Fresh feces, manure slurry (from earthen lagoons and/or concrete pits), and drinking and surface water samples were collected from 28 pig farms in the Midwestern United States. All samples were tested for hepatitis E virus (HEV) RNA by reverse transcription-PCR. Seven of 28 farms had fecal samples that contained HEV. Of 22 farms where pit samples were accessible, 15 contained HEV, and of 8 farms that had lagoons, 3 contained HEV. The highest virus titers were 10 and 10 genome equivalents per 60 ml of manure slurry in lagoon and pit samples, respectively. None of the water samples tested HEV positive. To determine the infectivity of the HEV found in the positive farm 19 lagoon (designated L19) or farm 12 pit (designated P12) samples, pigs were inoculated either intravenously (n = 3) or orally (n = 3) with the L19 or P12 manure slurry. Four pigs inoculated intravenously with prototype swine HEV served as positive controls. All positive-control pigs shed HEV in feces and 3 of 4 developed anti-HEV antibodies. Two pigs in the intravenously inoculated P12 group shed HEV in feces, and one of the pigs seroconverted to anti-HEV antibodies. None of the pigs in the negative-control, L19 oral, L19 intravenous, or P12 oral group shed HEV in feces. The findings indicate that HEV found in pig manure slurry was infectious when inoculated intravenously. Pit manure slurry is a potential source of HEV infection and for contamination of the environment. Contamination of drinking or surface water with HEV was not found on or near the pig farms.