Tetracyclines-induced phenotype and resistance mechanism in Aeromonas hydrophila

Abstract:

[Objective] The aim of this study is to explore the changes of sensitivity to tetracyclines and the resistance mechanism of Aeromonas hydrophila (Ah) after in vitro induction by exposure Ah to the increasing concentration of Doxycycline (DO). [Methods] The sensitive Ah isolates were first screened from clinical isolates and then incubated on TSA solid medium containing DO of 1/4×MIC value. The resistant strains were obtained through continuous subculture by increasing the concentration of DO in the medium for geometric series. The minimal inhibitory concentrations (MICs) of induced strains to DO and 16 other non-selected antimicrobials were determined. Meanwhile, MICs were also determined after adding efflux pump inhibitor 1-methyl-2-pyrrolidinone (NMP) to the medium. The relationship between sensitivity changes and efflux function were analyzed. Then five genes of tet from induced strains were amplified and sequenced. [Results] The MICs of induced strains to DO increased significantly after induction, whereas the MICs of strains to those non-selected tetracyclines also increased. The MICs of induced strains to fluoroquinolone increased much more than that of control. The induced strains exhibited a little higher sensitivity to aminoglycoside and rifampicin. However, the MICs of all induced strains to DO decreased after adding NMP to the medium. The detection of tet genes indicated that tetA and tetE were positive in No. 7 after induction and the tetC gene was positive in No. 2 before and after induction. The tetE gene was detected in strains No. 1, 3, 4, 5, 6 and 7 no matter whether it was induced or not. [Conclusion] This study suggested that tetE gene may be the predominant gene mediating tetracyclines-resistance of Ah, which would provide theoretical basis to clarify the resistance mechanism of Ah to tetracyclines.