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Severe clinical diseases associated to αCoronavirus (αCoV) infections were recently demonstrated for the first time in humans and a closely related but distinct canine CoV (CCoV) variant was identified in the nasopharyngeal swabs of children with pneumonia hospitalized in Malaysia, in 2017-2018. The complete genome sequence analysis demonstrated that the isolated strain, CCoV-HuPn-2018, was a novel canine-feline-like recombinant virus with a unique nucleoprotein. The occurrence of three human epidemics/pandemic caused by CoVs in the recent years and the detection of CCoV-HuPn-2018, raises questions about the ability of these viruses to overcome species barriers from their reservoirs jumping to humans. Interestingly, in this perspective, it is interesting to consider the report concerning new CCoV strains with a potential dual recombinant origin through partial S-gene exchange with porcine transmissible gastroenteritis virus (TGEV) identified in pups died with acute gastroenteritis in 2009. The significance of the ability of CCoVs to evolve is still unclear, but several questions arisen on the biology of these viruses, focusing important epidemiological outcomes in the field, in terms of both virus evolution and prophylaxis. The new CCoV-Hupn-2018 should lead researchers to pay more attention to the mechanisms of recombination among CoVs, rather than to the onset of variants as a result of mutations, suggesting a continuous monitoring of these viruses and in particular of SARS-CoV-2.

Feline infectious peritonitis (FIP) is a fatal systemic disease of felids caused by a Coronavirus (CoV) (FIPV). In spite of its clinical relevance and impact on feline health, currently the therapeutic possibilities for treatment of FIP in cats are limited. The emergence of the pandemic Severe Respiratory Syndrome (SARS) coronavirus (CoV) type 2 (SARS-CoV-2), etiological agent of the 2019 Coronavirus Disease (COVID-19), able to infect a broad spectrum of animal species including cats, triggered the interest for the development of novel molecules with antiviral activity for treatment of CoV infections in humans and animals. Essential oils (EOs) have raised significant attention for their antiviral properties integrating and, in some cases, replacing conventional drugs. Thymus vulgaris EO (TEO) has been previously shown to be effective against several RNA viruses including CoVs. In the present study the antiviral efficacy of TEO against FIPV was evaluated in vitro. TEO at 27 μg/ml was able to inhibit virus replication with a significant reduction of 2 log10 TCID50/50 μl. Moreover, virucidal activity was tested using TEO at 27 and 270 μg/ml, over the cytotoxic threshold, determining a reduction of viral titre as high as 3.25 log10 TCID50/50 μl up to 1 h of time contact. These results open several perspectives in terms of future applications and therapeutic possibilities for coronaviruses considering that FIPV infection in cats could be a potential model for the study of antivirals against CoVs.

Feline coronavirus (FCoV) exists as two different genotypes, FCoV type I and II, each including two biotypes, feline enteric coronavirus (FECV) and feline infectious peritonitis virus (FIPV), the latter being a virulent variant originating from the former virus. Recently, two amino acid substitutions, M1058L and S1060A, within the spike protein have been associated to the FECV/FIPV virulence change. In this study, we have analysed the frequency of detection of such mutations in FIPV and FECV strains circulating in Italian cats and obtained information about their evolutionary relationships with reference isolates. A total of 40 FCoV strains, including 19 strains from effusions or tissue samples of FIP cats and 21 strains from faecal samples of non-FIP cats, were analysed. Mutation M1058L was detected in 16/18 FCoV-I and 1/1 FCoV-II strains associated with FIP, while change S1060A was presented by two FIPV strains. By phylogenetic analysis, FCoV sequences clustered according to the genotype but not according to the biotype, with FECV/FIPV strains recovered from the same animal being closely related. Further studies are needed to better define the genetic signatures associated with the FECV/FIPV virulence shift.

Different strategies have been proposed to overcome maternally derived antibody (MDA) interference with canine parvovirus type 2 (CPV-2) immunisation, including intranasal vaccination, which presents some practical limitations. In the present study, the results of the oral administration of a commercial CPV-2b modified live virus (MLV) vaccine in pups with MDA are reported. The CPV-2b vaccine was orally administered to 14 6-week-old pups with a bait. Blood samples and rectal swabs were collected at different days post-vaccination (dpv) to determine CPV-2 antibody titres and DNA loads. Thirteen pups were positive to serological and virological tests after the first vaccination and one pup became positive after the second vaccine administration. The findings of this study suggest that systemic immunity against CPV-2 may be achieved by the use of an MLV CPV-2b vaccine administered orally even in the presence of MDA titres that usually interfere with vaccination.

