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Coronaviruses (CoVs) can cross species barriers and endanger public health. Despite reports on their circulation and evolution in companion animals during the pandemic, postpandemic surveillance remains crucial. Therefore, during the first postpandemic year, 309 samples from 263 companion animals (cats and dogs) in Chengdu, China, were detected for CoVs using a universal assay based on Rdrp genes combined with one‐generation sequencing. Four kinds of CoVs, including feline CoV (FCoV), canine CoV (CCoV), CRCoV, and SARS‐CoV‐2 (the first reported case of SARS‐CoV‐2 in a dog in mainland China, confirmed by viral nucleic acid detection and analysis), were detected with an overall positive rate of 21.7% (57/263); FCoV‐I and CCoV‐IIa were the dominant genotypes, and of these 57 positive cases, 71.9% (41/57) were in pets ≤12 months old. In CCoV‐positive dogs, 72.2% (13/18) were coinfected with other viruses (primarily canine parvovirus [CPV], 76.9%; 10/13), while 13.9% (5/36) codetection with feline parvovirus (FPV). A 21‐nt deletion in two FCoV S genes and a 145‐nt deletion in one FCoV ORF3abc gene were identified, and recombination events at positions 919 and 1639 nt in two S genes were noticed. Notably, the amino acid variations in FCoV and CCoV S genes revealed distinct regional adaptations: FCoV strains showed unique substitutions (e.g., Ala/Ser129Leu) and a shift from RSRR to RARR furin cleavage motifs; CCoV strains in China exhibited significant differences from those in other countries. Phylogenetic analysis demonstrated that the S genes of FCoV and CCoV were closely related to those of the prevalent strains in China, whereas the S genes of CRCoV were closely related to that of human CoV (HCoV) OC43. These findings highlight the need for continued surveillance of CoV infection in companion animals (especially ≤12 months old) in the postpandemic era.

Viral respiratory and intestinal infections are the most common causes of canine viral illness. Infection with multiple pathogens occurs in many cases. Rapid diagnosis of these multiple infections is important for providing timely and effective treatment. To improve diagnosis, in this study, two new multiplex polymerase chain reactions (mPCRs) were developed for simultaneous detection of canine respiratory viruses (CRV) and canine enteric viruses (CEV) using two separate primer mixes. The viruses included canine adenovirus type 2 (CAV-2), canine distemper virus (CDV), canine influenza virus (CIV), canine parainfluenza virus (CPIV), canine circovirus (CanineCV), canine coronavirus (CCoV) and canine parvovirus (CPV). The sensitivity of the mPCR results showed that the detection limit of both mPCR methods was 1×104 viral copies. Twenty nasal swabs (NS) and 20 anal swabs (AS) collected from dogs with symptoms of respiratory disease or enteric disease were evaluated using the novel mPCR methods as a clinical test. The mPCR protocols, when applied to these respiratory specimens and intestinal samples, could detect 7 viruses simultaneously, allowing rapid investigation of CRV (CAV-2, CDV, CIV and CPIV) and CEV (CAV-2, CanineCV, CCoV and CPV) status and prompt evaluation of coinfection. Our study provides an effective and accurate tool for rapid differential diagnosis and epidemiological surveillance in dogs.

Canine respiratory coronavirus (CRCoV) is a prevalent pathogen implicated in canine infectious respiratory disease, yet information on its genomic characteristics and pathogenicity remains scarce. To address this situation, we investigated the genetic evolution and pathogenic potential of CRCoV strains circulating in China. Five complete CRCoV genomes (GenBank: PQ725948-PQ725952) were obtained from clinical samples, and phylogenetic analysis showed these strains formed a distinct genetic branch. The evolutionary trees for ORF1ab, HE, and S genes closely mirrored the full genome tree, indicating key roles for these genes in CRCoV evolution. Multiple unique amino acid mutations were identified in the ORF1ab, HE, S, M, and N proteins. Notably, molecular docking analysis suggests that mutations S158F and L161F in the HE lectin domain are associated with improved docking scores, indicating a potential increase in receptor-binding affinity. Consecutive nucleotide deletions in two non-coding regions between non-structural protein genes-which were also identified in strains of a Thai lineage (OQ621707.1-OQ621727.1)-were observed. A CRCoV strain (10 6 TCID 50 /mL) was isolated, and experimental infection confirmed its ability to induce pneumonia and tracheal cilia loss in dogs. These findings reveal the emergence and unique genetic diversity of a novel CRCoV variant in China, highlighting the need for ongoing epidemiological surveillance.

