Characterization and phylogenetics of beta-lactamase Temoneira gene in Escherichia coli of the Bali cattle on Lombok island, Indonesia

Abstract

This study aims to determine the character and phylogenetics of the β-lactamase Temoneira (blaTEM) gene in Escherichia coli isolated from the reproductive tract of Bali cattle with repeat breeder cases. This study was conducted in June 2021 that used 16 female Bali cattle with repeat breeder cases to take their reproductive tract fluid using a plastic sheet gun. Isolation of Escherichia coli, using Eosin Methylene Blue Agar was identified by biochemical tests. Antibiotic susceptibility test on E. coli samples was carried out by disc diffusion method. The screening test for the presence of extended-spectrum β-lactamase was done using the double-disk approximation test method. Characterization of the blaTEM gene in E coli, using the PCR method and sequences of the blaTEM gene, were phylogenetically analyzed. The results showed that 4 E. coli isolates were obtained from 16 samples of reproductive tract fluid. The sensitivity test to antibiotics of 4 isolated samples of E. coli showed that 50% were resistant to penicillin and cefotaxime. Samples of E. coli resistant to penicillin and cefotaxime showed synergy results in the double-disk approximation test. The PCR results showed that the samples of E coli encoded the blaTEM gene which was located at the 560 bp position on gel electrophoresis. The phylogenetic tree analysis found that the blaTEM gene encoded by E. coli was related to E. coli encoding the blaTEM-206 gene. The character blaTEM gene in samples of E. coli showed the character of blaTEM-206 gene of extended-spectrum β-lactamase-producing E. coli. Escherichia coli from the reproductive tract of Bali cattle showed the character of E. coli S2.2-EK pEC-S2.2 blaTEM gene for class A broad-spectrum B-lactamase TEM-206 with a high degree of kinship.