Hepatitis E is an infectious disease caused by a small RNA virus (Hepatitis E Virus, HEV), which can cause acute hepatitis in humans. Four mammalian HEV genotypes are known, among which g3 and g4 are considered zoonotic. In developed countries, hepatitis E cases were mostly associated with travelling in endemic areas. However, recently autochthonous cases have been increasingly reported in developed country, including Italy. In Italy, g3 strains have been detected in swine, in both farms and slaughterhouses, with prevalence that can reach 50% (2,3). Swine strains often share a high nucleotide identity with HuHEV. During the last 10 years, hepatitis E cases have been correlated with consumption of swine, wild boar and deer meat or organs (particularly liver) consumed raw or undercooked, worldwide (1). In this study, we investigated the presence of HEV in liver sausage, which is often consumed raw or undercooked in Italy. In 2012, 4 packages (300 gr) of pork liver sausages were bought at a butcher shop. Sausages were chopped in 45 slices (250 mg each) and spiked with Murine Norovirus (MNV). After RNA extraction, samples were analyzed by RT‐PCR for MNV detection (extraction process control). Samples positive for MNV were then analyzed for HEV and Porcine Adenovirus (PAdV as index virus of fecal contamination) by real‐time RT‐PCR and real‐time PCR, respectively. Positive samples were further analyzed by RT‐PCR with specific primers for two different genomic regions (ORF1 and ORF2). Two out of 45 samples (4.4%) were positive for HEV. Moreover both samples were positive for HEV ORF1, and one was also confirmed in ORF2. This result suggests different sensitivity of the two PCR protocols. Sequence analysis confirmed the presence of swine g3 HEV strains in both samples. One sausage sample was positive for HEV and PAdV, suggesting that also HEV presence could be related to fecal contamination of pork liver. Attempts to infect A549 cells with a homogenate from a HEV and PAdV positive liver sausage were unsuccessful. This study confirms that pig liver sausage can be contaminated with HEV, which might at least partially reflect fecal contamination of pork meat and liver during improper slaughtering practice. Further studies to investigate residual HEV infectivity in pork are needed
Di Bartolo I., Ostanello F., Ruggeri F.M. (2013). DETECTION OF HEPATITIS E VIRUS IN SWINE LIVER SAUSAGE, IN ITALY. s.n. : (sine nomine).
DETECTION OF HEPATITIS E VIRUS IN SWINE LIVER SAUSAGE, IN ITALY
OSTANELLO, FABIO;
2013
Abstract
Hepatitis E is an infectious disease caused by a small RNA virus (Hepatitis E Virus, HEV), which can cause acute hepatitis in humans. Four mammalian HEV genotypes are known, among which g3 and g4 are considered zoonotic. In developed countries, hepatitis E cases were mostly associated with travelling in endemic areas. However, recently autochthonous cases have been increasingly reported in developed country, including Italy. In Italy, g3 strains have been detected in swine, in both farms and slaughterhouses, with prevalence that can reach 50% (2,3). Swine strains often share a high nucleotide identity with HuHEV. During the last 10 years, hepatitis E cases have been correlated with consumption of swine, wild boar and deer meat or organs (particularly liver) consumed raw or undercooked, worldwide (1). In this study, we investigated the presence of HEV in liver sausage, which is often consumed raw or undercooked in Italy. In 2012, 4 packages (300 gr) of pork liver sausages were bought at a butcher shop. Sausages were chopped in 45 slices (250 mg each) and spiked with Murine Norovirus (MNV). After RNA extraction, samples were analyzed by RT‐PCR for MNV detection (extraction process control). Samples positive for MNV were then analyzed for HEV and Porcine Adenovirus (PAdV as index virus of fecal contamination) by real‐time RT‐PCR and real‐time PCR, respectively. Positive samples were further analyzed by RT‐PCR with specific primers for two different genomic regions (ORF1 and ORF2). Two out of 45 samples (4.4%) were positive for HEV. Moreover both samples were positive for HEV ORF1, and one was also confirmed in ORF2. This result suggests different sensitivity of the two PCR protocols. Sequence analysis confirmed the presence of swine g3 HEV strains in both samples. One sausage sample was positive for HEV and PAdV, suggesting that also HEV presence could be related to fecal contamination of pork liver. Attempts to infect A549 cells with a homogenate from a HEV and PAdV positive liver sausage were unsuccessful. This study confirms that pig liver sausage can be contaminated with HEV, which might at least partially reflect fecal contamination of pork meat and liver during improper slaughtering practice. Further studies to investigate residual HEV infectivity in pork are neededI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
