Processing does not substantially abate endogenous virus.

1Hepatitis E is an acute disease of humans caused by a small RNA virus, Hepatitis E virus (HEV). In 2 recent years, an increasing number of autochthonous human infections have been reported in 3 industrialized countries. Genotype 3 is the main HEV type circulating in swine, and is also reported 4 in sporadic cases of hepatitis E in humans worldwide. To date one serotype has been described. We 5 have conducted a survey to detect antibodies against HEV in 48 swine at a slaughterhouse in 6Northern Italy, using ELISA test. Mean seroprevalence in the studied animal group was 87.%. Bile, 7 liver and feces from the 48 animals were also collected, and HEV RNA was detected by nested 8 reverse transcription-polymerase chain reaction, amplifying a fragment of the ORF2. HEV genome 9 was most frequently detected in bile samples (51.1%), followed by feces (33.3%) and liver (20.8%). 10Thirty-one out of 48 studied pigs (64.6%) were positive for HEV RNA in at least one sample. 11Overall, HEV RNA was found at a statistically higher rate in the 3-4 month-old than in 9-10 month-12 old animals (95.0% vs. 42.9%). Genetic characterization of swine strains identified was performed 13 by sequencing and database alignment. Phylogenetic analysis on the nucleotide sequences from 16 14 positive PCR products indicated that all strains belonged to genotype 3, clustering in two branches 15 subtypes g3c and g3f. 16 17

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25Hepatitis E virus (HEV) causes acute hepatitis in humans, and infects several animal species, 26 mostly asymptomatically. Swine and human HEV strains are genetically related suggesting 27 both a zoonotic and a possible foodborne transmission. The prevalence of swine HEV was 28 investigated in 274 randomly selected pigs from 6 different swine farms of Northern Italy, 29 testing viral RNA in stools by nested reverse-transcription-polymerase chain reaction. HEV 30 genome was detected in 115 stools (42%). All farms resulted positive for HEV, with a 31 prevalence ranging between 12.8% and 72.5%. HEV positive pigs were detected in all age 32 groups and production stages tested, although infection was more prevalent in weaners than in 33 the older fatteners (42.2% vs. 27.0%). Genetic characterization of swine strains identified was 34 performed by sequencing and database alignment. Phylogenetic analysis on the nucleotide 35 sequences from 16 positive PCR products indicated that all strains belonged to genotype 3. In 36 particular, one group of seven Italian strains clustered close (91.6% to 96.2% identity) to 37 human and swine European HEV strains. 38 39

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.