Prionailurus Bengalensis Euptilurus Research Articles

Metabarcoding of pathogenic parasites based on copro-DNA analysis of wild animals in South Korea

Four species of dominant wild animals, namely, Prionailurus bengalensis euptilurus, Nyctereutes procyonoides koreensis, Hydropotes inermis argyropus, and Sus scrofa coreanus, are hosts of potential infectious agents, including helminths and protozoa. Therefore, it is necessary to analyze the infectious agents present in these wild animals to monitor and control the spread of pathogens. In the present study, fecal samples from 51 wild animals were collected from the mountains of Yangpyeong, Hoengseong, and Cheongyang in South Korea and metabarcoding of the V9 region of the 18S rRNA gene was performed to identify various parasite species that infect these wild animals. Genes from nematodes, such as Metastrongylus sp., Strongyloides spp., Ancylostoma sp., and Toxocara sp., were detected in the fecal samples from wild animals. In addition, platyhelminthes, including Spirometra sp., Echinostomatidae gen. sp., Alaria sp., Neodiplostomum sp., and Clonorchis sp., and protozoa, including Entamoeba sp., Blastocystis sp., Isospora sp., Tritrichomonas sp., Pentatrichomonas sp., and Cryptosporidium sp., were detected. In the present study, various parasites infecting wild animals were successfully identified using metabarcoding. Our technique may play a crucial role in monitoring parasites within wild animals, especially those causing zoonoses.

New insights into predator-prey dynamics: First evidence of a leopard cat hunting coypus.

We present the first documented evidence of interactions between the leopard cat (Prionailurus bengalensis euptilurus) and the invasive coypu (Myocastor coypus) in South Korea, captured through camera traps in Hwapocheon wetland park from May 2015 to April 2017. Two interactions were recorded: one showing a leopard cat carrying a carcass of sub-adult coypu and the other a 4-min sequence of predation and defense between two species. The observed interactions indicate active predatory behavior by the leopard cat against coypus and cooperative defense by coypus. These findings shed new light on predator-prey dynamics, highlighting the leopard cat's potential role as a predator of coypus and coypus' defensive abilities. Understanding these relationships could facilitate more effective management of invasive species and offer broader implications for ecosystem dynamics and conservation strategies.

A cost-effective blood DNA methylation-based age estimation method in domestic cats, Tsushima leopard cats (Prionailurus bengalensis euptilurus) and Panthera species, using targeted bisulphite sequencing and machine learning models.

Individual age can be used to design more efficient and suitable management plans in both insitu and ex situ conservation programmes for targeted wildlife species. DNA methylation is a promising marker of epigenetic ageing that can accurately estimate age from small amounts of biological material, which can be collected in a minimally invasive manner. In this study, we sequenced five targeted genetic regions and used 8-23 selected CpG sites to build age estimation models using machine learning methods at only about $3-7 per sample. Blood samples of seven Felidae species were used, ranging from small to big, and domestic to endangered species: domestic cats (Felis catus, 139 samples), Tsushima leopard cats (Prionailurus bengalensis euptilurus, 84 samples) and five Panthera species (96 samples). The models achieved satisfactory accuracy, with the mean absolute error of the most accurate models recorded at 1.966, 1.348 and 1.552 years in domestic cats, Tsushima leopard cats and Panthera spp. respectively. We developed the models in domestic cats and Tsushima leopard cats, which were applicable to individuals regardless of health conditions; therefore, these models are applicable to samples collected from individuals with diverse characteristics, which is often the case in conservation. We also showed the possibility of developing universal age estimation models for the five Panthera spp. using only two of the five genetic regions. We do not recommend building a common age estimation model for all the target species using our markers, because of the degraded performance of models that included all species.

