Feline Coronavirus Research Articles

Intestinal injury and vasculitis biomarkers in cats with feline enteric coronavirus and effusive feline infectious peritonitis.

To investigate intestinal injury, repair and vasculitis biomarkers that may illuminate the progression and/or pathogenesis of feline infectious peritonitis (FIP) or feline enteric coronavirus (FECV) infection. A total of 40 cats with effusive FIP (30 with abdominal effusion, AE group; 10 with thoracic effusion, TE group) and 10 asymptomatic but FECV positive cats (FECV group), all were confirmed by reverse transcription polymerase chain reaction either in faeces or effusion samples. Physical examinations and effusion tests were performed. Trefoil factor-3 (TFF-3), intestinal alkaline phosphatase (IAP), intestinal fatty acid binding protein (I-FABP), myeloperoxidase-anti-neutrophilic cytoplasmic antibody (MPO-ANCA) and proteinase 3-ANCA (PR3-ANCA) concentrations were measured both in serum and effusion samples. Rectal temperature and respiratory rate were highest in the TE group (p<0.000). Effusion white blood cell count was higher in the AE group than TE group (p<0.042). Serum TFF-3, IAP and I-FABP concentrations were higher in cats with effusive FIP than the cats with FECV (p<0.05). Compared with the AE group, TE group had lower effusion MPO-ANCA (p<0.036), higher IAP (p<0.050) and higher TFF-3 (p<0.016) concentrations. Markers of intestinal and epithelial surface injury were higher in cats with effusive FIP than those with FECV. Compared to cats with abdominal effusions, markers of apoptosis inhibition and immunostimulation to the injured epithelium were more potent in cats with thoracic effusion, suggesting the possibility of a poorer prognosis or more advanced disease in these patients.

Prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and feline enteric coronavirus (FECV) in shelter-housed cats in the Central Valley of California, USA.

Non-human animals are natural hosts for the virus causing COVID-19 (severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2]) and a diversity of species appear susceptible to infection. Cats are of particular concern because of their close affiliation with humans and susceptibility to infection. Cats also harbour feline enteric coronavirus (FECV). Our objectives were to document the prevalence of SARS-CoV-2 and FECV in feline populations with high turnover and movement among households in the Central Valley of California, USA. A cross-sectional study of 128 shelter and foster cats and kittens in the Central Valley of California was performed from July to December 2020. PCR was performed on rectal and oropharyngeal samples to detect SARS-CoV-2 RNA and on rectal samples to detect FECV RNA. Among 163 rectal and oropharyngeal fluid samples gathered from sheltered and fostered cats and kittens in central California, SARS-CoV-2 nucleic acids were not detected from any cat or kitten. In contrast, FECV nucleic acids were detected in 18% of shelter-housed cats; 83% of these positive samples were collected from cats housed in adjacent cages. These data may be helpful when considering the allocation of resources to minimise the harm of FECV and SARS-CoV-2 in household pets and shelter environments.

Yeast Surface Dual Display Platform for Rapid Discovery of Shark VNAR from a Semi-Synthetic Library Followed by Next-Generation Sequencing

The shark-derived single-domain antibody VNAR (variable domain of new antigen receptor) has many advantageous features that make the VNAR suitable for improving current monoclonal antibody therapy deficiencies or disease diagnosis methods. In order to discover more VNARs, it is necessary to improve the efficiency of the isolation process. This research aims to enhance the VNAR discovery platform by dual displaying the semi-synthetic VNAR library and green fluorescent protein tag on the yeast surface. The GFP tag can be used to determine the degree of VNAR expression. The diversity of the semi-synthetic VNAR library constructed in this study is verified to be 1.97 × 109 by next-generation sequencing (NGS). We conveniently screened VNARs against the feline neonatal Fc receptor or feline infectious peritonitis virus nucleocapsid protein by sequential MACS and FACS. To find more diverse VNARs, we analyzed the NGS data of VNAR CDR3 genes before and after biopanning. By comparing the frequency change of each sequence, we found that the amplification factor of sequences was increased by biopanning. Four VNAR candidates selected by the high-frequency and high-amplification factor criteria showed an antigen-binding ability. The results demonstrate that biopanning from a yeast surface displaying a semi-synthetic VNAR library followed by the NGS assay can generate antigen binders rapidly without the need for shark rearing and long-term immunization.

