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In the present paper, we the report the first documented occurrence in the wild of Toxocara cati in the sole representative of the genus Otocolobus, the Pallas’ cat. The identity of the parasite was confirmed by morphological characteristics and by genetic barcoding of the second internal transcribed spacer of ribosomal DNA. The morphological measures of the T. cati specimens from the Kyrgyz Pallas’ cat were comparable to values reported. We discuss the conservation implication of our find.

We report new records of the trematode, Plagiorchis micracanthos from three North American bats. For the first time, the parasite is reported to occur in the hoary bat (Lasiurus cinereus), California myotis (Myotis californicus) and the pallid bat (Antrozous pallidus). The western small-footed bat (Myotis ciliolabrum), little brown bat (M. lucifugus) and the western pipistrelle (Parastellus hesperus) are confirmed as host species. The trematodes were isolated from the small intestines of the host individuals. For the first time, we identified P. micracanthos in bats originating from Canada and Mexico. The results presented here suggest that the trematode P. micracanthos has a much larger host range and geographic distribution than previously recognised. It is likely that further studies will confirm this results and extent both host and geographic ranges even further.

While the roe deer (Capreolus capeolus) is the most important game species in Germany and its venison is popular, there is limited knowledge about the prevalence of Toxoplasma gondii in this animal population in the country, and in wild ungulates in Germany generally. Between 2013 and 2015, we collected 295 blood samples from roe deer belonging to a central German population. Sera were analysed using a modified agglutination test (MAT, cut-off 1:20), and antibodies were detected in 86 of the 295 samples (29%). Seroprevalence values differed significantly between the different age classes, with antibodies more frequently observed in adults. In contrast, seroprevalence did not differ significantly between the sexes or collection years. Venison is frequently consumed raw or undercooked and may be a potential source of human infection with T. gondii.

Despite increasing consumption of mouflon (Ovis orientalis musimon) meat in Germany, there is currently no surveillance of Toxoplasma gondii infection in populations of these animals and generally little knowledge about the prevalence of this protozoan in German wild ungulates. Between 2011 and 2015, we collected 138 blood samples from a free-living mouflon population in central German and tested sera for the presence of T. gondii antibodies using a modified agglutination test (MAT, cut-off 1:20). Antibodies were detected in 31 of the 138 samples (22.46%). There was a significant difference in seroprevalence between the different age classes, with antibodies to T. gondii more frequent in adults. In contrast, there was no significant difference in seroprevalence depending on sex and year of sample collection. Game meat is frequently consumed as raw or undercooked meat and may therefore represent a potential source of human infection with T. gondii.

Raccoons (Procyon lotor) originated in North America and have been introduced to Europe. Due to their close contact with human settlements, they are important reservoirs for zoonotic pathogens, such as Baylisascaris procyonis. The relevance and prevalence of vector-borne pathogens have not yet been fully elucidated. In this study, we screened 285 spleen samples of raccoons, collected between 2019 and 2022 in Germany. The samples were analysed by PCR to detect Mycoplasma spp., Anaplasmataceae, Bartonella spp., Babesia spp., Rickettsia spp., Filarioidea, Trypanosomatida and Hepatozoon spp., and positive PCR products were sequenced. In total, 104 samples were positive for Mycoplasma spp. (36.49%), making this the first study to detect Mycoplasma spp. in raccoons outside of North America. Three samples were positive for Babesia spp. (1.05%) and two for Anaplasma phagocytophilum (0.7%). Phylogenetic analysis revealed that the Mycoplasma spp. detected all belong to the haemotrophic mycoplasmas cluster and were grouped within a single phylogenetic clade. Two different Babesia spp. were detected, one of which was closely related to Babesia canis, while the other was more closely related to Babesia sp. from ruminants. It is unclear whether the pathogens detected have an impact on the health of raccoons or whether they may serve as a reservoir for other animals., Competing Interests: Competing interests: The authors declare no competing interests. Ethics approval and consent to participate: The raccoon is on the list of invasive species in Germany and is not protected by law. The species can be hunted by licensed hunters outside the closed season and without a special permit. All animals were legally shot and made available to the authors. No animals were killed for the purpose of providing samples for this study., (© 2024. The Author(s).)

Background: Vector-borne pathogens (VBPs) are increasing in significance in veterinary medicine and public health settings, with wildlife playing a potentially crucial role in their transmission. Eurasian badgers (Meles meles) are widely distributed across Europe. However, information currently available on the prevalence of VBPs in badgers is limited. The objective of the current study was to investigate the occurrence of Anaplasmataceae, Bartonella spp., Mycoplasma spp., Rickettsia spp., Piroplasmida, Trypanosomatida and Filarioidea in badgers and subsequently, based on the results, assess the potential risk to domestic animals, other wildlife and humans., Methods: Between 2017 and 2021, blood or spleen samples from 220 badgers were collected in nine continental European countries: Austria (n = 7), Bosnia and Herzegovina (n = 2), Croatia (n = 22), France (n = 44), Germany (n = 16), Hungary (n = 7), Italy (n = 16), Romania (n = 80) and Serbia (n = 26). VBPs were identified by performing PCR analysis on the samples, followed by Sanger sequencing. Additionally, to distinguish between different Babesia lineages we performed restriction fragment length polymorphism (RFLP) analysis on piroplasm-positive samples, using HinfI as restriction enzyme. A phylogenetic analysis was performed on Mycoplasma spp., Results: The pathogens identified were Babesia sp. badger type A (54%), B (23%), and C (37%); Trypanosoma pestanai (56%); Mycoplasma sp. (34%); Candidatus Mycoplasma haematomelis (8%); Candidatus Mycoplasma haematominutum (0.5%); and Ehrlichia spp. (2%). Rickettsia spp., Bartonella spp. and filarioid nematodes were not detected among the tested samples., Conclusions: The large sample size and diverse study populations in this study provide valuable insights into the distribution and epidemiology of the analyzed pathogens. Some of the VBPs identified in our study show high similarity to those found in domestic animals, such as dogs. This finding suggests that badgers, as potential reservoirs for these pathogens, may pose a threat not only to other wildlife but also to domestic animals in close vicinity. Continuous surveillance is essential to monitor VBPs in wildlife as a means to enable the assessment of their impact on other wildlife species, domestic animals and human health., (© 2024. The Author(s).)

