evidence of H. felis and C. europaeus coinfection in felids in Europe, and of autochthonous H. felis infection in wild cats north of the Mediterranean Basin. New tick-host associations were also observed in this study. Based on the results, H. felis and H. martis might survive transstadially in I. ricinus and H. concinna, respectively., (© 2022. The Author(s).)

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., (Copyright © 2022. Published by Elsevier GmbH.)

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., (Copyright © 2021 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

All species of hard ticks associated with reptiles as hosts throughout their life cycle, are currently assigned to genera including Amblyomma and Africaniella. Among these species, based on literature data, Africaniella transversale has never been investigated for the presence of tick-borne pathogens. In this study, seven DNA extracts (two from A. transversale and five from Amblyomma exornatum) were screened for the presence of important tick-borne protozoa (piroplasms) and bacteria (Anaplasmataceae and Rickettsiaceae) with conventional PCRs and sequencing. A new heat shock protein chaperonin (groEL) gene-specific PCR was also developed to identify Occidentia spp. in these samples. In A. transversale, Occidentia massiliensis (previously detected in rodent-associated soft ticks) and Rickettsia hoogstraalii were present. While the latter was molecularly identical with formerly reported sequences of this rickettsia, the genotype of O. massiliensis was new based on sequence and phylogenetic analyses of its groEL gene. In A. exornatum, a Rickettsia genotype closely related to R. tamurae and R. monacensis, was detected. The ompA sequence of this genotype was identical to that of Rickettsia sp. Ae-8 reported from A. exornatum in a reptile breeding facility in the USA. These results show that A. transversale might carry O. massiliensis which (unless having a symbiotic nature in ticks) may originate either from the reptile host of this hard tick species or the rodent prey of reptiles. This is also the first detection of the reptile tick-associated Rickettsia sp. Ae-8 (phylogenetically aligning with R. tamurae, R. monacensis) in Africa, i.e. within the original geographical range of A. exornatum., (Copyright © 2021. Published by Elsevier GmbH.)

In this study, 263 tapeworms were collected from eight road-killed red foxes in Xinjiang Uygur Autonomous Region (XUAR, northwestern China). The tapeworms were analyzed based on morphological characters and mitochondrial cytochrome c oxidase subunit 1 ( cox1 ) gene sequences. Eighty-three Taenia and 180 Mesocestoides tapeworms were identified according to the presence or absence of rostellum, and the number, the length and the shape of the large rostellar hooks. The morphological and molecular analyses revealed that i ) 180 Mesocestoides tapeworms, here named as Mesocestoides sp . ( Vulpes vulpes ), showed 99.21% (378/381 bp) identity to Mesocestoides sp . reported from red fox in Mongolia; and ii) 83 Taenia tapeworms belonged to three species. The first Taenia species [n = 16, named as Taenia sp . ( Vulpes vulpes )], based on the length of large rostellar hooks (337-342 μm) and its cox1 sequence, was identified as a potentially novel species, which is phylogenetically close to Taenia laticollis . The second species [n = 54, named as Taenia sp. ( Vulpes vulpes & Rhombomys opimus )], was morphologically similar to Taenia endothoracicus according to the number (n = 52), the length (319-332 μm) and the shape of the large rostellar hooks. This species, infecting three red foxes, shared 100% cox1 sequence identity with Taenia sp. ( Rhombomys opimus ) genotype C found previously in great gerbils ( Rhombomys opimus ) in the same region. The third species (n = 13, named as Taenia polyacantha -like), had shorter large rostellar hooks (178-180 μm) and showed 96.27% (361/375 bp) sequence identity to Taenia polyacantha reported from red fox in Italy. The "great gerbil-red fox" life cycle of Taenia sp. ( Vulpes vulpes & Rhombomys opimus ), belonging to the mitochondrial lineage of T . endothoracicus , is confirmed. The T. polyacantha -like species was firstly found in red fox in China. Taenia sp . ( Vulpes vulpes ) is a potentially novel species, which is close to T . laticollis based on its phylogenetic properties., Competing Interests: No conflicts of interest exits in the submission of this manuscript, and the manuscript was approved for publication by all authors., (© 2021 The Authors. Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

