Molecular Characterization of CTX-M β-Lactamase and Associated Addiction Systems in Escherichia coli Circulating among Cattle, Farm Workers, and the Farm Environment

Abstract
A total of 84 extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolates from cattle, farm workers, and the farm environment isolated from February to September 2008 in the Republic of Korea were investigated. All 84 ESBL-producing isolates carried bla CTX-M genes that belonged to the CTX-M-1 ( n = 35) or CTX-M-9 ( n = 49) family. The most predominant CTX-M type identified was CTX-M-14 ( n = 49), followed by CTX-M-32 ( n = 26). The bla CTX-M genes were identified most commonly in E. coli isolates from feces ( n = 29), teats ( n = 25), and milk ( n = 14). A bla CTX-M-14 gene was also detected in an E. coli isolate from a farmer's hand. Transfer of the bla CTX-M gene from 60 bla CTX-M -positive E. coli isolates to the recipient E. coli J53 strain by conjugation was demonstrated. Plasmid isolation from bla CTX-M -positive transconjugants revealed a large (95- to 140-kb) conjugative plasmid. Almost all (82/84) bla CTX-M genes possessed an insertion sequence, IS Ecp1 , upstream of the bla CTX-M gene. Only in the case of the CTX-M-14 genes was IS 903 downstream of the gene. The bla CTX-M genes were associated with seven kinds of addiction systems. Among them, pndAC , hok-sok , and srnBC were the most frequently identified addiction systems in both wild strains and transconjugants. The spread of bla CTX-M genes was attributed to both clonal expansion and horizontal dissemination. Our data suggest that a combination of multiple addiction systems in plasmids carrying bla CTX-M genes could contribute to their maintenance in the host cells. To our knowledge, the bla CTX-M-32 gene has not previously been reported in animal isolates from the Republic of Korea.

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