Outbreak of Natural Severe Fever with Thrombocytopenia Syndrome Virus Infection in Farmed Minks, China.

We isolated severe fever with thrombocytopenia syndrome virus (SFTSV) from farmed minks in China, providing evidence of natural SFTSV infection in farmed minks. Our findings support the potential role of farmed minks in maintaining SFTSV and are helpful for the development of public health interventions to reduce human infection.

Genomic Context of SARS-CoV-2 Outbreaks in Farmed Mink in Spain during Pandemic: Unveiling Host Adaptation Mechanisms.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects various mammalian species, with farmed minks experiencing the highest number of outbreaks. In Spain, we analyzed 67 whole genome sequences and eight spike sequences from 18 outbreaks, identifying four distinct lineages: B.1, B.1.177, B.1.1.7, and AY.98.1. The potential risk of transmission to humans raises crucial questions about mutation accumulation and its impact on viral fitness. Sequencing revealed numerous not-lineage-defining mutations, suggesting a cumulative mutation process during the outbreaks. We observed that the outbreaks were predominantly associated with different groups of mutations rather than specific lineages. This clustering pattern by the outbreaks could be attributed to the rapid accumulation of mutations, particularly in the ORF1a polyprotein and in the spike protein. Notably, the mutations G37E in NSP9, a potential host marker, and S486L in NSP13 were detected. Spike protein mutations may enhance SARS-CoV-2 adaptability by influencing trimer stability and binding to mink receptors. These findings provide valuable insights into mink coronavirus genetics, highlighting both host markers and viral transmission dynamics within communities.

A Deletion Encompassing the Furin Cleavage Site in the Spike Encoding Gene Does Not Alter SARS-CoV-2 Replication in Lung Tissues of Mink and Neutralization by Convalescent Human Serum Samples.

SARS-CoV-2 has been shown to lose the furin polybasic cleavage site (FCS) following adaptation on cell culture. Deletion occurring in this region, which may include also the FCS flanking regions, seem not to affect virus replication in vitro; however, a chimeric SARS-CoV-2 virus without the sole FCS motif has been associated with lower virulence in mice and lower neutralization values. Moreover, SARS-CoV-2 virus lacking the FCS was shed to lower titers from experimentally infected ferrets and was not transmitted to cohoused sentinel animals, unlike wild-type virus. In this study, we investigated the replication kinetics and cellular tropism of a SARS-CoV-2 isolate carrying a 10-amino acid deletion in the spike protein spanning the FCS in lung ex vivo organ cultures of mink. Furthermore, we tested the neutralization capabilities of human convalescent SARS-CoV-2 positive serum samples against this virus. We showed that this deletion did not significantly hamper neither ex vivo replication nor neutralization activity by convalescent serum samples. This study highlights the importance of the preliminary phenotypic characterization of emerging viruses in ex vivo models and demonstrates that mink lung tissues are permissive to the replication of a mutant form of SARS-CoV-2 showing a deletion spanning the FCS. Notably, we also highlight the need for sequencing viral stocks before any infection study as large deletions may occur leading to the misinterpretation of results.

The basis of mink susceptibility to SARS-CoV-2 infection.

Of all known airborne diseases in the twenty-first century, coronavirus disease 19 (COVID-19) has the highest infection and death rate. Over the past few decades, animal origin viral diseases, notably those of bats-linked, have increased many folds in humans with cross-species transmissions noted and the ongoing COVID-19 pandemic has emphasized the importance of understanding the evolution of natural hosts in response to viral pathogens. Cross-species transmissions are possible due to the possession of the angiotensin-converting enzyme 2 (ACE2) receptor in animals. ACE2 recognition by SARS-CoV-2 is a critical determinant of the host range, interspecies transmission, and viral pathogenesis. Thus, the phenomenon of breaking the cross-species barrier is mainly associated with mutations in the receptor-binding domain (RBD) of the spike (S) protein that interacts with ACE2. In this review, we raise the issue of cross-species transmission based on sequence alignment of S protein. Based on previous reports and our observations, we can conclude that the occurrence of one of two mutations D614G or Y453F is sufficient for infection of minks by SARS-CoV-2 from humans. Unfortunately, D614G is observed in the world's most common line of virus B.1.1.7 and the latest SARS-CoV-2 variants B.1.617.1, B.1.617.2, and B.1.617.3 too.

