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Whole Genome Sequencing of a Citrobacter freundii Strain Isolated from the Hospital Environment: An Extremely
Raoudha Dziri1, Mert Ahmet Kuşkucu2, Amani Arfaoui1
1Laboratory of Microorganisms and Active Biomolecules, Department of Biology, Faculty of Sciences of Tunis, University of Tunis El Manar, Tunis, Tunisia.
Abstract:
Citrobacter freundii has acquired resistance to several antimicrobial drugs, including last-resort antibiotics affecting, therefore, clinical efficacy and causing high rates of mortality. In this study, we investigate the whole genome sequence of a carbapenem-resistant C. freundii strain isolated from the hospital environment in Tunisia. A total of 210 samples were taken using sterile swabs, from inanimate surfaces, medical devices, and care staff, during the period extended between March and April 2019. After the microbiological analysis of samples and antimicrobial susceptibility testing, only one strain identified as C. freundii showing resistance to carbapenems was selected for the whole genome sequencing. The genome analysis revealed a high-level resistance to most antibiotics. Interestingly, we have noted the coexistence of blaNDM-1 and blaVIM-48 metallo-β-lactamase (MBL) encoding genes conferring resistance to carbapenems. Other β-lactamases encoding genes have also been detected, including blaTEM-1, blaCMY-48, and blaOXA-1. Moreover, genes conferring resistance to aminoglycoside [aac(3)-IId, ant(3″)-Ia, aadA, aac(6')-Ib], macrolide [mph(A)], sulfonamide (sul1), trimethoprim (dfrA1), tetracycline [tet(D)], chloramphenicol [cat(B3)], rifamycin (arr-3), and quinolone (qnrB) have been revealed. The multi-locus sequence typing analysis showed that this isolate could not be assigned to an existing sequence type (ST), but it is almost identical to ST22. The plasmid investigation revealed the presence of five plasmids belonging to diverse incompatibility groups (IncFII, IncHI1A, IncHI1B, IncN, and IncX3). To the best of our knowledge, our findings report the first detection of NDM-1 and VIM-48 coproducing C. freundii in Tunisia and the second detection in the world of the blaVIM-48.
Insights
A carbapenem-resistant Citrobacter freundii strain from Tunisia harbors multiple resistance genes, including blaNDM-1 and blaVIM-48. This study highlights the urgent need for enhanced infection control measures against multidrug-resistant bacteria.
Area of Science:
- Microbiology and Infectious Diseases
- Genomics and Molecular Biology
- Antimicrobial Resistance
Background:
- Citrobacter freundii is a significant opportunistic pathogen.
- Acquired multidrug resistance, including to carbapenems, threatens clinical treatment efficacy.
- Hospital environments can act as reservoirs for resistant bacteria.
Purpose of the Study:
- To perform whole genome sequencing of a carbapenem-resistant C. freundii strain from a Tunisian hospital.
- To identify the genetic basis of its extensive antimicrobial resistance.
- To characterize the strain's genetic relatedness and plasmid content.
Main Methods:
- Sample collection from hospital surfaces, devices, and staff.
- Microbiological analysis and antimicrobial susceptibility testing.
- Whole genome sequencing and bioinformatic analysis.
- Multi-locus sequence typing (MLST) and plasmid profiling.
Main Results:
- Identification of a carbapenem-resistant C. freundii strain with high-level resistance to multiple antibiotics.
- Co-detection of blaNDM-1 and blaVIM-48 metallo-β-lactamase genes, alongside other resistance genes (blaTEM-1, blaCMY-48, blaOXA-1, aminoglycoside, macrolide, sulfonamide, trimethoprim, tetracycline, chloramphenicol, rifamycin, quinolone resistance genes).
- MLST analysis indicated the strain is closely related to ST22, but not assigned to an existing ST.
- Five plasmids from diverse incompatibility groups (IncFII, IncHI1A, IncHI1B, IncN, IncX3) were identified.
Conclusions:
- This study reports the first co-occurrence of NDM-1 and VIM-48 in C. freundii in Tunisia.
- The findings underscore the emergence of highly resistant Gram-negative bacteria in healthcare settings.
- Genomic surveillance is crucial for understanding and controlling the spread of antimicrobial resistance.
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