A Hobi-like pestivirus pair consisting of cytopathogenic (cp) and non-cytopathogenic (noncp) strains, Italy 83/10cp and Italy 83/10ncp, was isolated from the lung of a heifer that died of respiratory disease. The noncp and cp viruses were isolated on Madin–Darby bovine kidney cells and separated by plaque purification and end point dilution. Analysis of the nearly full-length genomes revealed that the two viruses were very closely related to each other and to the noncp Hobi-like strain Italy 1/10-1, which had been isolated a few weeks earlier from the same herd. One major difference between noncp and cp viruses concerned the presence of a cellular Jiv sequence in the 39 domain of the NS2-encoding region of the cp strain. This is the first study, to our knowledge, reporting the isolation and molecular characterization of a Hobi-like virus pair.

The clinical features and economic impact of the infection caused by an emerging group of pestiviruses, namely HoBi-like pestivirus, in a cattle herd of southern Italy are reported. In 2011, the virus was first associated with respiratory disease, causing an abortion storm after 1 year and apparently disappearing for the following 3 years after persistently infected calves were slaughtered. However, in 2014, reproductive failures and acute gastroenteritis were observed in the same herd, leading to a marked decrease of productivity. A HoBi-like strain closely related to that responsible for previous outbreaks was detected in several animals. Application of an intensive eradication programme, based on the detection and slaughtering of HoBi-like pestivirus persistently infected animals, resulted in a marked improvement of the productive performances.

Astroviruses are notable agents of gastroenteritis in many mammalian and avian hosts. Astroviruses are nonenveloped RNA small, round, viruses (SRVs) with a single-stranded, positive sense RNA of 6.1 to 7.9 kb (1). The genome contains 2 nonstructural genes, open reading frame (ORF) 1a and 1b, and a capsid gene, ORF2, with short 5′ and 3′ untranslated regions. Human astroviruses, a major cause of gastroenteritis, are classified in the human astrovirus species, comprising 8 serotypes (1). Recently, astroviruses genetically unrelated to canonical human astroviruses have been identified in human stools in several countries. These unusual astroviruses form 2 main genetic clades. One clade contains MLB1, MLB2, and MLB3 (2–4). The second clade contains VA1, VA2, VA3 (also known as HMO-C, HMO-A, and HMO-B, respectively) and VA4 (5,6). More recently, a VA1/HMO-C–like virus was detected in brain tissue from an immunocompromised child with encephalitis (7). The discoveries of these viruses provide novel candidate agents of human disease and raise concerns inherent of possible zoonotic implications. Here we describe the detection and genome characterization of MLB1-like astrovirus in a 4-year-old male child hospitalized with severe gastroenteritis during January 2007 at the University Hospital of Parma, Italy. Clinical signs included vomiting and severe diarrhea, with moderate dehydration. The child was treated with rehydration and maintenance therapy (balanced glucose-electrolyte solutions) and completely recovered after 3 days.

HoBi-like pestiviruses are emerging pestiviruses that infect cattle causing clinical forms overlapping to those induced by bovine viral diarrhea virus (BVDV) 1 and 2. As a consequence of their widespread distribution reported in recent years, molecular tools for rapid discrimination among pestiviruses infecting cattle are needed. The aim of the present study was to develop a multiplex real-time RT-PCR assay, based on the TaqMan technology, for the rapid and unambiguous characterisation of all bovine pestiviruses, including the emerging HoBi-like strains. The assay was found to be sensitive, specific and repeatable, ensuring detection of as few as 100-101 viral RNA copies. No cross-reactions between different pestiviral species were observed even in samples artificially contaminated with more than one pestivirus. Analysis of field samples tested positive for BVDV-1, BVDV-2 or HoBi-like virus by a nested PCR protocol revealed that the developed TaqMan assay had equal or higher sensitivity and was able to discriminate correctly the viral species in all tested samples, whereas a real-time RT-PCR assay previously developed for HoBi-like pestivirus detection showed cross-reactivity with few high-titre BVDV-2 samples.

We report detection and full-genome characterization of a novel orthopoxvirus (OPXV) responsible for a fatal infection in a cat. The virus induced skin lesions histologically characterized by leukocyte infiltration and eosinophilic cytoplasmic inclusions. Different PCR approaches were unable to assign the virus to a defined OPXV species. Large amounts of typical brick-shaped virions, morphologically related to OPXV, were observed by electron microscopy. This OPXV strain (Italy_09/17) was isolated on cell cultures and embryonated eggs. Phylogenetic analysis of 9 concatenated genes showed that this virus was distantly related to cowpox virus, more closely related to to ectromelia virus, and belonged to the same cluster of an OPXV recently isolated from captive macaques in Italy. Extensive epidemiologic surveillance in cats and rodents will assess whether cats are incidental hosts and rodents are the main reservoir of the virus. The zoonotic potential of this novel virus also deserves further investigation.