Abstract :
[en] Introduction
Complex oligosaccharides from human milk (HMO) promote growth of bifidobacteria such as Bifidobacterium bifidum [1]. Whey, a by-product of dairy-industry, contents complex oligosaccharides (BMO) similar to HMO, which are mainly represented in colostrum by 3’-sialyllactose (3’SL) [2]. Bifidobacterium crudilactis, a species of bovine origin, encodes for β galactosidases and α-glucosidases and could therefore be able to metabolise those BMO [3; 4; 5]. In addition, fermentation products from bifidobacteria can produce antivirulent activity against intestinal pathogenic bacteria [6; 7]. This study focused on capacity of bifidobacteria to metabolise BMO, more particularly 3’SL, and on potential antivirulent effect of cell-free spent media (CFSM) against virulence gene expression of pathogenic bacteria.
Material and methods
B. bifidum BBA1 and B. crudilactis FR/62/B/3 isolated respectively from breastfed children feces and from cow raw milk cheese were grown on media supplemented with BMO or 3’SL, as sole source of carbon. The CFSM were harvested after centrifugation of cells culture, freeze-dried and concentrated 10 fold. Next, their effects were tested against virulence gene expression using ler and hilA promoter activity of luminescent constructs of Escherichia coli 0157:H7 ATCC 43888 and Salmonella Typhimurium SA 941256, respectively. The effect was confirmed on wild type strains of E. coli O157:H7 ATCC 43890 and S. Typhimurium ATCC 14028 using RT-qPCR.
Results
Both strains were able to grow in presence of whey or 3’SL, but B. crudilactis showed the best growth compared to B. bifidum. The highest cell concentrations were observed with media containing whey (8.9 ± 0.6 log cfu/ml and 8.1 ± 0.3 log cfu/ml, respectively). CFSM from fermented media supplemented with 3’SL resulted in under-expression of hilA and ler genes for the luminescent constructs and in under-expression of ler (ratios of -15.4 and -8.1) and qseA (ratios of -2.1 and -3.1) genes for the wild type strain of E. coli O157:H7. No effect was observed for the wild type strain of S. Typhimurium.
Discussion
B. crudilactis presented the best growth potential probably because its genome encodes the enzymatic machinery to use BMO (β galactosidases and α-glucosidases) [3; 4; 5]. The positive effect of media supplemented with milk products on growth of probiotics has been demonstrated previously [8].
CFSM obtained from media supplemented with 3’SL down-regulate several virulence genes of E. coli O157:H7 and potentially S. Typhimurium. This effect has been observed previously with CFSM obtained from fermentation of lactic acid bacteria or bifidobacteria, by production of antivirulent metabolites [2; 3].
BMO combined with some bifidobacteria strains of bovine or human origin could therefore be an interesting synbiotic to maintain or restore the intestinal health of young children. These effects observed in vitro will be further investigated regarding the exact nature of the active molecules.
References
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