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Trichinella pseudospiralis belongs to the non-encapsulated clade of the genus and its epidemiology is influenced by various biotic and abiotic factors. The role of different animal species in the spread and epidemiology of the parasite is still not well understood and further research is needed in the areas where its occurrence has been recorded. In Slovakia, T. pseudospiralis was first documented in 2004 on a poorly maintained industrial pig farm where pigs, rats, and a domestic cat were found to be infected. In the following years, its occurrence was confirmed in foxes, wild boars, and three species of birds of prey. The objective of the present study was to investigate different micromammal and carnivore host species inhabiting the Tatra National Park in the north of Slovakia for the presence of Trichinella parasites. From 2018-2023, 715 small mammals belonging to 19 species and 92 muscle samples from 12 carnivorous species were individually examined for the presence of Trichinella muscle larvae using the artificial digestion method. The research brought new host records and completed the list of sylvatic hosts for T. pseudospiralis in Slovakia - the yellow-necked mouse (Apodemus flavicollis) and raccoon dog (Nyctereutes procyonoides) together with results of the genetic structure study conducted on a fragment of the 28S gene in T. pseudospiralis., 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: Sarcocystis includes a global group of apicomplexan parasites with two-host life cycle frequently circulating in wildlife and domestic hosts, including humans. Two of the most important wild terrestrial carnivores acting as definitive hosts are the red fox and raccoon dog, due to their wide distribution in Europe and usage of wild and farmed animals as prey. This study was conducted to determine the prevalence of Sarcocystis in hunted red foxes and raccoon dogs from nine regions of the Czech Republic and to identify isolated sporocysts by molecular techniques., Methods: Approximately 5 g of the contents of large intestine from 200 animals (197 red foxes and three raccoon dogs) were examined by flotation centrifugation coprological method. Only samples of 50 red foxes and one raccoon dog positive to Sarcocystis spp. were used for the nested PCR (nPCR) method to amplify a fragment or partial sequence on the cox1 gene. Ten species-specific primer pairs for detection of Sarcocystis spp. using farm animals as intermediate hosts were utilized., Results: In total, 38.1% of the red foxes and 66.7% of the raccoon dogs were positive to Sarcocystis by light microscopy. The molecular characterization resulted in the identification of five species in the red fox: S . arieticanis , S . capracanis , S. cruzi , S . miescheriana , and S. tenella , while the PCR was negative for the sole raccoon dog. The highest intraspecific variation was found for S . miescheriana , while S. tenella was the most prevalent. Co-infections occurred in the large intestine of the red fox. No zoonotic species were found in our samples., Conclusion: This is the first study where the potential role of the red fox and raccoon dogs as spreaders of Sarcocystis to farm animals in the Czech Republic is shown. The use of species-specific primers provides a fast and easy method for screening multiple samples for a particular Sarcocystis 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 Máca, Gudiškis, Butkauskas, González-Solís and Prakas.)

Highly pathogenic avian influenza (HPAI) has caused widespread mortality in both wild and domestic birds in Europe 2020-2023. In July 2023, HPAI A(H5N1) was detected on 27 fur farms in Finland. In total, infections in silver and blue foxes, American minks and raccoon dogs were confirmed by RT-PCR. The pathological findings in the animals include widespread inflammatory lesions in the lungs, brain and liver, indicating efficient systemic dissemination of the virus. Phylogenetic analysis of Finnish A(H5N1) strains from fur animals and wild birds has identified three clusters (Finland I-III), and molecular analyses revealed emergence of mutations known to facilitate viral adaptation to mammals in the PB2 and NA proteins. Findings of avian influenza in fur animals were spatially and temporally connected with mass mortalities in wild birds. The mechanisms of virus transmission within and between farms have not been conclusively identified, but several different routes relating to limited biosecurity on the farms are implicated. The outbreak was managed in close collaboration between animal and human health authorities to mitigate and monitor the impact for both animal and human health.

The diet of raccoon dogs, which live in close proximity to agricultural fields in Matsuyama, western Japan, was studied throughout the year by the fecal analysis method ( n = 114). Fruits were most important, accounting for 30-40% of the diet except in winter. Crops were next in importance, accounting for 10-30%. It was characteristic that the proportion of crops was greater and more stable than in other "satoyama" or rural areas in Japan. Crops included rice ( Oryza sativa ), wheat, sesame ( Sesamum indicum ), strawberries ( Fragaria × ananassa ), kiwi fruit ( Actinidia chinensis var. deliciosa ), ginkgo nuts ( Ginkgo biloba ), and persimmon ( Diospyros kaki ). Crops were important as the food for the raccoon dogs, but the use was not regarded as causing serious damage. Although wild boars ( Sus scrofa ), sika deer ( Cervus nippon ), and Japanese macaques ( Macaca fuscata ) were culled for pest control in the area, raccoon dogs were not culled. This may be a reason why the raccoon dog has coexisted with people in Japan.

Raccoon dogs ( Nyctereutes procyonoides ) and raccoons ( Procyon lotor ) are invasive alien species originating from East Asia and North America, respectively. They are discussed as vectors and reservoirs for various infectious diseases, including vector-borne and zoonotic pathogens, and are therefore a potential threat to human and domestic animal health, as well as to biodiversity and conservation. In the years 2021 and 2022, 110 raccoon dogs ( Nyctereutes procyonoides ) and 30 raccoons ( Procyon lotor ) were screened via qPCR for the presence of Leptospira spp., Rickettsia spp. and Borreliella spp. in the German federal state of Schleswig-Holstein as part of a health and risk assessment study. Borreliella spp. were confirmed in one raccoon dog and one raccoon, identified as Borreliella afzelii in the raccoon. Leptospira spp. were found in 21 (19.44%) raccoon dogs and 2 (6.90%) raccoons. In five raccoon dogs, Leptospira spp. were identified as Leptospira borgpetersenii , Leptospira kirschneri and Leptospira interrogans., Competing Interests: The authors declare no conflicts of interest.

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.

The raccoon dog ( Nyctereutes procyonoides ) is a typical omnivore possessing wide dietary adaptability and tolerance to rough feeding, which may be attributed to its intestinal microbiota. The study aimed to investigate the effect of dietary alfalfa meal levels on the growth performance, nutrient apparent digestibility, serum parameters, and intestinal microbiota of raccoon dogs. Sixty raccoon dogs were randomly divided into four dietary treatments containing 0% (AM0), 5% (AM5), 10% (AM10), and 15% (AM15) alfalfa meal for a 60-day experiment. The results showed that compared to raccoon dogs fed the AM0 diet, those fed the AM5 and AM10 diets had no significant difference in growth performance, while those fed the AM15 diet experienced a significant decrease. Raccoon dogs fed the AM5 diet had no significant effect on the nutrient apparent digestibility. Dietary supplementation with alfalfa meal significantly decreased serum urea levels and increased the antioxidant capacity of raccoon dogs. The intestinal microbiome analysis showed that the richness and diversity of colonic microbiota significantly increased in the AM15 group. With the increase in dietary alfalfa meal levels, the relative abundance of fiber-degrading bacteria in the colon of raccoon dogs, such as Treponema , Phascolarctobacterium , and Christensenellaceae R-7 group , increased. However, the relative abundance of pathogenic bacteria, including Anaerobiospirillum , decreased. In conclusion, the inclusion of 5% alfalfa meal in the raccoon dogs' diet had no effect on growth performance, but it exhibited the potential to improve serum antioxidant capacity and intestinal microbiota. This indicates that raccoon dogs have a certain tolerance to the addition of alfalfa meal in their diet.

