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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
Molecular Characterization of Pseudomonas putida Group Isolates Carrying blaVIM-2 Disseminated in a University
Jun Sung Hong1,2, Eun-Jeong Yoon2, Wonkeun Song3
11 Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine , Seoul, Republic of Korea.
Abstract:
Pseudomonas putida group are Gram-negative bacilli with polar flagellation, which are ubiquitous in the environment, although they are rarely involved in human infections. The aim of this study was to identify the dissemination of VIM-2-producing P. putida group in clinical isolates from a hospital in Korea. Thirteen strains were collected from 2014 to 2015 for the study. The isolates were recovered from urine cultures of both inpatients and outpatients at the hospital. Minimum inhibitory concentrations of antibiotics were determined by Etest. Carbapenemase genes were amplified by polymerase chain reaction and sequenced. Pulsed-field gel electrophoresis was performed for strain typing. Whole-genome sequencing was carried out randomly for two strains chosen from each year of the study to analyze the plasmid structure carrying the blaVIM-2 genes. The 13 isolates carried nine different class I integrons harboring VIM-2 and were resistant to meropenem and imipenem (minimum inhibitory concentrations, ≥32 μg/ml), thus exhibiting a multidrug-resistant phenotype. The blaVIM-2 gene was located on a plasmid in seven of the isolates and on the chromosome in six isolates. Each case of the blaVIM-2 gene was disseminated by clonal spread, horizontal transfer, and was mostly an occasional occurrence. In this study, we demonstrated that multidrug-resistant P. putida group carrying VIM-2 has reemerged in human specimens in Korea.
Insights
Multidrug-resistant Pseudomonas putida strains producing VIM-2 carbapenemase have reemerged in Korean clinical isolates. These bacteria, found in urine samples, show resistance to critical antibiotics like meropenem and imipenem.
Area of Science:
- Microbiology
- Infectious Diseases
- Genetics
Background:
- Pseudomonas putida group are common environmental Gram-negative bacteria.
- They are rarely implicated in human infections.
- The emergence of carbapenemase-producing strains poses a significant public health threat.
Purpose of the Study:
- To investigate the dissemination of VIM-2-producing Pseudomonas putida group in clinical isolates from a Korean hospital.
- To characterize the genetic mechanisms and spread of these multidrug-resistant strains.
Main Methods:
- Antibiotic susceptibility testing using Etest.
- Carbapenemase gene detection via PCR and sequencing.
- Strain typing using pulsed-field gel electrophoresis.
- Whole-genome sequencing to analyze plasmid structures carrying blaVIM-2.
Main Results:
- Thirteen multidrug-resistant P. putida isolates were identified from urine cultures (2014-2015).
- All isolates harbored VIM-2 carbapenemase genes within class I integrons, conferring resistance to meropenem and imipenem.
- The blaVIM-2 gene was found on plasmids in 7 isolates and on the chromosome in 6 isolates.
- Dissemination occurred through clonal spread and horizontal gene transfer.
Conclusions:
- Multidrug-resistant Pseudomonas putida group carrying VIM-2 has reemerged in human specimens in Korea.
- These findings highlight the need for continuous surveillance of carbapenemase-producing bacteria in clinical settings.
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