Introduction: Klebsiella pneumoniae is recognized as an urgent threat to human health. Development of novel treatment options is the need of the hour. One of the ways of achieving this goal is by conducting molecular characterization studies of antibiotic resistant bacteria. Hypothesis/Gap Statement: Efforts are being made by the Indian government to perform molecular characterization of MDR and potentially virulent bacteria by whole-genome sequencing (WGS). However, the data that we have at present is skewed as many parts of the country remain underrepresented. Uttarakhand is one such state, with relatively poor access to healthcare and a paucity of research. Aim: To generate WGS based preliminary data about the population structure, multi-locus sequence types (MLST), and virulence factors of CRKp isolates recovered from patients in a tertiary care teaching hospital located in Uttarakhand, India. Methodology: A cross-sectional study was conducted which included twenty-nine randomly selected and archived carbapenem resistant Klebsiella pneumoniae (CR-Kp) isolates obtained from various clinical samples submitted in the Bacteriology laboratory for culture and sensitivity testing, from July 2018 to August 2019. After preliminary identification (ID) and antibiotic susceptibility testing (AST), as per standard guidelines, these isolates were sent to Central Research Laboratory (CRL), Bengaluru, India, for further characterization & WGS. Results: 27 out of 29 test isolates were CRKp. Among the 27 CRKp isolates, ST14 was the most common sequence type (8, 29.6%), followed by ST231 (5, 18.5%) and ST147(3, 11.1%) respectively. KL2 (9/27, 33.3%) and KL51 (8/27, 29.6%) were dominant K loci types in this study. O1 and O2 together accounted for 88.9% (n=27) CRKp isolates. Yersiniabactin and Aerobactin were identified in 88.9% (24/27) & 29.6% (8/27) of the CRKp isolates of the isolate. The predominant plasmid replicons present were ColKP3 (55.5%), IncFII(K) (51.8%), IncFIB(pQil) (44.4%), IncFIB(K) (37%), IncR (33.3%) and Col44 0I (18.5%) respectively. As compared to phenotypic resistance, higher genotypic resistance for aminoglycosides (96.3%) and folate pathway inhibitors (92.6%) respectively, was observed. Conclusion: This study emphasizes the need for continued genomic surveillance of emerging CRKp and other MDR bacteria in Uttarakhand and neighbouring states of India.
Isolation and Molecular Characterization of Pesticide (Fenitrothion) Resistant Bacteria from Agricultural Field