To determine the prevalence of Hepatitis E virus (HEV) in industrialized nations, we analyzed the excretion of HEV strains by the populations of Spain, France, Greece, Sweden, and the United States. Twenty of 46 (43.5%) urban sewage samples collected in Barcelona from 1994 to 2002 tested positive for HEV. We identified 15 HEV strains, which were similar to two HEV isolates previously described in Barcelona in clinical samples and to strains from diverse geographic HEV-nonendemic areas. We also identified two HEV strains in sewage samples from Washington, D.C., and Nancy, France; these samples were also positive for Hepatitis A virus. In addition, we studied the role of pigs as a reservoir for HEV and identified one new swine HEV strain. Our results suggest that HEV may be more prevalent than previously considered in industrialized countries and that variants of the virus circulate simultaneously in one region.

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A study of the presence of human viruses (adenoviruses, enteroviruses, and hepatitis A viruses [HAVs]) in environmental and shellfish samples was carried out by applying DNA and cDNA amplification techniques by PCR. The detection of human adenoviruses by PCR was also examined as a potential molecular test to monitor viral pollution. The samples studied were urban and slaughterhouse sewage, river water, seawater, and shellfish. Enteroviruses were quantified by PFU in Buffalo green monkey kidney cells and fecal coliforms and phages of Bacteroides fragilis HSP40 were also evaluated in some of the samples. The amplification of viral DNA and cDNA has shown a high prevalence of human viruses that would not be detected by the use of classical techniques, such as the quantification of PFU in cell lines. The results of the analysis of slaughterhouse sewage samples together with the test of farm animal feces indicate that the adenoviruses and the HAVs detected in the environment are mostly of human origin. A significative correlation between the detection of human viruses by PCR and the values of bacteriophages of B. fragilis HSP40 in urban raw sewage was observed. Human adenoviruses were the viruses most frequently detected throughout the year, and all the samples that were positive for enteroviruses or HAVs were also positive for human adenoviruses. The results suggest that the detection of adenoviruses by PCR could be used as an index of the presence of human viruses in the environment where a molecular index is acceptable.

This is the first description, to our knowledge, of the distribution of human polyomavirus and simian virus 40 (SV40) in urban sewage. Using a nested-PCR procedure, we report the detection of human polyomaviruses JC virus (JCV) and BK virus (BKV) but not SV40 in a high percentage of urban sewage samples obtained from widely divergent geographical areas in Europe and Africa. For a total of 28 samples analyzed, JCV was detected in 26, BKV was detected in 22, and none was positive for SV40. All geographical areas showed a high prevalence of these viruses with mean estimated values of JC viral particles per ml on the order of 10 3 in Barcelona (Spain) and Nancy (France) and 10 2 in Pretoria (South Africa) and Umeå (Sweden) and mean values of BK viral particles on the order of 10 2 in Pretoria and Barcelona and 10 1 in Nancy and Umeå. This compares with estimated mean values of 10 2 to 10 3 for human adenovirus that was evaluated as a control. Nucleotide sequence analysis of the amplified DNA from some of the samples is also presented and represents the sequence of the most abundant JC and BK viral strains in these samples. The nucleotide sequence of the JCV detected was also analyzed in a phylogenetic study and for genomic characterization in the regulatory region. This study has shown that human polyomaviruses are spread in high concentrations in the sewage of different geographical areas and are present in contaminated environments. The frequency and concentration of JCV detected in the environment and the absence of described animal hosts suggest that JCV may be useful as a marker for fecal pollution of anthropogenic origin. The results also support the idea previously described that the strains of JCV are closely related to the ethnic origin of the population studied. The procedure applied should also be useful in future studies of population patterns of viral excretion and as a tool in epidemiological studies for the detection of changes in the prevalence of specific viral pathogens.

Please cite this article as: de Deus, N., Casas, M., Peralta, B., Nofrarías, M., Pina, S., Martín, M., Segalés, J., Hepatitis E virus infection dynamics and organic distribution in naturally infected pigs in a farrow-to-finish farm, Veterinary Microbiology (2007Microbiology ( ), doi:10.1016Microbiology ( /j.vetmic.2008 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. 53Moreover, the present study also indicates that the same pig can be infected with at 54 least two different strains of HEV during its productive life. This is the first study

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26In the present work, 262 serum samples and 29 faeces pools from chicken coming from 27 29 healthy flocks were analysed by RT-PCR for detection of avian HEV and by ELISA 28 using an aHEV derived antigen for detection of anti-HEV IgG. Additionally, other 300 29 randomly selected serum samples were also analysed by RT-PCR and by ELISA. Seven 30 serum samples were positive to RNA detection. Sequence analysis of both the helicase 31 and the capsid genes revealed that the Spanish isolates were clustered together and close 32 related to those strains from the United States isolated from farms with HSS. On the 33 serology study, 26/29 flocks had at least one positive animal (89.7%) and chickens older 34 than 40 weeks were found to have higher seropositivities compared to the rest of age 35 groups. Within positive farms, the proportion of positive animals ranged from 20% to 36 80%. This is the first report of aHEV sequences in chickens from Europe. Further studies are 37 needed to elucidate the clinical significance of avian HEV infections in Europe.