Characterisation of AmpC / ESBL genes in some pathogen gram-negatives isolated from clinical cases of livestock and companion animals

Abstract

This study was aimed to search and characterize the AmpC and/or ESBL genes of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa isolated from clinical cases of local livestock and companion animals between 2017 and 2019. A total of eight ceftiofur-resistant E. coli (n= 7) and ceftiofur-resistant K. pneumoniae (n= 1) and seven P. aeruginosa were isolated from different cases in local animals. By combination disc method, six E. coli isolates and one K. pneumoniae isolate were found to be ESBL producers. By combination of the disc method and double disc synergy test, no P. aeruginosa isolates were found as ESBL producers. In the agar disc diffusion test (ADDT) performed with cefoxitin and cefoxitin-boronic, only one E. coli was determined as AmpC producer. In ESBL-producing isolates, only the CTX-M class gene was detected by polymerase chain reaction (PCR) and subsequent sequence analysis revealed CTX-M-3 and CTX-M-15 variants. An AmpC positive E. coli isolate was found to carry plasmidic ampC gene in cmy-2 variant from CIT family. It was observed that P. aeruginosa isolates did not carry the plasmidic ampC gene. After the chromosomal ampC gene of one P. aeruginosa was amplified by PCR and sequenced, R79Q and T105A mutations in the chromosomal ampC gene was revealed. This showed that overproduction of the ampC enzyme is involved in the resistance to β-lactams in P. aeruginosa isolates in the study.