In March 2023, the Chinese CDC publicly released raw metagenomic sequencing data for environmental samples collected in early 2020 from the Huanan Seafood Market. Prior to that data release, some scientists had suggested that these samples could be informative for establishing if animals such as raccoon dogs had been infected with severe acute respiratory syndrome virus 2 (SARS-CoV-2). However, no one had analyzed how much SARS-CoV-2 was actually present in the metagenomic sequencing data. After the raw data became available, I fully analyzed the abundance of both viral and animal genetic material in the samples. That analysis, which was published in Virus Evolution , found that the SARS-CoV-2 content of most samples was very low and that the abundance of SARS-CoV-2 was most strongly associated with animals such as largemouth bass that are not plausible candidates for having been infected. Based on these results, I concluded that the metagenomic content of the samples was not informative for determining if any non-human animals in the market had been infected with SARS-CoV-2. One of the authors of an earlier study of these samples, Florence Débarre, recently submitted a response to my paper. Here, I reply in turn to explain why it is important to quantify the abundance of viral material before drawing conclusions from metagenomic sequencing. I also report new analyses of other animal coronaviruses in the samples and show that material from some other animal coronaviruses is much more abundant than SARS-CoV-2 in samples collected on the date when most wildlife stall sampling was performed. I further show that material from some of these animal coronaviruses is associated with the animals they probably infect but that no such association exists for SARS-CoV-2. Overall, these new analyses further emphasize the importance of quantifying the actual amount of viral material in metagenomic samples and underscore why the environmental samples from the Huanan Seafood Market are not informative for determining if any non-human animals were infected with SARS-CoV-2., Competing Interests: J.D.B. is on the scientific advisory boards of Apriori Bio, Aerium Therapeutics, Invivyd, and the Vaccine Company and receives royalty payments as an inventor on Fred Hutch–licensed patents related to deep mutational scanning of viral proteins., (© The Author(s) 2024. Published by Oxford University Press.)

Sarcoptic mange was detected in five free-ranging raccoon dogs ( Nyctereutes procyonoides ) in the federal state of Schleswig-Holstein, Germany, during a health assessment study of invasive species, including raccoon dogs, carried out between 2021 and 2022. Four raccoon dogs showed severe lesions, including extensive alopecia with thickening and hyperpigmentation of the skin (lichenification). The fifth animal was less affected, showing only thinning of the hair coat in multiple body locations. Skin scrapings were performed and confirmed the presence of Sarcoptes scabiei. Histopathology of the skin revealed diffuse epidermal hyperplasia and hyperkeratosis, mild eosinophilic dermatitis, and varying amounts of intralesional mites. Staphylococcus pseudintermedius and Corynebacterium auriscanis were detected in the skin samples of the affected animals, indicating a secondary bacterial infection. The source of sarcoptic mange remains unclear; interspecies transmission via direct or indirect contact seems likely. Raccoon dogs are therefore a potential vector for sarcoptic mange, and their behaviour could contribute to disease spread and persistence.

Platycladus orientalis leaves are rich in flavonoids and polysaccharides, which offer high medicinal and nutritional benefits. This study aimed to investigate the impact of P. orientalis leaf extract (PLE) on the growth performance, fur quality, serum parameters, and intestinal microbiota of raccoon dogs. Sixty healthy male black raccoon dogs, aged 85 (±5) days, were randomly assigned to four groups and fed a basal diet supplemented with 0, 0.25, 0.50, and 1.00 g/kg PLE for 125 days (designated as groups P0, P1, P2, and P3, respectively). The results revealed that the raccoon dogs in group P1 exhibited increased average daily gain and underfur length while showing a decreased feed/gain ratio compared to group P0 ( p < 0.05). However, the heart index in group P2 was significantly lower than in group P0 ( p < 0.05), and the kidney index and serum alanine aminotransferase activities in group P3 were higher than in groups P2 and P0 ( p < 0.05), suggesting potential adverse effects at higher PLE dosages. Notably, dietary PLE supplementation led to a reduction in serum glucose concentrations ( p < 0.05), which may have implications for glucose regulation. Furthermore, the study explored the impact of dietary supplementation with 0.25 g/kg PLE on the raccoon dogs' intestinal microbiota using high-throughput sequencing. The results showed significant alterations in the microbial community structure, with a notable decrease in the abundance of Prevotella copri in response to 0.25 g/kg PLE supplementation ( p < 0.05). In conclusion, supplementing raccoon dogs' diet with 0.25 g/kg PLE can lead to improved growth performance and a positive influence on the intestinal microbiota. However, caution should be exercised regarding higher dosages, as they may have adverse effects on certain parameters. As a result, PLE holds promise as a potential feed additive for fur animal production.

Originally from Asia, the raccoon dog Nyctereutes procyonoides is an invasive alien species in Europe, listed since 2019 on the List of invasive alien species of Union concern. The raccoon dog is considered to have negative impact on native biodiversity, as well as a crucial role in hosting and transmitting diverse parasites and pathogens of human and veterinary importance. In the present study, stomach content analyses and parasitological examinations were performed on 73 raccoon dogs from Germany. In addition, fecal samples were analyzed. The results of the study confirm the assumption that the examined raccoon dogs were infested with a various ecto- and endoparasite fauna. A total of 9 ecto- and 11 endoparasites were detected, with 6 of the endoparasites having human pathogenic potential. Trichodectes canis (P = 53.42%), Toxocara canis (P = 50.68%) and Uncinaria stenocephala (P = 68.49%) were the most abundant parasite species. The stomach contents consisted of approximately one-third vegetable and two-thirds animal components, composed of various species of amphibians, fish, insects, mammals and birds. Among them were specially protected or endangered species such as the grass frog Rana temporaria . The study shows that the raccoon dog exerts predation pressure on native species due to its omnivorous diet and, as a carrier of various parasites, poses a potential risk of infection to wild, domestic and farm animals and humans., Competing Interests: The authors declare no competing interests., (© 2023 The Authors.)

Microorganisms of the genus Eperythrozoon are a zoonotic chronic infectious disease with wide distribution. We found that raccoons infected with Eperythrozoon showed obvious stunting, which seriously affected the economic benefits of raccoon dogs. To investigate the pathogenesis of the raccoon dog, we used transcriptome and proteome sequencing to analyze the changes in mRNA, miRNA, and protein expression in raccoon dogs infected with Eperythrozoon and normal raccoons. The results showed that the expression levels of genes related to immunity, metabolism, and enzyme activity were significantly changed. Among these, ERLIN1, IGF1R, CREB3L1, TNS1, TENC1, and mTOR play key roles. Additionally, the miR-1268, miR-125b, miR-10-5p, and miR-10 as central miRNAs regulate the expression of these genes. Integrated transcriptomic and proteomic analyses revealed consistent trends in mRNA and protein changes in MYH9, FKBP1A, PRKCA, and CYP11B2. These results suggest that Eperythrozoon may contribute to the slow development of raccoons by affecting the expression of mRNAs and miRNAs, reducing their immunity and causing metabolic abnormalities.

Since mid-July 2023, an outbreak caused by highly pathogenic avian influenza A(H5N1) virus clade 2.3.4.4b genotype BB is ongoing among farmed animals in South and Central Ostrobothnia, Finland. Infections in foxes, American minks and raccoon dogs have been confirmed on 20 farms. Genetic analysis suggests introductions from wild birds scavenging for food in farm areas. Investigations point to direct transmission between animals. While no human infections have been detected, control measures are being implemented to limit spread and human exposure.

The present investigation endeavours to discern dental and non-dental pathologies affecting cranial structures of raccoon dogs, while focusing on cases of periodontitis. Furthermore, the study aims to conduct a comparative analysis based on sex and the nature of the pathologies encountered. The number of investigated skulls amounted to 126, including 76 males and 50 females. The predominant pathology identified was hypodontia, which accounted for 26.7% of males and 20% of females. Notably, the majority of hypodontia cases involved the absence of the mandibular third molar. Another noteworthy pathology was various stages of periodontitis, with rates ranging from 21.3% in males to 8% in females. Other pathologies, like tooth fractures and abrasion, were significantly less encountered. Excessive bone formation was relatively abundant and localized in specific areas-the parietal bone and the occipital regions. This tendency was observed in 8% of male cases and 6% of females. We found that the total number of dental and skull-bone pathologies is significantly more common in males than in females ( p = 0.003). Additionally, the total number of various cases of periodontitis is more common in males too ( p = 0.04).