Background Canine enteric coronavirus (CECoV) is a prevalent infectious disease among dogs worldwide, yet its epidemiology in mainland China remains poorly understood. This systematic review and meta-analysis aimed to assess the prevalence of CECoV in mainland China and identify factors influencing its prevalence. Methods A comprehensive literature search was conducted across multiple databases for studies regarding CECoV epidemiology of China. PubMed, CNKI, Wanfang, and CQVIP were searched to obtain the studies. Eligible studies were selected based on predefined criteria, and data were extracted and synthesized. The quality the studies was assessed using the JBI assessment tool. Heterogeneity was checked using I 2  test statistics followed by subgroup and sensitivity analysis. Subgroup analyses were performed to explore variations in CECoV prevalence by factors such as year, region, season, health status, social housing type, gender, age, and breed. Publication bias was assessed using a funnel plot and eggers test that was followed by trim and fill analysis. Results A total of 27 studies involving 21,034 samples were included in the meta-analysis. The overall pooled prevalence of CECoV in mainland China was estimated to be 0.30 (95% CI 0.24–0.37), indicating persistent circulation of the virus. Subgroup analyses revealed higher prevalence rates in younger dogs, multi-dog households, apparently healthy dogs, and certain regions such as southwest China. Seasonal variations were observed, with lower prevalence rates in summer. However, no significant differences in prevalence were found by gender. Conclusions This study provides valuable insights into the epidemiology of CECoV in mainland China, highlighting the persistent circulation of the virus and identifying factors associated with higher prevalence rates. Continuous monitoring and surveillance efforts, along with research into accurate detection methods and preventive measures, are essential for the effective control of CECoV and mitigation of its potential impact on animal and human health.

The aim of this study was to establish a multiplex PCR (mPCR) method that can simultaneously detect canine parvovirus (CPV-2), canine coronavirus (CCoV) and canine adenovirus (CAV), thereby eliminating the need to detect these pathogens individually. Based on conserved regions in the genomes of these three viruses, the VP2 gene of CPV-2, the endoribonuclease nsp15 gene of CCoV, and the 52K gene of CAV were selected for primer design. The specificity of the mPCR results showed no amplification of canine distemper virus (CDV), canine parainfluenza virus (CPIV), or pseudorabies virus (PRV), indicating that the method had good specificity. A sensitivity test showed that the detection limit of the mPCR method was 1 × 10 4 viral copies. A total of 63 rectal swabs from dogs with diarrheal symptoms were evaluated using mPCR and routine PCR. The ratio of positive samples to total samples for CPV-2, CCoV, and CAV was 55.6% (35/63) for mPCR and 55.6% (35/63) for routine PCR. Thirty-five positive samples were detected by both methods, for a coincidence ratio of 100%. This mPCR method can simultaneously detect CCoV (CCoV-II), CAV (CAV-1, CAV-2) and CPV-2 (CPV-2a, CPV-2b, CPV-2c), which are associated with viral enteritis, thereby providing an efficient, inexpensive, specific, and accurate new tool for clinical diagnosis and laboratory epidemiological investigations.

Although their pathogenicity is most often unclear, some canine viruses have been found to infect carnivores other than dogs. This study relies on the surveillance of coronaviruses in 206 saliva and fecal samples of huntable, sympatric canid and mustelid species captured in Hungary, such as the native red fox ( Vulpes vulpes ), European badger ( Meles meles ), golden jackal ( Canis aureus ), and stone marten ( Martes foina ), as well as the recently settled alien raccoon dog ( Nyctereutes procyonoides ). Metagenomics‐based and direct sequence analysis were deployed to determine the genome sequence of coronaviruses identified in two specimens collected from red foxes. Near‐complete genome sequences of two canine coronaviruses (CCoVs) were obtained, together with the complete genome sequence of a canine circovirus (CanineCV) and the near‐complete genome sequence of a canine picodicistrovirus (CPDV) from one of the samples. These provided the first fox origin CCoV and CPDV sequence data, and the first recorded appearance of the CPDV in Europe. The results suggested that recombination is of great importance in the evolution of CCoV, CanineCV, and CPDV infecting dogs and wild‐living carnivores, including the red fox and golden jackal. These are widespread in Central and Southeast Europe, and have large ranges, facilitating transmission of the multihost canine pathogens.

