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Scientific reports2025; 15(1); 1599; doi: 10.1038/s41598-024-84149-x

Emerging biofilm formation and disinfectant susceptibility of ESBL-producing Klebsiella pneumoniae.

Abstract: Klebsiella pneumoniae is an opportunistic pathogen responsible for various infections in humans and animals. It is known for its resistance to multiple antibiotics, particularly through the production of Extended-Spectrum Beta-Lactamases (ESBLs), and its ability to form biofilms that further complicate treatment. This study aimed to isolate and identify K. pneumoniae from animal and environmental samples and assess commercial disinfectants' effectiveness against K. pneumoniae isolates exhibiting ESBL-mediated resistance and biofilm-forming ability in poultry and equine farms in Giza Governorate, Egypt. A total of 320 samples, including nasal swabs from equine (n = 60) and broiler chickens (n = 90), environmental samples (n = 140), and human hand swabs (n = 30), were collected. K. pneumoniae was isolated using lactose broth enrichment and MacConkey agar, with molecular confirmation via PCR targeting the gyrA and magA genes. PCR also identified ESBL genes (bla, bla, bla, bla) and biofilm genes (luxS, Uge, mrkD). Antimicrobial susceptibility was assessed, and the efficacy of five commercial disinfectants was evaluated by measuring inhibition zones. Klebsiella pneumoniae was isolated from poultry (13.3%), equine (8.3%), wild birds (15%), water (10%), feed (2%), and human hand swabs (6.6%). ESBL and biofilm genes were detected in the majority of the isolates, with significant phenotypic resistance to multiple antibiotics. The disinfectants containing peracetic acid and hydrogen peroxide were the most effective, producing the largest inhibition zones, while disinfectants based on sodium hypochlorite and isopropanol showed lower efficacy. Statistical analysis revealed significant differences in the effectiveness of disinfectants against K. pneumoniae isolates across various sample origins (P < 0.05). The presence of K. pneumoniae in animal and environmental sources, along with the high prevalence of ESBL-mediated resistance and biofilm-associated virulence genes, underscores the zoonotic potential of this pathogen. The study demonstrated that disinfectants containing peracetic acid and hydrogen peroxide are highly effective against ESBL-producing K. pneumoniae. Implementing appropriate biosecurity measures, including the use of effective disinfectants, is essential for controlling the spread of resistant pathogens in farm environments.
© 2024. The Author(s).
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Publication Date: 2025-01-10 PubMed ID: 39794383PubMed Central: PMC11724021DOI: 10.1038/s41598-024-84149-xGoogle Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Overview

  • This study investigated the presence of Extended-Spectrum Beta-Lactamase (ESBL) producing Klebsiella pneumoniae in animals and the environment at poultry and equine farms in Egypt and tested the effectiveness of several commercial disinfectants against these resistant bacteria, particularly focusing on their biofilm-forming capabilities.

Background and Objectives

  • Klebsiella pneumoniae is a bacterial pathogen involved in various infections in humans and animals.
  • It is particularly dangerous due to its ability to resist multiple antibiotics by producing ESBL enzymes and forming biofilms that enhance survival and resistance.
  • This study aimed to isolate K. pneumoniae from different animal species, environmental samples, and human sources within farm settings.
  • It also aimed to identify genes responsible for antibiotic resistance (ESBL genes) and biofilm formation and to evaluate the effectiveness of commercial disinfectants against these resistant strains.

Sample Collection and Identification Methods

  • A total of 320 samples were collected:
    • 60 nasal swabs from equine (horses)
    • 90 nasal swabs from broiler chickens
    • 140 environmental samples including water, feed, and wild birds
    • 30 human hand swabs from farm workers
  • Isolation of K. pneumoniae was performed using lactose broth enrichment and plating on MacConkey agar.
  • Molecular confirmation employed PCR, targeting specific genes (gyrA and magA) to confirm K. pneumoniae.
  • ESBL genes (bla variants) and biofilm-associated genes (luxS, Uge, mrkD) were also analyzed using PCR.