Toxocara tanuki is a common large roundworm in raccoon dogs. Experimental infection studies of T. tanuki in mice were conducted to clarify the distribution and infectivity of larvae in tissue. Groups of BALB/c and C57BL/6 mice (n = 5 mice/group) were each inoculated with 1000 embryonated T. tanuki eggs and necropsied at 7, 31, 91, and 182 days post inoculation (dpi). The number of larvae in the central nervous system, heart, lungs, kidneys, spleen, gastrointestinal tract, liver, and carcass was examined. Larvae obtained from the aforementioned mice on different days of the necropsy were orally inoculated into four groups of ICR mice (n = 6 mice/group) that were then necropsied at 21 dpi. Larvae were recovered from all mice. In the BALB/c and C57BL/6 mice, most of the larvae (> 88.7%) were recovered from the liver and the remainder from other tissues. The total number of larvae recovered from C57BL/6 mice was significantly higher than that from BALB/c mice, but no difference in the relative larval distribution within the viscera between the two mouse strains was observed. The mean recovery percentage of larvae from ICR mice infected with 182-day-old tissue larvae was 3.3%. Our findings showed that T. tanuki larvae migrated predominantly to the liver of mice and that the larvae maintained their infectivity for at least half a year., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Dirofilaria repens is an expanding vector-borne zoonotic parasite of canines and other carnivores. Sub-clinically infected dogs constitute the most important reservoir of the parasite and the source of infection for its mosquito vectors. However, occurrence of D. repens infection in wild animals may contribute to the transmission of the parasite to humans and may explain the endemicity of filariae in newly invaded regions. The aim of the current study was to determine the occurrence of D. repens in 511 blood and spleen samples from seven species of wild carnivores (wolves, red foxes, Eurasian badgers, raccoons, raccoon dogs, stone martens, and pine martens) from different regions of Poland by means of a PCR protocol targeting the 12S rDNA gene. Dirofilaria repens-positive hosts were identified in seven of fourteen voivodeships in four of the seven regions of Poland: Masovia, Lesser Poland, Pomerania and Warmia-Masuria. The highest prevalence was found in Masovia region (8%), coinciding with the highest previously recorded prevalence in dogs in Central Poland. The DNA of Dirofilaria was detected in 16 samples of three species (total prevalence 3.13%). A low and similar percentage of positive samples (1.9%, 4.2% and 4.8%) was recorded among badgers, red foxes, and wolves, respectively. Dirofilaria repens-positive hosts were identified in seven of fourteen voivodships. Based on detection in different voivodeships, D. repens-positive animals were recorded in four out of the seven regions of Poland: in Masovia, Lesser Poland, Pomerania, and Warmia-Masuria. The highest prevalence of filariae was found in Masovia region (8%), reflecting the highest previously recorded prevalence in dogs (12-50%) in Central Poland. In summary, we conducted the first comprehensive study on the epidemiology of D. repens in seven species of wild hosts in all seven regions of Poland and identified the first case of D. repens infection in Eurasian badgers in Poland and the second in Europe., (© 2023. The Author(s).)

Background: The oriental eyeworm Thelazia callipaeda (Spirurida: Thelaziidae) is an emerging parasitic ocular nematode of carnivores and humans. In domestic animals and humans, the infection causes varying degrees of inflammation and lacrimation, and wild carnivores represent an important reservoir. In this study we examined the infection status and molecular characterization of T. callipaeda in two urban carnivores, raccoons Procyon lotor and wild Japanese raccoon dogs Nyctereutes viverrinus, in the Kanto region of Japan., Methods: From January 2020 to December 2021, 193 carcasses including 178 raccoons and 15 raccoon dogs were examined for the presence of worms in the eye. The worms from infected animals (one worm per host) were morphologically identified as T. callipaeda. Worms (1-5 worms per host) were subjected to genetic analysis using mitochondrial cytochrome c oxidase subunit I gene sequences., Results: The prevalence of T. callipaeda in raccoons and Japanese raccoon dogs was 20.2% (36/178) and 13.3% (2/15), respectively. The cox1 sequences from 56 worms from 38 animals revealed three haplotypes (h9, h10, and h12). Analysis of multiple worms for five raccoons showed co-infection of two different haplotypes (h9 and h10) in a single host. Comparing our data with published sequences, three sequences obtained from raccoons and raccoon dogs shared the same haplotypes as those reported in humans, dogs, and cats in Japan., Conclusions: Our findings show a high prevalence of T. callipaeda in raccoons, suggesting that this invasive carnivore species serves as an important natural reservoir of T. callipaeda in the Kanto region of Japan, an area with the highest human population of the country., (© 2023. The Author(s).)

Severe fever with thrombocytopenia syndrome (SFTS) is a fatal emerging tick-borne zoonotic disease caused by the SFTS virus (SFTSV). SFTSV infection in humans and companion animals is a matter of concern in endemic areas. Various wild animals are involved in the transmission cycle of SFTSV with vector ticks. Because the home range of medium-sized wild mammals commonly overlaps with humans' living spheres, this study aimed to reveal the endemicity of SFTSV in such mammals. This study investigated the prevalence of antibodies against SFTSV and viral RNA in medium-sized wild mammals in Miyazaki Prefecture, Japan where human cases have been most frequently reported in Japan and performed a phylogenetic analysis to compare the detected SFTSV with those previously reported. Forty-three of 63 (68%) Japanese badgers (Meles anakuma) and 12 of 53 (23%) Japanese raccoon dogs (Nyctereutes procyonoides viverrinus) had antibodies against SFTSV. Japanese marten (n = 1), weasels (n = 4), and Japanese red fox (n = 1) were negative. Two of 63 (3%) badgers tested positive for SFTSV RNA, whereas the other species were negative. Phylogenetic analysis of the partial nucleotide sequence of SFTSV revealed that viral RNA detected from badgers exhibited 99.8% to 100% similarity to SFTSV, as previously reported in humans, cat, and ticks in the study area. This study demonstrated high seropositivity of antibodies in medium-sized wild mammals and suggested that SFTSV could be shared among these mammals, humans, and companion animals in endemic areas., (Copyright © 2022. Published by Elsevier GmbH.)

Unlike farm animals, wild animals are not subject to continuous health surveillance. Individual projects designed to screen wildlife populations for specific pathogens are, therefore, also of great importance for human health. In this context, the possible formation of a reservoir for highly pathogenic zoonotic pathogens is a focus of research. Two of these pathogens that have received particular attention during the last years are the novel severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), due to its fast global spread and high impact to the human health, and, since its introduction into Germany, the flavivirus West Nile virus (WNV). Especially in combination with invasive vertebrate species (e.g., raccoons ( Procyon lotor ) and raccoon dogs ( Nyctereutes procyonoides ) in Germany), risk analysis must be done to enable health authorities to assess the potential for the establishment of new wild life reservoirs for pathogens. Therefore, samples were collected from raccoons and raccoon dogs and analyzed for the presence of SARS-CoV-2 and WNV infections in these populations. Molecular biological and serological data obtained imply that no SARS-CoV-2 nor WNV reservoir has been established in these two wild life species yet. Future investigations need to keep an eye on these invasive carnivore populations, especially since the close contact of these animals to humans, mainly in urban areas, would make animal-human transmission a challenge for human health.

Prion diseases are fatal degenerative encephalopathies caused by misfolded prion protein (PrP Sc ) converted from normal prion protein (PrP C ). Previous studies have reported that genetic polymorphisms of the prion protein gene ( PRNP ) play a critical role in susceptibility to prion diseases. In addition, prion disease-resistant animals showed unique structural features of prion protein (PrP) related to species-specific amino acids. However, investigations of genetic polymorphisms of the PRNP gene and structural characteristics of PrP have not been performed in raccoon dogs thus far. We investigated genetic polymorphisms of PRNP in 87 raccoon dogs using amplicon sequencing and analyzed the genotype, allele, haplotype frequencies, and linkage disequilibrium (LD) using Haploview version 4.2. In addition, we performed phylogenetic analysis and multiple sequence alignment (MSA) using MEGA X version 10.1.8 and Clustal X version 2.1, respectively. We estimated the impact of raccoon dog and Canidae family-specific amino acids using PolyPhen-2, PROVEAN, and AMYCO. Furthermore, we analyzed the effect of raccoon dog and Canidae family-specific amino acids using the AlphaFold2 and Swiss-PdbViewer programs. We found 4 novel single nucleotide polymorphisms (SNPs) of the raccoon dog PRNP gene. In addition, the raccoon dog PrP showed 99.61% identity and the closest genetic distance to dog PrP. Among the substitutions of Canidae-specific amino acids with interspecific amino acids, D163N showed increased amyloidogenic propensity, and R181H showed alterations of hydrogen bonds. Furthermore, electrostatic potentials were changed according to the substitutions of D163N and R181H. By comparing PrP between raccoon dogs and raccoons, R168K and K224R were found to be related to changes in hydrogen bonds, and K224R altered the electrostatic potential of raccoon dog PrP. In the present study, we first reported 4 novel synonymous SNPs of the raccoon dog PRNP gene. We also identified that the PrP of raccoon dog has high homology (99.61%) with PrP of dog, which is a prion-resistant animal. In addition, raccoon dog PrP-specific amino acids are related to low amyloid propensity and inherent characteristics of 3D structure of raccoon dog PrP compared to the PrP of prion-susceptible 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 © 2022 Jo, Kim, Oem and Jeong.)