GLOBAL RETROSPECTIVE REVIEW OF SEVERE ACUTE RESPIRATORY SYNDROME SARS COV-2 INFECTIONS IN NONDOMESTIC FELIDS: MARCH 2020-FEBRUARY 2021.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in nondomestic felids have been documented in North America, South America, Africa, Europe, and Asia. Between March 2020 and February 2021, at nine institutions across three continents, infection was confirmed in 16 tigers (Panthera tigris), 14 lions (Panthera leo), three snow leopards (Panthera uncia), one cougar (Puma concolor), and one Amur leopard cat (Prionailurus bengalensis euptilurus) ranging from 2 to 21 yr old (average, 10 yr). Infection was suspected in an additional 12 tigers, 4 lions, and 9 cougars. Clinical signs (in order of most to least common) included coughing, ocular and/or nasal discharge, wheezing, sneezing, decreased appetite, lethargy, diarrhea, and vomiting. Most felids recovered uneventfully, but one geriatric tiger with comorbidities developed severe dyspnea and neurologic signs necessitating euthanasia. Clinical signs lasted 1-19 d (average, 8 d); one tiger was asymptomatic. Infection was confirmed by various methods, including antigen tests and/or polymerase chain reaction (PCR) of nasal or oral swabs, tracheal wash, and feces, or virus isolation from feces or tracheal wash. Infection status and resolution were determined by testing nasal swabs from awake animals, fecal PCR, and observation of clinical signs. Shedding of fecal viral RNA was significantly longer than duration of clinical signs. Postinfection seropositivity was confirmed by four institutions including 11 felids (5 lions, 6 tigers). In most instances, asymptomatic or presymptomatic keepers were the presumed or confirmed source of infection, although in some instances the infection source remains uncertain. Almost all infections occurred despite using cloth facemasks and disposable gloves when in proximity to the felids and during food preparation. Although transmission may have occurred during momentary lapses in personal protective equipment compliance, it seems probable that cloth masks are insufficient at preventing transmission of SARS-CoV-2 from humans to nondomestic felids. Surgical or higher grade masks may be warranted when working with nondomestic felids.

A comparative profile of urinary scent signals of two endangered Japanese populations of leopard cat

Two subspecies of mainland leopard cats (Prionailurus bengalensis), Iriomote cats (Prionailurus bengalensis iriomotensis), and Tsushima leopard cats (Prionailurus bengalensis euptilurus), are small, endangered felids that are endemic to the Japanese islands, Iriomote-jima and Tsushima, respectively. Both subspecies have an estimated population size of only about 100 individuals each. Therefore, it is crucial to expand our understanding of these subspecies to successfully conserve them. While their morphology, anatomy, ecology, pathology, and genetics are well studied, little is known about their physiology regulated in subspecies-specific manners. This study characterized their urinary volatile and nonvolatile organic compounds, with which individuals express their physiological status and territorial ownership. We found significantly higher levels of 3-mercapto-3-methyl-1-butanol and its structural analogs, which are scent signals of domestic cats, in the urinary headspace of Iriomote cats than in that of Tsushima leopard cats. There were no differences in the urinary levels of felinine, an amino acid that acts as a precursor of 3-mercapto-3-methyl-1-butanol, suggesting the variation of catalytic activities for decomposing felinine to 3-mercapto-3-methyl-1-butanol between subspecies. Both subspecies exhibited physiological proteinuria due to the secretion of carboxylesterase 5A, also known as cauxin, for the production of felinine. In conclusion, while the two endangered Japanese subspecies of mainland leopard cats share similar genetic backgrounds, urinary contents of metabolites vary between the subspecies. We suggest that after the separation into two subspecies, the regulation of the metabolic pathway for 3-mercapto-3-methyl-1-butanol production likely evolved differently in each subspecies in response to their specific environments.

Neoplasia in captive Tsushima leopard cats (Prionailurus bengalensis euptilurus)

The Tsushima leopard cat (Prionailurus bengalensis euptilurus) is a subspecies of the mainland leopard cat that lives on the small island of Tsushima, Japan. Captive breeding has been attempted in zoos in Japan because only approximately 100 animals remain in the wild and the Tsushima leopard cat is an endangered species. There are very few reports on diseases, including tumours, of this species. We analysed the deaths of 58 Tsushima leopard cats and confirmed that nine had neoplastic disease. The average age at death of the animals with neoplasia was 14 years and tumours were the primary cause of death in all animals. Eight of the nine cases involved primary tumours of the pancreas, liver, gallbladder, tongue and salivary glands, suggesting that Tsushima leopard cats may have a predilection for digestive system tumours. This is the first report of neoplastic disease in the Tsushima leopard cat.

