Escherichia coli (STEC) in healthy dairy ruminants was investigated between 1996 and 1998 by a multiplex polymerase chain reaction (mPCR) technique. A total of 13 552 E. coli colonies from 726 cows, 28 sheep and 93 goats out of 112 randomly selected dairy farms in Hessia, Germany were analysed. STEC strains were recovered from 131 (18Á0%) cows, nine (32Á1%) sheep and 70 (75Á3%) goats. Further characterization of the STEC isolates showed that 89 (0Á66% of the investigated colonies) of animal ®eld strains carried stx1 gene, 64 (0Á47%) stx2 gene and 57 (0Á42%) stx1 and stx2 gene. Sixty (93Á8%) out of 64 stx2 ®eld strains were harboured by cows. In contrast, 74 (83Á1%) out of 89 stx1 dairy animal ®eld strains were from ovine or caprine origin. Only 17 (8Á1%) stxpositive isolates (13 from cattle, three from sheep and only one from goat) were positive for eaeA gene. Eight (9Á0%) of the stx1, ®ve (7Á8%) of the stx2 and four (7Á0%) of the stx1/stx2 gene-positive ®eld strains carried the eaeA gene. The prevalence of EHEC-haemolysin (EHEC-hlyA) gene sequence was 88Á8% (79 isolates) of the stx1 and 68Á8% (44 isolates) of the stx2 isolates. Out of 57 stx1-and stx2-positive ®eld-strains, 34 (59Á6%) carried the EHEC-hlyA gene. E. coli O serovars O:157 and O:111 were not found. Only one isolate was positive with O26 antiserum.