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Simple Summary: Hepadnaviruses, similar to human hepatitis B virus (HBV), have recently been discovered in several mammalian species, including domestic carnivores. Circulation of hepadnaviruses in cats (domestic cat hepadnavirus, DCH) has been documented in a number of molecular surveys, demonstrating a worldwide distribution. In contrast, studies on hepadnavirus epidemiology in dogs are still limited. In this study, we screened an age-stratified collection of 600 canine sera for the presence of antibodies against the recombinant DCH core antigen (DCHCAg) and DCH surface antigen (DCHSAg). DCHC antibodies were found with an overall prevalence of 10.0% (60/600). Out of 60 positive serum samples, 30.0% (18/60) also possessed IgG anti-DCHSAg. All samples were also assessed molecularly using a pan-hepadnavirus nested-PCR and a DCH-specific quantitative PCR to investigate the presence of viral DNA, revealing a prevalence rate of 0.7% (4/600). Our results demonstrated unequivocally that hepadnaviruses genetically and antigenically related to DCH circulate actively in the canine population, albeit with a lower prevalence than in cats. The discovery of hepadnaviruses in cats (domestic cat hepadnavirus, DCH) and of a DCH-like virus in dogs has raised several questions regarding the role of these viruses in pets, with particular emphasis on their potential impact on animal health and epidemiology, as well as possible zoonotic implications. In this study, by screening an age-stratified collection of 600 canine serum samples for DCH with an ELISA assay based on the recombinant core antigen (DCHCAg), specific antibodies were found with an overall prevalence of 10.0% (60/600), with a higher prevalence in younger and older dogs. By retesting the canine DCHCAbs-positive sera with an ELISA test based on the recombinant surface protein of DCH (DCHSAg), a total of 18 sera (30%, 18/60) also contained IgG anti-DCHSAg. All the sera were also assessed molecularly using either a consensus hepadnavirus PCR or a specific real-time PCR for DCH. Hepadnavirus DNA was detected in four seronegative dogs, with a prevalence rate of 0.7% (4/600). On sequence analysis of the polymerase region amplified with pan-hepadnavirus primers, the amplicons displayed the highest nucleotide identity (97.3–99.6%) to DCH sequences detected in cats and to the domestic dog hepadnavirus recently identified in a canine serum sample from Italy. [ABSTRACT FROM AUTHOR]

Highly divergent picornaviruses (PVs) classified in the genus Bopivirus have been recently discovered on faecal samples from sheep and goats in Hungary and from fallow and red deer in Australia. In this study, we investigated the epidemiology of these novel viruses in domestic and wild ruminants from Northwestern Italian Alps by testing archival faecal samples collected from 128 sheep, 167 goats, 61 red deer (Cervus elaphus), 77 roe deer (Capreolus capreolus), 43 chamois (Rupicapra rupicapra) and 32 Alpine ibex (Capra ibex). Bopivirus RNA was detected in a total of 19 animals, including 14 sheep (10.9%), 2 red deer (3.3%), 1 roe deer (1.3%), 1 chamois (2.3 %) and 1 Alpine ibex (3.3 %), but not in goats. Upon sequence analysis of the 3DRdRp region, the sequences generated from chamois, roe deer, Alpine ibex and ovine faecal samples showed the highest nucleotide identity (96.8–100%) to bopiviruses detected in goats and sheep from Hungarian farms, whereas strains found in red deer displayed the closest relatedness (90.8%–91.2%) to bopiviruses identified in fallow and red deer in Australia. The nearly complete genome sequence of strains 12/2020/ITA (ON497046) and 14–73/2020/ITA (ON497047) detected in an Alpine ibex and in a sheep, respectively, was determined by combining a modified 3'‐RACE protocol with Oxford Nanopore Technologies sequencing platform. On phylogenetic analysis based on the complete polyprotein, both strains segregated into the candidate species Bopivirus B along with ovine and caprine strains detected in Hungary (90.0–94.6% nucleotide and 94.6–98.0% amino acid identities). The findings of this study expand the host range of these novel viruses and hint to a possible virus circulation between domestic ruminants and wild animals. [ABSTRACT FROM AUTHOR]