Cryptosporidium spp. are common parasitic pathogens causing diarrhea in humans and various animals. Fur animals are widely farmed in Shandong Province, China, but the prevalence and genetic identity of Cryptosporidium spp. in them are unclear. In this study, 1,211 fecal samples were collected from 602 minks, 310 raccoon dogs and 299 foxes on two farms in Shandong and analyzed for Cryptosporidium spp. by nested PCR and sequence analyses of the small subunit rRNA gene. The overall infection rate of Cryptosporidium spp. was 31.5% (381/1,211), with a higher infection rate in raccoon dogs (37.7%, 117/310) than in foxes (32.4%, 97/299) and minks (27.7%, 167/602). By age, the highest infection rates of Cryptosporidium spp. were observed in raccoon dogs of 1-2 months, minks of 5-6 months, and foxes of > 12 months. Three Cryptosporidium species and genotypes were detected, including C. canis ( n = 279), C. meleagridis ( n = 65) and Cryptosporidium mink genotype ( n = 37). Among the three major host species, raccoon dogs were infected with C. canis only ( n = 117), while foxes were infected with both C. canis ( n = 32) and C. meleagridis ( n = 65), and minks with C. canis ( n = 130) and Cryptosporidium mink genotype ( n = 37). Subtyping of C. canis by sequence analysis of the 60 kDa glycoprotein gene identified eight subtypes. They belonged to two known subtype families, XXa and XXd, and two novel subtype families XXf and XXg, with host adaptation at the subtype family level. Notably, C. canis from foxes was genetically distant from those in other hosts. Further subtyping analysis identified three subtypes (IIIeA21G2R1, IIIeA19G2R1 and IIIeA17G2R1) of C. meleagridis and two novel subtype families Xf and Xg of the Cryptosporidium mink genotype. The presence of zoonotic C. canis subtypes in raccoon dogs and C. meleagridis subtypes in foxes suggests that these fur animals might be potential reservoirs for human-pathogenic Cryptosporidium spp., 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 Wang, Wei, Cao, Wu, Zhao, Guo, Xiao, Feng and Li.)

Over the last several decades, no emerging virus has had a profound impact on the world as the SARS-CoV-2 that emerged at the end of 2019 has done. To know where severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) originated from and how it jumped into human population, we immediately started a surveillance investigation in wild mammals in and around Wuhan when we determined the agent. Herein, coronaviruses were screened in the lung, liver, and intestinal tissue samples from fifteen raccoon dogs, seven Siberian weasels, three hog badgers, and three Reeves's muntjacs collected in Wuhan and 334 bats collected around Wuhan. Consequently, eight alphacoronaviruses were identified in raccoon dogs, while nine betacoronaviruses were found in bats. Notably, the newly discovered alphacoronaviruses shared a high whole-genome sequence similarity (97.9 per cent) with the canine coronavirus (CCoV) strain 2020/7 sampled from domestic dog in the UK. Some betacoronaviruses identified here were closely related to previously known bat SARS-CoV-related viruses sampled from Hubei province and its neighbors, while the remaining betacoronaviruses exhibited a close evolutionary relationship with SARS-CoV-related bat viruses in the RdRp gene tree and clustered together with SARS-CoV-2-related bat coronaviruses in the M, N and S gene trees, but with relatively low similarity. Additionally, these newly discovered betacoronaviruses seem unlikely to bind angiotensin-converting enzyme 2 because of the deletions in the two key regions of their receptor-binding motifs. Finally, we did not find SARS-CoV-2 or its progenitor virus in these animal samples. Due to the high circulation of CCoVs in raccoon dogs in Wuhan, more scientific efforts are warranted to better understand their diversity and evolution in China and the possibility of a potential human agent., (© The Author(s) 2022. Published by Oxford University Press.)

In recent decades, populations of the raccoon (Procyon lotor) and the raccoon dog (Nyctereutes procyonides) have increased and adapted to peri-urban and urban environments in many parts of the world. Their ability to rapidly colonize new territories, high plasticity and behavioral adaptation has enabled these two species to be considered two of the most successful invasive alien species. One of the major threats arising from continually growing and expanding populations is their relevant role in maintaining and transmitting various vector-borne pathogens among wildlife, domestic animals and humans. According to the WHO, over 17% of infectious diseases are vector-borne diseases, including those transmitted by ticks. Every year tick-borne pathogens (TBPs) create new public health challenges. Some of the emerging diseases, such as Lyme borreliosis, anaplasmosis, ehrlichiosis, babesiosis and rickettsiosis, have been described in recent years as posing important threats to global health. In this review we summarize current molecular and serological data on the occurrence, diversity and prevalence of some of the TBPs, namely Babesia, Theileria, Hepatozoon, Borrelia, Rickettsia, Bartonella, Anaplasma and Ehrlichia, that have been detected in raccoons and raccoon dogs that inhabit their native habitats and introduced areas. We draw attention to the limited data currently available on these invasive carnivores as potential reservoirs of TBPs in different parts of the world. Simultaneously we indicate the need for more research in order to better understand the epidemiology of these TBPs and to assess the future risk originating from wildlife., (© 2022. The Author(s).)

Paeoniae radix alba extract (PRA extract) has the functions of regulating immunity, resisting inflammation, and has antioxidant properties. However, current recommendations of dietary PRA extract levels for raccoon dogs were inadequate. The purpose of this experimental study was to gain information allowing for better estimating the effects of PRA extract on raccoon dogs, and their PRA requirements. Fifty healthy male raccoon dogs of (120 ± 5) days old were randomly divided into 5 groups (group PRA0, PRA1, PRA2, PRA4, PRA8) with 10 animals in each group and 1 in each replicate. Five kinds of experimental diets were prepared with five levels of Paeoniae radix alba extract (0, 1, 2, 4, 8 g/kg) in the basic diet. The prefeeding period was 7 days and the experimental period was 40 days. The results showed that the average daily feed intake in group PRA1 and PRA2 was significantly higher than that in other groups ( P < 0.01). The dry matter excretion in group PRA8 was significantly higher than that in other groups ( P < 0.01), while the dry matter digestibility and protein digestibility in group PRA8 were significantly lower than those in other groups ( P < 0.01). Nitrogen retention in group PRA1 and PRA2 was significantly higher than that in group PRA8 ( P < 0.05). With the increase of the content of Paeoniae radix alba extract in diet, the activity of alkaline phosphatase in group PRA2 was significantly higher than that in group PRA0 ( P < 0.05); The activity of serum SOD in group PRA4 was significantly higher than that in other groups ( P < 0.01). The content of serum IgA in group PRA2 was significantly higher than that in other groups ( P < 0.05). The content of TNF-α in intestinal mucosa in group PRA1 and group PRA2 was significantly lower than that in group PRA0 ( P < 0.05). In conclusion, we found that dietary Paeoniae radix alba extract intake significantly improved the feed intake and nitrogen deposition of Ussuri raccoon dog, increased the content of serum IgA and reduced the content of TNF-α in the small intestinal mucosa. We suggest that an estimated dietary Paeoniae radix alba extract level of 1 to 2 g/kg could be used as a guide to achieve the optimal performance of raccoon dogs., Competing Interests: XY was employed by the China Animal Husbandry Group. 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 © 2022 Wang, Li, Zhong, Zhang, Yang, Chen, Wang and Yang.)