Caprine alphaherpesvirus 1 (CpHV-1) induces genital lesions in its natural host similar to those caused by Human alphaherpesvirus 2 (HHV-2), commonly named herpes simplex virus 2 (HSV-2) in human patients. CpHV-1 infection in goats could represent a useful homologous animal model for the study of HSV-2 infection, chiefly for the assessment of antiviral drugs in in vivo studies. PHA767491 is a potent inhibitor of HSV-1 and HSV-2, being able to limit replication of HHVs both in vitro and in the mouse model. In the present study the antiviral efficacy of PHA767491 against CpHV-1 was evaluated in vitro in MDBK cells. PHA767491 inhibited significantly CpHV-1 replication in a dose-dependent fashion by up to 2.50 log10 TCID50/50 μl and was able to decrease viral DNA by nearly 8 log10. These findings confirm that PHA767491 is highly effective not only against simplexviruses (HSV-1 and HSV-2), but also against the varicellovirus CpHV-1. Experiments will be necessary to assess whether PHA767491 is suitable for treatment of vaginal lesions in CpHV-1-goat model. This could provide hints for the therapy of genital alphaherpesvirus infections in humans.

Background: Enteritis of an infectious origin is a major cause of productivity and economic losses to cattle producers worldwide. Several pathogens are believed to cause or contribute to the development of calf diarrhea. Astroviruses (AstVs) are neglected enteric pathogens in ruminants, but they have recently gained attention because of their possible association with encephalitis in humans and various animal species, including cattle. Objectives: This paper describes a large outbreak of neonatal diarrhea in buffalo calves (Bubalus bubalis), characterized by high mortality, which was associated with an AstV infection. Methods: Following an enteritis outbreak characterized by high morbidity (100%) and mortality (46.2%) in a herd of Mediterranean buffaloes (B. bubalis) in Italy, 16 samples from buffalo calves were tested with the molecular tools for common and uncommon enteric pathogens, including AstV, kobuvirus, and torovirus. Results: The samples tested negative for common enteric viral agents, including Rotavirus A, coronavirus, calicivirus, pestivirus, kobuvirus, and torovirus, while they tested positive for AstV. Overall, 62.5% (10/16) of the samples were positive in a single round reverse transcription polymerase chain reaction (PCR) assay for AstV, and 100% (16/16) were positive when nested PCR was performed. The strains identified in the outbreak showed a clonal origin and shared the closest genetic relationship with bovine AstVs (up to 85% amino acid identity in the capsid). Conclusions: This report indicates that AstVs should be included in a differential diagnosis of infectious diarrhea in buffalo calves.

Highly pathogenic bovine Delta papillomaviruses (δPVs) were detected and quantified for the first time using digital droplet polymerase chain reaction (ddPCR) by liquid biopsy in 103 clinically healthy sheep. Overall, ddPCR detected bovine δPVs in 68 blood samples (66%). Bovine papillomavirus (BPV) infection by a single genotype was revealed in 59% of the blood samples, and BPV coinfection by double, triple or quadruple genotypes was observed in 41% of liquid biopsies. The BPV-2 genotype was most frequently seen in sheep, whereas BPV-1 was the least common. Furthermore, ddPCR was very useful for detection and quantification; the BPV-14 genotype was observed for the first time in ovine species, displaying the highest prevalence in some geographical areas (Apulia). In 42 of the positive samples (61.8%), a single BPV infection was observed, 26 of which were caused by BPV-2 (61.9%) and 7 by BPV-13 (16.7%). BPV-14 was responsible for 7 single infections (16.7%) and BPV-1 for 2 single infections (4.7%). Multiple BPV coinfections were observed in the remaining 26 positive samples (38.2%), with dual BPV-2/BPV-13 infection being the most prevalent (84.6%). BPV infection by triple and quadruple genotypes was also observed in 11.5% and 3.8% of cases, respectively. The present study showed that ddPCR, a biotechnological refinement of conventional PCR, is by far the most sensitive and accurate assay for BPV detection. Therefore, ddPCR displayed diagnostic and epidemiological value resulting in the identification of otherwise undetectable BPV genotypes as well as their geographical distributions and suggesting that animal husbandry practices contribute to cross-species transmission of BPVs..