A novel orthohepadnavirus (domestic cat hepadnavirus [DCH]) similar to human hepatitis B virus has been recently detected in serum and liver samples from domestic cats with chronic hepatitis and hepatocellular carcinoma. Molecular investigations by independent research groups around the world have revealed positivity rates ranging from 6.5% to 12.5% in blood samples and up to 14.0% in liver tissue. In this study, we screened an age‐stratified collection of feline sera (n = 256) by using an antibody detection enzyme‐linked immunosorbent assay based on the recombinant core antigen of DCH (DCHc). Specific antibodies (DCHc Abs) were detected with a prevalence of 25.0%. The DNA of DCH was detected in 35.9% (23/64) of seropositive cats and only in 1.0% (2/192) of seronegative animals. Based on the serological (IgG and IgM anti‐DCHc) and virological status, the possible stages of DCH infection were predicted. [ABSTRACT FROM AUTHOR]

Simple Summary: Data on hepatitis E virus (HEV) strains circulating in carnivores are still limited. However, serological evidence suggests that their exposure to HEV infection is more common than expected. In addition, close relatives of Orthohepevirus C detected in rats and in human patients with persistent hepatitis have been detected in ferrets, minks, and foxes, providing a rationale for investigating the presence of HEV or HEV-like viruses in wild carnivores. Herewith, we report the identification and the nearly full-genome (7212 nucleotides) characterization of an HEV Gt3 strain detected in a faecal specimen collected from a wolf found dead in Northwestern Italy. This finding poses questions concerning the potential role of wolves as HEV mechanical, passive carriers, favouring the introduction and spread of HEV in the environment. Hepatitis E virus (HEV) infection is a major health problem worldwide. In developed countries, zoonotic transmission of HEV genotypes (Gt) 3 and 4 is caused by the ingestion of raw or undercooked meat of infected pigs and wild boars, the main reservoirs of HEV. However, additional animals may harbour HEV or HEV-related strains, including carnivores. In this study, we investigated the molecular epidemiology of orthohepeviruses in wild canids by screening a total of 136 archival faecal samples, collected from wolves (42) and red foxes (94) in Northwestern Italy. Orthohepevirus RNA was identified in a faecal specimen, collected from a wolf carcass in the province of La Spezia (Liguria Region, Italy). The nearly full-length (7212 nucleotides) genome of the strain HEV/81236/Wolf/2019/ITA (GenBank accession no. MZ463196) was determined by combining a sequence-independent single-primer amplification (SISPA) approach with the Oxford Nanopore Technologies sequencing platform. Upon phylogenetic analysis, the HEV detected in wolf was segregated into clade HEV-3.1, displaying the highest nucleotide (nt) identity (89.0–93.3%) to Gt3 strains belonging to subtype c. Interestingly, the wolf faecal sample also contained porcine astrovirus sequences, endorsing the hypothesis of a dietary origin of the HEV strain due to preying habits. [ABSTRACT FROM AUTHOR]

In recent years, advances in diagnostics and deep sequencing technologies have led to the identification and characterization of novel viruses in cats as protoparviruses and chaphamaparvoviruses, unveiling the diversity of the feline virome in the respiratory tract. Observational, epidemiological and experimental data are necessary to demonstrate firmly if some viruses are able to cause disease, as this information may be confounded by virus- or host-related factors. Also, in recent years, researchers were able to monitor multiple examples of transmission to felids of viruses with high pathogenic potential, such as the influenza virus strains H5N1, H1N1, H7N2, H5N6 and H3N2, and in the late 2019, the human hypervirulent coronavirus SARS-CoV-2. These findings suggest that the study of viral infections always requires a multi-disciplinary approach inspired by the One Health vision. By reviewing the literature, we provide herewith an update on the emerging viruses identified in cats and their potential association with respiratory disease. [ABSTRACT FROM AUTHOR]