Human infection by Baylisascaris procyonis can result in larva migrans syndromes, which can cause severe neurological sequelae and fatal cases. The raccoon serves as the definitive host of the nematode, harboring adult worms in its intestine and excreting millions of eggs into the environment via its feces. Transmission to paratenic hosts (such as rodents, birds and rabbits) or to humans occurs by accidental ingestion of eggs. The occurrence of B. procyonis in wild raccoons has been reported in several Western European countries. In France, raccoons have currently established three separate and expanding populations as a result of at least three independent introductions. Until now the presence of B. procyonis in these French raccoon populations has not been investigated. Between 2011 and 2021, 300 raccoons were collected from both the south-western and north-eastern populations. The core parts of the south-western and north-eastern French raccoon populations were free of B. procyonis . However, three worms (molecularly confirmed) were detected in a young raccoon found at the edge of the north-eastern French raccoon population, close to the Belgian and Luxemburg borders. Population genetic structure analysis, genetic exclusion tests and factorial correspondence analysis all confirmed that the infected raccoon originated from the local genetic population, while the same three approaches showed that the worms were genetically distinct from the two nearest known populations in Germany and the Netherlands. The detection of an infected raccoon sampled east of the northeastern population raises strong questions about the routes of introduction of the roundworms. Further studies are required to test wild raccoons for the presence of B. procyonis in the area of the index case and further east towards the border with Germany., Competing Interests: The authors declare no competing interests in association with this study., (© 2024 The Authors.)

The intracellular protozoan Toxoplasma gondii is distributed worldwide and infects many species of warm-blooded animals. Most mammals, including humans, can serve as intermediate hosts. This pathogen, with its zoonotic potential, causes toxoplasmosis, a condition that can range from subclinical to fatal in humans. It is therefore important to assess the occurrence of the pathogen, even if only indirectly through the detection of antibodies. Epidemiological data on the seroprevalence in wild animals, including invasive species, are rare in Poland. Therefore, we tested 197 wild raccoons ( Procyon lotor ) and 89 raccoon dogs ( Nyctereutes procyonoides ) from Zgorzelec County, southwestern Poland, for the presence of antibodies. Samples were collected between January 2019 and December 2020 and analysed using a commercial indirect modified agglutination test (MAT, cut-off 1:25). The statistical analysis revealed significant differences in seroprevalence between the two predatory species. Of the 197 surveyed raccoons, 96 (48.73%; 95% confidence interval (CI): 41.73-55.73%) tested positive, while 25 of the 89 raccoon dogs (28.09%; 95% CI: 18.70-37.48%) were positive. Regarding risk factors, body weight and sex influenced the presence of T. gondii antibodies in both the species, with a higher likelihood of seropositivity among heavier animals and females, respectively. For raccoon dogs, juveniles were more likely to be seropositive than adults at a given weight. Our results suggest that T. gondii infection is widespread in the regional raccoon and raccoon dog populations, indicating a high level of parasite circulation in the environment.

Toxoplasma gondii is an obligate intracellular protozoan that causes toxoplasmosis in warm-blooded animals. Although most infections in humans and animals are subclinical, an infection can nevertheless be fatal. One of the important characteristics in the epidemiology of this parasite is waterborne transmission. The American mink ( Neogale vison ), a mammal closely adapted to freshwater ecosystems, is a potential sentinel for T. gondii . We analysed meat juice from the heart of 194 wild minks collected between 2019 and 2022 in five study areas from Germany and Poland and tested for the presence of antibodies against T. gondii . The analysis was performed using a commercial enzyme-linked immunosorbent assay test (ELISA). Antibodies were detected in 45.36% (88/194, 95% confidence interval (CI): 38.39-52.41%) of the analysed animals. While the prevalence values ranged from 37.50% to 49.30%, there was no significant difference in seroprevalence between the study areas. Juveniles were less likely to carry T. gondii antibodies than adults (odds ratio: 0.216), whereas there was no significant difference in prevalence between the sexes (odds ratio: 0.933). The results of our study show that contact with T. gondii is widespread in minks, and the parasite is common in inland freshwater ecosystems in Germany and Poland. This indicates that watercourses play an important role in the spread of T. gondii oocysts.

