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Antimicrobial Agents and Chemotherapy, October 2002, p. 3156-3163, Vol. 46, No. 10
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.10.3156-3163.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

ß-Lactamases in Ampicillin-Resistant Escherichia coli Isolates from Foods, Humans, and Healthy Animals

Laura Briñas, Myriam Zarazaga, Yolanda Sáenz, Fernanda Ruiz-Larrea, and Carmen Torres*

Area de Bioquímica y Biología Molecular, Universidad de La Rioja, 26006 Logroño, Spain

Received 17 January 2002/ Returned for modification 23 April 2002/ Accepted 9 July 2002

TEM-, SHV-, and OXA-type ß-lactamases were studied by PCR with 124 ampicillin-resistant (AMPr) 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 blaTEM amplicons were sequenced. Different molecular variants of blaTEM-1 were found in 52 isolates: blaTEM-1a (n = 9), blaTEM-1b (n = 36), blaTEM-1c (n = 6), and blaTEM-1f (n = 1). Four inhibitor-resistant TEM (IRT) ß-lactamase-encoding genes were also detected: blaTEM-30c (IRT-2), blaTEM-34b (IRT-6), blaTEM-40b (IRT-11), and blaTEM-51a (IRT-15). A new blaTEM gene, named blaTEM-95b, which showed a mutation in amino acid 145 (P->A) was detected. It was found in a food isolate of chicken origin (AMPr, amoxicillin-clavulanic acid susceptible). The promoter region in 24 blaTEM amplicons was analyzed, and the weak P3 promoter was found in 23 of them (blaTEM-1 in 20 amplicons and blaTEM-51a, blaTEM-30c, and blaTEM-95b in 1 amplicon each). The strong Pa/Pb promoter was found only in the blaTEM-34b gene. No extended-spectrum ß-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 µg/ml. Different variants of blaTEM-1 and IRT blaTEM genes were found among the AMPr E. coli isolates from foods and the feces of humans and healthy animals, and a new gene, blaTEM-95b (P3), was detected.

* Corresponding author. Mailing address: Area de Bioquímica y Biología Molecular, Universidad de La Rioja, Madre de Dios, 51, 26006 Logroño, Spain. Phone: 34-941299750. Fax: 34-941299721. E-mail: carmen.torres{at}daa.unirioja.es.

Antimicrobial Agents and Chemotherapy, October 2002, p. 3156-3163, Vol. 46, No. 10
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.10.3156-3163.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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