High-resolution melting analysis for simultaneous detection and discrimination between wild-type and vaccine strains of feline calicivirus

High-resolution melting (HRM) analysis, a post-polymerase chain reaction (PCR) application in a single closed tube, is the straightforward method for simultaneous detection, genotyping, and mutation scanning, enabling more significant dynamic detection and sequencing-free turnaround time. This study aimed to establish a combined reverse-transcription quantitative PCR and HRM (RT-qPCR-HRM) assay for diagnosing and genotyping feline calicivirus (FCV). This developed method was validated with constructed FCV plasmids, clinical swab samples from living cats, fresh-frozen lung tissues from necropsied cats, and four available FCV vaccines. We performed RT-qPCR to amplify a 99-base pair sequence, targeting a segment between open reading frame (ORF) 1 and ORF2. Subsequently, the HRM assay was promptly applied using Rotor-Gene Q® Software. The results significantly revealed simultaneous detection and genetic discrimination between commercially available FCV vaccine strains, wild-type Thai FCV strains, and VS-FCV strains within a single PCR reaction. There was no cross-reactivity with other feline common viruses, including feline herpesvirus-1, feline coronavirus, feline leukemia virus, feline immunodeficiency virus, and feline morbillivirus. The detection limit of the assay was 6.18 × 101 copies/µl. This study, therefore, is the first demonstration of the uses and benefits of the RT-qPCR-HRM assay for FCV detection and strain differentiation in naturally infected cats.

Impact of the Application of Gaseous Ozone on Selected Pathogens Found in Animal Shelters and Other Facilities.

Correctly selecting disinfection procedures is crucial in facilities housing a high number of animals as it directly affects their health. The aim of this study was to verify the virucidal effect of gaseous ozone delivered by commercially available generators under controlled experimental conditions on a selection of viral pathogens (feline coronavirus, canine coronavirus, feline calicivirus, feline parvovirus) commonly found in shelters and other facilities. Two ozone generators with outputs of 3.5 g/h and 20 g/h were used to produce ozone. Virus viability after the application of ozone was evaluated by examining for typical pathogen-specific cytopathic effects on the CRFK (Crandell-Rees Feline Kidney) cell line post-incubation. No cytopathic effect was observed in feline coronavirus after the 2-h application of ozone; in canine coronavirus, the absence of a cytopathic effect was observed after the 4-h application of ozone. The absence of a cytopathic effect in feline calicivirus was observed after the 6-h application of ozone; the viability of feline parvovirus was not impaired even by the 6-h application of ozone. The results of the study confirm lower resistance to the application of gaseous ozone in enveloped viruses.

AN OUTBREAK OF FELINE INFECTIOUS PERITONITIS IN THREE RELATED SAND CATS (FELIS MARGARITA) IN HUMAN CARE.

Feline infectious peritonitis (FIP) is a systemic disease in felid species caused by infection with mutated forms of feline coronavirus (FCoV), and outbreaks can devastate exotic felid populations in human care. Feline infectious peritonitis was diagnosed in three of four related juvenile sand cats (Felis margarita) from a single institution over a 6-wk period. Case 1 was a 7-mon-old male found deceased with no premonitory signs. Case 2, an 8-mon-old male (littermate to Case 1), and Case 3, a 6-mon-old male (from a different litter with identical parentage), were evaluated for lethargy and anorexia 1 mon after Case 1. Both exhibited transient anisocoria and progressive lethargy, anorexia, and dehydration despite antibiotic and supportive treatment. Approximately 1 wk after initial presentation, Case 2 was humanely euthanized, and Case 3 was found deceased. Necropsy findings included intrathoracic and/or intra-abdominal lymphadenopathy (3/3 cases), bicavitary effusion (2/3), multifocal tan hepatic and intestinal nodules (1/3), and multifocal yellow renal nodules (1/3). Histologically, all cats had severe pyogranulomatous vasculitis in multiple organs, and the presence of FCoV antigen was confirmed using immunohistochemical staining. Next-generation sequencing of the virus from Case 3's affected kidney demonstrated ∼93% homology to the UG-FH8 virus, a serotype 1 feline alphacoronavirus isolated from Denmark. Future research will focus on comparative viral genomic sequencing with the goals of identifying potential sources of FCoV infection and identifying features that may have contributed to the development of FIP in this species.