Histopathology and localization of SARS-CoV-2 and its host cell entry receptor ACE2 in tissues from naturally infected US-farmed mink (Neovison vison).

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes respiratory disease in mink similar to human COVID-19. We characterized the pathological findings in 72 mink from US farms with SARS-CoV-2 outbreaks, localized SARS-CoV-2 and its host cellular receptor angiotensin-converting enzyme 2 (ACE2) in mink respiratory tissues, and evaluated the utility of various test methods and specimens for SARS-CoV-2 detection in necropsy tissues. Of SARS-CoV-2-positive animals found dead, 74% had bronchiolitis and diffuse alveolar damage (DAD). Of euthanized SARS-CoV-2-positive animals, 72% had only mild interstitial pneumonia or minimal nonspecific lung changes (congestion, edema, macrophages); similar findings were seen in SARS-CoV-2-negative animals. Suppurative rhinitis, lymphocytic perivascular inflammation in the lungs, and lymphocytic infiltrates in other tissues were common in both SARS-CoV-2-positive and SARS-CoV-2-negative animals. In formalin-fixed paraffin-embedded (FFPE) upper respiratory tract (URT) specimens, conventional reverse transcription-polymerase chain reaction (cRT-PCR) was more sensitive than in situ hybridization (ISH) or immunohistochemistry (IHC) for detection of SARS-CoV-2. FFPE lung specimens yielded less detection of virus than FFPE URT specimens by all test methods. By IHC and ISH, virus localized extensively to epithelial cells in the nasal turbinates, and prominently within intact epithelium; olfactory mucosa was mostly spared. The SARS-CoV-2 receptor ACE2 was extensively detected by IHC within turbinate epithelium, with decreased detection in lower respiratory tract epithelium and alveolar macrophages. This study expands on the knowledge of the pathology and pathogenesis of natural SARS-CoV-2 infection in mink and supports their further investigation as a potential animal model of SARS-CoV-2 infection in humans.

Lessons from Animal Culling during Human Pandemics: Is Vaccination a Viable Option for Animals?

Animal reservoirs for respiratory and coronavirus have been major health concerns. Zoonosis due to coronavirus involves bats, civet cat, camels, pangolins and now the minks. In the same vein, influenza pandemics occur when a new strain of the influenza virus is transmitted to humans from another animal species. Species thought to be of particular importance in the emergence of new human influenza strains are swine and poultry and these hosts are often culled during epidemics or pandemics. It is often too easy for humans to forget that millions of animals can die or be slaughtered in human pandemics, including the recent cull of minks in Europe and chickens in Asia. To co-exist with nature in a sustainable way, we must respect our animals by ensuring their welfare and immunizing them against pathogens where possible. Zoonotic diseases are here to stay and will continue to cause major epidemics and pandemics. The other side of the coin is that reverse zoonosis can also have devastating effects on animal populations if pandemics are not effectively prevented and controlled. Unfortunately, none of the COVID-19 vaccines in production are set aside to save the minks. We advocate that animals be immunized to save human lives.

Detection of Anti-Leishmania infantum Antibodies in Wild European and American Mink (Mustela lutreola and Neovison vison) from Northern Spain, 2014-20.