Piroplasmosis is an economically important tick-borne disease worldwide. However, little is known about the presence of Babesia spp. and Theileria spp. in ticks in Eastern and Southern Kazakhstan (ESK). During 2016 - 2019, adult ticks (at 26 sampling sites in 16 districts of 5 oblasts in ESK) were collected. Tick species were identified according to morphological and molecular characteristics. Two fragments (487 bp and 438 bp) of 18S ribosomal RNA (18S rRNA) were used to determine piroplasm species in representative 698 ticks. The genotype characteristics of Babesia caballi and Theileria equi were further analyzed by longer 18S rRNA gene fragments. A total of 6107 adult ticks (4558 parasitizing ticks and 1549 off-host ticks), including 4665 hard ticks and 1442 soft ticks, were collected from their natural hosts (cattle, horses, sheep, camels, shepherd dogs and hedgehogs) and the surrounding environment, respectively. Among the hard tick species, Dermacentor marginatus (62.59%, 2920/4665) was the most abundant, followed by Hyalomma asiaticum (19.36%, 903/4665) and Hyalomma detritum (9.95%, 464/4665). All soft ticks were identified as Argas persicus. 16S ribosomal DNA (16S rDNA) phylogenic analysis showed that several tick species in Kazakhstan, as exemplified by Haemaphysalis erinacei and D. marginatus, clustered together with conspecific ticks reported from China. Five species of piroplasms, i.e. Babesia occultans, Babesia caballi, Theileria ovis, Theileria annulata and Theileria equi, were detected in 698 representative ticks. Genotype E of T. equi in Almaty, and genotype A of B. caballi in Almaty and South Kazakhstan were identified., (Copyright © 2021. Published by Elsevier GmbH.)

The Eurasian lynx ( Lynx ) is a medium-sized wild cat species distributed throughout Eurasia. There has been no report on Taenia species (Cestoda: Cyclophyllidea) infecting this felid in China. In this study, 24 tapeworms were found in two Eurasian lynxes (#1 and #2) in Xinjiang Uygur Autonomous Region (XUAR), northwestern China. Based on the number, measurements and the shape of rostellar hooks, these tapeworms belong to two Taenia species. According to the number (n = 32) and length (185-194 μm) of small hooks, the first Taenia species (n = 1, found in #2 lynx) was identified as Taenia laticollis . Phylogenetically, this species was clustered with T . laticollis genotype C (JX860623) based on its cytochrome c oxidase subunit 1 ( cox1 ) and 16S r DNA sequences. The second Taenia species (n = 23, provisionally named as " Taenia sp.") may represent a potentially novel tapeworm species, because of its obvious differences in the shape and lengths (174-182 μm, 98-113 μm) of large and small rostellar hooks in comparison with ten taxonomically related species. Molecular and phylogenetic analyses of the cox1 gene revealed that " Taenia sp." has the highest rate of sequence identity (92.93%, 368/396 bp) with Taenia hydatigena reported from sheep ( Ovis aries ) in Slovakia. To sum up, a potentially novel tapeworm species, " Taenia sp.", is found in Eurasian lynx. In addition, T . laticollis was found for the first time in China., Competing Interests: The authors declare that they have no competing interests., (© 2021 The Authors.)

Babesia gibsoni is considered as an emerging protozoan parasite of dogs in North America and Europe. However, no data have been published on its prevalence, molecular-phylogenetic characteristics and associated co-infections in dogs used for illegal fighting (i.e. predisposed to acquiring this piroplasm via biting) in Europe. In this study, blood samples from 79 American Staffordshire Terrier dogs, confiscated for illegal dog fights, were molecularly analyzed for tick-borne pathogens. B abesia gibsoni was detected in 32 dogs, i.e. with a prevalence of 40.5%. In addition, Babesia vulpes was found in 8 samples (prevalence of 10.1%), for the first time in dogs in Hungary. Canine hemoplasmas were also identified in 49 samples (62%): only Mycoplasma haemocanis in 32 (40.5%) dogs, only " Candidatus Mycoplasma haematoparvum" in 9 (11.4%) dogs, and both hemoplasmas in 8 (10.1%) dogs. Thus, hemoplasma infections also showed a particularly high prevalence in this dog population. Based on a partial fragment of the 18S rRNA gene, B. gibsoni from Hungary exhibited complete sequence identity with conspecific strains reported from Europe and Asia. The cytochrome c oxidase subunit 1 ( cox 1) gene sequence of this isolate showed the closest identity with B. gibsoni reported from Japan but had a nonsynonymous mutation (M33I). Furthermore, the 11 B. gibsoni -positive samples analyzed for sequence variants of the cytochrome b ( cytb ) gene showed the presence of a common mutation (P310S). Most importantly, B. gibsoni had two further nonsynonymous mutations, M121I and F258L, in a dog with severe and relapsing anemia following atovaquone treatment. Phylogenetically, both cytb sequence variants clustered together, with a clear geographical pattern showing the closest relationship of both haplotypes identified in Hungary with those from China and Japan. To the best of our knowledge, this is the first cox 1 and cytb characterization of B. gibsoni in Europe, as well as the first report on the emergence of this piroplasm and hemoplasmas with high prevalence among "fighting dogs" north of the Mediterranean Basin., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Author(s).)