Detection of Feline Coronavirus, Feline Immunodeficiency Virus, Feline Leukemia Virus and Other Pathogen Genetic Material in Whole Blood from Domestic Cats in Türkiye

Detection of Feline Coronavirus, Feline Immunodeficiency Virus, Feline Leukemia Virus and Other Pathogen Genetic Material in Whole Blood from Domestic Cats in Türkiye

Clinical And Molecular Investigation in Feline Infectious Peritonitis Cat Treated with Gs-441524 Contained Drug

An immune-mediated disease caused by a highly virulent Feline coronavirus (FCoV), Feline infectious peritonitis, is one of the most challenging diseases in cats due to its difficulty in either diagnosis or treatment. However, the recent investigation has offered some hope with the discovery of adenosine nucleoside analog GS-441524, which has shown promising results in terms of survival rates and clinical recoveries. This study demonstrated the effectiveness of GS-441524 for FIP treatment. A-two-year-old male mix breed was admitted to Satwakita Animal Clinic, Yogyakarta and examined by responsible veterinarian. Abdominal effusion was aseptically collected for further examination in Veterinary Medicine Clinical Pathology Laboratory, Universitas Gadjah Mada. The analysis utilized RT-qPCR targeting 5’UTR confirmed FCoV infection in the cat. The cat was treated by daily administration of the nucleoside analog GS-441524 by subcutaneously injection route for 40 days. The result of this treatment proves that GS-441524 effectively cured FIP and eliminated FCoV infection in the cat. In this case report, we showed that GS-441524 was effective in this cat. Both clinical signs and FCoV RNA were significantly reduced. The results highlight the promising potential of GS-441524 for treating FIP that naturally occurs in cats

Epidemiological survey of feline viral infectious diseases in China from 2018 to 2020

AbstractTo analyze the prevalence of feline viral diseases in China, including feline panleukopenia virus (FPV), feline calicivirus (FCV), feline herpesvirus 1 (FHV‐1), and feline coronavirus (FCoV) infectious diseases from 2018 to 2020, swab samples from 304 cats and serum samples from 193 cats in 18 cities were collected. The etiological investigation results of 304 cats showed that 256 (84.21%) cats were positive, infected with at least one virus, and the positive rates for FPV, FCV, FHV‐1, and FCoV were 61.51%, 10.86%, 4.61%, and 55.92%, respectively. The mixed infection exhibited high complexity, and a total of eight mixed infection patterns were detected. The risk factor analysis of each pathogen in different clinical scenarios indicated that FPV positive status was significantly related to all the studied diseases, FCV positive status exhibited the most significant association with gingivostomatitis and conjunctivitis, and FHV‐1 positive status was significantly related to upper respiratory tract disease, but FCoV positive status was not significantly related to any disease. Additionally, the prevalence of FPV exhibited a strong seasonality and was related to age, while the prevalence of FCV, FHV‐1, and FCoV had nothing to do with season or age. FCV infection was sex related in cats, whereas the prevalence of FCV, FHV‐1, and FCoV was not sex related. FPV, FCV, FHV‐1, and FCoV were unrelated to breed or residential density. Antibody detection results of 193 serum samples by the virus neutralizing method indicated that the current commercial vaccines might not protect hosts against wild strains of FPV, FCV, and FHV‐1 in China. In general, this study enriches epidemiological survey data of common viral diseases in cats in China and provides a theoretical basis for further development of vaccines.