The European mink (Mustela lutreola) is listed as a critically endangered species because of ongoing population reduction from habitat degradation and the effects of introduced species, such as American mink (Neovison vison). This small, fragmented population becomes vulnerable to many other threats, including diseases. Leishmaniosis is a zoonotic disease caused by the protozoan parasite Leishmania infantum found in the Mediterranean area, which affects many mammals, including wild small mammals. Furthermore, clinical disease caused by L. infantum has recently been described in other mustelids. To assess the exposure to Leishmania sp. infection in mink species in northern Spain, blood samples from 139 feral American mink and 42 native European mink from north Spain were evaluated for Leishmania sp. infection using enzyme-linked immunosorbent assays against Leishmania spp. antibodies, with 52.4% of American mink and 45.3% of European mink being found seropositive. This finding raises questions regarding how the disease may affect these species and the potential repercussions for conservation efforts. Despite a high seroprevalence being observed in wild mink of both species in this study, association with clinical or pathologic signs of disease has yet to be elucidated.

Anthropogenic Infection of Cats during the 2020 COVID-19 Pandemic.

COVID-19 is a severe acute respiratory syndrome (SARS) caused by a new coronavirus (CoV), SARS-CoV-2, which is closely related to SARS-CoV that jumped the animal-human species barrier and caused a disease outbreak in 2003. SARS-CoV-2 is a betacoronavirus that was first described in 2019, unrelated to the commonly occurring feline coronavirus (FCoV) that is an alphacoronavirus associated with feline infectious peritonitis (FIP). SARS-CoV-2 is highly contagious and has spread globally within a few months, resulting in the current pandemic. Felids have been shown to be susceptible to SARS-CoV-2 infection. Particularly in the Western world, many people live in very close contact with their pet cats, and natural infections of cats in COVID-19-positive households have been described in several countries. In this review, the European Advisory Board on Cat Diseases (ABCD), a scientifically independent board of experts in feline medicine from 11 European Countries, discusses the current status of SARS-CoV infections in cats. The review examines the host range of SARS-CoV-2 and human-to-animal transmissions, including infections in domestic and non-domestic felids, as well as mink-to-human/-cat transmission. It summarises current data on SARS-CoV-2 prevalence in domestic cats and the results of experimental infections of cats and provides expert opinions on the clinical relevance and prevention of SARS-CoV-2 infection in cats.

Bats, pangolins, minks and other animals - villains or victims of SARS-CoV-2?

Coronavirus disease-19 (COVID-19) is caused by the severe acute Respiratory syndrome coronavirus-2 (SARS-CoV-2), which has become unstoppable, spreading rapidly worldwide and, consequently, reaching a pandemic level. This review aims to provide the information available so far on the likely animal origin of SARS-CoV-2 and its possible hosts/reservoirs as well as all natural animal infections and experimental evidence using animal models. Horseshoe bats from the species Rhinolophus affinis seem to be a natural reservoir and pangolins (Manis javanica) appear to be an intermediate host of SARS-CoV-2. Humans remain the most likely spreading source of SARS-CoV-2 to other humans and also to domestic, zoo and farm animals. Indeed, human-to-animal transmission has been reported in cats, dogs, tigers, lions, a puma and minks. Animal-to-human transmission is not a sustained pathway, although mink-to-human transmission remains to be elucidated. Through experimental infections, other animals seem also to be susceptible hosts for SARS-CoV-2, namely ferrets, some non-human primate species, hamsters and transgenic mice, while dogs, pigs and poultry are resistant. A One Health perspective must be implemented in order to develop epidemiological surveillance and establish disease control mechanisms to limit zoonotic transmission. Moreover, research in this field is important to better understand SARS-CoV-2 and to obtain the long-awaited vaccine and specific treatment.

SARS-CoV-2 infection in farmed minks, associated zoonotic concerns, and importance of the One Health approach during the ongoing COVID-19 pandemic.

