Feline Enteric Coronavirus Research Articles

Molecular epidemiology and risk analysis for asymptomatic infection with feline enteric coronavirus in domestic and stray cats in Japan.

Feline enteric coronavirus (FECoV) causes subclinical infection; therefore, asymptomatic cats can act as transmitters of FECoV. However, there have been few studies on the prevalence of FECoV in asymptomatic cats in Japan. In this study, we used a nested RT-PCR assay targeting the S gene of FECoV to test 319 normal fecal samples from stray cats and domestic cats in Japan between 2019 and 2020. Seventy-five samples tested positive, and the presence of FECoV in domestic cats was significantly associated with age (p<0.01), but not with gender, breed, or living in a multi-cat household. The results provide insights into the current prevalence of FECoV in asymptomatic cats in Japan.

Development and characterization of reverse genetics systems of feline infectious peritonitis virus for antiviral research

Feline infectious peritonitis (FIP) is a lethal, immune-mediated disease in cats caused by feline infectious peritonitis virus (FIPV), a biotype of feline coronavirus (FCoV). In contrast to feline enteric coronavirus (FECV), which exclusively infects enterocytes and causes diarrhea, FIPV specifically targets macrophages, resulting in the development of FIP. The transmission and infection mechanisms of this complex, invariably fatal disease remain unclear, with no effective vaccines or approved drugs for its prevention or control. In this study, a full-length infectious cDNA clone of the wild-type FIPV WSU79-1149 strain was constructed to generate recombinant FIPV (rFIPV-WT), which exhibited similar growth kinetics and produced infectious virus titres comparable to those of the parental wild-type virus. In addition, the superfold green fluorescent protein (msfGFP) and Renilla luciferase (Rluc) reporter genes were incorporated into the rFIPV-WT cDNA construct to generate reporter rFIPV-msfGFP and rFIPV-Rluc viruses. While the growth characteristics of the rFIPV-msfGFP virus were similar to those of its parental rFIPV-WT, the rFIPV-Rluc virus replicated more slowly, resulting in the formation of smaller plaques than did the rFIPV-WT and rFIPV-msfGFP viruses. In addition, by replacing the S, E, M, and ORF3abc genes with msfGFP and Rluc genes, the replicon systems repFIPV-msfGFP and repFIPV-Rluc were generated on the basis of the cDNA construct of rFIPV-WT. Last, the use of reporter recombinant viruses and replicons in antiviral screening assays demonstrated their high sensitivity for quantifying the antiviral effectiveness of the tested compounds. This integrated system promises to significantly streamline the investigation of virus replication within host cells, enabling efficient screening for anti-FIPV compounds and evaluating emerging drug-resistant mutations within the FIPV genome.

An Exploratory Bioinformatic Investigation of Cats' Susceptibility to Coronavirus-Deriving Epitopes.

Coronaviruses are highly transmissible and pathogenic viruses for humans and animals. The vast quantity of information collected about SARS-CoV-2 during the pandemic helped to unveil details of the mechanisms behind the infection, which are still largely elusive. Recent research demonstrated that different class I/II human leukocyte antigen (HLA) alleles might define an individual susceptibility to SARS-CoV-2 spreading, contributing to the differences in the distribution of the infection through different populations; additional studies suggested that the homolog of the HLA in cats, the feline leukocyte antigen (FLA), plays a pivotal role in the transmission of viruses. With these premises, this study aimed to exploit a bioinformatic approach for the prediction of the transmissibility potential of two distinct feline coronaviruses (FCoVs) in domestic cats (feline enteric coronavirus (FeCV) and feline infectious peritonitis virus (FIPV)) using SARS-CoV-2 as the reference model. We performed an epitope mapping of nonapeptides deriving from SARS-CoV-2, FeCV, and FIPV glycoproteins and predicted their affinities for different alleles included in the three main loci in class I FLAs (E, H, and K). The predicted complexes with the most promising affinities were then subjected to molecular docking and molecular dynamics simulations to provide insights into the stability and binding energies in the cleft. Results showed the FLA proteins encoded by alleles in the FLA-I H (H*00501 and H*00401) and E (E*01001 and E*00701) loci are largely responsive to several epitopes deriving from replicase and spike proteins of the analyzed coronaviruses. The analysis of the most affine epitope sequences resulting from the prediction can stimulate the development of anti-FCoV immunomodulatory strategies based on peptide drugs.

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.

