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Biofilm Formation and Associated Gene Expression in Multidrug-Resistant Klebsiella pneumoniae Isolated from Clinical Specimens

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Abstract

Biofilms reduce the bacterial growth rate, inhibit antibiotic penetration, lead to the development of persister cells and facilitate genetic exchange. The biofilm-associated Klebsiella pneumoniae infections have not been well studied, and their implications in overcoming the effects of antimicrobial therapy are yet to be fully understood. Hence this study evaluated the antibiotic resistance pattern, antibiotic resistance determinants of extended-spectrum beta-lactamase (ESBL) family. Biofilm-forming ability of seventy multidrug-resistant clinical isolates of K. pneumoniae and the biofilm-associated genes of representative biofilm formers from a tertiary care hospital were also assessed. The K. pneumoniae isolated from urine exhibited resistance towards ceftazidime, nalidixic acid and meropenem. Isolates from blood were resistant to cefuroxime. Higher rates of resistance were observed towards cefuroxime, nalidixic acid, and meropenem for the isolates from the endotracheal aspirate. Extended spectrum beta-lactamase production by CLSI’s disc diffusion-based confirmation test revealed all the K. pneumoniae to be as ESBL producers. Most of the isolates harboured the bla gene variants, blaSHV and blaTEM. Majority of the isolates were colistin sensitive. 97.1% of the K. pneumoniae produced biofilm. K. pneumoniae isolated from pus and blood produced fully established biofilms. Strong biofilm formers were sensitive to co-trimoxazole and ciprofloxacin. Moderate biofilm formers exhibited sensitivity towards meropenem and imipenem. Expression of the fimH gene was increased, while mrkD showed reduced expression among the strong biofilm formers. Moderate biofilm formers showed variable expression of the genes associated with the biofilm formation. The weak and non-biofilm formers showed reduced expression of both the fimbrial genes. Multidrug-resistant isolates produced ESBLs and formed well-established biofilms.

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Fig. 1

Source-wise representation of antibiotic resistance pattern of all the clinical isolates of K. pneumoniae against nine antibiotics. CIP ciprofloxacin, CAZ ceftazidime, CXM cefuroxime, C chloramphenicol, COT co-trimoxazole, CTX cefotaxime, IPM imipenem, MRP meropenem, NA nalidixic acid

Fig. 2
Fig. 3

source and the biofilm-forming ability of the clinical isolates of K. pneumoniae

Fig. 4
Fig. 5

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Funding

We would like to acknowledge the Nitte (Deemed to be University) for the Financial support received from (Grant no. NUFR1/2018/10/17), Mangalore, towards this study.

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Authors and Affiliations

  1. Nitte (Deemed to be University), Nitte University Centre for Science Education & Research, Deralakatte, Mangalore, 575018, India

    Priyanka Ashwath, Vijaya Kumar Deekshit, Indrani Karunasagar & Dharnappa Sannejal Akhila

  2. Department of Microbiology, Madras Medical Mission, Chennai, India

    Anusha Rohit & I. Dhinakaran

  3. Nitte (Deemed to be University), University Enclave, Medical Sciences Complex, Mangalore, India

    Anusha Rohit, Iddya Karunasagar & Indrani Karunasagar

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  1. Priyanka Ashwath
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PA—AM and BB. DVK—AM and GR. AR—AM and GR. ID – AM. IK – BB. IK – BB. ADS—AM and DB.

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Correspondence to Dharnappa Sannejal Akhila.

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Ashwath, P., Deekshit, V.K., Rohit, A. et al. Biofilm Formation and Associated Gene Expression in Multidrug-Resistant Klebsiella pneumoniae Isolated from Clinical Specimens. Curr Microbiol 79, 73 (2022). https://doi.org/10.1007/s00284-022-02766-z

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