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.

The one-step polymerase chain reaction (one-step PCR) detection assay is an innovative PCR detection method, eliminating nucleic acid extraction steps, in which samples can be directly added to PCR reagents for testing. For simultaneous detection of CDV and CCoV, a sensitive and specific one-step duplex PCR (one-step dPCR) assay was developed with two pairs of primers that were designed based on H and M gene sequences of CDV and CCoV, respectively. The one-step dPCR with optimized detection conditions has high specificity and sensitivity; independent sequencing assays further verified these results.

Canine coronavirus (CCoV) is a common agent of gastroenteritis in dogs, although some variants have been found associated with systemic and often fatal diseases. Distinct genotypes (CCoV-I and CCoV-II) and subgenotypes (CCoV-IIa and CCoV-IIb) are worldwide distributed. In Italy, CCoV infections have been occasionally evaluated, but information about the molecular epidemiology and the genomic features of currently circulating strains is limited. This study reports the detection and molecular characterization of CCoV strains from samples collected from 284 dogs in Italy between 2019 and 2021. CCoV RNA was detected in 39 (13.7%) dogs, as a single viral agent (5 animals, 12.8%) or with other viral pathogens (canine parvovirus types 2a/2b/2c; canine adenovirus type 1; norovirus GIV.2) (34 animals, 87.2%). A total of 48 CCoV strains were detected either alone (CCoV-I: 51.3%, CCoV-IIa: 20.5%) or in copresence (CCoV-I and CCoV-IIa, 23.1%); surprisingly, CCoV-IIb was not identified in this study. Five clusters of CCoV-I were detected, and their spike gene sequences showed the highest nucleotide identities with CCoV-I strains collected from Greece in 2008/2009 and from China in 2021. CCoV-IIa spike gene sequences (three variants) had the highest nucleotide identities with CCoV-IIa strains collected in Greece in 2008/2009 and in Italy in 2009/2011. Given the high CCoV diversity and the variable pathogenicity potential, we underline the need of further surveillance studies to increase our understanding of the epidemiology and evolution of these viruses.

Viral and chemical analyses were performed on 80 dead cats and 51 dead dogs from the Campania Region (Southern Italy), with the aim of evaluating in vivo the potential correlation between coronavirus (CoV) infections and levels of environmental pollutants such as dioxins and PCSs (PCDD/F, DL-PCB and NDL-PCB). The overall viral prevalence was 16.3% in cats and 23.5% in dogs. Both feline coronavirus (FCoV) and canine coronavirus (CCoV) were identified, with variable detection rates in all the other organs investigated, supporting studies that provide evidence of systemic viral spread. The highest prevalence of coronaviruses (CoVs) was observed in Naples (19.2% for FCoV; 30.7% for CCoV) and Caserta (11.1% for FCoV; 50.0% for CCoV), areas that include municipalities with the highest Municipality Index of Environmental Pressure (MIEP) scores. Chemical analyses showed that DL-PCBs were present at more elevated concentrations in CoV-infected dogs and cats than in non-infected animals, whereas ∑NDL-PCB and ∑PCDD/F were detected in greater amounts in non-infected subjects. Among PCDDs, the congener 2,3,7,8-TCDD displayed different distribution patterns between infected and non-infected animals. In cats, 70.0% of FCoV-positive individuals had 2,3,7,8-TCDD levels above the limit of quantification (LOQ), compared with 38.0% of FCoV-negative cats. In dogs, 78.0% of CCoV-infected animals exceeded the LOQ, compared with 20.0% of non-infected ones; this difference was statistically significant. The results of the study suggest that elevated levels of 2,3,7,8-TCDD may be associated with CCoV infection and replication in dogs, suggesting a possible relationship between environmental pollution and susceptibility to coronavirus infections.