Feline calicivirus (FCV) is a contagious viral pathogen that usually causes a mild, self-limiting respiratory disease. More recently, highly virulent FCV strains have emerged and have been associated with severe systemic infection, referred to as virulent systemic disease (VSD). The objective of this study is to report VSD cases in Italian cats along with the molecular characterization of two detected FCV strains. Three client-owned cats showed clinical signs resembling to those described for VSD cases. The cats were subjected to molecular investigations for detection of FCV and other feline pathogens. Histopathology and immunohistochemistry were performed on internal organs of one cat; molecular characterization of two detected FCV strains was obtained through sequence and phylogenetic analyses. Putative VS-FCV strains were detected in all three cats, which were co-infected with feline panleukopenia virus. The cat submitted to histopathology and immunohistochemistry displayed severe histological changes and FCV antigens in internal organs. Two Italian FCV strains, for which amplification of ORF2 was successful, were strictly related and formed a unique phylogenetic cluster. These viruses did not show consistent changes in the amino acid sequences with respect to reference VS-FCVs. The results of our study confirm that VS-FCV strains are circulating in Italy and that VSD diagnosis is complicated since both genetic and clinical markers have not been identified so far.

BRD is associated with infectious agents, but management and transport-stress are trigger factors. Metaphylactic administration of antimicrobial reduces colonization of respiratory tract by pathogens, but the development of antibiotic-resistance raises public health concerns leading to propose new control strategies. The study analyzed nasopharyngeal swabs of 231 imported cattle, 10% of 49 trucks, transported from France to southern Italy and, through Real-time PCR identified the prevalence of the involved pathogens speculating on strategies to reduce the impact of BRD. The samples were tested by Real-time PCR, for the detection of bovine coronavirus (BCoV), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus (BPiV), bovine adenovirus (BAdV), Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. Yates-corrected chi squared, or Fisher's exact test were used to compare both animal-health status and positivity/negativity to pathogens, and the relationship between presence/absence of clinical signs and Real-time PCR-positivity. H. somni and BCoV were the most frequently identified pathogens. In BRD-diagnosed cattle, BAdV was detected in 13.8% (19/138), BRSV in 14.5% (20/138) and BPiV in 4.3% (6/138). Healthy cattle were mostly positive for H. somni (89.2%, 83/93). A statistically significant association was observed between clinical signs and positivity to M. haemolytica (p value = 0.016). Although mass-medication and vaccination are used for BRD control, it still remains a primary health problem. Our results highlight that the nasopharyngeal microbiota could be affected by transport and that strategies to enhance calf immunity for reducing BRD-risk development would be more effective if applied at farm of origin prior to loading.

Hepatitis E virus (HEV) typically causes self-limiting acute viral hepatitis, however chronic infection and extrahepatic manifestations have increasingly become a significant health problem. Domestic pigs and wild boars are the main reservoirs of HEV genotype 3 and genotype 4 for human infections in industrialized countries, although molecular and serological evidence suggest that several additional animal species may act as HEV hosts. In this study, by assessing serologically and molecularly the sera of 324 household cats from Apulia region (Italy), HEV antibodies were detected with an overall prevalence of 3.1%. Viral RNA was not detected in the sera of the animals using both HEV-specific assays and a pan-hepevirus broadly reactive set of primers for Hepeviridae. These findings document a low seroprevalence to HEV in cats in the investigated geographical setting. The exact nature of the HEV-like strains circulating in feline population remains to be established.

Bovine Pestivirus heterogeneity is a major challenge for vaccines against bovine viral diarrhea (BVD). In breeding herds, fetal protection is a high priority issue. To some degree, fetal infections in vaccinated heifers have been attributed to the antigenic diversity of bovine Pestiviruses. The purpose of this study was to assess fetal protection against a divergent bovine Pestivirus (Hobi-like Pestivirus, HoBiPeV) with a commercially available modified live vaccine (MLV) claiming fetal protection against BVDV 1 and BVDV 2 up to one year after the first inoculation. Five vaccinated and four unvaccinated heifers were challenged by intranasal inoculation with the HoBiPeV Italy-1/10-1 strain between 82 and 89 days after insemination, i.e. between 4 and 6 months after vaccination. At challenge, neutralizing antibody titers to HoBiPeV in vaccinated heifers were low or even undetectable. Of the four unvaccinated heifers, one control animal aborted (fetus not available) and the remaining three gave birth to HoBiPeV positive calves. Among the heifers of the vaccinated group, one aborted the fetus in the sixth month of pregnancy, which tested Pestivirus negative, while three others gave birth to healthy, HoBiPeV negative calves; the remaining heifer delivered one HoBiPeV positive calf. The results suggest that the BVDV vaccine might be able to elicit a partial fetal protection against HobiPeV, even in absence of a strong specific antibody response.