Feline Foamy Virus Transmission in Tsushima Leopard Cats (Prionailurus bengalensis euptilurus) on Tsushima Island, Japan.

Tsushima leopard cats (TLC; Prionailurus bengalensis euptilurus) only inhabit Tsushima Island, Nagasaki, Japan and are critically endangered and threatened by infectious diseases. The feline foamy virus (FFV) is widely endemic in domestic cats. Therefore, its transmission from domestic cats to TLCs may threaten the TLC population. Thus, this study aimed to assess the possibility that domestic cats could transmit FFV to TLCs. Eighty-nine TLC samples were screened, and FFV was identified in seven (7.86%). To assess the FFV infection status of domestic cats, 199 domestic cats were screened; 14.07% were infected. The phylogenetic analysis revealed that the FFV partial sequence from domestic cats and TLC sequences clustered in one clade, suggesting that the two populations share the same strain. The statistical data minimally supported the association between increased infection rate and sex (p = 0.28), indicating that FFV transmission is not sex dependent. In domestic cats, a significant difference was observed in FFV detection in feline immunodeficiency virus (p = 0.002) and gammaherpesvirus1 infection statuses (p = 0.0001) but not in feline leukemia virus infection status (p = 0.21). Monitoring FFV infection in domestic cats and TLC populations is highly recommended as part of TLC surveillance and management strategies.

Quality of cauda epididymal sperm immediately after collection and after freezing-thawing from Amur leopard cats (Prionailurus bengalensis euptilurus) and a local population of the subspecies Tsushima leopard cats.

In this study, cauda epididymal sperm were collected from Amur leopard cats with various causes of death as well as Tsushima leopard cats that underwent castration surgery, and sperm quality was compared with that in domestic cats. A sufficient number of sperm similar to those in domestic cats could be collected from the cauda epididymis of Amur leopard cats. However, in old leopard cats, no or very few cauda epididymal sperm were recovered, although there were no differences in sperm motility and sperm abnormality. There were no significant differences in sperm quality immediately after collection and after freezing-thawing of cauda epididymal sperm compared with corresponding estimates in domestic cats.

Virome Profiling of an Amur leopard cat Reveals Multiple Anelloviruses and a Bocaparvovirus.

As a small top predator, Amur leopard cat (Prionailurus bengalensis euptilurus) is widely distributed in northeast Asia and plays an important role in the control of small rodent populations and in the maintenance of ecological equilibrium. However, the viruses harbored by this creature have been rarely investigated. Here, we report the DNA and RNA eukaryotic virome profiling of an injured Amur leopard cat followed by PCR validation, which revealed diverse anelloviruses in multiple organs and a bocaparvovirus in the lymph, but no RNA viruses. These anelloviruses have diverse genomic structures and are classified into four phylogroups with viruses of various felines, while the bocaparvovirus is extremely similar to those recovered from diarrheal domestic cats, illustrating the transmission of the virus between domestic animals and wildlife. These data provide the first insight into the genetic diversity of Amur leopard cat viruses, highlighting the need for further investigation of wild animals.

Comparison of the Genetic Diversity of the Captive and Wild Populations of the Tsushima Leopard Cat Using a GRAS-Di Analysis.

