Detection of hepatitis E virus RNA in rats caught in pig farms from Northern Italy

Hepatitis E virus (HEV) strains belonging to the Orthohepevirus genus are divided into four species (A-D). HEV strains included in the Orthohepevirus A species infect humans and several other mammals. Among them, the HEV-3 and HEV-4 genotypes are zoonotic and infect both humans and animals, of which, pigs and wild boar are the main reservoirs. Viruses belonging to the Orthohepevirus C species (HEV-C) have been considered to infect rats of different species and carnivores. Recently, two studies reported the detection of HEV-C1 (rat HEV) RNA in immunocompromised and immunocompetent patients, suggesting a possible transmission of rat HEV to humans. The role of rats and mice as reservoir of HEV and the potential zoonotic transmission is still poorly known and deserves further investigation. To this purpose, in this study, the presence of HEV RNA was investigated in the intestinal contents and liver samples from 47 Black rats (Rattus rattus) and 21 House mice (Mus musculus) captured in four pig farms in Northern Italy. The presence of both Orthohepevirus A and C was investigated by the real-rime RT-PCR specific for HEV-1 to HEV-4 genotypes of Orthohepevirus A species and by a broad spectrum hemi-nested RT-PCR capable of detecting different HEV species including rat HEV. The intestinal content from two Black rats resulted positive for HEV-C1 RNA and for HEV-3 RNA, respectively. None of the House mice was HEV RNA positive. Sequence analyses confirmed the detection of HEV-C1, genotype G1 and HEV-3 subtype e. The viral strain HEV-3e detected in the rat was identical to swine HEV strains detected in the same farm. Liver samples were negative for the detection of either rat HEV or HEV-3.