Using TnphoA transposon insertion mutagenesis, we found a porcine EPEC (PEPEC) mutant demonstrating as inability to induce AE lesions. The insertion was identified in a gene designated paa (porcine attaching and effacing associated). The distribution of paa in PEPEC O45 strains revealed that it was associated with presence of the eae and its AE phenotype in vivo. On examination of enteric E. coli isolates from humans and various animal species, a strong correlation with the presence of paa was found in EHEC O157:H7 and O26, and dog, rabbit, and pig eae-positive isolates, and to a lesser extent in human EPEC eae-positive isolates. Also, among porcine ETEC isolates, a strong association was found with the presence of LT encoded genes. In contrast, paa sequence was rarely found in enteric E. coli isolates lacking ETEC and AEEC virulence determinants. Thus, our results suggest that Paa could play a role in the AE mechanism and other mechanisms of enteric disease.