The raccoon roundworm (Baylisascaris procyonis), a gastrointestinal nematode of the raccoon (Procyon lotor), may cause a severe form of larva migrans in humans, which can lead to death or permanent neurological damage. Although roundworms were inadvertently introduced to Europe alongside their raccoon hosts, the parasite is not present in every raccoon population. It is important to understand the geographic distribution of B. procyonis, as early and rapid treatment can prevent severe pathologies in humans. We present evidence for the roundworm spreading into a naive raccoon population through natural dispersal of infected raccoons. We sampled 181 raccoons from Saxony-Anhalt, a German federal state containing contact zones of different raccoon populations, two of which were previously free of the parasite. We screened the raccoons for roundworms and used microsatellite-based assignment tests to determine the genetic origin of the raccoons and their parasites. We detected roundworms in 16 of 45 raccoons sampled in a previously roundworm-free area in the northern part of the state. The largest proportion of the genetic ancestry (≥ 0.5) of the 16 raccoon hosts was assigned to the previously naive raccoon population. Conversely, the genetic ancestry of almost all the roundworms was assigned to the nearest roundworm population in the southern part of the state. Infected raccoons have, therefore, spread to the north of the state, where they interbred with and infected local raccoons. It seems likely that the roundworms will continue to spread. Health authorities should consider continuous surveillance programmes of naive populations and raise public awareness., (© 2023. The Author(s).)

Giardia , Cryptosporidium , Cyclospora , and microsporidia are gastrointestinal pathogens that can cause various disease symptoms in both animals and humans. Numerous studies worldwide have confirmed the presence of these eukaryotic pathogens in nesting and migrating wild geese, ducks, and swans. Migration spreads these zoonotic enteric pathogens to distant locations, which could have public health implications. Soils and water bodies (lakes, ponds, rivers and wetlands) in urban and suburban areas have been shown to be vulnerable to contamination by waterfowl droppings. This review addresses the epidemiology of these enteric pathogens in wild migratory bird species (Anatidae) and some consequences of their spread in the environment. To date, both zoonotic pathogens and genotypes restricted to avian hosts have been found in faecal samples from 21 anatid species worldwide. One of the routes of infection for these zoonotic gastrointestinal micropathogens is the indirect route. For example, shared water bodies (e.g., for drinking or recreational purposes) previously contaminated by birds during the migratory season may facilitate infections of humans through water. However, it is unclear how much wild waterfowl contribute to the transmission of giardiasis, cryptosporidiosis, cyclosporosis, and microsporidiosis in many regions through contaminated environmental sources. Comprehensive epidemiological surveillance based on molecular data on gastrointestinal pathogens is crucial to take measures to control infections in the future.

Background: Important information on movement pathways and introduction routes of invasive parasites can be obtained by comparing the genetic makeup of an invader with its spatial genetic structure in other distribution areas. Sometimes, the population genetic structure of the host might be more informative than that of the parasite itself, and it is important to collect tissue samples of both host and parasite. However, host tissue samples are frequently not available for analysis. We aimed to test whether it is possible to generate reliable microsatellite profiles of host individuals by amplifying DNA extracted from a nematode parasite, using the raccoon (Procyon lotor) and the raccoon roundworm (Baylisascaris procyonis) as a test case., Methods: Between 2020 and 2021, we collected tissue as well as a single roundworm each from 12 raccoons from central Germany. Both the raccoon and the roundworm DNA extracts were genotyped using 17 raccoon-specific microsatellite loci. For each roundworm DNA extract, we performed at least eight amplification reactions per microsatellite locus., Results: We extracted amplifiable raccoon DNA from all 12 roundworms. We obtained at least two amplification products for 186 of the 204 possible genotypes. Altogether 1077 of the 1106 genotypes (97.4%) matched the host-DNA derived reference genotypes and thus did not contain genotyping errors. Nine of the 12 roundworm-derived genetic profiles matched the reference profiles from the raccoon hosts, with one additional genetic profile containing genotyping errors at a single locus. The remaining two genetic profiles were deemed unsuitable for downstream analysis because of genotyping errors and/or a high proportion of missing data., Conclusions: We showed that reliable microsatellite-based genetic profiles of host individuals can be obtained by amplifying DNA extracted from a parasitic nematode. Specifically, the approach can be applied to reconstruct invasion pathways of roundworms when samples of the raccoon hosts are lacking. Further research should assess whether this method can be replicated in smaller species of parasitic nematodes and other phyla of parasites more generally., (© 2023. The Author(s).)

The urinary bladder and lower urinary tract of domestic and wild carnivores can be parasitised by filamentous nematodes from the genus Pearsonema (syn. Capillaria ). Infestations are often asymptomatic, but severe courses in dogs and cats have been described. Hosts are infested through the ingestion of earthworms (Lumbricidae) which act as intermediate hosts. Epidemiological studies of Pearsonema in raccoons ( Procyon lotor ) in North America are scarce and previous studies of urinary bladder parasites of European raccoons did not provide evidence of infestation. We examined urine sediment or rinse water from urinary bladders of 499 wild raccoons from Luxembourg, Poland and five study sites in Germany. Pearsonema eggs were found in the urine sediment of 31 (6.2%) raccoons. Infested animals were found in all study areas with prevalence values ranging from 3.7% to 8.7%. No significant difference in prevalence was found either between animals in urban and rural areas or between sexes and age classes. Based on their morphology, the eggs were likely to be P. plica . Considering their increasing density in Central Europe, raccoons may play a previously overlooked role in environmental contamination with Personema eggs., Competing Interests: There is not any conflict of Interest., (© 2022 The Authors.)