Multiplex One-Step RT-qPCR Assays for Simultaneous Detection of SARS-CoV-2 and Other Enteric Viruses of Dogs and Cats.

The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) was transmitted from humans to dogs and cats (reverse zoonosis) during the COVID-19 pandemic. SARS-CoV-2 has been detected in fecal samples of infected dogs and cats, indicating potential fecal-oral transmission, environmental contamination, and zoonotic transmission (i.e., spillback). Additionally, gastrointestinal viral infections are prevalent in dogs and cats. In this study, we developed and validated a panel of multiplex one-step reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays for the simultaneous detection of SARS-CoV-2 and common canine enteric viruses: Canine Enteric Assay_1 (CEA_1) for the detection of canine adenovirus-1, canine enteric coronavirus, canine distemper virus, and canine parvovirus, and CEA_2 for the detection of rotavirus A (RVA), and SARS-CoV-2); or common feline enteric viruses (Feline Enteric Assay_1 (FEA_1) for the detection of feline enteric coronavirus, feline panleukopenia virus, RVA, and SARS-CoV-2). All assays demonstrated high analytical sensitivity, detecting as few as 5-35 genome copies/µL in multiplex format. The repeatability and reproducibility of the multiplex assays were excellent, with coefficient of variation <4%. Among the 58 clinical samples tested, 34.5% were positive for at least one of these viruses, and SARS-CoV-2 was detected in two samples collected from one dog and one cat, respectively. In conclusion, these newly developed one-step multiplex RT-qPCR assays allow for rapid diagnosis of enteric viral infections, including SARS-CoV-2, in dogs and cats.

Immunohistochemical investigation of FIPV3-70 antigen expression in the ileum of cats with effusive feline infective peritonitis

Abstract One of the most common infectious causes of cat mortality is feline infective peritonitis (FIP), along with panleukopenia and viral upper respiratory tract infections. FIP is more likely to affect cats whose immune system is weak or suppressed. It is thought that the infection of macrophages and monocytes plays a major role in the pathogenic process. In order to set a definitive diagnosis for this infectious disease, a histopathological examination of tissues, and feline coronavirus (FCoV) detection by immunohistochemistry (IHC) is necessary. In this investigation, 15 cats between the ages of 5 and 24 months with clinical suspicion of FIP, underwent post-mortem necropsy, pathohistological and immunohistochemical examination. The results showed that all the cats had abdominal effusion with pyogranulomas throughout the abdominal serosa. Ten out of fifteen cats were FIP positive using immunohistochemical methods. This method also showed the antigen deposition in the macrophages thus confirming their role in the pathogenesis of FIP.

Retrospective study and outcome of 307 cats with feline infectious peritonitis treated with legally sourced veterinary compounded preparations of remdesivir and GS-441524 (2020-2022).

Feline infectious peritonitis (FIP) is a serious disease that arises due to feline coronavirus infection. The nucleoside analogues remdesivir and GS-441524 can be effective in its treatment, but most studies have used unregulated products of unknown composition. The aim of the present study was to describe the treatment of FIP using legally sourced veterinary-prescribed regulated veterinary compounded products containing known amounts of remdesivir (injectable) or GS-441524 (oral tablets). Cats were recruited via email advice services, product sales contacts and study publicity. Cats were excluded if they were deemed unlikely to have FIP, were not treated exclusively with the veterinary compounded products, or if there was a lack of cat and/or treatment (including response) data. Extensive cat and treatment data were collected. Among the 307 cats recruited, the predominant type of FIP was most commonly abdominal effusive (49.5%) and then neurological (14.3%). Three treatment protocols were used; remdesivir alone (33.9%), remdesivir followed by GS-441524 (55.7%) and GS-441524 alone (10.4%). The median (range) initial treatment period duration and longest follow-up time point after starting treatment were 84 (1-330) days and 248 (1-814) days, respectively. The most common side effect was injection pain (in 47.8% of those given subcutaneous remdesivir). Of the 307 cats, 33 (10.8%) relapsed, 15 (45.5%) during and 18 (54.5%) after the initial treatment period. At the longest follow-up time point after completion of the initial treatment period, 84.4% of cats were alive. The cats achieving a complete response within 30 days of starting treatment were significantly more likely to be alive at the end of the initial treatment period than those cats that did not. Legally sourced remdesivir and GS-441524 products, either alone or used sequentially, were very effective in the treatment of FIP in this group of cats. Variable protocols precluded statistical comparison of treatment regimens.