The coronavirus disease 2019 (COVID-19) pandemic has now affected over 72.5 million people worldwide, with nearly 1.6 million deaths reported globally as of December 17, 2020. SARS-CoV-2 has been implicated to have originated from bats and pangolins, and its intermediate animal hosts are being investigated. Crossing of the species barrier and exhibition of zoonosis have been reported in SARS-CoV-2 in farm (minks), domesticated (cats and dogs), and wild animals (tigers, puma, and lions). Recently, the rapid spread of SARS-CoV-2 infection was reported in mink farms, which led to the death of a myriad minks. The clinical and pathological findings of SARS-CoV-2 infection and the rapid animal-to-animal transmission in minks are almost similar to the findings observed in patients with COVID-19. Additionally, the rapid virus transmission among minks and the associated mutations resulted in a new mink-associated variant that was identified in both minks and humans, thereby providing evidence of mink-to-human transmission of SARS-CoV-2. The new mink-associated SARS-CoV-2 variant with a possible reduced sensitivity to neutralizing antibodies poses serious risks and is expected to have a direct effect on the diagnostic techniques, therapeutics, and vaccines that are currently under development. This article highlights the current evidence of SARS-CoV-2 infection in farmed minks, and provides an understanding of the pathogenesis of COVID-19 in minks and the associated zoonotic concerns of SARS-CoV-2 transmission from minks to humans with an emphasis on appropriate mitigation measures and on the necessity of adopting the One Health approach during the COVID-19 pandemic.

Establishment and Application of Multiplex PCR for Simultaneously Detecting Escherichia coli, Salmonella, Klebsiella pneumoniae, and Staphylococcus aureus in Minks.

To establish a multiplex PCR for simultaneous detection of Escherichia coli (E. coli), Salmonella, Klebsiella pneumoniae (K. pneumoniae), and Staphylococcus aureus (S. aureus), four pairs of specific primers were designed according to the conservative regions of phoA gene for E. coli, invA gene for Salmonella, khe gene for K. pneumoniae, nuc gene for S. aureus. The quadruple PCR system was established through optimization of multiplex PCR and detection of specificity, sensitivity, and stability. The results showed that target gene bands of E. coli (622 bp), Salmonella (801 bp), K. pneumoniae (303 bp), and S. aureus (464 bp) could be amplified by this method specifically and simultaneously from the same sample containing the four pathogens, with a detection sensitivity of 100 pg/µL. Meanwhile, no bands of common clinical bacteria, including Clostridium perfringens, Pseudomonas aeruginosa, Pasteurella multocida, Streptococcus pneumoniae, Streptococcus pneumoniae, Proteus mirabilis, Staphylococcus sciuri, Staphylococcus pseudintermedius, Acinetobacter baumannii, Enterococcus faecalis, and Bacillus subtilis were amplified. In addition, 380 tissue samples were detected by multiplex and single PCR established in current study, respectively. Among the 368 carcass samples, positive detection rates of E. coli, K. pneumoniae, Salmonella, and S. aureus were 33.7, 12.0, 10.6, and 13.9%. Among the 12 visceral tissue samples, positive detection rates of E. coli, K. pneumoniae, Salmonella, and S. aureus were 41.7, 25.0, 16.7, and 8.3%, respectively. Positive detection rates of multiplex PCR were consistent with that of single PCR. Compared with single PCR, the multiplex PCR method had the advantages of time-saving, high specificity and high sensitivity. The results showed that the minks in these farms had mixed infection of these four pathogens, and the method established in this study could be applied to the rapid and accurate detection and identification of these four bacteria. In conclusion, the multiplex PCR method has stable detection results, good repeatability, and short detection time. It is suitable for the rapid and accurate detection of four kinds of bacteria above the carcass of fur animals, which could be suitable in microbial epidemiology investigation. It can provide a reliable technical reference for rapid clinical diagnosis and detection.

Evaluation of Collagen and Elastin Content in Skin of Multiparous Minks Receiving Feed Contaminated with Deoxynivalenol (DON, Vomitoxin) with or without Bentonite Supplementation.

