Barbara Di Martino

Detection and molecular characterization of a canine norovirus.

We identified a novel calicivirus in a pup with enteritis. The isolate was related genetically (90.1% aa identity in the capsid protein) to a lion norovirus strain.

Canine kobuviruses in diarrhoeic dogs in Italy

Abstract Canine kobuviruses (CaKVs) are newly recognized picornaviruses recently detected in dogs in the US. By molecular analysis of the whole genome, CaKV that appeared genetically closest to the murine kobuvirus (MuKV) and to the human Aichi virus (AiV), may be classified in the Kobuvirus genus as new genotype (CaKV type 1) within the species Aichivirus A. To date, there are no information on the epidemiology of these novel viruses in other continents. In this study, by screening a collection of 256 dog fecal samples either from diarrhoeic or asymptomatic animals, CaKV was identified in six specimens with an overall prevalence of 2.34% (6/256). All the positive dogs presented diarrhea and were found to be infected by CaKV alone or in mixed infections with canine coronavirus (CCoV) and/or canine parvovirus type 2 (CPV-2). By molecular analysis of the partial 3D gene, all the strains detected displayed a close relatedness with the CaKVs recently identified in the US. This study provides evidence that CaKVs circulate in diarrhoeic dogs in Italy and are not geographically restricted to the North American continent, where they were first signaled.

Detection of antibodies against norovirus genogroup GIV in carnivores.

ABSTRACT Noroviruses (NoVs) resembling human NoV genotype GIV (Alphatron-like) have recently been detected in carnivores. By using an enzyme-linked immunosorbent assay based on baculovirus-expressed capsid protein VP1 of lion strain GGIV.2/Pistoia/387/06/ITA, NoV-specific antibodies were detected in cats (16.11%) and dogs (4.8%), demonstrating that these animals are exposed to infections caused by NoVs.

Detection of hepatitis E virus in slaughtered pigs in Italy

The stools of slaughtered pigs were screened for hepatitis E virus (HEV). HEV RNA was detected in 7.3% of the samples. HEV strains were characterized as genotype 3 subtype c, a cluster previously not described in Italy. These findings provide evidence that slaughterhouse workers may be exposed to HEV infection.

Molecular detection of porcine kobuviruses in Italian swine.

In this study, fecal specimens (n=260) collected from Italian healthy pigs aged between 6 and 10 months were screened by RT-PCR using generic kobuvirus primers. Porcine kobuviruses (PKVs) were detected in 3.85% of the samples tested. Based on the analysis of the partial 3D gene, the Italian sequences identified here were more closely related to PKVs previously identified in Japan, Thailand, Korea, China and Brazil than to the European PKVs recently detected in Hungary and in the Netherlands.

Seroprevalence of Norovirus Genogroup IV Antibodies among Humans, Italy, 2010-2011

Antibodies specific to genogroup IV identified in human specimens suggest zoonotic exposure.

Characterization of a strain of feline calicivirus isolated from a dog faecal sample.

Abstract To expand the epidemiological understanding of feline calicivirus (FCV) in dogs, genotypic and phenotypic characterizations of a FCV strain isolated from a puppy showing enteritis were performed. After isolation in cell culture, the novel isolate was analysed by RT-PCR and the amplicons obtained were sequenced. In order to characterize the growth properties of the isolate, the size of the plaques, the temperature of inactivation and the kinetics of growth were evaluated. Moreover, the novel strain was used to perform a serological study on 86 canine serum samples and 81 feline sera by virus-neutralization assay. The comparative analysis of nucleotide and amino acid sequences of the isolate, named FCV-Te/10/07, revealed the highest identity to strain FCV-F65. The growth kinetic revealed that strain Te/10/07 grew more rapidly than F9 strain. By virus-neutralization assay, dogs from the same region of the isolate showed antibodies against the FCV-F9 vaccinal strain in 63.9% (55/86) of sera, while antibodies against the Te/10/07 were found in seven sera (8.13%). In cats neutralizing antibodies against Te/10/07 strain were recovered in 50.62% (41/81) of samples tested, even if 38 sera were positives for F9 strain with similar titres or higher. In three cats neutralization to Te/10/07 alone was seen. Our results confirmed the interspecific circulation of FCV strains among different animal species, but new investigations are needed to establish whether FCV is pathogenic in the dog and the role of interspecies circulation in pathogen spreading.

Diagnosis of ovine chlamydial abortions by PCR-RFLP performed on vaginal swabs.

Ovine enzootic abortion is an infectious and contagious disease clinically characterized by abortion and weak neonates, affecting sheep and goats. The etiological agent is Chlamydophila (C.) abortus, which is considered one of the most common animal pathogens of small ruminants; it has important economic implications and represents a significant zoonotic risk. Clinical diagnosis is often difficult because the clinical signs and the pathological lesions are not specific for C. abortus infection, in fact they can also be observed as a result of infections with other abortifacient agents. Moreover, the involvement of the laboratory is necessary to perform the definitive diagnosis. One hundred and seventeen vaginal swabs from sheep with clinical signs related to chlamydial infection were examined by a PCR-RFLP assay that demonstrated high specifity and sensitivity. Six samples were positive for C. abortus. Vaginal swabs are easy to handle and allow to deal with biohazardous material in safety conditions.

Molecular evidence of kobuviruses in free-ranging red foxes (Vulpes vulpes).

Abstract Red foxes (Vulpes vulpes) are susceptible to viral diseases of domestic carnivores. In this study, by screening rectal swabs collected from 34 red foxes in Italy, we identified kobuvirus RNA in five samples. Based on analysis of partial RdRp and full-length VP1 genes, all of the strains shared the highest identity with canine kobuviruses (CaKVs) recently detected in the US, the UK and Italy. These findings provide the first evidence of the circulation of these novel viruses in foxes.

Detection of hepatitis E virus (HEV) in goats

Hepatitis E virus (HEV) is a major cause of acute hepatitis worldwide. Genotypes 1 and 2 are restricted to humans, whereas genotypes 3 and 4 also occur in animals and are recognized as zoonotic pathogens. In this study, by screening goat faecal samples collected from six small farms located in the province of Teramo (Abruzzo region, Italy), HEV RNA was found with an overall prevalence of 9.2% (11/119). Upon sequence analysis of a 0.8kb portion of the ORF2 gene, four strains were grouped with animal and human genotype 3 HEVs, subtype c, with the highest match (94.2-99.4% nt identity) to a wild boar strain, WB/P6-15/ITA, identified in the same geographical area in which the six goat farms were located. Further investigations are needed in order to assess if goat may represent an additional active host for HEV.