Hepatitis B virus (HBV) is a major cause of liver disease in humans including chronic hepatitis and hepatocellular carcinoma. Domestic cat hepadnavirus (DCH), a novel HBV-like hepadnavirus, was identified in domestic cats in 2018. From 6.5 %-10.8 % of pet cats are viremic for DCH and altered serological markers suggestive of liver damage have been identified in 50 % of DCH-infected cats. DCH DNA has been detected in association with characteristic lesions of chronic hepatitis and with hepatocellular carcinoma in cats, suggesting a possible association. In this study longitudinal molecular screening of cats infected with DCH was performed to determine if DCH can cause chronic infections in cats. Upon re-testing of sera from five DCH-positive animals, 2-10 months after the initial diagnosis, three cats tested negative for DCH on two consecutive occasions using quantitative PCR. Two other cats remained DCH-positive, including an 8-month-old female cat re-tested four months after the initial positive result, and a 9-year-old male cat, which tested positive for DCH on six occasions over an 11-month period. The latter had a history of chronic hepatopathy with jaundice, lethargy and elevated serum alanine transaminase levels (ALT). During the period of observation, DCH titers ranged between 1.64 × 105 and 2.09 × 106 DNA copies/mL and ALT was persistently elevated, suggesting chronic infection. DCH DNA was not detected in oral, conjunctival, preputial and rectal swabs from the two animals collected at several time points. Long-term (chronic) infection would be consistent with the relatively high number of viremic cats identified in epidemiological investigations, with the possible association of DCH with chronic hepatic pathologies and with what described with HBV in human patients.

Ready-to-eat (RTE) salads have recently been associated with food-borne norovirus outbreaks, although these infections are mainly related to shellfish and berry consumption in the EU. A total of 135 bagged RTE vegetables were analyzed in order to investigate the occurrence of norovirus (NoV) genotype I (GI) and II (GII) RNA and to differentiate between infectious and non-infectious viruses by using propidium monoazide (PMAxx) coupled with the real time Reverse Transcription (RT) PCR method. Initially, the PMAxx real time RT-PCR assay was optimized on NoV GI and GII suspensions, and proved capable of detecting significant (p &lt, 0.05) differences between infectious and inactivated viruses. Our analysis conducted on RTE salads samples showed the presence of norovirus GII in 74.8% of samples, of which 37.6% were infectious. The samples tested for viral contamination came from only two RTE vegetable-processing plants. The findings in this study could also be due to virally-contaminated water used in food production, processing, or preparation. This study stresses the need for effective real-time RT-PCR tools capable of qualitative and quantitative detection of NoV RNA, as well as being able to measure virus infectivity, for risk assessment, which is crucial in several public health measures and food regulations.

Highlights • Despite extensive vaccination, canine parvovirus (CPV) still represents one of the major causes of pups’ mortality. • CPV immunisation failures occur frequently and recognize different reasons. • Interference by maternally-derived antibodies is the main cause of CPV immunisation failures. • Eradication of CPV infection is a challenge for the future, but it will not be achieved in a short time, Despite extensive vaccination, canine parvovirus (CPV) remains a leading infectious cause of canine mortality, especially among juveniles. This review provides an update on CPV vaccine types and vaccination protocols. The design of CPV prevention strategies and vaccination programs with a goal of herd immunity has been hampered by deficiencies of studies that model companion animal viral infections and inform an understanding of the basic reproduction number. However, the most important issue in eradication of CPV disease is represented by immunisation failures including: i) the presence of interfering titres of maternally-derived antibodies; ii) the presence of non-responders; and iii) possible reversion to virulence. In contrast, the role of the CPV variants in immunisation failures is widely debated. Taking into account the reduced circulation of canine distemper virus and canine adenovirus type 1 in countries where extensive vaccination is carried out, more effort should be made to aim for CPV eradication, including antibody testing to determine the optimal time for vaccinations of pups and adults and homogeneous vaccine coverage of dog population.