European wildcats ( Felis silvestris silvestris ) have not been investigated in large numbers for blood-associated pathogens in Germany, because wildcats, being a protected species, may not be hunted, and the collection of samples is therefore difficult. Thus, spleen tissue and whole blood from 96 wildcats from Germany found as roadkill or dead from other causes in the years 1998-2020 were examined for the prevalence of blood associated pathogens using molecular genetic tools. PCR was used to screen for haemotrophic Mycoplasma spp., Hepatozoon spp., Cytauxzoon spp., Bartonella spp., Filarioidea, Anaplasmataceae, and Rickettsiales, and positive samples were subsequently sequenced. Phylogenetic analyses were performed for Mycoplasma spp. and Hepatozoon spp. by calculating phylogenetic trees and DNA haplotype networks. The following pathogens were found: Candidatus Mycoplasma haematominutum (7/96), Mycoplasma ovis (1/96), Hepatozoon silvestris (34/96), Hepatozoon felis (6/96), Cytauxzoon europaeus (45/96), and Bartonella spp. (3/96). This study elucidates the prevalence of blood-associated pathogens in wildcats from Germany., Competing Interests: None., (© 2022 The Authors.)

Multiple helminth species commonly co-occur within mammals and their interactions may negatively affect the survival and breeding success of their hosts. However, it has been difficult to prove competition or mutualism between co-infesting helminths in field studies of wild mammals. The sinus cavities of European polecats ( Mustela putorius ) can be parasitised by the trematode Troglotrema acutum and the nematode Skrjabingylus nasicola and both helminths can co-occur within hosts. While both parasites can damage the host's bone structure and cause severe pathologies, their impact on host body condition is unclear. It is also unknown whether both parasites interact and how this might affect cranial damage and host body condition. We examined 515 fresh polecat skulls for the presence of both helminths and measured the hosts' amount of kidney perirenal fat as a measure of body condition. Our results demonstrated that, in addition to a host-intrinsic fixed factor (sex) and random factors accounting for spatial and temporal stochasticity, the helminths influenced each other's presence. Infestation with T. acutum increased the probability of catching S. nasicola with increasing age of the host, while males already infested with S. nasicola were more likely to become infested with T. acutum than females infested with the nematode. While we speculate that both effects resulted from parasite-induced behavioural alterations (increased foot consumption), it is not clear why, in the latter case, this effect would be stronger in males than females. We showed that the abundances of both parasites had significant positive effect on the likely presence of skull damage and a significant negative effect on the predicted presence of kidney fat. Given the evolutionary arms race that both host-parasite systems have undergone, it appears unlikely that either helminth played a significant factor in the population decline of the polecat in Europe., 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., (© 2022 The Author(s).)

The geographic range of the zoonotic raccoon roundworm (Baylisascaris procyonis) is expanding together with the range of its host, the raccoon (Procyon lotor). This creates a new public health risk in parts of Europe where this parasite was previously absent. In the Netherlands, a raccoon population is becoming established and incidental findings of B. procyonis have been reported. To assess the risk to public health, the prevalence of B. procyonis was determined in the province of Limburg, where currently the largest Dutch raccoon population is present, as well as in the adjoining region of southern Belgium. Furthermore, genetic methods were employed to assess invasion pathways of both the raccoon and B. procyonis to aid in the development of control measures. Macroscopic analysis of intestinal content and testing of faecal samples were performed to detect B. procyonis adults and eggs. The population genetics of both B. procyonis and its raccoon host were analysed using samples from central and northwestern Europe. B. procyonis was found in 14/23 (61%, 95% CI: 41%-78%) raccoons from Limburg, but was not detected in 50 Belgian raccoons. Genetic analyses showed that the majority of the Dutch raccoons and their roundworms were introduced through ex-captive individuals. As long as free-living raccoon populations originate from captivity, population control methods may be pursued. However, natural dispersal from the border regions will complicate prolonged population control. To reduce the public health risk posed by B. procyonis, public education to increase awareness and adapt behaviour towards raccoons is key., (© 2021 Wiley-VCH GmbH.)

To provide the most comprehensive picture of species phylogeny and phylogeography of European roe deer ( Capreolus capreolus ), we analyzed mtDNA control region (610 bp) of 1469 samples of roe deer from Central and Eastern Europe and included into the analyses additional 1541 mtDNA sequences from GenBank from other regions of the continent. We detected two mtDNA lineages of the species: European and Siberian (an introgression of C .  pygargus mtDNA into C .  capreolus ). The Siberian lineage was most frequent in the eastern part of the continent and declined toward Central Europe. The European lineage contained three clades (Central, Eastern, and Western) composed of several haplogroups, many of which were separated in space. The Western clade appeared to have a discontinuous range from Portugal to Russia. Most of the haplogroups in the Central and the Eastern clades were under expansion during the Weichselian glacial period before the Last Glacial Maximum (LGM), while the expansion time of the Western clade overlapped with the Eemian interglacial. The high genetic diversity of extant roe deer is the result of their survival during the LGM probably in a large, contiguous range spanning from the Iberian Peninsula to the Caucasus Mts and in two northern refugia., Competing Interests: The authors declare that they have no conflict of interest., (© 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