Deoxynivalenol (DON, vomitoxin) is considered one of the most dangerous mycotoxins contaminating cereal products for food and feed. One of the protective methods against the adverse effect of DON on mink health is to use a component such as bentonite as a feed supplement to allow toxins absorption. The aim of this study was to determine the effect of DON, administered alone or with bentonite, on the histological structure of the skin and the content of collagen and elastin. A multiparous minks from control group (not exposed to DON) and a study groups receiving fed with DON-containing wheat for seven months: I: at a concentration of 1.1 mg/kg of feed, II: at a concentration of 3.7 mg/kg; III: DON at a concentration of 3.7 mg/kg and bentonite at a concentration of 0.5 kg/1000 kg of feed (0.05%); and IV: DON at a concentration of 3.7 mg/kg and bentonite at a concentration 2 kg/1000 kg (0.2%). After performing euthanasia and before pelting, skin samples of 2 cm in diameter were drawn from the multiparous minks from the lateral surface of the right anterior limb. Our obtained results clearly indicate that DON administered for a period of seven months at a dose of 1.1 mg/kg significantly changes the thickness of skin of a multiparous mink. It causes an increase in the percentage of elastin from 5.9% to 9.4% and a decrease in immature collagen, which results in a change in the collagen/elastin ratio from 10/1 to 5/1. A dose of 3.7 mg/kg DON in feed without or with 0.05% bentonite causes the absence of immature collagen in the dermis, but the addition of 0.2% bentonite in the feed reveals the presence of immature collagen and increase the percentage of the elastin.

Leishmania infection in lagomorphs and minks in Greece.

Greece is an endemic country for human and canine leishmaniosis. Studies about the role of lagomorphs and minks in the epidemiology of the diseases are, so far, limited. The aim of the present study was to investigate the prevalence of Leishmania infection in these animals, in different areas of the country. Samples from 393 domestic and wild rabbits, 90 hares and 200 minks were collected and examined by cytology (spleen imprints) and serology (ELISA), while spleen samples of 116, 56 and 95 of the rabbits, hares and minks, respectively, were examined by a PCR assay targeting the ITS1 region. For every animal examined a form was created, recording information like date, area, animal species, sex, etc. All imprint smears examined were negative, while serology revealed infection in 7.6% (C.I. 5.0-10.3%) rabbits, 6.7% (C.I. 1.5-11.8%) hares and 20% (C.I. 14.5-25.5%) minks. Infection was confirmed by molecular methods in 2.6% (C.I. 0.0-5.5%), 3.6% (C.I. 0.0-8.4%) and 2.1% (C.I. 0.0-5.0%) of the animals, respectively. The statistical analysis showed that minks are most likely to be seropositive and that in rabbits, the breeding method (i.e. homestead reared animals) was associated with infection. Because of the proximity of lagomorphs and minks to humans and dogs it is necessary to further elucidate their role in the epidemiology of leishmaniosis.

Enterocytozoon bieneusi in Minks (Neovison vison) in Northern China: A Public Health Concern.

Enterocytozoon bieneusi is the most important causative agent of microsporidiosis and can infect almost all vertebrate and invertebrate hosts, including minks (Neovison vison). In the present study, a total of 298 feces samples (including 79 from Heilongjiang province, 31 from Hebei province, 67 from Jilin province, 90 from Liaoning province, and 31 from Shandong province, Northern China) were examined by nested PCR amplification of the internal transcribed spacer (ITS) region of the rRNA gene. The overall prevalence of E. bieneusi in minks was 10.1%, with 10.5% in Jilin province, 32.3% in Hebei province, 8.9% in Liaoning province, 0% in Shandong province, and 6.3% in Heilongjiang province. Furthermore, multiple logistic regression analysis revealed that region was only risk factors associated with E. bieneusi infection in the investigated minks. Five E. bieneusi ITS genotypes (three known genotypes, namely D, Peru11, and EbpC; two novel genotypes, namely, NCM-1 and NCM-2) were found in the current study. Importantly, genotypes D, Peru11 and EbpC, previously identified in humans, were also found in minks, which suggested that minks are the potential sources of human microsporidiosis. To the best of our knowledge, this is the first report of E. bieneusi infection in minks worldwide. The results of the present survey have implications for the controlling E. bieneusi infection in minks, other animals and humans.