Simple SummaryThe Tsushima leopard cat (Prionailurus bengalensis euptilurus) is exclusively found on the Tsushima Island in Japan; however, its population has been declining in recent years and is in danger of extinction. Hence, effort is underway for both in situ and ex situ conservation. Genetic management is also important in the management of captive populations, and various studies are being conducted regarding the same. In our previous study, we developed a reference genome of the Tsushima leopard cat and used GRAS-Di analysis, a genome-wide analysis, to genetically evaluate the wild populations. In this study, we attempted to improve the Tsushima leopard cat reference genome and compared the captive and wild populations by GRAS-di analysis. The results showed that the captive population had almost the same genetic diversity as the wild population and managed to remain in good condition.The Tsushima leopard cat (Prionailurus bengalensis euptilurus) (TLC) is a regional population of the Amur leopard cat (P. bengalensis euptilurus) that lives only on the Tsushima Island in Japan and is threatened with extinction. Because the TLC population is small, genetic management is important. In this study, we obtained the draft genome of the TLC and identified single-nucleotide polymorphism (SNP) markers using a genotyping by random amplicon sequencing-direct (GRAS-Di) analysis. We genotyped 31 captive individuals and 50 wild individuals, of which 48 were from a previous study. The identified SNPs were used to clarify the genetic diversity and genetic structure of the wild and captive populations of the TLC. The size of the genome was estimated to be about 2.42 Gb. The number of SNP markers developed was 139, and although PID and probability of exclusion obtained using these SNP markers were not as high as those reported in the studies of other wild species, these SNP markers could be used to identify individuals and parentage. Moreover, the genetic diversity indices of the captive population were similar to those of the wild population. These SNP markers will be useful for understanding the ecology of the TLC and planning conservation strategies.

Fecal Glucocorticoid Metabolites as a Noninvasive Indicator of Stress in the Tsushima Leopard Cats (Prionailurus bengalensis euptilurus): Application to Health Care.

Simple SummaryThe Tsushima leopard cat (Prionailurus bengalensis euptilurus) and the Iriomote cat (P. b. iriomotensis) are the only two wild felids living in Japan. Both species are endangered. The Tsushima leopard cat is undergoing captive breeding (ex situ conservation) in zoos with the aim of returning it to the wild, in addition to conservation activities in the habitat (in situ conservation). In order to promote captive breeding, it is important to elucidate the reproductive physiology and to evaluate stress conditions. We investigated the possibility of measuring glucocorticoid metabolites (GCMs), which are stress hormones, using feces in Tsushima leopard cats. We examined changes in fecal GCM in individuals who underwent a health examination under anesthesia as an unavoidable stressful condition during normal health care. Fecal GCM concentrations increased 1–2 days after the health examination. Fecal GCM concentrations were higher in diseased individuals than in apparently healthy individuals. Individuals diagnosed with disk herniation also showed a decrease in fecal GCM concentrations after treatment with therapeutics. These results indicate that the measurement of fecal GCMs is useful for improving the husbandry and health management of the Tsushima leopard cat.This study investigates whether the measurement of glucocorticoid metabolites (GCMs) in feces is a useful method for the noninvasive evaluation of stress in the endangered Tsushima leopard cats (Prionailurus bengalensis euptilurus). Feces were collected from six seemingly healthy and five diseased (renal dysfunction, adrenal tumor, hernia, feline immunodeficiency virus (FIV), feline leukemia virus (FeLV)) Tsushima leopard cats in captivity. Fecal GCMs were measured by enzyme immunoassay (EIA) for cortisol. Individuals that experienced a physical examination under anesthesia showed increased fecal GCMs 1–2 days after the event. An individual diagnosed with disk herniation showed decreased fecal GCMs after medical administration. The mean fecal GCM concentrations for six healthy animals and five diseased animals were 0.66 ± 0.08 and 2.65 ± 0.76 μg/g, respectively, which was significantly different. Cortisol and corticosterone were not clearly detected in the feces examined by the use of the HPLC-EIA analysis. GCMs may be excreted in the feces; however, the exact identification of these substances is not achieved. The results suggest that the measurement of fecal GCMs is useful for the husbandry and health management of this species.

Successful Laparoscopic Oviductal Artificial Insemination in the Endangered Tsushima Leopard Cat (Prionailurus bengalensis euptilurus)