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.

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.

Exploring the Enteric Virome of Cats with Acute Gastroenteritis.

Viruses are a major cause of acute gastroenteritis (AGE) in cats, chiefly in younger animals. Enteric specimens collected from 29 cats with acute enteritis and 33 non-diarrhoeic cats were screened in PCRs and reverse transcription (RT) PCR for a large panel of enteric viruses, including also orphan viruses of recent identification. At least one viral species, including feline panleukopenia virus (FPV), feline enteric coronavirus (FCoV), feline chaphamaparvovirus, calicivirus (vesivirus and novovirus), feline kobuvirus, feline sakobuvirus A and Lyon IARC polyomaviruses, was detected in 66.1% of the samples.. Co-infections were mainly accounted for by FPV and FCoV and were detected in 24.2% of the samples. The virome composition was further assessed in eight diarrhoeic samples, through the construction of sequencing libraries using a sequence-independent single-primer amplification (SISPA) protocol. The libraries were sequenced on Oxford Nanopore Technologies sequencing platform. A total of 41 contigs (>100 nt) were detected from seven viral families infecting mammals, included Parvoviridae, Caliciviridae, Picornaviridae, Polyomaviridae, Anelloviridae, Papillomaviridae and Paramyxoviridae, revealing a broad variety in the composition of the feline enteric virome.

Disease Surveillance of Cougars (Puma concolor) in Utah, USA.

Adult, free-ranging cougars (Puma concolor) were sampled in three regions of Utah, US, from 2018 to 2021. A total of 68% (23/34) of the sampled cougars had antibodies to feline parvovirus, 15% (5/33) to canine distemper virus, 18% (6/34) to calicivirus, and 22% (8/37) to Yersinia pestis. Forty-one percent (13/32) had IgG antibodies to Toxoplasma gondii and 6% (2/33) to feline immunodeficiency virus, and 3% (1/32) were positive for Dirofilaria immitis (heartworm) antigen. All were seronegative for Toxoplasma gondii IgM, feline enteric coronavirus, SARS-CoV-2, feline leukemia virus, feline herpesvirus, and Francisella tularensis. Tapeworms and Toxascaris leonina eggs were detected in the feces. The disease exposures detected were similar to what has been reported from cougar populations in other western US states, and the current level of exposures is unlikely to have a negative impact on the state's population.

Epidemiology and Molecular Characterizations of Coronavirus from Companion Animals Living in Chengdu, Southwest China

The recent COVID-19 pandemic has once again caught the attention of people on the probable zoonotic transmission from animals to humans, but the role of companion animals in the coronavirus (CoV) epidemiology still remains unknown. The present study was aimed to investigate epidemiology and molecular characterizations of CoVs from companion animals in Chengdu city, Southwest China. 523 clinical samples from 393 animals were collected from one veterinary hospital between 2020 and 2021, and the presence of CoVs was detected by end-point PCR using pan-CoV assay targeting the RdRp gene. Partial and complete S genes were sequenced for further genotyping and genetic diversity analysis. A total of 162 (31.0%, 162/523) samples and 146 (37.2%, 146/393) animals were tested positive for CoVs. The positive rate in rectal swabs was higher than that in eye/nose/mouth swabs and ascitic fluid but was not statistically different between clinically healthy and diseased ones. Genotyping identified twenty-two feline enteric coronavirus (FCoV) I, four canine enteric coronavirus (CECoV) I, fourteen CECoV IIa, and one CECoV IIb, respectively. Eight complete S genes, including one canine respiratory coronavirus (CRCoV) strain, were successfully obtained. FCoV strains (F21071412 and F21061627) were more closely related to CECoV strains than CRCoV, and C21041821-2 showed potential recombination event. In addition, furin cleavage site between S1 and S2 was identified in two strains. The study supplemented epidemiological information and natural gene pool of CoVs from companion animals. Further understanding of other functional units of CoVs is needed, so as to contribute to the prevention and control of emerging infectious diseases.