Paragonimiasis is a zoonotic trematode infection caused by Paragonimus spp. To determine the recent status of Paragonimus infections in wild animals, this study investigated Paragonimus spp. in 39 raccoon dogs and 54 Japanese badgers from March 2019 to January 2021 in Miyazaki Prefecture, and examined metacercariae in freshwater crabs. Triploid P. westermani was found in one raccoon dog (2.6%), and metacercariae were recovered from Eriocheir japonica captured near the infected animal collected. One Japanese badger (1.9%) harbored P. skrjabini miyazakii; this prevalence was lower than the approximately 30% that was reported in the 1970s. Results indicated that zoonotic Paragonimus was sporadically prevalent in wild animals. Further investigation in various animals is awaited to elucidate current wildlife reservoirs for those Paragonimus.

Alaria alata is an emerging parasite that poses a potential risk for those consuming game, pork, snails and frogs. One paratenic host of A. alata that is known to play an important role in its spread through its feeding habitats is the wild boar. However, no statistical analysis of the influence of aquatic environments and carnivores on the occurrence of A. alata in wild boars has yet been performed. The present study combines a small-scale analysis based on hunting districts in the Mazowieckie province with a large-scale analysis based on data for all provinces in Poland. We applied various modeling approaches, including logistic regression and a generalized linear model in order to determine the presence, intensity and prevalence of A. alata. We used the Alaria mesocercariae migration technique (AMT) to estimate the risk of A. alata among wild boar in a given hunting district or province. The small-scale analysis found that mesopredators (red fox ( Vulpes vulpes )) and racoon dog ( Nyctereutes procyinoides ) were likely to influence A. alata infestation of wild boar; however, the effect was weak, probably as a result of the large home range size of these animals. The large-scale analysis found that wetlands influence the prevalence of A. alata in wild boar, with the estimated risk increasing in the north of the country; this finding is consistent with other studies. Our findings indicate that the occurrence of A. alata in wild boar requires analysis on many levels, and environmental factors play a key role in risk assessment., 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 Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

The aim of the study was to determinate the prevalence and intensity of infection of raccoon dogs with internal parasites, with a particular emphasis on particular species of helminths known to be dangerous to humans. A total of 96 raccoon dogs were obtained from hunters from September 2018 to October 2021. The digestive tract was taken for examination. The parasitological examination was performed using the dissection methods. The extensity of infection with all internal parasites was 60.3%. The following parasites were found in the tested animals: Echinococcus multilocularis (in 10.42% of animals), Toxocara canis (18.75%), Alaria alata (25.0%), Taenia spp. (19.79%), Uncinaria stenocephala (27.08%), Mesocestoides spp. (54.17%) and Dipylidium caninum (6.25%). The highest mean intensity of infection was demonstrated by A. alata and E. multilocularis then by Mesocestoides spp. This study showed that the raccoon dog from northern Poland is a reservoir host of zoonotic pathogens, such as E. multilocularis , Toxocara canis and Alaria alata . Although the role of the racoon dog as a final host of the life cycle of E. multilocularis is considered of less importance than that of the red fox, this species may increase the risk of echinococcosis in humans, mainly due to its growing population in northern Poland.

The molecular background of disorders of sex development (DSD) in dogs is poorly understood. Several copies of the SRY genes have been reported in the dog genome. We used droplet digital PCR with the aim of determining variability in SRY copy number and its association with DSD in dogs. Altogether 19 DSD male dogs (XY DSD) of 10 breeds and 87 control dogs of eight breeds were analyzed. Moreover, we performed a comparative analysis of SRY copy number in other canids: wolves (3), red foxes (16), and Chinese raccoon dogs (10). We found that the modal number of SRY copies in dogs, wolves, red foxes, and Chinese raccoon dogs was 3, 3, 1, and 3 respectively. Variability of copy number was only observed in Yorkshire Terriers (two or three copies) and red foxes (one or two copies). An analysis of six DSD Yorkshire Terriers and 38 control males of this breed showed that 50% of the DSD dogs had two copies, while the incidence of this variant was significantly lower in the control dogs (10.5%). Searching for the copy number of the coding and 5'-flanking fragments revealed full concordance with the copy number. These fragments were also sequenced in DSD (19) and control (24) dogs, and no DNA variants were found. We conclude that, in the dog, two or three functional copies of the SRY gene are present, and a smaller number of copies showed an association with the risk of DSD phenotype in Yorkshire Terriers., (© 2021 Stichting International Foundation for Animal Genetics.)

The raccoon ( Procyon lotor ) and the raccoon dog ( Nyctereutes procyonoides ) were introduced to Europe and, in the past decades, their populations have increased and adapted to synanthropic environments across Europe. In view of their possible further spread in Europe, the invasive species should be examined as potential reservoirs for helminths, including tapeworms. This study aims to investigate the prevalence and diversity of tapeworms in introduced wild carnivores in Poland by identifying cestode species based on copro-DNA analysis. A total of 214 individual fecal samples from non-native invasive carnivores, i.e., raccoons and raccoon dogs, and additionally 47 samples from native carnivores, i.e., European badgers ( Meles meles ), were analyzed for the presence of cestodes. PCR analysis of fecal samples targeting a fragment of mitochondrial (mt) 12S rRNA gene revealed the presence of cestode DNA in 19 of 103 (18.4%) raccoons, in 13 of 111 (11.7%) raccoon dogs and in 23 of 47 (48.9%) badgers. Sequence analysis demonstrated the presence of Mesocestoides litteratus in raccoons and raccoon dogs, while Mesocestoides lineatus was identified only in two samples derived from raccoon dogs. Moreover, in this study, Atriotaenia incisa was for the first time molecularly characterized by using fragments of mt 12S rRNA gene, and the DNA of this cestode species was detected in the fecal samples of all the examined host species.

Urbanisation and invasion of wildlife into urban areas as well as human leisure activities create diverse wildlife-domestic animal-human interfaces, increasing the risk of (zoonotic) parasite spillover from sylvatic to domestic and synanthropic cycles. This study investigated the endo- and ectoparasite fauna, emphasising on parasites of One Health Concern, of the most common predators in northern Germany between November 2013 and January 2016. Eighty red foxes (Vulpes vulpes), 18 stone martens (Martes foina) and nine raccoon dogs (Nyctereutes procyonoides) were available for the study. Overall, 79 (73.8%) of the examined predators (n=107) harboured at least one endoparasite. The most frequently detected endoparasites in red foxes were Toxocara canis (43.8% positive individuals), Capillaria spp. (36.3%), Alaria alata (25.0%), Echinococcus multilocularis (26.3%) and Uncinaria stenocephala (25.0%). Furthermore, Toxascaris leonina, Trichuris vulpis, Taenia ssp., Mesocestoides spp. and coccidian oocysts were observed. The endoparasite species richness in raccoon dogs was comparable to red foxes, while in stone martens, only Capillaria spp. were found. Muscle digestion for detection of Trichinella spp. and antigen testing for Giardia spp. did not show positive results. Ectoparasite analyses revealed infestations with ticks species of the genus Ixodes as well as Dermacentor reticulatus. Scabies mites were not present in digested skin samples, while Demodex spp. mites were observed by faecal flotation in one red fox. Furthermore, fleas (Archaeopsylla erinacei and Chaetopsylla globiceps) were observed in the fur of red foxes, while lice were not present in any predator species. However, infestation frequency with ectoparasites was with 19.2% generally low in available predator skins (n=99). Overall, the present study showed that predators in northern Germany serve as reservoirs for parasites of One Health concern, with four of the five most frequent endoparasites being zoonotic, highlighting the need of parasite surveillance in wildlife predators in order to implement measures avoiding spillovers to domestic animals and humans., (© 2021. The Author(s).)