Simple SummaryThe Tsushima leopard cat is an endangered wild felid that lives solely on Tsushima Island, Nagasaki, Japan. Approximately, only 100 Tsushima leopard cats can be found in the wild, and there are concerns that the population will further reduce due to habitat degradation and traffic accidents. In 1994, Japan’s Ministry of the Environment (MOE) developed a conservation and breeding project for the Tsushima leopard cat. The MOE is working with the Japanese Association of Zoo and Aquariums for ex situ conservation of this species. However, considering genetic diversity, it is difficult to conduct captive breeding programs using only natural breeding; hence, assisted reproductive technologies are required. This study aimed to breed Tsushima leopard cats using artificial insemination (AI) by depositing sperms into the oviducts. Ovulation was artificially induced in two females, laparoscopically inseminated with fresh sperms into the oviducts. The pregnancies were monitored via fecal levels of progestogens and radiography. One female had spontaneous delivery of a female kitten 66 days post-AI. This is the first successful case of AI in a Tsushima leopard cat.The Tsushima leopard cat (Prionailurus bengalensis euptilurus) is an endangered wild felid that lives solely on Tsushima Island, Nagasaki, Japan. Japan’s Ministry of the Environment is working with the Japanese Association of Zoo and Aquariums for ex situ conservation of this species. However, considering genetic diversity, it is difficult to conduct captive breeding programs by natural breeding alone; hence, assisted reproductive technologies are required. This study aimed to breed Tsushima leopard cats, which otherwise cannot be paired, using laparoscopic oviductal artificial insemination (AI). Female Tsushima leopard cats (female 1, aged 7 years; female 2, aged 6 years) were treated with 200 IU equine chorionic gonadotropin, followed by administration of 1000 IU porcine luteinizing hormone (pLH) after 96 h to induce ovulation. Laparoscopic AI was performed 32 h post-pLH administration. Females 1 and 2 were inseminated in the oviduct with 2.4 × 10⁶ and 3.3 × 10⁶ motile spermatozoa, respectively, collected from two males. Pregnancy was confirmed by radiography 45 and 51 days post-AI in females 1 and 2, respectively; one fetus was found in female 2. Moreover, female 2 had spontaneous delivery of a female kitten 66 days post-AI. This is the first successful case of AI in a Tsushima leopard cat.

Genetic Diversity and Genetic Structure of the Wild Tsushima Leopard Cat from Genome-Wide Analysis.

Simple SummaryThe Tsushima leopard cat, Prionailurus bengalensis euptilurus, is a small regional population of the Amur leopard cat and is only found on Tsushima Island in Japan. A breeding program will require adequate information on parentage, kinship, and inbreeding for this population. Hence, there is an urgent need to develop this information in order to conserve the population and its genetic diversity. We performed GRAS-Di analysis to investigate the genetic diversity and genetic structure of the Tsushima leopard cat. We identified between 133 and 158 single-nucleotide polymorphism (SNP) markers in three different genotyping methods. These SNP markers can be used in identification of individuals and parentage. In addition, structure analysis using these markers demonstrated the similar genetic composition of the samples from 48 Tsushima leopard cats, and indicated Tsushima leopard cats have no subpopulations. We have provided genetic markers that are useful for conservation of the Tsushima leopard cat, such as individual identification and parentage. Moreover, we have also clarified units for conservation of the Tsushima leopard cat population from structure analysis.The Tsushima leopard cat (Prionailurus bengalensis euptilurus) lives on Tsushima Island in Japan and is a regional population of the Amur leopard cat; it is threatened with extinction. Its genetic management is important because of the small population. We used genotyping by random amplicon sequencing-direct (GRAS-Di) to develop a draft genome and explore single-nucleotide polymorphism (SNP) markers. The SNPs were analyzed using three genotyping methods (mapping de novo, to the Tsushima leopard cat draft genome, and to the domestic cat genome). We examined the genetic diversity and genetic structure of the Tsushima leopard cat. The genome size was approximately 2.435 Gb. The number of SNPs identified was 133–158. The power of these markers was sufficient for individual and parentage identifications. These SNPs can provide useful information about the life of the Tsushima leopard cat and the pairings and for the introduction of founders to conserve genetic diversity with ex situ conservation. We identified that there are no subpopulations of the Tsushima leopard cat. The identifying units will allow for a concentration of efforts for conservation. SNPs can be applied to the analysis of the leopard cat in other regions, making them useful for comparisons among populations and conservation in other small populations.

