Antibacterial effects of Anodonta cygnea fluids on Escherichia coli and enterococci multi-drug-resistant strains: environmental implications

Abstract

The persistence of bacteria in Anodonta cygnea largely depends on the hemolymph bactericidal activity against aquatic micro-organisms.The aim of this study was to assess the in vitro bactericidal activity of whole hemolymph against multi-drug-resistant Escherichia coli and enterococci strains. Four mussels were injected with multi-resistant strains of E. coli in order to determine their in vivo bactericidal reactivity. In vitro experiments showed that the hemocytes viability decreased almost 70% in 4 h, the same happened in the control hemolymph. Enterococci tested in in vitro experiments were more susceptible than E. coli to whole hemolymph and hemocytes fractions. None of the enterococci bacteria was detected in the hemolymph fractions from 24 to 72 h, while E. coli was still detectable 72 h after inoculation both in the control and respective plasma fractions. The microbial inhibition on both hemolymph and cell pellets suggest that hemocytes may be the main responsibles for that process.In vivo experiments showed that hemolymph inhibited the E. coli load injected and it was correlated with a high tendency for the increase in hemocytes counts, after 72 h. High adherence of E. coli and an intense pseudopods manifestation, mainly to the granulocytes, suggest a clear phagocytosis process.