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

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

Susmita Kuinkel1, Jyoti Acharya2, Binod Dhungel1, Sanjib Adhikari1, Nabaraj Adhikari1, Upendra Thapa Shrestha1, Megha Raj Banjara1, Komal Raj Rijal1, * and Prakash Ghimire1

 

1Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu, Nepal

2National Public Health Laboratory, Teku, Kathmandu, Nepal

 

* Correspondence: rijalkomal@gmail.com or komal.rijal@cdmi.tu.edu.np

 

ABSTRACT

Klebsiella spp. are associated with several nosocomial and opportunistic infections. Increasing antimicrobial resistance of Klebsiellaspecies 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.

 

Keywords: Antimicrobial resistance; Multi drug resistant (MDR); Extended-spectrum β-lactamase (ESBL); AmpC β-lactamase (ABL); Carbapenemase; Biofilm

 

Citation: Kuinkel, S.; Acharya, J.; Dhungel, B.; Adhikari, S.; Adhikari, N.; Shrestha, U.T.; Banjara, M.R.; Rijal, K.R.; Ghimire, P. 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. Microbiol. Res. 2021, 12, 683–697. https://doi.org/10.3390/microbiolres12030049, www.mdpi.com/journal/microbiolres

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