Vaccine resistant pseudorabies virus causes mink infection in China.

BACKGROUND: Pseudorabies, a highly contagious infectious disease of swine is caused by pseudorabies virus (PRV). PRV can cause fatal infection in other animal species. RESULTS: We report a deadly outbreak of pseudorabies that killed 87.2% (3522/4028) minks in a farm in 2014 in Shandong Province, China. PRV was isolated by using Vero cell culture and detected in mink samples by PCR from minks died during the outbreak. Epidemiological analysis indicated that 5.8% of minks (33/566) were PCR positive to PRV in Shandong Province. Phylogenetic analysis indicated that the PRV strains isolated from minks in this study were in the same clade with the Chinese porcine PRV isolates, which are resistant to the PRV vaccine. CONCLUSIONS: We demonstrated that pseudorabies virus caused an outbreak of minks in a farm in Shandong Province of China and the virus has a very high infection rate in minks in Shandong Province, which is a challenge for the fur industry in China.

Molecular detection and genetic characterizations of Cryptosporidium spp. in farmed foxes, minks, and raccoon dogs in northeastern China.

Cryptosporidium spp. are common intestinal protozoa causing diarrhea in humans and a variety of animal species. With the recent development of fur industry, a large number of fur animals are farmed worldwide, especially in China. The existence of identical Cryptosporidium species/genotypes in humans and fur animals suggests zoonotic potential. In order to assess the presence of zoonotic Cryptosporidium species and/or genotypes in farmed fur animals, 367 fecal specimens were collected from 213 foxes, 114 minks and 40 raccoon dogs farmed in Heilongjiang, Jilin, and Liaoning provinces, northeastern China, during the period from June 2014 to October 2016. By PCR and sequencing of the partial small subunit (SSU) rRNA gene of Cryptosporidium, 20 of 367 (5.4%) animal samples were found to be infected, corresponding to 12 of 213 fox samples (5.6%) and 8 of 114 mink samples (7.0%) screened. Three Cryptosporidium species/genotypes were identified: C. canis (n = 17), C. meleagridis (n = 1) and Cryptosporidium mink genotype (n = 2). Two host-adapted C. canis types (C. canis dog genotype and C. canis fox genotype) were found. By PCR and sequencing of the partial 60 kDa glycoprotein (gp60) encoding gene, one mink genotype isolate was successfully subtyped as XcA5G1R1. The three Cryptosporidium species/genotypes identified in this study have been previously reported in humans suggesting that fur animals infected with Cryptosporidium spp. may pose a risk of zoonotic transmission of cryptosporidiosis, especially for the people working in fur animal farming and processing industry.

Severe Fever with Thrombocytopenia Syndrome Virus Infection in Minks in China.

We analyzed the seroprevalence of tick-borne severe fever with thrombocytopenia syndrome virus (SFTSV) in farm-raised minks using double antigen ELISA (enzyme-linked immunosorbent assay) kit and indicated that 8.4% (15/178) of the minks had antibodies to the nucleoprotein of SFTSV and 72.7% (8/11) of mink farms had minks positive to SFTSV. The ELISA results were further confirmed by presence of neutralization to SFTSV in the mink sera. Our results suggested that minks were widely infected with SFTSV in China.

Study on Antiemetic Effect of Capsicum frutescens / 中国药房

OBJECTIVE: To study the antiemetic effects of Capsicum frutescens and its mechanism.METHODS: Novel vomiting model of minks were established.Inhibition effect of C.fruescens(capsicin as main component) on vomiting behavior induced by cisplatin and apomorphine were observed.The mechanism was investigated through the immunohistochemistry test and the specific 5-HT3-receptor agonist 1-phenylbiguanide hydrochloride(PBG).RESULTS: C.frutescens inhibited vomiting behavior of minks induced by cisplatin and apomorphine(P