Food Habit Analysis of the Amur Leopard Cat Prionailurus bengalensis euptilurus in the Russian Far East

The ecology of the Amur leopard cat (Prionailurus bengalensis euptilurus) has not been studied enough, especially in the Russian Far East. This study presents the food habits of the Amur leopard cat, based on the analysis of scat samples collected in the Primorsky Region of Russia. Mammals and birds had the highest contribution in the diet (89.3 and 30.6% frequency of occurrence, respectively). Rodents had an important role in the diet of the leopard cat throughout the year. The proportion of birds and poikilothermic animals increased during the snow-free season. The total list of animals recorded to be consumed by the leopard cat is presented, consisting of 63 species (seven fishes, eight amphibians, one reptile, 21 birds, and 26 mammals).

A Clonorchiasis Case of a Leopard Cat, Prionailurus bengalensis euptilurus, Diagnosed by Ultrasonography and Egg Detection in Republic of Korea

A clonorchiasis case in a captive leopard cat, Prionailurus bengalensis euptilurus, was confirmed by ultrasonographic findings and egg morphologies found in the bile juice sample in the Korea. The leopard cat was introduced from the wild habitat of Gyeongsangnam-do, to Cheongju Zoo in Cheongju-si, Chungcheongbuk-do, Korea in August 2014. Physical examinations were basically performed for quarantine and check-up health. The cat was comparatively good in health except anorexia. The cyst-like bile duct dilation and the increased echogenicity of gall bladder wall and hepatic parenchyma were observed by ultrasonography. Ultrasound-guided needle biopsy was conducted for collecting bile juice and the specimens were observed under light microscope. The numerous small trematode eggs were detected in the bile juice sample of the light microscopy. The eggs were 25–33 (28±3) μm by 18–22 (20±1) μm in size and showed typical characteristics of Clonorchis sinensis egg, i.e., a dominantly developed operculum, shoulder rim and dust-like wrinkles in surface. To treat the liver fluke infection, 20 mg/kg of praziquantel was orally administered only once to the case. Follow-up studies including fecal examinations were conducted during 2 years after treatment. But no more eggs were detected from the case. In the present study, we described the first clonorchiasis case of leopard cat, which was confirmed by ultrasonographic findings and egg morphologies from the bile juice sample in Korea.

Sex identification using ZFX and ZFY genes in leopard cats (Prionailurus bengalensis euptilurus) in Korea.

In this study, we used multiplex polymerase chain reaction (PCR) to determine the sex of leopard cats (Prionailurus bengalensis euptilurus) in Korea. When we applied PCR using primers based on the intronic region between ZFX and ZFY, only one DNA band of 143 bp was detected in females, compared with two specific bands of 143 and 310 bp in males, indicating that the band patterns were clearly different between females and males. Since the set of primers also proved compatible with tissue and fecal samples, the results of the present study demonstrate that the present PCR could be a convenient tool for identifying the sex of leopard cats.

Efficient immortalization of cells derived from critically endangered Tsushima leopard cat (Prionailurus bengalensis euptilurus) with expression of mutant CDK4, Cyclin D1, and telomerase reverse transcriptase

Tsushima leopard cat is the subspecies of Amur cats, and it is classified as the most highest class of critically endangered animals. Although the protection activity is highly recognized, the number of animals is decreasing due to the human activity and invasion of domestic cats and infectious disease. In this study, we succeeded primary culture of normal fibroblasts derived from the Tsushima leopard cat (Prionailurus bengalensis euptilurus). Furthermore, we introduced the human derived mutant Cyclin Dependent Kinase 4, Cyclin D1, and telomere reverse transcriptase. We showed that the expression of these three genes efficiently immortalized cells derived from Tsushima leopard cat. Furthermore, we showed that the chromosome pattern of the established cells is identical with the original one. These data indicate that our method of immortalization is useful to establish cell lines from critically endangered cats, which potentially contributes to the re-generation of critically endangered animals from cultured cell with reproductive technique, such as somatic cloning.