Findings on Prevalence

  • K. pneumoniae was found in:
    • 13.3% of poultry samples
    • 8.3% of equine samples
    • 15% of wild bird samples
    • 10% of water samples
    • 2% of feed samples
    • 6.6% of human hand swabs
  • Majority of these isolates carried ESBL genes, indicating multi-drug resistance.
  • Biofilm formation genes were also prevalent, suggesting enhanced virulence and survival.

Antimicrobial Susceptibility and Resistance

  • Phenotypic testing showed significant resistance of the isolates to multiple antibiotics.
  • This resistance complicates treatment and control efforts on farms and potentially threatens public health through zoonotic transmission.

Disinfectant Efficacy Testing

  • Five commercial disinfectants with different active ingredients were tested for their ability to inhibit K. pneumoniae growth by measuring zones of inhibition.
  • Disinfectants containing peracetic acid and hydrogen peroxide showed the largest inhibition zones, indicating the highest effectiveness.
  • Disinfectants based on sodium hypochlorite and isopropanol demonstrated lower efficacy against the resistant isolates.
  • There were statistically significant differences (P < 0.05) in disinfectant effectiveness depending on the sample origin of the bacteria.

Significance and Recommendations

  • The study highlights the presence of multi-drug resistant, biofilm-forming K. pneumoniae in farm animals, wildlife, environment, and human handlers, emphasizing its zoonotic potential.
  • The identification of effective disinfectants such as those containing peracetic acid and hydrogen peroxide is crucial for formulating control strategies against such resistant pathogens on farms.
  • Biosecurity measures including proper use of effective disinfectants are recommended to limit the spread of resistant bacteria in agricultural environments.
  • Continuous monitoring and molecular testing can help track resistance and virulence factors, aiding timely interventions.

Cite This Article

APA
Khalefa HS, Arafa AA, Hamza D, El-Razik KAA, Ahmed Z. (2025). Emerging biofilm formation and disinfectant susceptibility of ESBL-producing Klebsiella pneumoniae. Sci Rep, 15(1), 1599. https://doi.org/10.1038/s41598-024-84149-x

Publication

Scientific reports
ISSN: 2045-2322
NlmUniqueID: 101563288
Country: England
Language: English
Volume: 15
Issue: 1
Pages: 1599
PII: 1599

Researcher Affiliations

Khalefa, Hanan S
  • Department of Veterinary Hygiene and Management, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.
Arafa, Amany A
  • Department of Microbiology and Immunology, Veterinary Research Institute, National Research Centre, Dokki, Egypt.
Hamza, Dalia
  • Department of Zoonoses, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt. daliahamza@cu.edu.eg.
El-Razik, Khaled A Abd
  • Department of Animal Reproduction, Veterinary Research Institute, National Research Centre, Dokki, Egypt.
Ahmed, Zeinab
  • Department of Zoonoses, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.

MeSH Terms

  • Klebsiella pneumoniae / drug effects
  • Klebsiella pneumoniae / genetics
  • Klebsiella pneumoniae / physiology
  • Klebsiella pneumoniae / isolation & purification
  • Biofilms / drug effects
  • Biofilms / growth & development
  • Disinfectants / pharmacology
  • beta-Lactamases / genetics
  • beta-Lactamases / metabolism
  • Animals
  • Humans
  • Klebsiella Infections / microbiology
  • Klebsiella Infections / drug therapy
  • Microbial Sensitivity Tests
  • Horses
  • Chickens / microbiology
  • Anti-Bacterial Agents / pharmacology
  • Egypt

Conflict of Interest Statement

Declarations. Competing interests: The authors declare no competing interests. Ethics approval and consent to participate: The research was conducted according to the guidelines of the Ethical Committee of the Faculty of Veterinary Medicine, Cairo University, Egypt, and approved by the Institutional Animal Care and use committee, Vet Cu ((VET. CU. IACUC; approval no. Vet CU 18042024926). All experimental protocols were approved by the Institutional Animal Care and Use Committee (IACUC) of the Faculty of Veterinary Medicine, Cairo University, Egypt. The study was carried out in compliance with the ARRIVE guidelines. The oral consent of the humans involved in the study was obtained from all owners after they had been informed of the use of nasal swab samples. Ethical clearance to use human subjects was obtained from the designated health facility (National Research Centre, Giza, Egypt). Consent for publication: All authors have read the manuscript and consent to publish.

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