Wild carnivores, both introduced and native species, are able to adapt well to peri-urban environments, facilitating cross-species pathogen transmission with domestic animals, and potentially humans. The role of wild living reservoir hosts cannot be ignored because of their known carriage of E. bieneusi zoonotic genotypes. In the past decades, populations of wild living carnivores, i.e., native, such as red foxes, and invasive, such as raccoon dogs and raccoons, have increased and adapted to synanthropic environments across Europe, including Poland. The knowledge concerning E. bieneusi genotype identification and distribution in wild carnivores is limited worldwide. A total of 322 individual fecal samples from six carnivore species, i.e., raccoon, raccoon dog, red fox, European badger, pine and beech martens, were collected and then analysed for the presence of E. bieneusi using the nested PCR method. Overall prevalence of the pathogen was estimated to be as high as 27.3%. The infection rates for E. bieneusi varied between the carnivore species, from 13.7% in beech martens to 40.4% in raccoon dogs. Based on sequence analysis of the ITS region of the rRNA gene marker, we detected five known genotypes of E. bieneusi in examined animals. In the invasive species, E. bieneusi NCF2 and D genotypes have been identified, whereas in the native ones, E. bieneusi NCF2, D, C, EbCar2 and Type IV genotypes were identified. All E. bieneusi genotypes recorded in this survey clustered in Group 1, showing their zoonotic potential. Our results provide the first description of the occurrence and genotypes of the microsporidian E. bieneusi in wild living population of raccoon dogs in Europe. Our findings are important for the study of pathogen epidemiology and emphasize the fact that the invasive and the native wild living carnivores, both widely distributed, should be considered more seriously as significant sources of zoonotic pathogens hazardous to domestic and farmed animals and humans.

Raccoon dogs have successfully invaded Europe, including Denmark. Raccoon dogs are potential vectors and reservoir hosts of several zoonotic pathogens and thus have the potential for posing a threat to both human and animal health. This study includes analysis of four zoonotic parasites, 16 tick-borne pathogens and two pathogen groups from 292 raccoon dogs collected from January 2018 to December 2018. The raccoon dogs were received as a part of the Danish national wildlife surveillance program and were hunted, found dead or road killed. The raccoon dogs were screened for Alaria alata and Echinococcus multilocularis eggs in faeces by microscopy and PCR, respectively, Trichinella spp. larvae in muscles by digestion, antibodies against Toxoplasma gondii by ELISA and screening of ticks for pathogens by fluidigm real-time PCR. All raccoon dogs tested negative for E. multilocularis and Trichinella spp., while 32.9% excreted A. alata eggs and 42.7% were T. gondii sero-positive. Five tick-borne pathogens were identified in ticks collected from 15 raccoon dogs, namely Anaplasma phagocytophilum (20.0%), Babesia venatorum (6.7%) , Borrelia miyamotoi (6.7%) , Neoehrlichia mikurensis (6.7%) and Rickettsia helvetica (60.0%). We identified raccoon dogs from Denmark as an important reservoir of T. gondii and A. alata infection to other hosts, including humans, while raccoon dogs appear as a negligible reservoir of E. multilocularis and Trichinella spp. infections. Our results suggest that raccoon dogs may be a reservoir of A. phagocytophilum ., Competing Interests: None., (© 2021 The Authors. Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

Hydrocephalus is one of the most common central nervous system malformations in domestic dogs, yet they are poorly documented and studied in wild carnivoran mammals. A pup of raccoon dog (Nyctereutes procyonoides) was rescued and brought to Wildlife Center. The pup showed generalized ataxia, a domed skull, and an open bregmatic fontanelle. Ultrasound and MRI showed severe enlargement of the lateral ventricle with the loss of septum pellucidum resulting in a single large ventricle and cervical syringohydromyelia. Although treatment was attempted, the animal was euthanized due to poor prognosis. At necropsy, macroscopic findings were identical to the diagnostic imaging, where marked enlargement of the calvarium, and attenuated gyri and sulci were observed. Finally, hydrocephalus was confirmed. Here, we describe a case of hydrocephalus in a raccoon dog (Nyctereutes procyonoides).

This study reports outbreak of a new disease caused by Staphylococcus pseudintermedius (S. pseudintermedius) in raccoon dogs. The disease occurred in a breeding farm of raccoon dogs in Guan County of Shandong Province in China in August of 2019. 47% (425/896) of the raccoon dogs showed some abnormal symptoms; 17.6% (75/425) of which had severe skin and soft tissue infections (SSTIs), dyspnoea and severe pathological lesions in lungs, livers, etc; and 4.2% (18/425) of which died within 4 weeks. The pathogen of the disease was identified as S. pseudintermedius by mass spectrometer detection, animal pathogenicity tests, microscopic examination and biochemical reaction tests. Its nucleotide homology of 16S rRNA gene was 100% with that of other published strains, and its genotype was between the American and Brazilian strains from other animals. The isolated S. pseudintermedius strain from the diseased raccoon dogs could cause ulceration and suppuration in the skins and severe pathological lesions not only in raccoon dogs, but also in mice; and it is confirmed as a methicillin-resistant S. pseudintermedius (MRSP) strain by the amplification of mecA gene; and 12 sensitive drugs were screened by drug sensitivity tests. Full attention should be paid to the great economic loss and the potential zoonotic risk caused by the S. pseudintermedius in raccoon dogs, and this study can provide a reference for the clinical diagnosis and treatment of this new disease., (© 2020 Wiley-VCH GmbH.)

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic with millions of infected humans and hundreds of thousands of fatalities. As the novel disease - referred to as COVID-19 - unfolded, occasional anthropozoonotic infections of animals by owners or caretakers were reported in dogs, felid species and farmed mink. Further species were shown to be susceptible under experimental conditions. The extent of natural infections of animals, however, is still largely unknown. Serological methods will be useful tools for tracing SARS-CoV-2 infections in animals once test systems are evaluated for use in different species. Here, we developed an indirect multi-species ELISA based on the receptor-binding domain (RBD) of SARS-CoV-2. The newly established ELISA was evaluated using 59 sera of infected or vaccinated animals, including ferrets, raccoon dogs, hamsters, rabbits, chickens, cattle and a cat, and a total of 220 antibody-negative sera of the same animal species. Overall, a diagnostic specificity of 100.0% and sensitivity of 98.31% were achieved, and the functionality with every species included in this study could be demonstrated. Hence, a versatile and reliable ELISA protocol was established that enables high-throughput antibody detection in a broad range of animal species, which may be used for outbreak investigations, to assess the seroprevalence in susceptible species or to screen for reservoir or intermediate hosts., (© 2020 The Authors. Transboundary and Emerging Diseases published by Wiley--VCH GmbH.)

The definitive hosts of Metagonimus hakubaensis are reported to be hamsters, rats, mice, dogs, cats, chickens, and quails in experimental infection and Japanese water shrews in natural infection. Here we report that raccoon dogs are new natural definitive hosts of M. hakubaensis, based on morphological and molecular analyses of Metagonimus flukes collected from the host species from Aomori Prefecture, Japan. Moreover, M. hakubaensis recovered from raccoon dogs showed higher fecundity than those recovered from Japanese water shrews. Therefore, raccoon dogs were considered as a more suitable natural definitive host of M. hakubaensis than Japanese water shrews.

Background: Rabies vaccination of wildlife carnivores is a powerful tool to prevent, control and eliminate rabies. The presence of neutralizing rabies antibodies in blood is considered a reliable indicator of adequate vaccination. The main purpose of the present study was to analyze the seroprevalence of specific antibodies in target populations of Lithuanian red fox (RF) and raccoon dog (RD) during the oral rabies vaccination (ORV) campaigns during the 2010-2019 period., Results: Over the ten-year period, 7,261 RF and 2,146 RD sera samples were collected post-mortem in field conditions and tested using a commercial standardized enzyme-linked immunosorbent assay (ELISA) kit in Lithuania. In the ORV spring and autumn vaccination periods, 31.8% (20.3-43.4 95% CI - 95% confidence interval) and 31.7% (21.2-42.1 95% CI) of RF, and 34.1% (22.5-45.7 95% CI) and 34.7% (22.7-46.7 95% CI) of RD sera samples, respectively, were identified as ELISA-positive (seroconversion ≥ 0.5 EU/mL-Equivalent Units per Millilitre). The seroprevalence analysis in adult/ juvenile animal subpopulations indicated that 34.9% (27.2-42.5 95% CI) and 29.2% (20.3-37.9 95% CI) of RF, and 35.6% (25.2-46.0 95% CI) and 30.6% (20.2-40.9 95% CI) of RD sera samples, respectively, were identified as ELISA-positive (seroconversion ≥ 0.5 EU/mL). Statistically strong determinate correlations (r) between the serological results (pos.%) in RF adult/juvenile animal subpopulations (r = 0.937) and between RF and RD positive seroconvert (pos.%) sera samples during the spring vaccinations (r = 0.864) were demonstrated. In different ORV periods, 14-29% of RF and 7-25% of RD sera samples were identified as ELISA-negative (seroconversion < 0.5 EU/mL), but with low (0.125 < 0.49 EU/mL) antibody (Abs) titres., Conclusions: The 2010-2019 ORV programme has been an effective tool in both RF and RD populations in Lithuania. The rabies-free status of Lithuania was self-declared in 2015 with only three rabies cases identified in buffer zones since then. The percentage of ELISA-positive serum samples (seroconversion ≥ 0.5 EU/mL) during the different periods of vaccination was similar in RF and RD populations-32% and 34% respectively. The identified seroconversion average of 21.5% in RF and 16% in RD sera samples were officially identified as ELISA-negative (seronversion < 0.5 EU/mL), but with low 0.125 < 0.49 EU/mL Abs titres. That low, but positive seroconversion participated in the formation of populations overall immune status and can influence the interpretation of oral vaccination efficacy.