Renal Ultrasonography Findings in Cats with Feline Infectious Peritonitis

Feline infectious peritonitis (FIP) is a fatal disease caused by a mutated feline enteric coronavirus (FECV) that causes a wide diversity of clinical findings. Antemortem diagnosis may be challenging as the non-effusive form causes pyogranulomatous inflammation in various organs including the eye, brain, omentum, liver and kidney compared to the effusive form. Since it has been discussed that the kidney is the organ most susceptible to FIP-related lesion development, this study aimed to evaluate the renal ultrasonography findings in cats with naturally developed non-effusive FIP. Clinical and renal ultrasonographic examinations of 17 cats with compatible clinical findings that would suggest the presence of non-effusive FIP were performed with the appropriate protocol. Both cats’ kidneys were evaluated for echogenicity, size (longitudinal length), shape, presence of free fluid, if any, and echogenicity of this fluid. As a result of renal ultrasonography, it was observed that the most prominent abnormal ultrasonographic findings were cortical hyperechogenicity (11 out of 17 cats), medullary rim sign (11 out of 17 cats), renomegaly (10 out of 17 cats), pyelectasis (5 out of 17 cats), loss of corticomedullary differentiation (4 out of 17 cats) and distortion of internal architecture (4 out of 17 cats). In conclusion, it was observed that morphological and parenchymal alterations occur in the renal ultrasonographic evaluation in cats with non-effusive FIP, and renal ultrasonography could provide useful clinical information in evaluating the clinical reflection of vasculitis due to FIP. Although these abnormal renal ultrasonography findings were not specific for FIP, it was concluded that the combination of the observed ultrasonographic findings and other compatible clinical findings and their evaluation together can be used to increase the index of suspicion for antemortem FIP infection.

Natural selection differences detected in key protein domains between non-pathogenic and pathogenic feline coronavirus phenotypes.

Feline coronaviruses (FCoVs) commonly cause mild enteric infections in felines worldwide (termed feline enteric coronavirus [FECV]), with around 12 per cent developing into deadly feline infectious peritonitis (FIP; feline infectious peritonitis virus [FIPV]). Genomic differences between FECV and FIPV have been reported, yet the putative genotypic basis of the highly pathogenic phenotype remains unclear. Here, we used state-of-the-art molecular evolutionary genetic statistical techniques to identify and compare differences in natural selection pressure between FECV and FIPV sequences, as well as to identify FIPV- and FECV-specific signals of positive selection. We analyzed full-length FCoV protein coding genes thought to contain mutations associated with FIPV (Spike, ORF3abc, and ORF7ab). We identified two sites exhibiting differences in natural selection pressure between FECV and FIPV: one within the S1/S2 furin cleavage site (FCS) and the other within the fusion domain of Spike. We also found fifteen sites subject to positive selection associated with FIPV within Spike, eleven of which have not previously been suggested as possibly relevant to FIP development. These sites fall within Spike protein subdomains that participate in host cell receptor interaction, immune evasion, tropism shifts, host cellular entry, and viral escape. There were fourteen sites (twelve novel sites) within Spike under positive selection associated with the FECV phenotype, almost exclusively within the S1/S2 FCS and adjacent to C domain, along with a signal of relaxed selection in FIPV relative to FECV, suggesting that furin cleavage functionality may not be needed for FIPV. Positive selection inferred in ORF7b was associated with the FECV phenotype and included twenty-four positively selected sites, while ORF7b had signals of relaxed selection in FIPV. We found evidence of positive selection in ORF3c in FCoV-wide analyses, but no specific association with the FIPV or FECV phenotype. We hypothesize that some combination of mutations in FECV may contribute to FIP development, and that it is unlikely to be one singular 'switch' mutational event. This work expands our understanding of the complexities of FIP development and provides insights into how evolutionary forces may alter pathogenesis in coronavirus genomes.

Comparison of Clinical and Laboratory Findings at Different Clinical Stages in Cats Naturally Infected with Feline Coronavirus

