In Vitro Susceptibility and Florfenicol Resistance inCitrobacterIsolates and Whole-Genome Analysis of Multidrug-ResistantCitrobacter freundii

Abstract

The genusCitrobacteris an opportunistic pathogen causing infections in animals, and the published data for its resistance to florfenicol are scarce. In this study, we investigated the antimicrobial susceptibility and molecular characteristics of florfenicol resistance genes amongCitrobacterisolates from animal and relevant environmental samples and conducted a comparative analysis of a multidrug-resistantCitrobacter freundiistrain isolated from a rabbit. Among 20Citrobacterstrains isolated from animal samples, resistance was most commonly observed to ampicillin (100%), tetracycline (75%), streptomycin (65%), florfenicol (60%), chloramphenicol (60%), and aztreonam (50%), while all the strains found in environmental samples were resistant to few antibiotics. The florfenicol resistance genefloRwas detected in 12 isolates (48%, 12/25) from animal samples, and all of thefloR-positive isolates were resistant to florfenicol with minimum inhibitory concentration (MIC) values ≥256 μg/mL. Sequencing and comparative analysis of the plasmids from a multidrug-resistantC. freundiiisolate named R47 showed that thefloR-containing region in the plasmid pR47-54 was a truncated transposon-like structure and could be found on both plasmids and chromosomes of bacteria of either animal or human origin. Furthermore, a range of antimicrobial and metal resistance genes associated with mobile genetic elements could be identified in pR47-54 and the other plasmid pR47-309 ofC. freundiiR47. These results provide in-depth views into the phenotypic and molecular characteristics ofCitrobacterisolates recovered from animal and relevant environmental samples, as well as highlight the role horizontal gene transfer plays in the dissemination of plasmid-encoded resistance genes.