The field distribution of the oral rabies vaccine is effective in controlling the spread of rabies. The present study aimed to investigate efficient distribution locations based on the environment, contact rate, and consumption by target wildlife species in South Korea. The target species (Korean raccoon dogs, domestic dogs, and feral cats) accounted for 945 contacts (52.2%), in total 1,808 contacts. There were 863 (47.8%) contacts by non-target species. Raccoon dogs, a main reservoir of rabies in South Korea, had the highest contact rate (34.1%) among all species. The contact rate by target species was highest at riparian sites and bushy mountainous vegetation, where raccoon dogs are abundant. There was remarkable contact by raccoon dogs in mountainous areas below 150 m with bushy vegetation. Our results indicate that these locations are efficient areas for vaccine distribution, especially targeting the raccoon dog. Vaccines were continuously contacted with intervals ranging from one hour to one day. Vaccines at 94.4% of the distribution points were completely consumed within two weeks. The mean consumption rate was 95.2 ± 1.93% during the overall study period. These findings suggest that the oral rabies vaccine attracts wildlife including domestic dogs and feral cats. Our results suggest that low sections of mountainous areas with bushy vegetation and/or neighboring riparian areas are rich in target wildlife species (especially raccoon dogs) and are efficient locations for vaccine distribution to control rabies in South Korea.

Fungal organisms of the genus Pneumocystis may cause Pneumocystis pneumonia (PCP) in humans, but also domestic and wild mammals. Almost every animal species hosts its own genetically distinct Pneumocystis species, however information is sparse. In this study, 62 red foxes (Vulpes vulpes) and 37 raccoon dogs (Nyctereutes procyonoides) were collected in North-East Germany. The lung tissues of the animals were analysed by a new designed specific pan-Pneumocystis mtLSU rRNA gene PCR and sequencing. With this PCR, detection and discrimination of all known Pneumocystis spp. in a single step should be possible. This first detection of Pneumocystis spp. in 29/62 (46.8%) red foxes and 29/37 (78.4%) raccoon dogs indicated, that they harbour two dissimilar strains, as seen by specific single nucleotide position changes (SNPs). Nevertheless, five samples with contrary SNPs showed a probable inter-species transmission., (Copyright © 2020. Published by Elsevier Ltd.)

Background: Spirometra erinaceieuropaei is a diphylobothriid tapeworm with a complex life-cycle including definitive, intermediate and paratenic (transport) hosts. Multiple routes of parasite transmission often make it impossible to determine what type of host a specific infected animal is considered to be. Spargana larvae cause sparganosis, a severe food- and water-borne disease mainly found in Asia. In Poland, Spirometra sp. was reported in large carnivores in Białowieża Primeval Forest for the first time in the 1940s and was recently confirmed as S. erinaceieuropaei in several mammals and snakes using molecular methods., Methods: In total, 583 carcasses of 9 carnivore species were necropsied between 2013 and 2019 in north-eastern (NE) Poland. The larvae of S. erinaceieuropaei (spargana) were isolated from subcutaneous tissue, counted, and preserved for genetic analyses. We calculated the prevalence and intensity of infection. To assess spatial variation in S. erinaceieuropaei infection probability in NE Poland, we applied a generalized additive model (GAM) with binomial error distribution. To confirm the species affiliation of isolated larvae, we amplified a partial fragment of the 18S rRNA gene (240 bp in length)., Results: Spirometra larvae were found in the subcutaneous tissue of 172 animals of 7 species and confirmed genetically as S. erinaceieuropaei. The overall prevalence in all studied hosts was 29.5% with a mean infection intensity of 14.1 ± 33.8 larvae per individual. Native European badgers and invasive raccoon dogs were characterized by the highest prevalence. An analysis of parasite spread showed a spatially diversified probability of infection with the highest values occurring in the biodiversity hot spot, Białowieża Primeval Forest., Conclusions: Our study revealed that various mammal species (both native and non-native) can serve as S. erinaceieuropaei reservoirs. The frequency and level of infection may differ between selected hosts and likely depend on host diversity and habitat structure in a given area. Further studies are needed to assess the distribution of the parasite throughout Europe and the environmental and biological factors influencing infection severity in wild mammals.

The objective of this study was to determine the infection rate and genetic diversity of Cryptosporidium spp. in minks, foxes, and raccoon dogs, farmed in the Xinjiang Uygur Autonomous Region, Northwest China. Fresh fecal specimens were collected from individual cages of farmed minks (n = 214), blue foxes (n = 35), and raccoon dogs (n = 39) and examined using nested PCR based on the Cryptosporidium spp. small subunit rRNA gene. Cryptosporidium spp. was detected in 35 cages (12.2%, 35/288), with a higher infection rate detected in raccoon dogs (20.5%) compared with minks (12.1%) and blue foxes (2.9%). Sequence analysis showed that Cryptosporidium canis was the only species identified in blue foxes and raccoon dogs, while in the 26 Cryptosporidium-positive mink specimens, Cryptosporidium mink genotype (n = 17), C. canis (n = 7), and Cryptosporidium parvum (n = 2) were identified. Further analysis based on the 60-kDa glycoprotein (gp60) gene determined that both C. parvum isolates belonged to the subtype IIdA15G1, while eight of the 17 Cryptosporidium mink genotype isolates were a novel subtype that we have named XeA5G1. To the best of our knowledge, this is the first report of C. parvum subtype IIdA15G1 infection in minks. Since all the Cryptosporidium species/genotypes identified in minks, foxes, and raccoon dogs from Xinjiang have been previously found in humans, our results suggest that these fur animals may play a role in the transmission of zoonotic Cryptosporidium.

Although kinship (parent-offspring or siblings) contact has been suggested as a driving factor for sarcoptic mange epizootic in raccoon dogs (Nyctereutes procyonoides), no effect has been reported. In contrast, habitat fragmentation caused by urbanization may result in a high occurrence of sarcoptic mange, because habitat fragmentation may promote contact infection by increasing the population density of raccoon dogs. The habitat distribution of raccoon dogs may therefore influence epizootic sarcoptic mange. The genetic relationship between raccoon dogs was analyzed to examine Sarcoptes scabiei transmission between kin. The relationship between S. scabiei infection and the habitat of raccoon dogs was also investigated. Seventy-five raccoon dogs from Takasaki, Gunma prefecture, were examined from 2012 to 2018; 23 were infested with S. scabiei. The genotypes were determined using 17 microsatellite loci, and the relationships were categorized into four patterns by the ML-Relate software. There was no significant difference between infested pairs and other two pairs (Chi- squared test: χ 2 =0.034, df=1, P=0.85). Although it was difficult to predicate because the mortality rate was unclear in this study, kinship contact does not seem to be an important factor for sarcoptic mange epizootic. S. scabiei infection rates were significantly associated with the location of village sections (OR=1.55, 95% CI=1.11-2.17, P=0.011). It is suggested that direct/indirect contact between individuals living closely together is an important factor for the transmission of S. scabiei.

