Identification of two divergent swine Noroviruses detected at the slaughterhouse in North East Italy

Norovirus (NoV) has emerged as one of the major causative agents of non-bacterial, food- and water-borne gastroenteritis in humans, with the main genogroup involved in human outbreaks (GII), which has been detected worldwide in different animal species including swine. A four-month investigation at the slaughterhouse aiming to examine the presence of NoV in the swine in North-Eastern Italy, enabled the detection of two divergent Noroviruses (NoVs) (GII.P11) in two swine farms. This represents the first study in the swine population of North-Eastern Italy, which has paved the way for future integrated virological and epidemiological investigations on swine NoVs

Molecular evolution of Feline parvovirus (FPV)

Feline panleukopenia is a viral disease known since the beginning of the 20th century that occurs in cats, causing severe leukopenia, gastro-enteritis and nervous signs. Feline panleukopenia is caused by feline panleukopenia virus (FPV), a DNA virus belong to the family Parvoviridae, genus Parvovirus, subgroup of feline parvovirus. Also the new antigenic variants of canine parvovirus (CPV) type 2a-2b-2c have gained the feline host range and can infect, replicate and cause disease in cats with clinical signs similar to feline panleukopenia. In this report, we performed a molecular study on feline parvovirus strains isolated from domestic cats to investigate which parvoviruses strains are spread in the feline population and to monitor the evolution of FPV in Italy. Twenty-one viral isolates from clinical samples of diseased cats collected between 2000-2007 were examined. Phylogenetic analysis of the VP2 gene sequences was performed using the PHYLIP version 3.6 and TREE PUZZLE version 5.0 softwares. The reliability of the phylogenetic trees were assessed by bootstrap resampling (1000 datasets). Nineteen strains were found to be FPV and 2 strains were CPV-2. These CPV-2 strains show high genetic complexity with the presence of more variants in the same animal. Analysis of the VP2 protein of Italian FPV strains did not show specific amino acid changes in antigenic epitopes so we did not observe new variants for FPV. The phylogenetic trees constructed with VP2 sequences of reference strains show the Italian FPV strains clustering with the other FPV strains except for the more recent isolates that form a separate virus lineage

Identificazione di microRNA nel genoma del parvovirus canino di tipo 2 (CPV-2).

Precedenti studi hanno evidenziato la presenza di breve regioni geniche omologhe tra i parvovirus e le rispettive specie ospiti, ed \ue8 stata avanzata l'ipotesi che queste regioni potrebbero fungere da microRNA virali. In questo studio si \ue8 dimostrato la presenza di brevi regioni omologhe tra il genoma del parvovirus canino di tipo 2 (CPV-2) e il genoma canino. Queste regioni potrebbero essere coinvolte nel fenomeno dell'RNA silencing facilitando la persistenza virale, modulando i fattori di trascrizione ed eludendo il sistema immunitario

First detection of porcine circovirus type 2e in Europe

Porcine circovirus 2 (PCV-2) infections are among the most economically important in swine farming. Because of the high evolutionary rate of PCV-2, several variants have emerged and are currently classified into several genotypes. However, only three (i.e. PCV-2a, PCV-2b, and PCV-2d) have a worldwide distribution, with other genotypes restricted to certain geographical regions and/or for limited time periods. Underdiagnosis or underreporting of these genotypes cannot be excluded. This is the first report of the detection and genetic characterisation of the PCV-2e genotype in Europe, from sows on a farm in Italy showing no clinical evidence of porcine circovirus disease. A follow-up study demonstrated persistent subclinical evidence of PCV-2e on the farm, at low frequency and viral load. This incidental finding emphasises the need for more intensive routine monitoring activities involving asymptomatic animals, coupled with DNA sequencing and data sharing. Considering the relevant genetic and phenotypic divergence of such genotypes, the actual efficacy of currently applied vaccines and diagnostic assays should be further evaluated

NOROVIRUS IN FAECES OF HEALTHY PIGS IN NORTH-EAST ITALY

Introduction Norovirus (NoV) has emerged has one of the major causative agents of non-bacterial, food- and water-borne gastroenteritis in humans all over the world. NoVs are classified into six groups (G), from GI to GVI, which are further subdivided into 30 genotypes. NoVs identified in human gastroenteritis cases are only GI, GII, and GIV. NoVs have also been isolated from several animal species, including pigs, dogs, cattle, rodents and lions. The detection of GII NoV from pigs in Japan and Europe, and GII NoV antibodies in US swine have raised public health concerns about the zoonotic potential of porcine NoVs. Material & Methods Faeces were collected at slaughterhouse in 2017 in two regions of North-East Italy. Forty-six samples originated from Veneto and thirty-three from Friuli Venezia Giulia regions, covering seven and three provinces, respectively, were analysed for presence of Calicivirus. A two-step RT-PCR targeting the RdRP gene using the p290-p110 primer pairs was used. Sanger sequence was conducted on samples presenting enough amount of the target amplified DNA. Phylogenetic analysis was carried out using the neighbour-joining method and Kimura two-parameter substitution model using MEGA7 software. Results Fourteen samples collected in Veneto region, were PCR positive. Nucleotide sequence of about 300bp were obtained from only two samples. BLAST analysis showed a homology between 89 and 92% with swine NoV detected in Europe. Phylogenetic analysis showed that Italian strains belong to the GII.11 and cluster with other swine NoV from USA and Asia. Discussion and Conclusion This study identified GII.11 NoVs in the swine population of North-East Italy, similarly to a previous report in 2011. Further molecular analysis on the VP1 gene are ongoing on positive samples. The real distribution and the role of NoVs in swine needs to be further investigated by proper sampling approach and full genome analysis

Environmental exposure of wild carnivores to zoonotic pathogens: Leptospira infection in the first free living wolf (canis lupus linnaeus, 1758) found dead in the friuli venezia giulia region

Leptospirosis is a worldwide-spread zoonosis causing disease and death in dogs and in humans. A Leptospiral infection has been recorded in several wild carnivore species in Europe, but tissue pathological changes were not commonly described. The Grey wolf (Canis lupus) has been expanding its distribution range in north-eastern Italy during the last decade. A young wolf, repre-senting the first individual handled in the region, was found road-killed and then submitted to necropsy. Pathological changes included erosive lesions of gingival mucosa, mild liver enlargement, and multifocal degenerative-necrotic areas along with hyperemic reactive lesions; multifocal inter-stitial nephritis and multifocal lung hemorrhages were observed. A Polymerase Chain Reaction (PCR) able to detect pathogenic species of Leptospira performed on a kidney sample was positive. Serological reactions for serogroup Gryppotyphosa (1:6400), Pomona (1:800), and Icterohaemor-rhagiae (1:200) were evidenced by MAT. Genotyping by Multilocus Sequence Typing (MLST) performed on detected Leptospira characterized it as belonging to Sequence Type (ST) 117, which refers to L. kirschneri, serogroup Pomona, serovar Mozdok. Regardless of the role of Leptospira infection as an eventual predisposing factor to the road killing of this wolf, to the best of the authors\u2019 knowledge, this is the first report of Leptospira-induced pathology in a wolf in Europe. Surveys on Leptospira infection in free-ranging wildlife species should be pursued in order to achieve further epidemio-logical knowledge on the circulation of the Leptospira strain