Antibacterial activity of exopolysaccharide produced by bee gut-resident Enterococcus sp. BE11 against marine fish pathogens

Abstract

Background: In recent years, the demand for innovative antimicrobial agents has grown, considering the growing problem of antibiotic resistance in aquaculture. Adult Apis mellifera honeybees' gut represents an outstanding habitat to isolate novel lactic acid bacteria (LAB) able to produce prominent antimicrobial agents.

Methods: In the current study, twelve LAB were isolated and purified from the gut of adult Apis mellifera. The isolates were screened for exopolysaccharide (EPS) production. The most promising isolate BE11 was identified biochemically and molecularly using 16 S rRNA gene sequence analysis as Enterococcus sp. BE11 was used for the mass production of EPS. The partially purified BE11-EPS features were disclosed by its physicochemical characterization. Moreover, the antimicrobial activity of BE11 cell free supernatant (CFS) and its EPS was investigated against some fish pathogens namely, Pseudomonas fluorescens, Streptococcus agalactiae, Aeromonas hydrophila, Vibrio sp. and Staphylococcus epidermidis using well-cut diffusion method.

Results: The physicochemical characterization of BE11-EPS revealed that the total carbohydrate content was estimated to be ~ 87%. FTIR and NMR analysis ascertained the presence of galactose and glucose residues in the EPS backbone. Moreover, the GC-MS analysis verified the heterogeneous nature of the produced BE11-EPS made up of different monosaccharide moieties: galactose, rhamnose, glucose, arabinose sugar derivatives, and glucuronic acid. BE11 CFS and its EPS showed promising antimicrobial activity against tested pathogens as the inhibition zone diameters (cm) ranged from 1.3 to 1.7 and 1.2-1.8, respectively.

Conclusion: The bee gut-resident Enterococcus sp. BE11, CFS, and EPS were found to be promising antimicrobial agents against fish pathogens and biofilm producers affecting aquaculture. To the best of our knowledge, this is the first study to purify and make a chemical profile of an EPS produced by a member of the bee gut microbiota as a potential inhibitor for fish pathogens.

Keywords: Bees; Enterococcus sp; Exopolysaccharides; Fish pathogens; Lactic acid bacteria.