Shift in the seasonality of ixodid ticks after a warm winter in an urban habitat with notes on morphotypes of Ixodes ricinus and data in support of cryptic species within Ixodes frontalis.

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.

Long term evaluation of factors influencing the association of ixodid ticks with birds in Central Europe, Hungary.

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.

Molecular investigation of endoparasites of marine mammals (Cetacea: Mysticeti, Odontoceti) in the Western Mediterranean.

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.

East-to-west dispersal of bird-associated ixodid ticks in the northern Palaearctic: Review of already reported tick species according to longitudinal migratory avian hosts and first evidence on the genetic connectedness of Ixodes apronophorus between Siberia and Europe.

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.

Investigations of the tick burden on passeriform, water-associated and predatory birds reveal new tick-host associations and habitat-related factors of tick infestation.

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.

A morpho-phylogenetic update on ixodid ticks infesting cattle and buffalos in Vietnam, with three new species to the fauna and a checklist of all species indigenous to the country.

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.

Contributions to our knowledge on avian louse flies (Hippoboscidae: Ornithomyinae) with the first European record of the African species Ornithoctona laticornis.

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.

An update on the occurrence of flies (Diptera: Muscidae, Calliphoridae) and sucking lice (Phthiraptera: Anoplura) of veterinary importance in Malta: First record of Lucilia cuprina and Linognathus africanus.

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.

Molecular-phylogenetic analyses of Ixodes species from South Africa suggest an African origin of bird-associated exophilic ticks (subgenus Trichotoixodes).

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.

Ornithological and molecular evidence of a reproducing Hyalomma rufipes population under continental climate in Europe.

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.

Hard ticks (Acari: Ixodidae) associated with birds in Europe: Review of literature data.

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.

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.