There is now considerable evidence that in Europe, babesiosis is an emerging infectious disease, with some of the causative species spreading as a consequence of the increasing range of their tick vector hosts. In this review, we summarize both the historic records and recent findings on the occurrence and incidence of babesiosis in 20 European countries located in southeastern Europe (Bosnia and Herzegovina, Croatia, and Serbia), central Europe (Austria, the Czech Republic, Germany, Hungary, Luxembourg, Poland, Slovakia, Slovenia, and Switzerland), and northern and northeastern Europe (Lithuania, Latvia, Estonia, Iceland, Denmark, Finland, Sweden, and Norway), identified in humans and selected species of domesticated animals (cats, dogs, horses, and cattle). Recorded cases of human babesiosis are still rare, but their number is expected to rise in the coming years. This is because of the widespread and longer seasonal activity of Ixodes ricinus as a result of climate change and because of the more extensive use of better molecular diagnostic methods. Bovine babesiosis has a re-emerging potential because of the likely loss of herd immunity, while canine babesiosis is rapidly expanding in central and northeastern Europe, its occurrence correlating with the rapid, successful expansion of the ornate dog tick ( Dermacentor reticulatus ) populations in Europe. Taken together, our analysis of the available reports shows clear evidence of an increasing annual incidence of babesiosis across Europe in both humans and animals that is changing in line with similar increases in the incidence of other tick-borne diseases. This situation is of major concern, and we recommend more extensive and frequent, standardized monitoring using a "One Health" approach.

Background: Skrjabingylus spp. are nematodes that parasitize in the frontal and nasal sinuses of small mustelids. In Europe, the two species S. nasicola and S. petrowi are known, although records of S. petrowi are extremely rare, except in the former part of the USSR. The aim of the present study was to screen pine martens (Martes martes) from Poland for the first time for the presence of S. petrowi., Methods: Three road-killed pine martens were collected in 2018 and 2020 in the province Lower Silesian in southwestern Poland. A complete necropsy was performed on the fresh pine marten heads and especially the frontal sinuses and paranasal sinuses were examined for the presence of Skrjabingylus spp. The species identity of the recovered nematodes was determined by morphological measurements and genetic barcoding., Results: One of three pine martens examined showed infection with 12 Skrjabingylus spp. in the right frontal sinus. Measurements of the spicule length of the males revealed a range of 480-521 µm which is characteristic of S. petrowi. There was a close match between the COI sequence reported here and of S. petrowi sequences reported from Germany., Conclusion: This is the first report of infection of a pine marten with the sinus worm S. petrowi in Poland., (© 2021. Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Baylisascaris procyonis is a common gastrointestinal parasite of raccoons (Procyon lotor) in their native range, and both have been introduced to Europe. Humans may ingest ascarid eggs shed via the racoons' faeces, and this could lead to severe infections affecting the central nervous system. Here, we report the first occurrence of B. procyonis in Austria. The parasite was detected in a two-year-old male raccoon that was road-killed in November 2019 near Hittisau (Vorarlberg). Genetic profiling provided strong evidence that the raccoon (and its parasite) originated from the nearest German raccoon population. The first finding in Austria highlights the need for monitoring the parasite and information of the public and practitioners., (© 2020 The Authors. Transboundary and Emerging Diseases published by Wiley-VCH GmbH.)

In the past few decades, the relevance of Dirofilaria immitis and Dirofilaria repens , causing cardiopulmonary and subcutaneous dirofilariosis in dogs and cats, and of Angiostrongylus vasorum , causing canine angiostrongylosis, has steadily increased in Central and Northern Europe. In this review, a summary of published articles and additional reports dealing with imported or autochthonous cases of these parasites is provided for Central (Austria, Czechia, Germany, Hungary, Luxemburg, Poland, Slovakia, Slovenia, and Switzerland) and Northern (Denmark, Finland, Iceland, Norway, and Sweden) Europe. Research efforts focusing on Dirofilaria spp. and A. vasorum have varied by country, and cross-border studies are few. The housing conditions of dogs, pet movements, the spread of competent vectors, and climate change are important factors in the spread of these nematodes. Dogs kept outside overnight are a major factor for the establishment of Dirofilaria spp. However, the establishment of invasive, diurnal, synanthropic, competent mosquito vectors such as Aedes albopictus may also influence the establishment of Dirofilaria spp. The drivers of the spread of A. vasorum remain not fully understood, but it seems to be influenced by habitats shared with wild canids, dog relocation, and possibly climatic changes; its pattern of spreading appears to be similar in different countries. Both Dirofilaria spp. and A. vasorum merit further monitoring and research focus in Europe.