Abstract Feline coronavirus (FCoV) infections occur commonly in cats, with entrocyte and monocyte-macrophage tropism. Most FCoV-infected cats remain asymp tomatic, but up to 10% develop fatal feline infectious peritonitis (FIP). This study aims to investigate the diagnostic utility of clinical and laboratory examinations including serum and effusion AGP levels in cats either with symptomatic effusive FIP or asymptomatic feline enteric coronavirus (FECV). The study included 40 cats with effusive FIP and 10 cats with FECV infection. The FIP group was divided into two subgroups: abdominal (AE; n=30) and thoracic effusion (TE; n=10). Clinical and laboratory examinations, including serum or effusion AGP measurement, were performed. Among all the groups, TE group had higher body temperature, heart and respiratory rates (P&lt;0.000). Compared with the FECV group, the FIP group had lower pH and HCO3 levels and higher base excess and lactate levels (P&lt;0.05). The leukocyte and lymphocyte counts were higher and the hematocrit was lower in the AE group among all the groups (P&lt;0.023). MCV was lower in the FIP group compared to the FECV group (P&lt;0.002). In the AE group, total protein level was the lowest and the AST, GGT, total bilirubin and cholesterol levels were the highest (P&lt;0.032) among all the groups. Magnesium level was lower in the FIP group compared to the FECV group (P&lt;0.044). Although the serum AGP level was highest in the TE group among all groups (P&lt;0.004), the AGP levels of cats with FECV were similar to the AE group (P&gt;0.05). Since FECV-positive cats will likely develop FIP, differences in clinical and laboratory findings in FECV-positive cats were identified. Among them, pH, HCO3, base excess, lactate, MCV and magnesium were found to be important in the course of the disease, and AGP in the evaluation of the presence of an inflammatory state. It was concluded that clinical, laboratory and serum AGP evaluation could be used in the index of suspicion of development of FIP and FECV.

Investigation of some acute phase proteins, cytokines and hepcidin values in feline enteric corona virus antibodies and feline infectious peritonitis antigen positive cats

Feline Infectious Peritonitis (FIP) is a fatal disease caused by feline coronaviruses. The causative agent is Feline Infectious Peritonitis Virus, a mutation of Feline Enteric Coronavirus. Feline Corona Virus is very common in the cat population. In Feline Corona Virus infected cats, the development of FIP depends on the cat's immune response. FIP disease is more common in young and old cats because young and old animals have a weaker immune system. The acute phase response is a complex systemic reaction that occurs as a response to acute or chronic inflammatory processes such as infection, neoplasia or immunological disorders, tissue damage, trauma and surgery. The study material included 15 cat with presumptive FIP (study group) and 10 healthy cats (control group). Of the 15 cats, 9 (60%) showed dry and 6 (40%) effusive form. Serum amyloid A (SAA), haptoglobin (Hp), α1-acid glycoprotein (AGP), albumin, interleukin-6 (IL-6), hepcidin, alanine-amino transferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), blood urea nitrogen(BUN), and creatinine levels were measured in the serum collected from both groups. There were no difference between the wet and dry FIP in albumin values; haptoglobin, α1-acid glycoprotein, SAA and hepcidin values were significantly different between the two groups. IL-6, showed significant difference in both wet and dry FIP groups. It was also concluded that hepcidin may be used as a biomarker in Feline Infectious Peritonitis disease like other acute phase proteins.

Phylogenetic analysis of feline infectious peritonitis virus, feline enteric coronavirus, and severe acute respiratory syndrome coronavirus 2 of cats in Surabaya, Indonesia.

Questions about the origin of coronavirus and its introduction to human beings have persisted. The detection of a variety of coronavirus related to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in bats and pangolins led to the widespread belief that SARS-CoV-2 originated from wild ani-mals and was introduced to humans through an inter-mediate animal. Thus, coronaviruses from animals, especially those in close contact with humans, have attracted particular attention. This study aimed to phylogenetically analyze feline enteric coronavirus (FECV), feline infectious peritonitis virus (FIPV), and SARS-CoV-2 found in cats in Surabaya amid the COVID-19 pandemic. The results will provide a basis for developing basic preventive and pet healthcare strategies. Samples were collected on physical examinations of domestic and Persian cats (males and females) from March 2020 to March 2022. Samples were collected if there were clinical signs of FECV and FIP based on a veterinarian's diagnosis in several clinics in Surabaya. Laboratory examinations in this study were performed by reverse-transcription-polymerase chain reaction (RT-PCR) with primers for conserved regions of FIP and FECV, DNA sequencing was performed with Applied Biosystem Genetic Analyzer protocol, homology analysis was performed using Basic Local Alignment Search Tool NCBI, phylogenetic analysis was carried out with BioEdit 7.2 software, and sequences were compared with references from GenBank. Samples were collected from ten cats showing clinical signs of FECV and FIP, based on a veterinarian's diagnosis. On RT-PCR examinations performed with specifically designed primers for detecting FIPV in blood, peritoneal fluid, and feces, only one sample showed positivity for FIPV (1/10), namely, a peritoneal sample from a domestic cat in Surabaya. Homology analysis of the FIPV Surabaya isolate showed 98% similarity with FECV and FIPV reported in GenBank (MT444152 and DQ010921, respectively). In phylogenetic analysis, the FIPV Surabaya isolate was clustered together with SARS-CoV-2 of Clade A (MT198653) from Spain, SARS-CoV-2 Clade A (MT192765) from the USA, SARS-CoV-2 Clade D (039888) from the USA, and SARS-CoV-2 Clade F (MT020781) from Finland. This study revealed a relationship between the SARS-CoV-2 viruses that infect humans and cats (FECV), which is an important finding for those keeping cats at home. However, this finding requires further comprehensive support from laboratory studies.

