Detection And Molecular Characterization Of Klebsiella Pneumoniae From Raw Milk Meat And Water Samples In And Around Tirupati

Abstract

The present study was undertaken to isolate the Klebsiella pneumoniae from raw milk, chicken and mutton samples collected from local vendors and retail outlets and water samples collected from different farms and nearby water bodies of Tirupati. Klebsiella species were detected from the samples by conventional methods and they were confirmed by biochemical tests. Molecular characterization of K. pneumoniae was done by targeting rpoB gene and their virulence genes were detected by using the primers targeting rmpA, fimH and entB genes, respectively. The study also carried out the detection of antimicrobial resistance pattern of the K. pneumoniae isolates and also phenotypic detection of ESBLs. Further, the genes responsible for extended spectrum beta lactamase activity were detected by using the primers targeting TEM, SHVand CTX-M1 genes, respectively. newlineOut of 103 milk, 117 chicken, 107 mutton and 112 water samples collected from different sources, ten (9.7%) isolates from milk, one (0.8%) isolate from chicken, eight (7.5%) isolates from mutton and 16 (14.3%) isolates from water were positive for Klebsiella by conventional methods. All the 35 isolates were found positive for biochemical tests such as catalase test, Voges-Proskauer test, citrate utilization test, triple sugar iron test, nitrate reduction test, urease test and negative for xix the oxidase, motility, indole, methyl red tests which were specific for the confirmation of K. pneumoniae. The further confirmation of K. pneumoniae was done by PCR targeting for rpoB gene, which encodes for RNA polymerase B. The PCR targeting rpoB gene with amplification size of 108bp was standardized. Out of 103 milk, 117 chicken, 107 mutton and 112 water samples, ten (9.7%) isolates from milk, one (0.8%) isolate from chicken, eight (7.5%) isolates from mutton and 16 (14.3%) isolates from water were positive for species-specific rpoB gene, confirming K. pneumoniae by PCR method. Biofilm producing ability of K. pneumoniae was detected by Congo red agar method. In this study, ou