Beta-lactamases in ampicillin-resistant Escherichia coli isolates from foods, humans, and healthy animals

Antimicrob Agents Chemother. 2002 Oct;46(10):3156-63. doi: 10.1128/AAC.46.10.3156-3163.2002.

Abstract

TEM-, SHV-, and OXA-type beta-lactamases were studied by PCR with 124 ampicillin-resistant (AMP(r)) Escherichia coli isolates recovered from foods of animal origin (n = 20) and feces of humans (n = 49) and healthy animals (n = 55). PCR showed that 103 isolates were positive for TEM and negative for SHV and OXA. Three E. coli isolates showed a positive reaction for OXA, and one showed a positive reaction for SHV. The remaining 17 E. coli isolates were negative for the three enzymes by PCR. Fifty-seven of the 103 bla(TEM) amplicons were sequenced. Different molecular variants of bla(TEM-1) were found in 52 isolates: bla(TEM-1a) (n = 9), bla(TEM-1b) (n = 36), bla(TEM-1c) (n = 6), and bla(TEM-1f) (n = 1). Four inhibitor-resistant TEM (IRT) beta-lactamase-encoding genes were also detected: bla(TEM-30c) (IRT-2), bla(TEM-34b) (IRT-6), bla(TEM-40b) (IRT-11), and bla(TEM-51a) (IRT-15). A new bla(TEM) gene, named bla(TEM-95b), which showed a mutation in amino acid 145 (P-->A) was detected. It was found in a food isolate of chicken origin (AMP(r), amoxicillin-clavulanic acid susceptible). The promoter region in 24 bla(TEM) amplicons was analyzed, and the weak P3 promoter was found in 23 of them (bla(TEM-1) in 20 amplicons and bla(TEM-51a), bla(TEM-30c), and bla(TEM-95b) in 1 amplicon each). The strong Pa/Pb promoter was found only in the bla(TEM-34b) gene. No extended-spectrum beta-lactamases were detected. Mutations at position -42 or -32 in the ampC gene promoter were demonstrated in 4 of 10 E. coli isolates for which the cefoxitin MIC was >/=16 micro g/ml. Different variants of bla(TEM-1) and IRT bla(TEM) genes were found among the AMP(r) E. coli isolates from foods and the feces of humans and healthy animals, and a new gene, bla(TEM-95b) (P3), was detected.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ampicillin / pharmacology
  • Animals
  • Animals, Domestic*
  • Cattle
  • Chickens
  • Dogs
  • Escherichia coli / drug effects*
  • Escherichia coli / enzymology
  • Escherichia coli / isolation & purification
  • Escherichia coli Infections / microbiology*
  • Feces / microbiology*
  • Humans
  • Meat Products / microbiology*
  • Microbial Sensitivity Tests
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Swine
  • beta-Lactam Resistance
  • beta-Lactamases / genetics*
  • beta-Lactamases / metabolism
  • beta-Lactams / pharmacology

Substances

  • beta-Lactams
  • Ampicillin
  • beta-Lactamases

Associated data

  • GENBANK/AJ308558