Feline Infectious Peritonitis: European Advisory Board on Cat Diseases Guidelines.

Feline coronavirus (FCoV) is a ubiquitous RNA virus of cats, which is transmitted faeco-orally. In these guidelines, the European Advisory Board on Cat Diseases (ABCD) presents a comprehensive review of feline infectious peritonitis (FIP). FCoV is primarily an enteric virus and most infections do not cause clinical signs, or result in only enteritis, but a small proportion of FCoV-infected cats develop FIP. The pathology in FIP comprises a perivascular phlebitis that can affect any organ. Cats under two years old are most frequently affected by FIP. Most cats present with fever, anorexia, and weight loss; many have effusions, and some have ocular and/or neurological signs. Making a diagnosis is complex and ABCD FIP Diagnostic Approach Tools are available to aid veterinarians. Sampling an effusion, when present, for cytology, biochemistry, and FCoV RNA or FCoV antigen detection is very useful diagnostically. In the absence of an effusion, fine-needle aspirates from affected organs for cytology and FCoV RNA or FCoV antigen detection are helpful. Definitive diagnosis usually requires histopathology with FCoV antigen detection. Antiviral treatments now enable recovery in many cases from this previously fatal disease; nucleoside analogues (e.g., oral GS-441524) are very effective, although they are not available in all countries.

Membrane-Targeting Perylenylethynylphenols Inactivate Medically Important Coronaviruses via the Singlet Oxygen Photogeneration Mechanism.

Perylenylethynyl derivatives have been recognized as broad-spectrum antivirals that target the lipid envelope of enveloped viruses. In this study, we present novel perylenylethynylphenols that exhibit nanomolar or submicromolar antiviral activity against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) and feline infectious peritonitis virus (FIPV) in vitro. Perylenylethynylphenols incorporate into viral and cellular membranes and block the entry of the virus into the host cell. Furthermore, these compounds demonstrate an ability to generate singlet oxygen when exposed to visible light. The rate of singlet oxygen production is positively correlated with antiviral activity, confirming that the inhibition of fusion is primarily due to singlet-oxygen-induced damage to the viral envelope. The unique combination of a shape that affords affinity to the lipid bilayer and the capacity to generate singlet oxygen makes perylenylethynylphenols highly effective scaffolds against enveloped viruses. The anticoronaviral activity of perylenylethynylphenols is strictly light-dependent and disappears in the absence of daylight (under red light). Moreover, these compounds exhibit negligible cytotoxicity, highlighting their significant potential for further exploration of the precise antiviral mechanism and the broader scope and limitations of this compound class.

Prevalence and genetic variation of the M, N, and S2 genes of feline coronavirus in Shandong Province, China.

Feline coronavirus (FCoV) is the causative agent of feline infectious peritonitis and diarrhoea in kittens worldwide. In this study, a total of 73 feline diarrhoeal faecal samples were collected from animal hospitals and pet markets in ShanDong province from 2017 to 2019. FCoV was detected in 58.23% (46/73) of the samples, using the RT-PCR method. The results showed that the detection rate of FCoV in healthy cats and sick cats was 41.7% (10/24) and 81.6% (40/49), respectively. Full gene amplification and sequencing of the N, M, and S2 genes of FCoV isolates were performed. An amino acid mutation (M1058L) in the S2 gene was found that can be used as a marker for distinguishing feline enteric coronavirus (FECV) from feline infectious peritonitis virus (FIPV). This study provides new epidemiological information about FCoV that will aid in the prevention of FCoV in China.

