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Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes
Published on: March 3, 2023
Widespread Distribution of Carbapenem-Resistant Klebsiella spp. in Clinical and Environmental Settings
Karla Vanessa Molina Maldonado1,2, Julia Marchese Pereira1, Tamires Nascimento da Costa1,2
1Laboratório de Imunologia e Microbiologia, Escola de Ciências da Saúde e da Vida, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre 90619-900, RS, Brazil.
Abstract:
Background/Objectives:Klebsiella spp., particularly K. pneumoniae, are major opportunistic pathogens in healthcare settings driven by carbapenemase- and ESBL-producing strains. We assessed antimicrobial resistance and biofilm formation abilities in Klebsiella spp. from a Brazilian tertiary hospital and related environments and characterized capsular types. Methods: Over six months (July-December 2023), 303 carbapenem-resistant Klebsiella spp. were collected from clinical specimens (n = 198), ICU/non-ICU surfaces (n = 79), hospital sewage (n = 22), and stream water (n = 4). Species were identified by MALDI-TOF. Susceptibility testing covered eight antibiotic classes, focusing on carbapenems and polymyxin B. Biofilm formation was quantified by crystal violet, and capsular typing used wzi/K-locus approaches. Results: Most isolates (70.95%) had meropenem MICs ≥ 128 μg/mL, while 77.6% (n = 235) remained susceptible to polymyxin B. Resistance profiles largely consisted of extensive drug resistance (95.4%), with 1.3% exhibiting pandrug resistance, including isolates from bed rails. Biofilm formation was detected in 96.7% of isolates, mainly weak (67.6%) or moderate (28%), with 4.4% being strong producers. Among the Klebsiella isolates analyzed, 21 K types were identified with an uneven distribution dominated by K64, followed by K24, K173, and K50. K75 was the only K type detected across all sources-clinical isolates, bed-rail surfaces (non-ICU), wastewater, and fluvial water. Conclusions: Carbapenem-resistant Klebsiella spp. exhibited widespread resistance, with residual susceptibility to aminoglycosides, ceftazidime-avibactam, and polymyxins. Environmental reservoirs-hospital surfaces, sewage, and stream water-harbored resistant biofilm producers, reinforcing their role in persistence and dissemination. K-typing revealed concentrated distribution (predominantly K64) and cross-source K75. These findings underscore the urgency of integrated strategies combining molecular surveillance, antimicrobial stewardship, and environmental control.
Insights
Carbapenem-resistant Klebsiella, a major hospital pathogen, shows widespread antimicrobial resistance and biofilm formation. Environmental sources like surfaces and water harbor these resistant strains, highlighting the need for integrated control strategies.
Area of Science:
- Clinical Microbiology
- Antimicrobial Resistance
- Environmental Microbiology
Background:
- Klebsiella spp., particularly K. pneumoniae, are significant opportunistic pathogens in healthcare settings.
- Carbapenemase- and ESBL-producing strains drive the prevalence of antimicrobial resistance in Klebsiella.
- Understanding resistance and biofilm formation is crucial for controlling Klebsiella infections.
Purpose of the Study:
- To assess antimicrobial resistance and biofilm formation in Klebsiella spp. from a Brazilian tertiary hospital and related environments.
- To characterize the capsular types of carbapenem-resistant Klebsiella isolates.
- To identify potential environmental reservoirs for Klebsiella dissemination.
Main Methods:
- Collected 303 carbapenem-resistant Klebsiella spp. from clinical specimens, hospital surfaces, sewage, and stream water.
- Identified species using MALDI-TOF and performed antimicrobial susceptibility testing against eight antibiotic classes.
- Quantified biofilm formation using crystal violet and characterized capsular types via wzi/K-locus approaches.
Main Results:
- Most isolates showed high meropenem resistance (70.95% MICs ≥ 128 μg/mL), but 77.6% remained susceptible to polymyxin B.
- 95.4% of isolates exhibited extensive drug resistance, with biofilm formation detected in 96.7% (mostly weak or moderate).
- 21 K types were identified, dominated by K64; K75 was found across all sources (clinical, surfaces, sewage, water).
Conclusions:
- Carbapenem-resistant Klebsiella spp. display extensive resistance but retain susceptibility to certain agents like polymyxins.
- Environmental reservoirs harbor resistant biofilm-producing Klebsiella, facilitating persistence and dissemination.
- Integrated strategies including surveillance, antimicrobial stewardship, and environmental control are essential.
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