Gastrointestinal survival and adaptation of antibiotic-resistant enterococci subjected to an in vitro digestion model

Abstract The ability of four enterococcal strains to survive human digestion and maintain their antibiotic resistance (AR) traits was investigated to determine the health risk posed by seafood-borne streptomycin-, erythromycin-, tetracycline- and gentamycin-resistant enterococci. After demonstrating ant(6)-I, ermB, tetO and aac(6')-aph(2'') gene transferability by mating assays, strains were inoculated into a mussel homogenate which underwent in vitro digestion. Digestion reduced plate counts by 2-3 log; qPCR counts decreased by 1.5 log (E. faecium 22571/2), 2.5 log (E. faecalis 113324) or did not decrease (E. faecium 125745 and 6767/2). The much lower plate than qPCR counts seen in E. faecium suggested a viable-but-non-culturable (VBNC) subpopulation. Replica plating on antibiotic-supplemented agar and qPCR demonstrated that maintenance of AR traits was antibiotic-dependent, with a variable amount of erythromycin-, tetracycline- and gentamycin-resistant colonies and no streptomycin-resistant colonies. The seeking of unexpressed/lost AR genes in cells turned antibiotic-susceptible after digestion, identified aac(6')-aph(2'') in all colonies of E. faecalis 113324 and E. faecium 6767/2, in the 40% of 125745 and in none of the E. faecium 22571/2. In conclusion, after digestion the AR traits of antibiotic-resistant (also VBNC) enterococci contained in seafood may be maintained and transferred to the human microbiota and from it to intestinal pathogens.