Giardia duodenalis is a cosmopolitan flagellate that causes giardiasis, one of the most significant gastrointestinal diseases in humans. This parasite can be a serious threat to public health because it can cause waterborne outbreaks as well as sporadic infections in humans. Invasive raccoons (Procyon lotor) may play a role in disseminating Giardia into the environment and transmitting it to humans and domestic animals because they live in high densities and deposit their faces in latrines near areas used by humans. While Giardia infections have been reported from raccoons in North America, it is unknown whether they carry G. duodenalis with zoonotic assemblage A and B, which have the potential to cause illness in humans. We collected faecal samples from 66 legally harvested raccoons in Germany and Luxembourg and examined for Giardia using molecular techniques. Using a quantitative PCR based on primers specific to Giardia genetic assemblages A and B, we detected the presence of zoonotic assemblage B in 27% (95% CI, 17.0-39.6) of all examined faecal samples from raccoons, including animals sampled in buildings. We did not detect genetic assemblage A in any of the samples. Sequences obtained from the glutamate dehydrogenase and beta-giardin gene fragments from a selection of three of the positive samples showed that raccoons carried a zoonotic G. duodenalis genotype belonging to sub-assemblage BIV, which is commonly found in humans and animals worldwide. Our results suggest that free-ranging raccoons have the potential to play an increasingly important role in the epidemiology of Giardia and pose a threat to public health in Europe and other regions where this species is common and lives in close association with humans., (© 2021 Wiley-VCH GmbH.)

We report the first record of the trematode Troglotrema acutum in skulls of European polecats (Mustela putorius) from Luxembourg. Thirty-one road-killed polecats from the northern and eastern parts of the country were examined. We found the trematode in three polecats from the administrative districts of Clervaux and Vianden in the northeast of the country. The parasites were detected in the frontal sinuses of one juvenile male as well as one adult of each sex. The animals were infected with six, nine and 13 adult trematodes, respectively, and we recovered 24 trematode eggs from one polecat. While the two adult hosts showed the lesions and deformations of the skull that are typical of T. acutum infections in polecats, the skull of the juvenile was not deformed. The prevalence of 9.7% reported here was lower than the values reported from Central European studies. Our findings indicate that the distribution range of T. acutum in western Central Europe extends to the Ardennes low mountain range.

Protists of the genus Cytauxzoon infect a wide variety of wild and domestic felids worldwide. While the American Cytauxzoon felis has been well described, data on the European isolates of Cytauxzoon are still scant. The aim of the current study was to determine the genetic diversity of European Cytauxzoon spp. in wild felids across Europe by analyzing one nuclear and two mitochondrial genes, along with representative complete mitochondrial genomes. Overall, 106 biological samples from wild felids (92 from Felis silvestris and 14 from Lynx lynx) from Germany, Romania, Czech Republic, and Luxembourg were collected and screened for the presence of Cytauxzoon spp. using nested PCR protocols, targeting the highly conserved 18S rDNA, mitochondrial cytochrome b (CytB) and cytochrome c oxidase subunit I (COI) genes. Furthermore, 18 previously confirmed wild felid biological samples from Europe, and comparative material from USA positive for C. felis, were included in the study. In 18S rDNA sequences analyses, Cytauxzoon spp. from felids formed two separate clades of New World and Old World isolates, with a low inner diversity of the European clade. In contrast to 18S rDNA, the phylogenetic analyses of CytB and COI genes affirmatively revealed three highly supported clades, resulting in three defined genotypes. Similar intra- and interspecific variability of CytB and COI genes was observed in the case of different Babesia spp. Considering geography, host species and analyses of three genes, we conclude that the three detected genotypes of Cytauxzoon in European wild felids represent three new species, which we herein describe., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Cryptosporidium spp. are protozoan parasites that cause enteric infection in a wide range of mammals, including humans. The raccoon (Procyon lotor) is an invasive species in many parts of the world and studies have shown that they can be infected with Cryptosporidium spp. both outside and in their original distribution area. The aim of the present study was to determine the presence of Cryptosporidium spp. antigens in the faeces of raccoons in Luxembourg., Methods: Using an enzyme-linked immunosorbent assay (ELISA), we tested 81 faeces samples, collected between 2014 and 2018, for the presence of Cryptosporidium spp. coproantigens. Samples with an optical density equal to or greater than 0.15% were considered positive., Results: Antigens were detected in 12.35% (10/81; 95% CI 6.68-21.26) of the tested samples. There was no significant difference in the prevalence of Cryptosporidium spp. infection between the sexes and age categories. Cryptosporidium spp.-positive raccoons were found in 7 of the 12 Luxembourg administrative districts (Clervaux, Diekirch, Echternach, Mersch, Remich, Vianden and Wiltz)., Conclusion: The results show that Cryptosporidium infections are not uncommon in Luxembourg raccoons and suggest possible transmission of Cryptosporidium by raccoons.