Simple Summary: Feline calicivirus (FCV) is a common viral pathogen affecting domestic cats, which is responsible for diverse clinical presentations, commonly including upper respiratory tract signs, oral ulcerations, and a fever. In addition, FCV infection can be associated with severe pneumonia, lameness, and virulent systemic disease. In this study, we clinically and pathologically describe a rare case of FCV-associated paw and mouth disease in a febrile household cat. The FCV strains detected in the animal and in an overtly healthy cohabiting cat were analyzed to assess the phenotype and antigenic properties. Feline calicivirus (FCV) infection in cats can led to several diverse clinical presentations, ranging from mild upper respiratory signs to virulent systemic disease. Herein, we report a paw and mouth disease case in a 7-year-old household cat due to an FCV infection. An asymptomatic cat living in the same household was also infected with FCV. Clinical and pathological investigations were combined with the molecular and phenotypical characterization of the FCV strains. The RNA of the FCV was detected using qualitative and quantitative reverse transcription (RT)-PCR assays, and FCV antigen was confirmed by immunohistochemistry. After the whole genome analysis, the strains detected in the two cats appeared to be genetically diverse from FCVs previously detected in association with paw and mouth disease and with virulent systemic disease. Interestingly, the isolates obtained in this study were resistant to low pH conditions and slightly susceptible to bile salts, but they were susceptible to a trypsin treatment, revealing a phenotype pattern that is different from that which has been observed for respiratory FCVs. [ABSTRACT FROM AUTHOR]

Feline chaphamaparvovirus (FeChPV) is a novel parvovirus, first discovered in a multi‐facility feline shelter in Canada in 2019, during an outbreak of acute gastro‐enteritis (AGE) in cats, and detected at high prevalence (47.0%) in faecal samples. Whether this finding was anecdotal or similar viruses are common components of feline virome is still unclear. Also, the potential impact of this virus on feline health is uncertain. Herewith, a case–control study was performed to investigate whether this novel parvovirus may play a role as enteric pathogen, screening samples collected from cats with and without AGE signs. Furthermore, we extended the research by testing archival paired oropharyngeal and ocular samples collected from cats with or without upper respiratory tract disease (URTD). FeChPV DNA was detected at high prevalence rate (36.8%, 14/38) in clinical cases, representing the most frequently identified enteric virus, followed by feline panleukopenia parvovirus (23.7%, 9/38), feline coronavirus (5.3%, 2/38), feline kobuvirus (5.3%, 2/38) and noroviruses (5.3%, 2/38). The different prevalence rates of FeChPV between the case and control group were statistically significant, suggesting a possible association of the virus with acute gastro‐enteric disease. The virus was also detected at low rate in the respiratory samples of cats with (3.3%, 6/183) or without URTD (4.3%, 6/140), although there was no significant association between FeChPV and URTD. The complete VP encoding gene was determined for five viruses and the nearly full‐length genome was reconstructed for three viruses, namely 313R/2019/ITA, 284R/2019/ITA and 49E/2019/ITA. In the NS1‐based tree, the Italian strains clustered tightly with the two FeChPV prototypes detected in Canada, within a monophyletic cluster related to but clearly distinct from canine chaphamaparvovirus, currently classified in the species Carnivore chaphamaparvovirus 1 (CaChPV‐1). [ABSTRACT FROM AUTHOR]

Summary: Canine bufavirus (CBuV) is a protoparvovirus, genetically related to human and non‐human primate bufaviruses and distantly related to canine parvovirus type 2 (CPV‐2). CBuV was initially identified from young dogs with respiratory signs but subsequent studies revealed that this virus is also a common component of the canine enteric virome. In this survey, by assessing archival and recent collections of dogs faecal samples, CBuV DNA was detected with a higher prevalence rate (8.8%) in animals with enteritis than in control animals (5.0%), although this difference was not statistically significant. The rate of co‐infections with other enteric viruses in diarrhoeic dogs was high (84.6%), mostly in association with canine parvovirus CPV‐2 (90.1%). The complete ORF2 gene was determined in five samples, and the nearly full‐length genome was reconstructed for three strains, 62/2017/ITA, 9AS/2005/ITA and 35/2018/ITA. Upon sequence comparison, the viruses appeared highly conserved in the NS1 (97.2%–97.9% nt and 97.5%–98.1% aa identities). In the complete VP2 coding region, three strains were similar to the prototype viruses (99.7–99.8 nt and 99.6%–99.8% aa) whilst strains 9AS/2005/ITA and 35/2016/ITA were distantly related (87.6%–89.3% nt and 93.9%–95.1% aa identities). Interestingly, genetic diversification occurred downstream conserved regions such as the VP1/VP2 splicing signals and/or the G‐rich motif in the N terminus of the VP2, suggesting a potential recombination nature. Upon phylogenetic analysis, the two divergent CBuV strains formed a distinct cluster/genotype. [ABSTRACT FROM AUTHOR]