Primary study of seroprevalence to virus pathogens in wild felids of South Primorie, Russia

Seroprevalence to nine different virus pathogens was estimated for Russian big cats (Amur tiger (Panthera tigris altaica Temminck, 1844) and far-eastern leopard (Panthera pardus orientalis (Schiegel, 1857))) in Southern Primorie, Russia (n = 25), in 2008–2016. Serum samples from smaller cats (Eurasian lynx (Lynx lynx (Linnaeus, 1758)) and far-eastern wildcat (leopard cat) (Prionailurus bengalensis euptilurus (Elliot, 1871))) were also tested for these pathogens (n = 19) during the same period. Felids of Russian Southern Primorie showed seroprevalence to eight out of nine tested pathogens, including highly dangerous feline immunodeficiency virus, feline leukemia virus, and canine distemper virus. Antibodies to feline panleukopenia virus were found to be much more widespread in cats (45%) than antibodies to any other virus. They were detected in samples taken from tigers, leopards, and far-eastern wildcats but not lynxes. Antibodies to pseudorabies virus were detected only in Amur tiger (29%), whose main prey is the most common carrier of the virus (wild boar), unlike for the other studied cats’ species.

Identification of Felis catus Gammaherpesvirus 1 in Tsushima Leopard Cats (Prionailurus bengalensis euptilurus) on Tsushima Island, Japan.

Felis catus gammaherpesvirus 1 (FcaGHV1) is a widely endemic infection of domestic cats. Current epidemiological data identify domestic cats as the sole natural host for FcaGHV1. The Tsushima leopard cat (TLC; Prionailurus bengalensis euptilurus) is a critically endangered species that lives only on Tsushima Island, Nagasaki, Japan. Nested PCR was used to test the blood or spleen of 89 TLCs for FcaGHV1 DNA; three (3.37%; 95% CI, 0.70–9.54) were positive. For TLC management purposes, we also screened domestic cats and the virus was detected in 13.02% (95% CI, 8.83–18.27) of 215 cats. Regarding phylogeny, the partial sequences of FcaGHV1 from domestic cats and TLCs formed one cluster, indicating that similar strains circulate in both populations. In domestic cats, we found no significant difference in FcaGHV1 detection in feline immunodeficiency virus-infected (p = 0.080) or feline leukemia virus-infected (p = 0.163) cats, but males were significantly more likely to be FcaGHV1 positive (odds ratio, 5.86; 95% CI, 2.27–15.14) than females. The higher frequency of FcaGHV1 detection in domestic cats than TLCs, and the location of the viral DNA sequences from both cats within the same genetic cluster suggests that virus transmission from domestic cats to TLCs is likely.

Characterization and cryopreservation of Amur leopard cats (Prionailurus bengalensis euptilurus) semen collected by urethral catheterization

The Amur leopard cat (Prionailurus bengalensis euptilurus) is a globally endangered species, and there is thus an urgent need to increase its population. The objectives of this study were to: (1) confirm the utility of urethral catheterization (UC) for semen collection from Amur leopard cats; (2) investigate proper dilution media for fresh semen; and (3) investigate the effectiveness of sperm cryopreservation, including examining the effect of glycerol concentration. Six adult males in captivity (mean weight 5.03 ± 0.44 kg, aged 2–6 years) were included. All study procedures were performed during the breeding season (February to April) over two consecutive years. Semen samples (n = 28) were collected four or five times from each animal (four times for two animals and five times for four animals) by UC under general anaesthesia, and their characteristics (including sperm motility) were evaluated. First, the sperm motility in semen diluted in Ham's F-10 or phosphate buffer saline (PBS) was compared. Next, semen diluted with TEST-yolk buffer containing 2%, 4%, or 6% glycerol was frozen in a liquid nitrogen tank, and sperm motility and acrosome integrity were evaluated after thawing. No difference in motility was observed between sperm diluted in Ham's F-10 and PBS. The percentages of sperm motility and kinetic values in semen frozen in 2% or 4% glycerol were higher than those in semen stored in 6% glycerol. In conclusion, the UC method for semen collection is recommendable for Amur leopard cats and should be useful for artificial insemination. Although sperm motility decreased after thawing, samples thus preserved may be usable for advanced reproductive techniques, such as intracytoplasmic sperm injection or in-vitro fertilization.