Background: The raccoon dog (Nyctereutes procyonoides), endemic to East Asia, is classified as six subspecies according to their geographical distribution including a population introduced to Europe. Studies on phylogenetic relationship or population genetics in both native and introduced areas have been carried out recently. Lately, opinions that Japanese raccoon dogs should be classified as a different species were asserted based on several studies using karyotypes, morphometric characters, mtDNA, and microsatellites analysis. However, no data pertaining to the nuclear DNA (nDNA) or Y chromosome are available., Objective: To estimate the relationship among the species using different genes is necessary in understanding of the history of this species., Method: Therefore, we investigated nDNA and Y chromosomes in our study to define relationships: (1) between continental raccoon dog populations, (2) between original and introduced groups, and (3) between continental and Japanese groups., Results: The analysis of four nuclear (CHRNA1, VTN, TRSP, WT1) and ZFY genes indicated that there had been no genetic differentiation among the continental populations. However, significant differences were observed between continental and Japanese raccoon dogs in VTN and ZFY genes implying genetic differentiation has been going between them., Conclusion: To better understand the phylogenetic relationship among raccoon dog populations, further study will be necessary.

Wild carnivores serve as reservoirs of several zoonotic Bartonella species such as Bartonella henselae , Bartonella vinsonii subsp. berkhoffii , and Bartonella rochalimae . The raccoon dog ( Nyctereutes procyonoides viverrinus ) is the most common native carnivore in Japan, but epidemiologic studies of Bartonella infections have not been performed in this animal species yet. Here, we report a molecular survey of B. rochalimae prevalence in 619 wild raccoon dogs captured from 2009 to 2017 in western Japan. Bartonella rochalimae DNA was detected in 7.1% (44/619) of the raccoon dogs examined by PCR targeting the rpoB and ssrA genes. All of the sequences obtained were identical in each of the genes. The prevalence of B. rochalimae by sex of the animals was 6.1% (21/344) in male and 8.4% (23/275) in female. The prevalence by year varied from 2% (1/45) in 2011 to 14% (4/28) in 2016. The prevalence (7.9%) of B. rochalimae in the raccoon dogs with sarcoptic mange tended to be higher than the prevalence (4.0%) in the animals without the infestation of mites, although the differences were not significant. Sequence analysis indicated that Japanese raccoon dogs in the area examined were infected with B. rochalimae carrying identical sequences in the rpoB and ssrA genes. In addition, the raccoon dog strain had few sequence variations in both genes compared to other known B. rochalimae strains detected in other parts of the world.

Background: Apicomplexan parasites of the genus Sarcocystis have an obligate two-host life-cycle and comprise about 200 species, which infect different cold- and warm-blooded hosts, including humans. Recently, morphological and molecular studies of sarcocysts in broadly spread carnivore hosts have been on the rise. The description of muscular tissues infection by Sarcocystis in the raccoon dog and the common raccoon from the Czech Republic is herein presented., Methods: During January-August 2019, 15 raccoon dogs and 1 common raccoon were examined from 5 districts (Česká Lípa, Liberec, Mladá Boleslav, Trutnov and Ústí nad Labem) of the Czech Republic. Muscle parts (diaphragm, forearm, hind-limb, tongue and heart) were examined in wet preparations under compression by light microscopy. After finding Sarcocystis sp., morphological characteristics and molecular analyses of 18S rRNA, 28S rRNA, ITS1 and cox1 loci were used to identify the species., Results: Sarcocysts were detected and identified in 1 out of 15 raccoon dogs and in the single common raccoon. Preferential infection sites were diaphragm and tongue, followed by forearm and hind limb. To our knowledge, this is the first identification of microscopic sarcocysts by multi-locus genetic analysis from both host species. Molecular analyses revealed 100% similarity at 18S rRNA, 28S rRNA, and cox1 genes with S. lutrae for both hosts and 98-100% identity at the ITS1 region of the isolate from the common raccoon., Conclusions: Both widely distributed non-indigenous wild carnivores represent new intermediate host records for S. lutrae and the first report of this parasite in a member of the family Procyonidae, but still with an unknown natural definitive host. Molecular data revealed that this parasite species appears more closely related to the Sarcocystis spp. using raptorial birds as definitive hosts. Therefore, further studies aimed at its identification, including the complete life-cycle, remain necessary.

We analyzed the genotypes of three pregnant females and their litters to investigate the phenomenon of multiple paternity in wild raccoon dogs (Nyctereutes procyonoides) using 17 microsatellite markers. If a female has mated with only one male during estrus, then the maximum number of paternal alleles will not exceed two among littermates with the same father. The results revealed two out of three litters had three or four paternal alleles at one or five microsatellite loci. Therefore, the female had mated with more than one male during estrus. To the best of our knowledge, the present study is the first to report the possibility of multiple paternity in wild raccoon dogs.

Raccoon dogs ( Nyctereutes procynoides ) are canids indigenous to Eastern Asia being one of the most invasive non-native animals in Europe and potential vectors for several hazardous parasitic and viral diseases. To present, there is a lack of studies regarding objective techniques used to appreciate the integrity and functionality of the nervous system in this species. Brainstem auditory evoked potentials (BAER) is a cost-effective, quick and noninvasive technique to assess the functionality of nervous system. The aim of the study is to provide reference values and evaluates the reliability of BAER recording with needle electrodes in clinically healthy raccoon dogs. Nine individuals were investigated for BAER under xylazine and ketamine general anesthesia. Four waves (I, II, III and V) were identified and evaluated for latencies, morphologically similarity to those previously reported for other species (dogs, horses, cats, and ferret). Our data can be used in further studies to asses objectively the auditory system function in raccoon dogs., Competing Interests: The authors declare no conflict of interest.

The goal of this study is to compare the gut microbiota of domestic blue fox (Alopex lagopus) and raccoon dog (Nyctereutes procyonoides) to provide better understanding of their intestinal gut microbiota. We analyzed the structure of fecal microbes in 40 blue foxes and 40 raccoon dogs that were raised under same conditions, using high-throughput Illumina sequencing targeting the V3-V4 region of the 16S rRNA gene. In total, 295,146 sequence reads were obtained. The average number of operational taxonomical units in the two group samples was 194 to 286. Firmicutes (blue fox 73.40%, raccoon dog 46.90%) and Bacteroidetes (blue fox 21.92%, raccoon dog 44.25%) were the most abundant phyla in the gut of blue fox and raccoon dog. At the genus level, Prevotella (blue fox 16.89%, raccoon dog 36.22%), Blautia (blue fox 9.02%, raccoon dog 13.72%), and Peptostreptococcaeae&#95;incertae&#95;sedi (blue fox 22.41%, raccoon dog 2.84%) were commonly presented in the gut of two kinds of animal. Principal coordinates analysis showed that the microbial communities were different between blue fox and raccoon dog. The Firmicutes-to-Bacteroidetes ratio was higher in blue foxes (3:1) than in raccoon dogs (1:1). Moreover, Peptostreptococcaeae&#95;incertae&#95;sedi and Prevotella, were more abundant in the gut of blue fox, whereas the abundance of Prevotella and Blautia were higher in the gut of raccoon dog. In conclusion, the present study revealed the difference of the gut microbial composition between blue fox and raccoon dog under the same diet conditions.

Sarcoptes scabiei (Acari: Sarcoptidae) causes a common contagious skin disease that affects many mammals. Here, the complete mitochondrial genome of a mite, S. scabiei var. nyctereutis, from Japanese wild raccoon dogs was analyzed. The 13,837bp circular genome contained 13 protein-coding genes, two rRNA genes, and 22 tRNA genes. For the first time, two tRNAs (alanine and tyrosine), that were thought to be absent in scabies mites from other animals, were predicted to have short, non-cloverleaf structures by in silico annotation and detected by RT-PCR, sequencing, and northern analysis. The mitochondrial genome structure of S. scabiei is similar to that of Psoroptes cuniculi and Dermatophagoides farinae. While small and unusual tRNA genes seem to be common among acariform mites, further experimental evidence for their presence is needed. Furthermore, through an analysis of the cox1 gene, we have provided new evidence to confirm the transmission of this mite between different animal hosts., (Copyright © 2019 Elsevier Inc. All rights reserved.)