A Retrospective Study of Viral Molecular Prevalences in Cats in Southern Italy (Campania Region).

From 2019 to 2021, a retrospective molecular study was conducted in the Campania region (southern Italy) to determine the prevalence of viral diseases in domestic cats. A total of 328 dead animals were analyzed by Real-Time PCR for the presence of feline panleukopenia virus (FPV), feline leukemia virus (FeLV), feline enteric coronavirus (FCoV), rotavirus (RVA), feline herpesvirus type 1 (FHV-1), and feline calicivirus (FCV). The possible presence of SARS-CoV-2 was also investigated by Real-Time PCR. The cats included in this study were specifically sourced and referred by local veterinarians and local authorities to the Zooprofilactic Experimental Institute of Southern Italy (IZSM) for pathological evaluation. The samples consisted of owners, catteries, and stray cats. Results revealed: 73.5% positive cats for FPV (189/257), 23.6% for FeLV (21/89), 21.5% for FCoV (56/266), 11.4% for RVA (16/140), 9.05% for FeHV-1 (21/232), and 7.04 for FCV (15/213). In contrast, SARS-CoV-2 was never detected. FPV was more prevalent in winter (p = 0.0027). FCoV FHV-1, FCV, and RVA predominated in autumn, whereas FeLV predominated in summer. As expected, viral infections were found more frequently in outdoor and shelter cats than in indoor ones, although no statistical association was found between animal lifestyle and viral presence. The study showed a high prevalence of FPV, FeLV, and FCoV and a moderate prevalence of RVA, FHV-1, and FCV. Moreover, the prevalence of these pathogens varied among the cat populations investigated.

Применение Фоспренила в составе комплексной терапии хронической коронавирусной инфекции кошек, осложненной токсоплазмозом

Feline enteric coronavirus (FECV) infects enterocytes and causes inflammation of the intestinal mucosa in young cats, resulting in diarrhea. In adult cats the infection is usually asymptomatic, but the constant circulation of FECV in the feline population increases the likelihood of mutation, as a result of which about 5-10% of infected cats develop a virulent strain of the virus that causes usually lethal feline infectious peritonitis – FIP. The purpose of this study was to study the effectiveness of Phosprenyl (PP), an immunomodulator with antiviral activity (active substance - polyprenyl phosphate), in the treatment of chronic coronavirus infection of cats, partially complicated by toxoplasmosis.&#x0D; A total of 117 cats were under observation, 10 of which also had toxoplasmosis. The diagnosis was confirmed by the determination of antibodies by the methods of IHA and ELISA, as well as by ultrasound examination of the abdominal organs. Prescribed treatment: Phosprenyl per os, 1 ml , twice a day, course 2-4 months; Gamavit daily, s/c, course 10 days, with a repeated course after 3 weeks; 3-4 times. As symptomatic therapy, drugs were used primarily to improve the condition of the stool, with toxoplasmosis - Fansidar. At the end of treatment symptoms of gastrointestinal tract damage disappeared in all the patients. If before the start of therapy, the majority of patients demonstrated changes characteristic of anemia (a significant decrease in the number of red blood cells and hemoglobin levels), then after 2-4 months, the indices of erythropoiesis completely normalized. In patients with coronavirus infection complicated by toxoplasmosis, anemia was detected by CBC test, as well as an increase in the number of neutrophils. In the biochemical profile of these patients: increased activity of the enzymes aspartate aminotransferase (AST) and alanine aminotransferase (ALT) was found as well as hyperbilirubinemia, which indicates greater liver damage and a decrease in its detoxification function during the course of the disease caused simultaneously by coronavirus and toxoplasmas. At the end of therapy, these indices also returned to normal. &#x0D; Thus, the use of Phosprenyl for 2-4 months reduces the duration of the disease of coronavirus infection in cats (including cases complicated by toxoplasmosis), prevents its exacerbation and possible fatal outcome. Long-term use of PP in cats with a chronic form of coronavirus infection is recommended.