The family Cimicidae includes obligate hematophagous ectoparasites (bed bugs and their relatives) with high veterinary/medical importance. The evolutionary relationships of Cimicidae and their hosts have recently been reported in a phylogenetic context, but in the relevant study, one of the six subfamilies, the bat-specific Latrocimicinae, was not represented. In this study the only known species of Latrocimicinae, i.e., Latrocimex spectans, was analyzed with molecular and phylogenetic methods based on four (two nuclear and two mitochondrial) genetic markers. The completed subfamily-level phylogeny of Cimicidae showed that Latrocimicinae is most closely related to Haematosiphoninae (ectoparasites of birds and humans), with which it shares systematically important morphologic characters, but not hosts. Moreover, in the phylogenetic analyses, cimicid bugs that are known to infest phylogenetically distant bat hosts clustered together (e.g., Leptocimex and Stricticimex within Cacodminae), while cimicid subfamilies (Latrocimicinae, Primicimicinae) that are known to infest bat hosts from closely related superfamilies clustered distantly. In conclusion, adding Latrocimicinae significantly contributed to the resolution of the phylogeny of Cimicidae. The close phylogenetic relationship between Latrocimicinae and Haematosiphoninae is consistent with long-known morphologic data. At the same time, phylogenetic relationships of genera within subfamilies are inconsistent with the phylogeny of relevant hosts., (© 2021. The Author(s).)

Ixodes simplex is a bat tick species, a common parasite of the Schreibers' bent-winged bat, Miniopterus schreibersii. Its distribution is linked to the range of its host, free stages occurring exclusively inside the underground bat shelters. Here we present the first case of human infestation with I. simplex. An adult female tick was found attached to the upper limb after a visit to the underground shelter of a large bat colony. This unusual host selection is a likely consequence of the reduction of suitable hosts, as the number of bats was much lower at the time of the visit than in previous years. Bat ticks rarely feed on humans, with soft ticks (Argasidae) being more commonly involved. In the light of the potential vectorial capacity of I. simplex, the incidence and potential future risks are discussed., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Babesia species (Apicomplexa: Piroplasmorida) are tick-borne protozoan hemoparasites, which pose a significant threat to domestic animals, wildlife and humans. This study aimed to determine and characterize Babesia species in red foxes (Vulpes vulpes), Asian badgers (Meles leucurus) and their ticks. Blood, heart, liver, spleen, lung, kidney, large intestine and small intestine were collected from 19 wild carnivores (12 red foxes and 7 Asian badgers). All ticks were removed from these animals and identified according to morphological and molecular characteristics. The samples were tested for the presence of Babesia species using the 18S rRNA gene. Molecular analyses showed that the DNA of Babesia vogeli and Babesia vulpes was present in red fox organs/tissues and blood samples. A total of 54 hard ticks (38 Ixodes canisuga, 6 Haemaphysalis erinacei, 9 Ixodes kaiseri and 1 Dermacentor marginatus) were collected from red foxes and 12 (I. kaiseri) from Asian badgers. All ticks were adults. Among them, one I. kaiseri parasiting a red fox contained the DNA of B. vulpes while one I. canisuga was positive for Babesia sp. belonging to the clade "Babesia sensu stricto". Molecular and phylogenetic analyses indicated the presence of a novel genotype, Babesia sp. "badger China". Babesia sp. badger type A and type B from Asian badgers were different from those in European badgers. Co-infection with three Babesia genotypes was found in one Asian badger. This study provides the first data on Babesia infection in red foxes, Asian badgers and their ticks in China. Babesia vogeli was detected for the first time in red foxes in Asia. Co-infection and genetic diversity of Babesia genotypes in Asian badgers were also demonstrated., (Copyright © 2021 Elsevier GmbH. All rights reserved.)