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Updated: May 3, 2026

Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes
Published on: March 3, 2023
Antibiotic resistance determinants in a Pseudomonas putida strain isolated from a hospital
Lázaro Molina1, Zulema Udaondo2, Estrella Duque2
1Laboratorio de Investigación y Control Agroalimentario, Universidad de Huelva, Huelva, Spain ; Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Granada, Spain ; Centro de Investigación en Química Sostenible, Universidad de Huelva, Huelva, Spain.
Abstract:
Environmental microbes harbor an enormous pool of antibiotic and biocide resistance genes that can impact the resistance profiles of animal and human pathogens via horizontal gene transfer. Pseudomonas putida strains are ubiquitous in soil and water but have been seldom isolated from humans. We have established a collection of P. putida strains isolated from in-patients in different hospitals in France. One of the isolated strains (HB3267) kills insects and is resistant to the majority of the antibiotics used in laboratories and hospitals, including aminoglycosides, ß-lactams, cationic peptides, chromoprotein enediyne antibiotics, dihydrofolate reductase inhibitors, fluoroquinolones and quinolones, glycopeptide antibiotics, macrolides, polyketides and sulfonamides. Similar to other P. putida clinical isolates the strain was sensitive to amikacin. To shed light on the broad pattern of antibiotic resistance, which is rarely found in clinical isolates of this species, the genome of this strain was sequenced and analysed. The study revealed that the determinants of multiple resistance are both chromosomally-borne as well as located on the pPC9 plasmid. Further analysis indicated that pPC9 has recruited antibiotic and biocide resistance genes from environmental microorganisms as well as from opportunistic and true human pathogens. The pPC9 plasmid is not self-transmissible, but can be mobilized by other bacterial plasmids making it capable of spreading antibiotic resistant determinants to new hosts.
Insights
A Pseudomonas putida strain isolated from a French hospital exhibits extensive antibiotic resistance due to chromosomal and plasmid-borne genes. This strain can transfer resistance genes to other bacteria, posing a public health risk.
Area of Science:
- Microbiology
- Genomics
- Antimicrobial Resistance
Background:
- Environmental microbes possess a vast reservoir of antibiotic resistance genes.
- Horizontal gene transfer can spread these genes to human and animal pathogens.
- Pseudomonas putida is common in the environment but rarely found in clinical settings.
Purpose of the Study:
- To investigate the genomic basis of broad-spectrum antibiotic resistance in a clinical isolate of Pseudomonas putida.
- To understand the origin and potential dissemination of resistance determinants.
Main Methods:
- Isolation and characterization of Pseudomonas putida strains from hospital patients.
- Whole-genome sequencing and analysis of a highly resistant strain (HB3267).
- Plasmid analysis to identify the location of resistance genes.
Main Results:
- The isolated strain HB3267 displayed resistance to numerous antibiotic classes.
- Resistance determinants were found on both the chromosome and the pPC9 plasmid.
- The pPC9 plasmid acquired genes from environmental microbes and human pathogens.
- The pPC9 plasmid is mobilizable, enabling gene transfer to other bacteria.
Conclusions:
- Clinical Pseudomonas putida strains can harbor extensive antibiotic resistance.
- The pPC9 plasmid acts as a significant vehicle for spreading antibiotic resistance genes.
- This highlights the potential for environmental resistance genes to impact clinical settings.
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