Surface Display of Peptides Corresponding to the Heptad Repeat 2 Domain of the Feline Enteric Coronavirus Spike Protein on Bacillus subtilis Spores Elicits Protective Immune Responses Against Homologous Infection in a Feline Aminopeptidase-N-Transduced Mouse Model.

Although feline coronavirus (FCoV) infection is extremely common in cats, there are currently few effective treatments. A peptide derived from the heptad repeat 2 (HR2) domain of the coronavirus (CoV) spike protein has shown effective for inhibition of various human and animal CoVs in vitro, but further use of FCoV-HR2 in vivo has been limited by lack of practical delivery vectors and small animal infection model. To overcome these technical challenges, we first constructed a recombinant Bacillus subtilis (rBSCotB-HR2P) expressing spore coat protein B (CotB) fused to an HR2-derived peptide (HR2P) from a serotype II feline enteric CoV (FECV). Immunogenic capacity was evaluated in mice after intragastric or intranasal administration, showing that recombinant spores could trigger strong specific cellular and humoral immune responses. Furthermore, we developed a novel mouse model for FECV infection by transduction with its primary receptor (feline aminopeptidase N) using an E1/E3-deleted adenovirus type 5 vector. This model can be used to study the antiviral immune response and evaluate vaccines or drugs, and is an applicable choice to replace cats for the study of FECV. Oral administration of rBSCotB-HR2P in this mouse model effectively protected against FECV challenge and significantly reduced pathology in the digestive tract. Owing to its safety, low cost, and probiotic features, rBSCotB-HR2P is a promising oral vaccine candidate for use against FECV/FCoV infection in cats.

Crystallization of Feline Coronavirus Mpro With GC376 Reveals Mechanism of Inhibition.

Coronaviruses infect a variety of hosts in the animal kingdom, and while each virus is taxonomically different, they all infect their host via the same mechanism. The coronavirus main protease (Mpro, also called 3CLpro), is an attractive target for drug development due to its essential role in mediating viral replication and transcription. An Mpro inhibitor, GC376, has been shown to treat feline infectious peritonitis (FIP), a fatal infection in cats caused by internal mutations in the feline enteric coronavirus (FECV). Recently, our lab demonstrated that the feline drug, GC373, and prodrug, GC376, are potent inhibitors of SARS-CoV-2 Mpro and solved the structures in complex with the drugs; however, no crystal structures of the FIP virus (FIPV) Mpro with the feline drugs have been published so far. Here, we present crystal structures of FIPV Mpro-GC373/GC376 complexes, revealing the inhibitors covalently bound to Cys144 in the active site, similar to SARS-CoV-2 Mpro. Additionally, GC376 has a higher affinity for FIPV Mpro with lower nanomolar Ki values compared to SARS-CoV and SARS-CoV-2 Mpro. We also show that improved derivatives of GC376 have higher potency for FIPV Mpro. Since GC373 and GC376 represent strong starting points for structure-guided drug design, determining the crystal structures of FIPV Mpro with these inhibitors are important steps in drug optimization and structure-based broad-spectrum antiviral drug discovery.

ERDRP-0519 inhibits feline coronavirus in vitro

BackgroundCoronaviruses (CoVs) are major human and animal pathogens and antiviral drugs are pursued as a complementary strategy, chiefly if vaccines are not available. Feline infectious peritonitis (FIP) is a fatal systemic disease of felids caused by FIP virus (FIPV), a virulent pathotype of feline enteric coronavirus (FeCoV). Some antiviral drugs active on FIPV have been identified, but they are not available in veterinary medicine. ERDRP-0519 (ERDRP) is a non-nucleoside inhibitor, targeting viral RNA polymerase, effective against morbilliviruses in vitro and in vivo.ResultsThe antiviral efficacy of ERDRP against a type II FIPV was evaluated in vitro in Crandell Reese Feline Kidney (CRFK) cells. ERDRP significantly inhibited replication of FIPV in a dose-dependent manner. Viral infectivity was decreased by up to 3.00 logarithms in cell cultures whilst viral load, estimated by quantification of nucleic acids, was reduced by nearly 3.11 logaritms.ConclusionsThese findings confirm that ERDRP is highly effective against a CoV. Experiments will be necessary to assess whether ERDRP is suitable for treatment of FIPV in vivo.