Vector-Borne Pathogens in Stray Cats in Eastern Germany (Thuringia).

Bacterial, protozoal, and viral vector-borne pathogens (VBPs) can cause infections in cats. There is little information on feline VBP prevalence in Germany. Stray cats are frequently exposed to vectors but receive no veterinary care. The aim of this study was to determine the prevalence of selected VBPs in stray cats. EDTA blood and serum samples were taken from apparently healthy stray cats during a spay/neuter campaign in the federal state of Thuringia. Overall, 11/50 (22%) and 32/50 (64%) cats tested positive for at least one VBP by direct and indirect detection methods, respectively. PCR testing of EDTA blood detected hemotropic Mycoplasma spp. in 12% of cats, Hepatozoon spp. in 10%, and Anaplasma phagocytophilum in 4%. PCR testing for Rickettsia spp. and piroplasms was negative. IFAT on serum samples showed 46% of cats had detectable antibodies for Bartonella spp., 30% for Rickettsia spp., and 16% for A. phagocytophilum. The cats were additionally tested for feline coronavirus, FIV, and FeLV to identify potential risk factors for pathogen contact and/or infections. No correlation between FIV and FeLV status and VBP positivity was detected. Anaplasma phagocytophilum, Rickettsia spp., and Bartonella spp. have zoonotic potential, and surveillance is recommended in the context of the One Health approach.

Comparative Evaluation of GS-441524, Teriflunomide, Ruxolitinib, Molnupiravir, Ritonavir, and Nirmatrelvir for In Vitro Antiviral Activity against Feline Infectious Peritonitis Virus.

Feline infectious peritonitis (FIP), caused by feline coronavirus (FcoV), is considered one of the most enigmatic diseases in cats. Developing effective drugs for FIP is crucial due to its global prevalence and severity. In this study, six antiviral drugs were tested for their cytotoxicity, cell viability, and antiviral efficacies in Crandell-Reese feline kidney cells. A cytotoxicity assay demonstrated that these drugs were safe to be used with essentially no cytotoxicity with concentrations as high as 250 µM for ruxolitinib; 125 µM for GS441524; 63 µM for teriflunomide, molnupiravir, and nirmatrelvir; and 16 µM for ritonavir. GS441524 and nirmatrelvir exhibited the least detrimental effects on the CRFK cells, with 50% cytotoxic concentration (CC50) values of 260.0 µM and 279.1 µM, respectively, while ritonavir showed high toxicity (CC50 = 39.9 µM). In the dose-response analysis, GS441524, nirmatrelvir, and molnupiravir demonstrated promising results with selectivity index values of 165.54, 113.67, and 29.27, respectively, against FIPV. Our study suggests that nirmatrelvir and molnupiravir hold potential for FIPV treatment and could serve as alternatives to GS441524. Continued research and development of antiviral drugs are essential to ensure the well-being of companion animals and improve our preparedness for future outbreaks of coronaviruses affecting animals and humans alike.

Long-term follow-up of cats in complete remission after treatment of feline infectious peritonitis with oral GS-441524.

Feline infectious peritonitis (FIP), a common disease in cats caused by feline coronavirus (FCoV), is usually fatal once clinical signs appear. Successful treatment of FIP with oral GS-441524 for 84 days was demonstrated recently by this research group. The aim of this study was to evaluate the long-term outcome in these cats. A total of 18 successfully treated cats were followed for up to 1 year after treatment initiation (9 months after completion of the antiviral treatment). Follow-up examinations were performed at 12-week intervals, including physical examination, haematology, serum biochemistry, abdominal and thoracic ultrasound, FCoV ribonucleic acid (RNA) loads in blood and faeces by reverse transciptase-quantitative PCR and anti-FCoV antibody titres by indirect immunofluorescence assay. Follow-up data were available from 18 cats in week 24, from 15 cats in week 36 and from 14 cats in week 48 (after the start of treatment), respectively. Laboratory parameters remained stable after the end of the treatment, with undetectable blood viral loads (in all but one cat on one occasion). Recurrence of faecal FCoV shedding was detected in five cats. In four cats, an intermediate short-term rise in anti-FCoV antibody titres was detected. In total, 12 cats showed abdominal lymphadenomegaly during the follow-up period; four of them continuously during the treatment and follow-up period. Two cats developed mild neurological signs, compatible with feline hyperaesthesia syndrome, in weeks 36 and 48, respectively; however, FCoV RNA remained undetectable in blood and faeces, and no increase in anti-FCoV antibody titres was observed in these two cats, and the signs resolved. Treatment with GS-441524 proved to be effective against FIP in both the short term as well as the long term, with no confirmed relapse during the 1-year follow-up period. Whether delayed neurological signs could be a long-term adverse effect of the treatment or associated with a 'long FIP syndrome' needs to be further evaluated.

