Adaptive laboratory evolution of Vibrio cholerae to doxycycline associated with spontaneous mutation.

Cholera, caused by the Gram-negative bacterium Vibrio cholerae, remains a serious threat in underdeveloped countries. Although rehydration therapy has been the mainstay of disease management, antibiotics are also being used as an adjunct treatment, resulting in an increase in the circulation of antimicrobial-resistant V. cholerae strains. In the present study, adaptive laboratory evolution, whole-genome sequencing and molecular docking studies were performed to identify putative mutations related to doxycycline resistance in V. cholerae isolates. The V57L mutation in the RpsJ protein was identified to be important in conferring doxycycline resistance. As revealed by molecular docking studies, the mutation was identified to alter the ribosome structure near the doxycycline binding site. Doxycycline stress also induced co-resistance to colistin, a last-resort antibiotic to treat extensively drug-resistant bacteria. This study illustrates for the first time a possible mechanism of doxycycline-selected resistance in V. cholerae as well as doxycycline-selected co-resistance, warranting strict restrictions on the indiscriminate use of antibiotics.