In the present study, we investigated blood samples of 196 invasive Reeve's muntjac (Muntiacus reevesi) and 91 native roe deer (Capreolus capreolus) originating from the same area in Thetford Forest in Eastern England for the occurrence of blood pathogens such as Anaplasmatacae, Rickettsiales and Piroplasmida (Babesia spp., Theileria spp.) by using PCR. Babesia spp., Rickettsia spp. and Theileria spp. were not detected. Only two male (1%) Reeve's muntjacs and six (6.6%) roe deer were positive for Anaplasma phagocytophilum with 100% identity among their sequences. However, it is not clear whether Reeve's muntjac is less susceptible to infection, less susceptible to infestation by I. ricinus, or an infection in Reeve's muntjac is more lethal and therefore less positive animals are taken during hunting events., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Wild carnivores are important hosts for ixodid ticks and presumed reservoirs for several tick-borne pathogens (TBPs) of medical and veterinary importance. However, little is known about the role that the European grey wolf (Canis lupus) plays in the enzootic cycle of TBPs. The recent recolonization of Central European lowland, including some regions in Germany, by the grey wolf, opened up an excellent opportunity for studying the impact of the wolf population on pathogen diversity and transmission dynamics. Therefore, in the present study, we investigated spleen samples collected from 276 grey wolves in 11 federal states in Germany for common TBPs by molecular methods. In total, 127 grey wolves (46.0 %) were found to be positive for Hepatozoon canis. Only two genetic variants of this protozoan parasite (herein designated as G1 and G2) were found to circulate among the grey wolves, which can be potentially shared between populations of domestic and other wild carnivores in the country. Two grey wolves were positive for Anaplasma phagocytophilum (0.7 %), and both were co-infected with H. canis G1 genotype. The presence of other pathogens could not be confirmed by PCR and sequencing. This study represents the first one reporting H. canis in a grey wolf population worldwide, and it provides highly relevant information, which may contribute to a better understanding of the epidemiology of TBPs and the pathogen transmission dynamics among the reintroduced population of grey wolves and other carnivores., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Infestation with Baylisascaris procyonis, a gastrointestinal nematode of the raccoon, can cause fatal disease in humans. We found that the parasite is widespread in central Germany and can pose a public health risk. The spread of B. procyonis roundworms into nematode-free raccoon populations needs to be monitored.

We examined 438 fresh skulls of pine marten ( Martes martes ) from Germany to check for the presence of the cranial trematode Troglotrema acutum . The parasite was detected in the frontal sinuses of one 3-year-old road-killed male from the Rhoen Mountains (Thuringia). We detected 19 adult T. acutum and 34 eggs. The infested pine martens showed no deformations or lesions of the skull. This is the first time that specimens of the trematode were recovered from a pine marten from Germany, but the low prevalence of 0.23% confirms that the pine marten is only a rare host of T. acutum ., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Indian Society for Parasitology 2019.)

Urbanization affects key aspects of wildlife ecology. Dispersal in urban wildlife species may be impacted by geographical barriers but also by a species' inherent behavioural variability. There are no functional connectivity analyses using continuous individual-based sampling across an urban-rural continuum that would allow a thorough assessment of the relative importance of physical and behavioural dispersal barriers. We used 16 microsatellite loci to genotype 374 red foxes (Vulpes vulpes) from the city of Berlin and surrounding rural regions in Brandenburg in order to study genetic structure and dispersal behaviour of a mobile carnivore across the urban-rural landscape. We assessed functional connectivity by applying an individual-based landscape genetic optimization procedure. Three commonly used genetic distance measures yielded different model selection results, with only the results of an eigenvector-based multivariate analysis reasonably explaining genetic differentiation patterns. Genetic clustering methods and landscape resistance modelling supported the presence of an urban population with reduced dispersal across the city border. Artificial structures (railways, motorways) served as main dispersal corridors within the cityscape, yet urban foxes avoided densely built-up areas. We show that despite their ubiquitous presence in urban areas, their mobility and behavioural plasticity, foxes were affected in their dispersal by anthropogenic presence. Distinguishing between man-made structures and sites of human activity, rather than between natural and artificial structures, is thus essential for better understanding urban fox dispersal. This differentiation may also help to understand dispersal of other urban wildlife and to predict how behaviour can shape population genetic structure beyond physical barriers., (© 2019 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

In the framework of a mosquito-monitoring program conducted from 2014 to 2018, non-culicid dipteran bycatch was identified to species-level with a focus on Diptera of medical and veterinary importance as part of a biodiversity initiative and barcoding project ("Austrian Barcode of Life"). Two species hitherto not known from Austria, the regularly sampled synanthropic moth fly Clogmia albipunctata (Psychodidae) and a single specimen of the louse fly Ornithoica turdi (Hippoboscidae), were collected in Vienna and Lower Austria. We confirmed identification results using a barcoding approach and provide the first reference sequence for O. turdi.

This study examined the concentration of total mercury (THg) and selenium (Se), as well as the molar ratio of Se:THg in hair samples of terrestrial animals. THg and Se concentrations were measured from the hair of raccoons (Procyon lotor) and European wildcats (Felis s. silvestris) from Germany and Luxembourg. Median THg concentrations in hair from raccoons and wildcats were 0.369 and 0.273 mg kg -1 dry weight (dw), respectively. Se concentrations were higher in the hair of raccoons than of wildcats (0.851 and 0.641 mg kg -1 dw, respectively). Total mercury concentration in hair of raccoons from Luxembourg was almost 5× higher that found in hair of raccoons from Germany; however, Se concentration was similar. Thus, molar ratio of Se:THg was ~4× higher in the hair of raccoons from Germany than those from Luxembourg. Significant negative correlation was found between THg concentration and Se:THg molar ratio in both wildcats and raccoons.

European populations of free-living wildcats have been shown to be exposed to cat viruses. Luxembourg has a high degree of habitat fragmentation, and hybridisation rates between domestic cats and wildcats are high. We therefore assessed the seroprevalence of six viruses in 34 serum samples collected between 2001 and 2016 from wildcats in Luxembourg. The values for feline leukemia virus (FeLV; 52.9%) and feline coronavirus (FCoV; 47.1%) were amongst the highest reported for wildcats. We found evidence for the cumulative likelihood of exposure to FCoV affecting its seroprevalence. Routine monitoring of viral agents in this strictly protected species should be considered.