Molecular epidemiology and genetic diversity of canine coronavirus from domestic dogs in Chengdu, China from 2020 to 2021 using a multiplex RT-PCR.

Recent reports on identification of canine coronavirus (CCoV) in humans have emphasized the urgency to strengthen surveillance of animal CoVs. The fact that recombinations between CCoV with feline, porcine CoVs brought about new types of CoVs indicated that more attention should be paid to domestic animals like dogs, cats and pigs, and the CoVs they carried. However, there are about ten kinds of CoVs that infect above animals, and thus representative CoVs with zoonotic potentials were considered in this study. Multiplex RT-PCR against CCoV, Feline coronavirus (FCoV), porcine deltacoronavirus and porcine acute diarrhea syndrome coronavirus was developed to investigate the prevalence of CoVs from domestic dogs in Chengdu, Southwest China. Samples from a total of 117 dogs were collected from a veterinary hospital, and only CCoV (34.2%, 40/117) was detected. Therefore, this study focused on CCoV and its characteristics of S, E, M, N and ORF3abc genes. Compared with CoVs that are capable of infecting humans, CCoV strains showed highest nucleotide identity with the novel canine-feline recombinant detected from humans (CCoV-Hupn-2018). Phylogenetic analysis based on S gene, CCoV strains were not only clustered with CCoV-II strains, but also closely related to FCoV-II strains ZJU1617 and SMU-CD59/2018. As for assembled ORF3abc, E, M, N sequences, CCoV strains had the closest relationship with CCoV-II (B203_GZ_2019, B135_JS_2018 and JS2103). What's more, specific amino acid variations were found, especially in S and N proteins, and some mutations were consistent with FCoV, TGEV strains. Altogether, this study provided a novel insight into the identification, diversification and evolution of CoVs from domestic dogs. It is of top priority to recognize zoonotic potential of CoVs, and continued comprehensive surveillance will help better understand the emergence, spreading, and ecology of animal CoVs.

Cross-sectional and histopathological studies of Feline Coronavirus infections in stray cats in Kuwait.

Feline Coronavirus (FCoV) is a worldwide viral infection of felids. The disease is usually asymptomatic, but it can cause mild diarrhoea; however, few numbers of cases may develop a severe systemic disease known as feline infectious peritonitis (FIP). This study aims to determine the prevalence of FCoV shedding in the faeces of stray cats in Kuwait and detect antibodies against FCoV in their serum. Histopathological analyses and RT‑PCR were used to prove cases of FIP. A total of 178 cats were examined for the presence of FCoV in their faeces using a rapid immunochromatography (IC) test. Anti‑FCoV Antibody (Anti‑FCoV Ab) was detected in their serum using ELISA. Eleven samples were tested using RT‑PCR to confirm positive cases. The prevalence of FCoV faecal antigen in stray cats was 32.6%. The overall detection rate of Anti‑FCoV Ab in stray cats was 44.9%. Nine cats tested positive using the RT‑PCR test. Six out of those nine were confirmed to be FIP positive through gross and histopathological examination. The characteristic uveitis and discoloration of the irises were seen. The present study is the first report confirming FCoV infection in stray cats in Kuwait. Postmortem and histopathological lesions in cases of FIP were recorded.