Simple Summary: Hepatitis E virus (HEV) infection can cause both acute and chronic hepatitis in humans and represents an emerging public health concern worldwide. In developed countries, zoonotic transmission of HEV genotypes 3 and 4 is caused by ingestion of raw or undercooked meat of infected swine or wild boars, the main reservoirs of HEV. However, in the last few years, molecular and serological evidence seem to indicate that several other animal species may act as HEV host, including domestic and wild ruminants. In this study, serum and fecal specimens from sheep, goats, red deer, roe deer, chamois, and Alpine ibex collected in two northwestern Italian regions (Piemonte and Valle d'Aosta) were screened molecularly and serologically. With the exception of chamois, HEV antibodies were found both in the domestic and wild ruminant species investigated with the highest rates in sheep and goats. These findings demonstrate that wild also domestic ruminants may be implicated in the viral cycle transmission. In industrialized countries, increasing autochthonous infections of hepatitis E virus (HEV) are caused by zoonotic transmission of genotypes (Gts) 3 and 4, mainly through consumption of contaminated raw or undercooked pork meat. Although swine and wild boar are recognized as the main reservoir for Gt3 and Gt4, accumulating evidence indicates that other animal species, including domestic and wild ruminants, may harbor HEV. Herein, we screened molecularly and serologically serum and fecal samples from two domestic and four wild ruminant species collected in Valle d'Aosta and Piemonte regions (northwestern Italy. HEV antibodies were found in sheep (21.6%), goats (11.4%), red deer (2.6%), roe deer (3.1%), and in Alpine ibex (6.3%). Molecular screening was performed using different primer sets targeting highly conserved regions of hepeviruses and HEV RNA, although at low viral loads, was detected in four fecal specimens (3.0%, 4/134) collected from two HEV seropositive sheep herds. Taken together, the data obtained document the circulation of HEV in the geographical area assessed both in wild and domestic ruminants, but with the highest seroprevalence in sheep and goats. Consistently with results from other studies conducted in southern Italy, circulation of HEV among small domestic ruminants seems to occur more frequently than expected. [ABSTRACT FROM AUTHOR]

• In Italy some geographical areas are considered at high risk of HEV infection. • Here, sewage samples from Abruzzo Region (South-Italy) were assessed for HEV RNA. • We detected an overall prevalence of 23.2%, the highest to date reported in Italy. • HEVs closely related to human and animal strains from the same Region were found. Hepatitis E virus (HEV) infections constitute a significant health problem worldwide. The burden of hepatitis E in Italy seems low when compared with other European countries. In recent years, improved surveillance activities in Italy have revealed marked geographical differences in HEV epidemiology, with some regions characterised by higher seroprevalence rates. Abruzzo Region (Southern Italy) is currently recognised as a high-risk area for HEV infection. In this study, we investigated the epidemiology of HEV in Teramo Province by monitoring four wastewater treatment plants (WWTPs). Out of 56 influent sewage specimens collected during 2016–2017, HEV RNA was detected in 13/56 (23.2%) sewage samples from all the four WWTPs. Upon sequence analysis of the partial ORF2 gene, four strains showed the highest nucleotide identity to Gt3 subtype c, being more closely related to other HEVs previously identified in human and animal hosts in Abruzzo. For one strain, sequence data were generated only for a short region of the ORF1 gene, revealing the highest identity to HEVs Gt3 of subtype f. Altogether, the findings of this study confirm that HEV largely circulates in the setting investigated. [ABSTRACT FROM AUTHOR]