Background Bovine alphaherpesvirus type 2 (BoHV-2) belongs to family Herpesviridae, subfamily Alphaherpesviridae and can cause two distinct, well-defined conditions: a generalized benign skin infection that somewhat mimics lumpy skin disease (LSD), referred to as Pseudo-Lumpy Skin Disease (PSLD) and a localized ulcerative mammillitis, referred to as Bovine Herpetic Mammillitis (BHM). BHM is a localized form of BoHV-2 infection that causes erosive-ulcerative self-limiting lesions on breast and nipples. BHM is chiefly a disease of lactating dairy cows and has been described sporadically in several countries. In this study we describe an outbreak of bovine herpetic mammillitis caused by BoHV-2 occurred in a dairy farm in Southern Italy. Clinical signs were observed in 26/59 lactating cows with the age ranging between 2 and 6 years. The affected animals were afebrile, showed lesions on the skin of nipples, breast and ventral surface of the abdomen, near the mammary veins and spontaneously recovered within 2 months. Results BoHV-2 DNA was detected in the crust samples by pan-herpes PCR and real-time quantitative PCR. The virus was isolated on bovine kidney cells and was characterised by deep sequencing technologies. The nucleotide identity to BoHV-2 of the strain ITA/2018/468 retrieved in this study ranged from 98.83 to 100%. Phylogenetic analyses based on three full-length gene (glycoprotein B, thymidine kinase and glycoprotein G) sequences confirmed the close relatedness of the strain ITA/2018/468 to BoHV-2 sequences. Conclusions The report represents a significant outbreak of BHM in a dairy farm 50 years after the last description in Italy. However, outbreaks of PLSD have been described in Europe recently, indicating that the virus is present in European territories. Improving the diagnostic algorithms and enacting specific surveillance plans could be useful to understand better the epidemiological and pathogenetic patterns of BoHV-2 infection in livestock animals, and to develop, eventually, effective prophylaxis plans.

Multiple, epizootic outbreaks of feline panleukopenia (FPL) caused by feline parvovirus(FPV) occurred in eastern Australia between 2014 and 2018. Most affected cats were unvaccinated.We hypothesised that low population immunity was a major driver of re‐emergent FPL. The aim ofthis study was to (i) determine the prevalence and predictors of seroprotective titres to FPV amongshelter‐housed and owned cats, and (ii) compare the prevalence of seroprotection between a regionaffected and unaffected by FPL outbreaks. FPV antibodies were detected by haemagglutinationinhibition assay on sera from 523 cats and titres &ge, 1:40 were considered protective. Socioeconomicindices based on postcode and census data were included in the risk factor analysis. The prevalenceof protective FPV antibody titres was high overall (94.3%), even though only 42% of cats wereknown to be vaccinated, and was not significantly different between outbreak and non‐outbreakregions. On multivariable logistic regression analysis vaccinated cats were 29.94 times more likelyto have protective FPV titres than cats not known to be vaccinated. Cats from postcodes of relativelyless socioeconomic disadvantage were 5.93 times more likely to have protective FPV titres. Thepredictors identified for FPV seroprotective titres indicate targeted vaccination strategies in regionsof socioeconomic disadvantage would be beneficial to increase population immunity. The criticallevel of vaccine coverage required to halt FPV transmission and prevent FPL outbreaks should bedetermined.

Feline calicivirus (FCV) is a major pathogen of cats associated with either respiratory disease or systemic disease, but its possible role as an enteric pathogen is neglected. Using RT-PCR, the RNA of FCV was identified in 25.9% (62/239) of stools of cats with enteritis and in 0/58 (0%) of cats without diarrhoea or other clinical signs. Isolates of enteric origin were obtained and a large 3.2-kb portion of the genome was sequenced, encompassing the 3' end of the RNA polymerase, the capsid protein precursor and the minor capsid protein. Also, the complete genome sequence of one such strain, the 160/2015/ITA, was determined. Upon sequence analysis, the enteric viruses were found to be genetically heterogeneous and to differ from each other and from isolates of respiratory origin. The enteric isolates were found to be more resistant to low pH conditions, to trypsin and to bile treatment than respiratory isolates. Overall, these findings are consistent with the hypothesis that some FCVs may acquire enteric tropism and eventually act as enteric pathogens. Whether this enteric tropism is maintained stably and whether it may affect, to some extent, the ability of the virus to trigger the classical and/or hypervirulent forms of disease should be assessed. Also, FCV should be included in the diagnostic algorithms of enteric diseases of cats to gain further information about FCV strains displaying enteric pathotype.

