Biofilm Formation and Phenotypic Detection of ESBL, MBL, KPC and AmpC Enzymes and Their Coexistence in Klebsiella spp. Isolated at the National Reference Laboratory, Kathmandu, Nepal

Susmita Kuinkel,Megha Raj Banjara,Binod Dhungel,Nabaraj Adhikari,Komal Raj Rijal,Sanjib Adhikari,Prakash Ghimire,Upendra Thapa Shrestha,Jyoti Acharya

doi:10.3390/microbiolres12030049

Susmita Kuinkel, Megha Raj Banjara + Show 7 more

Open Access

https://doi.org/10.3390/microbiolres12030049

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Abstract

Klebsiella spp. are associated with several nosocomial and opportunistic infections. Increasing antimicrobial resistance of Klebsiella species is aggravated by a number of intrinsic and extrinsic factors. The main aim of this study is to determine antimicrobial resistance due to production of β-lactamase enzymes, extended spectrum beta-lactamase (ESBL), metallo-beta-lactamase (MBL) and AmpC and Klebsiella pneumoniae carbapenemase (KPC) and biofilm formation in Klebsiella isolates. A total of 2197 non-duplicate specimens of urine, sputum and pus were obtained from the National Public Health Laboratory (NPHL), Kathmandu, Nepal, between February and August 2019. Klebsiella species were isolated, identified and screened for antimicrobial susceptibility testing with the disk diffusion method. Phenotypic detection of ESBL, MBL, KPC and AmpC production was observed and biofilm production was detected by the microtiter plate method. Out of a total of 2197 clinical specimens, bacterial growth was detected in 8% (175/2197) of the specimens. Of the total isolates, 86.3% (151/175) were Gram-negative bacteria and 37.7% (57/151) were Klebsiella spp. Of the total Klebsiella spp., 56% (32/57) were multi drug resistant (MDR), 16% (9/57) were ESBL, 26% (15/57) were MBL, 4% (2/57) were KPC (class A carbapenemase), 16% (9/57) were AmpC producers and 95% (54/57) were biofilm producers. Gentamicin was the most effective antibiotic, followed by cotrimoxazole, as 68% (39/57) and 47% (27/57) of the Klebsiella isolates were susceptible towards these drugs, respectively. The study results show evidence of β-lactamase production, high prevalence of MDR and biofilm producing Klebsiella species. Integrating the test parameters for phenotypic confirmation of ESBL, MBL, AmpC β lactamase and KPC in routine diagnostic procedures can help in the early detection and management of these resistant strains.

Highlights

The main objective of this study is to explore the burden of extended spectrum beta-lactamase (ESBL), AmpC, MBL, Klebsiella pneumoniae carbapenemase (KPC) and biofilm producing Klebsiella species, in addition to assessing co-existence of these factors and their role in development of drug resistance
A total of 2197 different clinical specimens were processed for microbial culture, of which 175 (8.3%) showed growth of bacteria
One-third of the isolated Gram-negative bacteria was constituted by Klebsiella spp

Summary

Introduction

Klebsiella species are Gram-negative, non-motile, encapsulated, lactose-fermenting and facultative anaerobic bacteria that cause several diseases, especially in countries with a poor health system [1]. K. rhinoscleromatis) and K. oxytoca are the frequently isolated and medically important members of this genus [2]. Despite being normal flora organisms of the human gastrointestinal tract, K. pneumoniae is the most common pathogenic agent of nosocomial infections among. Urinary tract infection (UTI), blood stream and surgical wound infections, peritonitis, septicemia, meningitis [4] and pyogenic liver abscess [5]

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