Probiotic Potential and Effects on Gut Microbiota Composition and Immunity of Indigenous Gut Lactobacilli in Apis cerana.

This study aimed to investigate the probiotic potential of gut indigenous lactic acid bacteria (LAB) originated from Apis cerana. Six Limosilactobacillus reuteri and one Lactobacillus helveticus were isolated from gut samples of A. cerana adult worker bee. All isolates antagonized the growth of pathogens including Salmonella typhimurium, Escherichia coli, Shigella flexneri, and Flavobacterium frigidimaris, and L. helveticus KM7 showed the greatest antimicrobial activity among them. All strains were sensitive to cefotaxime, amoxicillin, cephalothin, penicillin G, kanamycin, and vancomycin, moderately sensitive to novobiocin and resistant to gentamicin. Six out of seven strains were sensitive to ampicillin. L. helveticus KM7 was chosen to evaluate in vivo probiotic effect of adult worker bees of A. cerana through fed sucrose syrup supplemented with KM7. Administration of KM7 increased survival rate and gut LAB but decreased gut fungi and Enterococcus in honeybees. Expressions of genes related to antimicrobial peptides (AMPs) including Abaecin and Defensin were also induced in the gut of honeybees. The results suggested that L. helveticus KM7 with greater probiotic properties could improve the survival rate of adult worker honeybees of A. cerana through regulating gut microbiota and AMPs genes expression.

In vitro Effects of Prebiotics and Synbiotics on Apis cerana Gut Microbiota.

This study aimed to investigate in vitro effects of the selected prebiotics alone, and in combination with two potential probiotic Lactobacillus strains on the microbial composition of Apis cerana gut microbiota and acid production. Four prebiotics, inulin, fructo-oligosaccharides, xylo-oligosaccharides, and isomalto-oligosaccharides were chosen, and glucose served as the carbon source. Supplementation of this four prebiotics increased numbers of Bifidobacterium and lactic acid bacteria while decreasing the pH value of in vitro fermentation broth inoculated with A. cerana gut microbiota compared to glucose. Then, two potential probiotics derived from A. cerana gut at different dosages, Lactobacillus helveticus KM7 and Limosilactobacillus reuteri LP4 were added with isomalto-oligosaccharides in fermentation broth inoculated with A. cerana gut microbiota, respectively. The most pronounced impact was observed with isomalto-oligosaccharides. Compared to isomalto-oligosaccharides alone, the combination of isomalto-oligosaccharides with both lactobacilli strains induced the growth of Bifidobacterium, LAB, and total bacteria and reduced the proliferation of Enterococcus and fungi. Consistent with these results, the altered metabolic activity was observed as lowered pH in in vitro culture of gut microbiota supplemented with isomalto-oligosaccharides and lactobacilli strains. The symbiotic impact varied with the types and concentration of Lactobacillus strains and fermentation time. The more effective ability was observed with IMO combined with L. helveticus KM7. These results suggested that isomalto-oligosaccharides could be a potential prebiotic and symbiotic with certain lactobacilli strains on A. cerana gut microbiota.

In vitro probiotic properties of Pediococcus pentosaceus L1 and its effects on enterotoxigenic Escherichia coli-induced inflammatory responses in porcine intestinal epithelial cells.

This study aimed to evaluate in vitro probiotic characteristics of Pediococcus pentosaceus strain L1 from pickled radish and investigate its impacts on inflammatory responses in porcine intestinal epithelial cells (IEC) to enterotoxigenic Escherichia coli (ETEC) F4+. The abilities of P. pentosaceus L1 to tolerate gastrointestinal conditions and to antagonize ETEC F4+ growth were determined. Adhesion of P. pentosaceus L1 and its effect on ETEC F4+ adhesion to porcine IPEC-J2 IEC were evaluated. Furthermore, the effects of this strain on proinflammatory gene expression and cytokines/chemokine production in porcine IPEC-J2 IEC induced by ETEC F4+ were determined. P. pentosaceus L1 showed good tolerance to the medium adjusted at pH 2.5 and consequently supplemented with 0.3% oxgall. Reduction of ETEC F4+ growth in co-culture with L1 was found. Effective adhesion of L1 to porcine. IPEC-J2 IEC was observed under these conditions. P. pentosaceus L1 decreased the adhesion of ETEC F4+ to IPEC-J2 IEC and the extent of inhibition of ETEC F4+ adhesion depended on the timing of L1 addition. Further analysis revealed down-regulation of expression of ETEC F4+-induced proinflammatory genes encoding interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-8 (IL-8) in IPEC-J2 IEC. Expression of the genes involved in NF-κB pathway, including RELA and NFKB1, were also repressed, as was production of IL-6, TNF-α, and IL-8. These results indicate that P. pentosaceus L1 may have potential as a probiotic for control of ETEC infection in pigs.