Aims To develop a protocol for environmental sampling to detect parvoviruses of dogs and cats in the environment. Methods and results Environmental contamination was carried out using different dilutions of parvovirus-contaminated materials; further field samplings were performed in areas in which clinical cases of parvovirus infections were present. Sterile cotton swabs and sponges for microbial surface sampling were used. Viruses were detected in these samples with different methods: conventional PCR, nested PCR and real-time PCR, detecting viral DNA; virus isolation, detecting infectious virus; and a commercial rapid enzyme immunoassay, detecting viral antigen. No substantial differences were observed in the two sampling methods, although the sponge was more convenient for sampling rough surfaces. Molecular assays were the most sensitive methods, identifying even very low amounts of viral DNA (up to 10 copies of viral DNA/10 µl of sample). Virus isolation and the rapid test detected the viruses only at the highest viral concentrations, both in the experimental setting and field conditions. Conclusions Environmental sampling and molecular protocols were effective in detecting environmental contamination with parvoviruses. Significance and impact of the study The protocol will be useful to identify possible sources of infection and to assess the efficacy of disinfection protocols in the environment.

The prevalence data of Leishmania infantum infection in cats are characterized by a large variability mainly attributed to the differences in diagnostic techniques. In the absence of consensus about the method of choice for diagnosing feline leishmaniosis, the performance of a new immunofluorescence antibody test (IFAT) was herein analytically described by the comparison with IFAT commonly used for the diagnosis of canine leishmaniosis (i.e., IFAT-OIE) and a laboratory enzyme-linked immunosorbent assay (ELISA). Sera of cats living in visceral leishmaniosis–endemic (n = 105) and visceral leishmaniosis–non-endemic (n = 50) areas were tested by the above methodologies and real-time PCR (qPCR). The most frequent result was represented by triple negativity to the three tests (IFAT-OIE, ELISA, and qPCR) in 42.9% and 80% cats from endemic and non-endemic areas, respectively. Bayes latent class analysis gave an output probability of 34.1% (posterior standard deviation, psd = 5.4%) of true L. infantum cases (TCL) which represent the true estimated prevalence of infection. The sensitivity of each variable contributing to define the TCL was 24% (psd = 6.3%) for qPCR, 78.8% (psd = 8.7%) for ELISA and 91.8% (psd = 5.2%) for IFAT-OIE. The probability to be a TCL was 94.5% for the sample from an endemic area. The cross-validation of the new IFAT by a logistic model correctly identified as positive 80.7% of subjects defined as TCL and negative 89.9% as not TCL, respectively, by the Bayesian model. The study results estimate a good accuracy of the IFAT in predicting cats exposed to L. infantum. Therefore, this procedure may be beneficial for screening cat populations for a better understanding of the epidemiology of feline leishmaniosis.

Astroviruses (AstVs) have been identified in the stools of calves with enteritis and in the brain tissues of bovines with encephalitis but their pathogenic role has not been clarified. In this study, we report the detection and characterization of bovine and water buffalo AstV strains identified in young bovine and buffalo calves with enteritis in Italy between 2012 and 2015. By negative staining transmission electron microscopy (TEM) observation, AstV-like particles were identified in the stools of the animals and AstV RNA was confirmed molecularly. The sequence (~3.2-kb) at the 3′ end of the genome was determined for two bovine and two buffalo AstVs. Sequence and phylogenetic analysis on the partial ORF1b and full-length ORF2 revealed a marked genetic diversity although the viruses were distantly related to other AstV identified from ruminants. Gathering sequence information on ruminant AstVs is important to understand the extent of inter-species circulation and for the development of reliable, specific diagnostic tools., Highlights • Astroviruses in ruminants are neglected enteric viruses. • Recently, neurotropic strains have been identified in ruminants. • Astrovirus was detected in outbreaks of enteritis in cattle and buffalo in Italy. • Genome sequencing of the viruses unveiled marked genetic diversity. • Implementing the diagnostics is pivotal to conceive surveillance studies.

A novel protoparvovirus species, related genetically to human bufaviruses, was identified in dogs with respiratory signs. The canine bufavirus was distantly related to the well-known canine protoparvovirus, canine parvovirus type 2, sharing low amino acid identities in the nonstructural protein 1 (40.6%) and in the capsid protein 1 (33.4%). By screening collections of fecal, nasal, and oropharyngeal samples obtained from juvenile dogs (