Animal parasitic nematodes can cause serious diseases and their emergence in new areas can be an issue of major concern for biodiversity conservation and human health. Their ability to adapt to new environments and hosts is likely to be affected by their degree of genetic diversity, with gene flow between distinct populations counteracting genetic drift and increasing effective population size. The raccoon roundworm (Baylisascaris procyonis), a gastrointestinal parasite of the raccoon (Procyon lotor), has increased its global geographic range after being translocated with its host. The raccoon has been introduced multiple times to Germany, but not all its populations are infected with the parasite. While fewer introduced individuals may have led to reduced diversity in the parasite, admixture between different founder populations may have counteracted genetic drift and bottlenecks. Here, we analyse the population genetic structure of the roundworm and its raccoon host at the intersection of distinct raccoon populations infected with B. procyonis. We found evidence for two parasite clusters resulting from independent introductions. Both clusters exhibited an extremely low genetic diversity, suggesting small founding populations subjected to inbreeding and genetic drift with no, or very limited, genetic influx from population admixture. Comparison of the population genetic structures of both host and parasite suggested that the parasite spread to an uninfected raccoon founder population. On the other hand, an almost perfect match between cluster boundaries also suggested that the population genetic structure of B. procyonis has remained stable since its introduction, mirroring that of its raccoon host., (Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Infections with the tick-borne pathogens Anaplasma phagocytophilum and Babesia spp. can cause febrile disease in several mammalian species, including humans. Wild ruminants in Europe are suggested to serve as reservoir hosts for particular strains or species of these pathogens. The aims of this study were to investigate the occurrence of A. phagocytophilum and Babesia spp. in roe deer (Capreolus capreolus), fallow deer (Dama dama) and mouflon (Ovis musimon orientalis) in Germany, and the diversity and host association of genetic variants of A. phagocytophilum and Babesia species. From 2009 to 2010, 364 spleen samples from 153 roe deer, 43 fallow deer and 168 mouflon from 13 locations in Germany were tested for DNA of A. phagocytophilum and Babesia spp. by real-time PCR or conventional PCR, respectively. Variants of A. phagocytophilum were investigated with a nested PCR targeting the partial 16S rRNA gene, and species of piroplasms were identified by sequencing. DNA of A. phagocytophilum was detected in 303 (83.2%) samples: roe deer, 96.1% (147/153); fallow deer, 72.1% (31/43); and mouflon, 74.4% (125/168). Sequence analysis of 16S rRNA-PCR products revealed the presence of nine different genetic variants. DNA of Babesia spp. was found in 113 (31.0%) samples: roe deer, 62.8% (96/153); fallow deer, 16.3% (6/43); and mouflon, 6.5% (11/168). Babesia capreoli, Babesia sp. EU1 (referred to also as B. venatorum), B. odocoilei-like and a Theileria species were identified. Co-infections with A. phagocytophilum and Babesia spp. were detected in 30.0% of the animals which were tested positive for A. phagocytophilum and/or Babesia spp. Roe deer had a significantly higher percentage of co-infections (60.8%), followed by fallow deer (14.0%) and mouflon (6.5%). Thus, the results suggest that roe deer plays a key role in the endemic cycles of the pathogens investigated., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

While genetic profiling can be a powerful tool to solve wildlife crime, comparably few examples of individual identification in wildlife forensics are available in the literature. Here, we report a case of an accidental shooting of a hunting dog during a wild boar drive hunt. The market value of trained hunting dogs can reach several thousand euro. No one admitted to killing the dog. Wild boar hairs were found in the dog's wound, suggesting that the bullet first hit a wild boar and then the dog. Since it was known who harvested each boar, we aimed to use individual-specific genetic profiles to link these hairs to a bagged animal and to identify the culprit. We genotyped 19 harvested boar and the unknown hair sample using 13 STRs. In the case of the hair sample, we performed multiple genotyping to ensure the reliability of the genetic profile. We showed that we genotyped sufficient loci to distinguish between separate individuals with certainty. While the three most informative loci were enough to differentiate the 19 reference individuals, we did find a perfect match at all 13 STRs between the hair DNA and one tissue sample. Since our methods were reliable and reproducible, we passed the relevant information on to forestry officials who will use the information we have provided to attempt to find an amicable solution.

Biological invasions provide excellent study systems to understand evolutionary, genetic and ecological processes during range expansions. There is strong evidence for positive effects of high propagule pressure and the associated higher genetic diversity on invasion success, but some species have become invasive despite small founder numbers. The raccoon (Procyon lotor) is often considered as a typical example for such a successful invasion resulting from a small number of founders. The species' largest non-native population in Germany is commonly assumed to stem from a small number of founders and two separate founding events in the 1930s and 1940s. In the present study we analyzed 407 raccoons at 20 microsatellite loci sampled from the invasive range in Western Europe to test if these assumptions are correct. Contrary to the expectations, different genetic clustering methods detected evidence for at least four independent introduction events that gave rise to genetically differentiated subpopulations. Further smaller clusters were either artifacts or resulted from founder events at the range margin and recent release of captive individuals. We also found genetic evidence for on-going introductions of individuals. Furthermore a novel randomization process was used to determine the potential range of founder population size that would suffice to capture all the alleles present in a cluster. Our results falsify the assumption that this species has become widespread and abundant despite being genetically depauperate and show that historical records of species introductions may be misleading.