Eleonora Ponterio

Detection of hepatitis E virus in pork liver sausages

Hepatitis E infection is regarded as an emerging public-health concern. The disease is normally self-limiting (mortality rate 1%), but chronic infections have recently been observed in transplanted patients. The etiological agent HEV is a small RNA virus infecting both humans and animals. In humans, the disease may be food-borne and pig is a main reservoir for zoonotic strains. In the present study, we evaluated the presence of HEV and swine fecal cross-contamination in pork liver sausages sold at a grocery store in Italy. HEV genome detection was performed by RT-qPCR, using harmonized protocols that included a process control (murine norovirus) and an internal amplification control. Swine fecal cross-contamination was assessed by determination of the ubiquitous porcine adenovirus. Overall, HEV genome belonging to genotype 3 was detected in both raw (10 out of 45 slices, 250 mg each, 22.2%) and dry (1 of 23 slices, 4.3%) liver sausages, but infectivity of the virus was not demonstrated. This pilot study fosters more investigations on HEV presence in pork-derived food, to assess the possible risk for the consumers.

Presence of Hepatitis E Virus in a RED Deer (Cervus elaphus) Population in Central Italy.

&NA; Hepatitis E is an acute human disease caused by the hepatitis E virus (HEV). In addition to humans, HEV has been detected in several animal species and is recognized as a zoonotic pathogen. Pigs, wild boar and deer can be reservoir. In this study, we evaluated HEV prevalence in a free‐living red deer (Cervus elaphus) population in central Italy by detecting virus‐specific antibodies and RNA in sera. A total of 35 of 251 red deer sera were positive for anti‐HEV IgG. HEV RNA was detected in 10 of 91 sera examined. Two genomic fragments targeted by diagnostic PCRs in the capsid region were sequenced, both matching with genotype 3 HEV. Overall results confirmed the occurrence of HEV infection in deer also in Italy.

Detection of serum antibodies to hepatitis E virus in domestic pigs in Italy using a recombinant swine HEV capsid protein

BackgroundThe hepatitis E virus (HEV) has been detected in both humans and animals, particularly pigs, worldwide. Several evidences, including human infection following consumption of raw contaminated meat, suggest a zoonotic transmission of HEV. In Italy, large circulation of genotype 3 HEV has been reported in swine, and recent studies have confirmed the involvement of this genotype in autochthonous human cases.ResultIn this study 111 sera collected from healthy pigs in two Italian regions were tested for anti-HEV IgG antibodies. For specific HEV antibody detection in swine, we developed ELISA and Western blotting methods, using a truncated capsid (ORF2) protein lacking the first 111 amino acids of a swine HEV genotype 3 strain. The ORF2-based ELISA revealed anti-HEV antibodies in 104 out of 111 pigs compared with 102 detected with a commercial ELISA kit. A lower number of sera reacted with the recombinant ORF2 protein in a Western blotting format (81/111). Using a Latent class analysis (LCA), the estimated sensitivities for ELISA-ORF2 and ELISA-kit tests were 0.961 and 0.936, respectively, whereas specificities were 0.599 and 0.475. The estimated sensitivity of Western blotting was 0.775, and the specificity was 0.944.ConclusionsThe overall results confirm the high prevalence of HEV seropositive healthy pigs in Italy. Through comparisons with a commercial ELISA test, the swine genotype 3 HEV antigen produced in this study was proven suitable to detect anti-HEV antibodies in pig sera by both ELISA and Western Blotting.

A pilot survey of bovine norovirus in northern Italy.

NOROVIRUS has a positive-sense single-stranded RNA genome and belongs to the family Caliciviridae. Members of the family are assigned to four genera, named Vesivirus , Lagovirus , Norovirus and Sapovirus ,on the basis of their genomic organisation and phylogenetic pattern (Mayo 2002). Two virus strains detected in faecal samples of cattle called Newbury agent-1 (Bridger and others 1984) and Nebraska (Smiley and others 2002) cluster together in a separate group for which the genus names Becovirus or Nabovirus have been proposed (Oliver and others 2006). Norovirus and sapovirus are an important cause of acute gastroenteritis in human beings (Koo and others 2010) and may also infect herds of pigs and cattle (van der Poel and others 2000, Reuter and others 2010) and pet animals (Martella and others 2008). Norovirus infection in animals has been either associated with diarrhoea or found to be asymptomatic (van der Poel and others 2000, Mijovski and others 2010). Bovine norovirus prototype strains identified so far are the Newbury2 strain, previously known as Newbury agent 2, and the Jena virus. All bovine norovirus strains fall into genogroup III (GIII) of Norovirus , which includes two genotypes. Viruses genetically related to Jena virus were assigned to genotype 1 (GIII.1) and viruses genetically related to Newbury2 were assigned to genotype 2 (GIII.2) (Ando and others 2000). This short communication describes the identification of bovine norovirus strains belonging to either GIII.1 or GIII.2 in a sample of calves from Italy presenting …

Pattern of activation of human antigen presenting cells by genotype GII.4 norovirus virus-like particles

BackgroundVirus-like particles (VLPs) from an Italian GII.4 norovirus strain were used to investigate activation and maturation of circulating antigen presenting cells (APCs) of human origin.MethodsPeripheral blood mononuclear cells (PBMCs) isolated from five healthy subjects were pulsed ex vivo with VLPs, and stained with a set of monoclonal antibodies (MAbs) for phenotypic analysis by flow cytometry. Cytokine release in cell supernatants was investigated by ELISA.ResultsNorovirus VLPs induced activation and maturation of circulating APCs derived from the five donors, as well as production of IL-6, IFN-γ and TNF-α cytokines.ConclusionsThe present results suggest that VLPs can activate antigen presenting cells for an efficient induction of the adaptive immune response.