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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
Hospital sink traps as a potential source of the emerging multidrug-resistant pathogen Cupriavidus pauculus:
James Butler1, Sean D Kelly2, Katie J Muddiman2
1School of Engineering, Computing and Mathematics, Faculty of Science and Engineering, University of Plymouth, Plymouth PL4 8AA, UK.
Abstract:
Introduction. Cupriavidus pauculus is historically found in soil and water but has more recently been reported to cause human infection and death. Hospital sink traps can serve as a niche for bacterial persistence and a platform for horizontal gene transfer, with evidence of dissemination of pathogens in hospital plumbing systems driving nosocomial infection.Gap Statement. This paper presents the first C. pauculus strain isolated from a hospital sink trap. There are only six genome assemblies available on NCBI for C. pauculus; two of these are PacBio/Illumina hybrids. This paper presents the first ONT/Illumina hybrid assembly, with five contigs. The other assemblies available consist of 37, 38, 111 and 227 contigs. This paper also presents data on biofilm formation and lethal dose in Galleria mellonella; there is little published information describing these aspects of virulence.Aim. The aims were to identify the isolate found in a hospital sink trap, characterize its genome, and assess whether it could pose a risk to human health.Methodology. The genome was sequenced, and a hybrid assembly of short and long reads produced. Antimicrobial susceptibility was determined by the broth microdilution method. Virulence was assessed by measuring in vitro biofilm formation compared to Pseudomonas aeruginosa and in vivo lethality in Galleria mellonella larvae.Results. The isolate was confirmed to be a strain of C. pauculus, with a 6.8 Mb genome consisting of 6468 coding sequences and an overall G+C content of 63.9 mol%. The genome was found to contain 12 antibiotic resistance genes, 8 virulence factor genes and 33 metal resistance genes. The isolate can be categorized as resistant to meropenem, amoxicillin, amikacin, gentamicin and colistin, but susceptible to cefotaxime, cefepime, imipenem and ciprofloxacin. Clear biofilm formation was seen in all conditions over 72 h and exceeded that of P. aeruginosa when measured at 37 °C in R2A broth. Lethality in G. mellonella larvae over 48 h was relatively low.Conclusion. The appearance of a multidrug-resistant strain of C. pauculus in a known pathogen reservoir within a clinical setting should be considered concerning. Further work should be completed to compare biofilm formation and in vivo virulence between clinical and environmental strains, to determine how easily environmental strains may establish human infection. Infection control teams and clinicians should be aware of the emerging nature of this pathogen and further work is needed to minimize the impact of contaminated hospital plumbing systems on patient outcomes.
Insights
A multidrug-resistant Cupriavidus pauculus strain was found in a hospital sink trap, a potential reservoir for nosocomial infections. This study characterized its genome and virulence, highlighting concerns for patient safety.
Area of Science:
- Clinical Microbiology
- Genomics
- Infectious Diseases
Background:
- Cupriavidus pauculus, typically found in soil and water, is increasingly recognized as a cause of human infections.
- Hospital plumbing systems, including sink traps, can act as reservoirs for bacterial persistence and dissemination, contributing to nosocomial infections.
Purpose of the Study:
- To identify and characterize a Cupriavidus pauculus strain isolated from a hospital sink trap.
- To assess the potential human health risks posed by this environmental isolate through genomic and virulence factor analysis.
Main Methods:
- Whole-genome sequencing using Oxford Nanopore Technology (ONT) and Illumina platforms for hybrid assembly.
- Determination of antimicrobial susceptibility using the broth microdilution method.
- Assessment of in vitro biofilm formation and in vivo lethality in Galleria mellonella larvae.
Main Results:
- A multidrug-resistant Cupriavidus pauculus strain was identified, possessing genes for antibiotic, virulence, and metal resistance.
- The isolate exhibited significant biofilm formation, exceeding that of Pseudomonas aeruginosa under specific conditions.
- Antimicrobial susceptibility testing revealed resistance to meropenem, amoxicillin, amikacin, gentamicin, and colistin, but susceptibility to cefotaxime, cefepime, imipenem, and ciprofloxacin.
Conclusions:
- The isolation of a multidrug-resistant C. pauculus strain from a hospital sink trap is a significant concern for infection control.
- Further research is needed to compare virulence and biofilm formation between clinical and environmental strains to understand their potential for causing human infections.
- Awareness among healthcare professionals and enhanced infection control measures are crucial to mitigate the impact of contaminated hospital plumbing on patient outcomes.
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