Swine as a potential reservoir of shiga toxin-producing Escherichia coli O157:H7 in Japan.

To the Editor: Shiga toxin-producing Escherichia coli (STEC) O157:H7 has become a major meat safety issue worldwide. Cattle, an important reservoir of human infection (1), may not be the only source of this organism (2,3). In a survey of pigs in England (4), non-STEC O157 was isolated from four (0.4%) fecal samples collected (after slaughter) from 1,000 pigs. We found that, although an unlikely source of infection for humans, pigs are a potential reservoir of STEC O157:H7 in Japan. In 1997, there were 14,400 pig farms and 9,823,000 pigs (average 682 per farm) in Japan. Thirty-five (0.24%) of these farms were randomly selected for study, and rectal swabs were taken from 221 healthy pigs during May and June 1997. The average number of animals examined on each farm was 6.3. Fecal samples were dipped into test tubes containing Cary-Blair transport medium (Nissui, Japan) and kept refrigerated until processing (usually within 48 hours). Swabs were then incubated overnight at 42°C in 10 ml of mEC broth (Kyokuto, Japan) containing 20 μg/ml of novobiocin (Sigma, USA), after which one loop of the broth was spread onto MacConkey sorbitol agar medium (Difco, USA). After overnight incubation at 37°C, sorbitol-negative colonies from the agar plates were tested by slide agglutination with E. coli O157-latex test (Oxoid, UK). Strains that agglutinated were confirmed as E. coli by using the API 20E system (BioMerieux, France). Strains confirmed as E. coli O157 were subcultured in a motility medium for 3 to 4 days to enhance development of flagella, then they were tested by tube agglutination with E. coli H7 antiserum (Denkaseiken, Japan). The swine E. coli O157:H7 isolates were examined by polymerase chain reaction for the presence of Shiga-toxin genes stx1 and stx2 and to elucidate intimin (eaeA) DNA sequences (5), for a plasmid of 92 kb (pO157) by agarose gel electrophoresis (6), and for phage type by the previously described method (7). Although the numbers sampled were too small to allow comparisons between farms, samples from three (1.4%) apparently healthy pigs (ages: 2, 6, and 9 months) from three farms (8.6%) were positive for STEC O157:H7. The three strains from the pigs were biochemically typical of STEC O157:H7 that did not ferment sorbitol and lacked ß-glucuronidase; agglutinated with E. coli O157-latex and with H7 antiserum; possessed stx1, stx2, and eaeA genes; and harbored pO157 plasmid characteristic of STEC O157:H7. The strains belonged to phage type 21, 37, or 43. The 1.4% carriage rate of STEC O157:H7 in pigs in this investigation is almost the same as that in cattle in Japan (8), which suggests that STEC O157:H7 strains are probably widespread in Japanese pig populations. The STEC O157positive pigs were each housed in a concretefloored pen and kept separate from cattle. Whether these pig isolates are the same as cattle or human isolates needs to be clarified; however, they had the same biochemical and genetic markers as STEC O157:H7 isolated from cattle and humans (6,9). The phage type 21 that we found among pig isolates was also observed in bovine and human STEC O157:H7 isolates in Japan (7). These results suggest that common vehicles for dissemination of the organism may exist. So far, pork has not been identified as a source of human STEC O157:H7 illness in industrialized countries, but our results indicate that eating pork, contact with pigs, and contamination with pig feces should be considered potential sources of this pathogen. This is the first isolation of naturally occurring STEC O157:H7 in pigs in Japan.