Virulence genes of O149 enterotoxigenic Escherichia coli from outbreaks of postweaning diarrhea in pigs.

The goal of this research was to determine whether isolates of O149 porcine enterotoxigenic Escherichia coli (ETEC) recovered from recent outbreaks of severe diarrhea in weaned pigs in Ontario, Canada, had virulence attributes different from those of isolates of the same serogroup from diarrhea of pigs in the 1970s and 1980s. Polymerase chain reaction amplification was used to determine the distribution of 11 virulence-associated genes in recent (100 isolates) and old (35 isolates) Ontario O149 porcine ETEC. These tests demonstrated that 92% of the recent isolates possessed the estA gene for STa enterotoxin, whereas none of the old isolates had this gene. H antigen determination showed that all the isolates which lacked the estA gene (all 35 old isolates plus 8 recent isolates) were H43, whereas isolates which had the estA gene were H10. The astA gene for enteroaggregative heat-stable enterotoxin (EAST1) and the K88ac antigen were present in all 135 isolates. Plasmid analyses identified a cryptic 5.1kb plasmid in 99% of recent and 60% of old isolates. Suppressive subtractive hybridization associated several types of DNA fragments with the recent O149 ETEC, namely, fragments with no homology to DNA in databases, fragments of LPS biosynthesis genes, and F plasmid DNA. We conclude that the recent outbreaks of PWD in Ontario pigs were associated primarily with a new serotype of O149 ETEC and that isolates of this serotype possessed the estA gene that was not present in old O149 ETEC isolated from pigs in Ontario.