Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicity

Carbapenem drugs have the potential to prevent disease caused by both gram-positive and gram-negative bacteria. This beta-lactam family drug prevents the cell wall biosynthesis of pathogens, but the pathogen hydrolyzes it using the enzymes carbapenemase and beta-lactamase. In this study, we analyzed...

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Main Authors: V. Samyuktha, C.R. Pauline, G. Ragul, P. Sivaperumal, K. Kamala
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:The Microbe
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Online Access:http://www.sciencedirect.com/science/article/pii/S2950194625001396
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author V. Samyuktha
C.R. Pauline
G. Ragul
P. Sivaperumal
K. Kamala
author_facet V. Samyuktha
C.R. Pauline
G. Ragul
P. Sivaperumal
K. Kamala
author_sort V. Samyuktha
collection DOAJ
description Carbapenem drugs have the potential to prevent disease caused by both gram-positive and gram-negative bacteria. This beta-lactam family drug prevents the cell wall biosynthesis of pathogens, but the pathogen hydrolyzes it using the enzymes carbapenemase and beta-lactamase. In this study, we analyzed the biochemical characterization, biofilm formation, hemolytic activity, and biofilm-forming ability of carbapenemase-producing Klebsiella pneumoniae under various conditions. The Gram-negative rod-shaped bacteria showed positive results in the methyl red and citrate tests, and lactose fermentation in sugar tests; other biochemical characteristics identified the isolate as Klebsiella pneumoniae. The well developed biofilm has developed after five days (120 hours) of incubation at ambient temperature (35 C°). The hemolytic activity has shown 100 % cell lysis at 1 mg/mL (1000μg/mL) which retained the toxicity after sterilization indicates the virulence of pathogen. The viability of pathogens under thermal stress has revealed substantial survival at 100°C. The growth declined at 150°C, and the cells death at 200°C and above. Antibiofilm assay using carbapenemase at a concentration of 1000 µg/mL resulted in the reduction of cell density, and the biofilm has a layer of live active cells marked by acridine orange. It showed the stress adaptation, higher level of tolerance, and cytotoxicity of the carbapenemase-producing pathogen Klebsiella pneumoniae. Overall, it can cause difficult challenges in medical and clinical practices and treatments.
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spelling doaj-art-7a9aa1c3eb894daeb84490ff7ca94dbd2025-08-20T03:48:46ZengElsevierThe Microbe2950-19462025-06-01710037110.1016/j.microb.2025.100371Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicityV. Samyuktha0C.R. Pauline1G. Ragul2P. Sivaperumal3K. Kamala4Saveetha Dental College and Hospitals, Saveetha Institute of Medical & Technical Sciences, Chennai, Tamil Nadu 600077, IndiaMarine Biomedical Research Lab & Environmental Toxicology Unit, Cellular and Molecular Research Centre, Saveetha Dental College and Hospitals, Saveetha Institute of Medical & Technical Sciences, Saveetha University, Chennai, Tamil Nadu 600077, India; Centre for Marine and Aquatic Research (CMAR), Saveetha Institute of Medical & Technical Sciences, Saveetha University, Chennai 602105, IndiaSaveetha Dental College and Hospitals, Saveetha Institute of Medical & Technical Sciences, Chennai, Tamil Nadu 600077, India; Centre for Marine and Aquatic Research (CMAR), Saveetha Institute of Medical & Technical Sciences, Saveetha University, Chennai 602105, IndiaMarine Biomedical Research Lab & Environmental Toxicology Unit, Cellular and Molecular Research Centre, Saveetha Dental College and Hospitals, Saveetha Institute of Medical & Technical Sciences, Saveetha University, Chennai, Tamil Nadu 600077, India; Centre for Marine and Aquatic Research (CMAR), Saveetha Institute of Medical & Technical Sciences, Saveetha University, Chennai 602105, IndiaSaveetha Dental College and Hospitals, Saveetha Institute of Medical & Technical Sciences, Chennai, Tamil Nadu 600077, India; Centre for Marine and Aquatic Research (CMAR), Saveetha Institute of Medical & Technical Sciences, Saveetha University, Chennai 602105, India; Corresponding author at: Centre for Marine and Aquatic Research (CMAR), Saveetha Institute of Medical & Technical Sciences, Saveetha University, Chennai 602105, India.Carbapenem drugs have the potential to prevent disease caused by both gram-positive and gram-negative bacteria. This beta-lactam family drug prevents the cell wall biosynthesis of pathogens, but the pathogen hydrolyzes it using the enzymes carbapenemase and beta-lactamase. In this study, we analyzed the biochemical characterization, biofilm formation, hemolytic activity, and biofilm-forming ability of carbapenemase-producing Klebsiella pneumoniae under various conditions. The Gram-negative rod-shaped bacteria showed positive results in the methyl red and citrate tests, and lactose fermentation in sugar tests; other biochemical characteristics identified the isolate as Klebsiella pneumoniae. The well developed biofilm has developed after five days (120 hours) of incubation at ambient temperature (35 C°). The hemolytic activity has shown 100 % cell lysis at 1 mg/mL (1000μg/mL) which retained the toxicity after sterilization indicates the virulence of pathogen. The viability of pathogens under thermal stress has revealed substantial survival at 100°C. The growth declined at 150°C, and the cells death at 200°C and above. Antibiofilm assay using carbapenemase at a concentration of 1000 µg/mL resulted in the reduction of cell density, and the biofilm has a layer of live active cells marked by acridine orange. It showed the stress adaptation, higher level of tolerance, and cytotoxicity of the carbapenemase-producing pathogen Klebsiella pneumoniae. Overall, it can cause difficult challenges in medical and clinical practices and treatments.http://www.sciencedirect.com/science/article/pii/S2950194625001396CarbapenemaseKlebsiella pneumoniaeNosocomial infectionTolerance
spellingShingle V. Samyuktha
C.R. Pauline
G. Ragul
P. Sivaperumal
K. Kamala
Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicity
The Microbe
Carbapenemase
Klebsiella pneumoniae
Nosocomial infection
Tolerance
title Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicity
title_full Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicity
title_fullStr Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicity
title_full_unstemmed Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicity
title_short Virulence and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae: Insights into biofilm formation and cytotoxicity
title_sort virulence and resistance mechanisms of carbapenemase producing klebsiella pneumoniae insights into biofilm formation and cytotoxicity
topic Carbapenemase
Klebsiella pneumoniae
Nosocomial infection
Tolerance
url http://www.sciencedirect.com/science/article/pii/S2950194625001396
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