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See detailImpact of 3’-sialyllactose and Bifidobacterium crudilactis on infant feces microbial composition and virulence modulation of Escherichia coli O157:H7, using the SHIME® gastrointestinal model
Bondue, Pauline ULiege

Conference (2017, June 15)

Bifidobacterium crudilactis, a bacterial species from bovine origin, growths on bovine milk oligosaccharides (BMO). Cell free spent media (CFSM) from B. crudilactis and 3’-sialyllactose (3’SL), a major ... [more ▼]

Bifidobacterium crudilactis, a bacterial species from bovine origin, growths on bovine milk oligosaccharides (BMO). Cell free spent media (CFSM) from B. crudilactis and 3’-sialyllactose (3’SL), a major BMO, modulated Escherichia coli O157:H7 virulence gene expressiona. In this study, the SHIME® gastrointestinal model was inoculated with 4 different treatments: 3’SL (ttm1), B. crudilactis (ttm2), 3’SL and B. crudilactis (ttm3) and CFSM from 3’SL and B. crudilactis culture (ttm4). In each section of the colon, samples were collected and analysed for short chain fatty acid (SCFA) concentration using HPLC, microbial populations using 16S rDNA metagenetic analysis and correlated with E. coli O157:H7 virulence gene expression. The results showed that SCFA levels were stable during the experiments. Metagenetic analysis showed a microbial diversity in transverse (TC) and descending colon (DC) close to feces, dominated by Bacteroides, Prevotella and Fusobacterium, while the ascending colon (AC) showed a different microbial diversity dominated by Veillonella. Ttm4 and ttm2 induced mainly a down-regulation of virulence genes: fliC in DC with ttm4, and luxS, stx1, qseA in AC, DC or TC with ttm2. Ttm1 also showed a down-regulation of fliC in DC, similar to the one observed with ttm4, but this was associated with an up-regulation of fliC and stx1 in AC or TC. Finally, ttm3 showed slight upregulation of ler, fliC and qseA in AC. These results show that ttm4 and ttm2 might have a positive effect against virulence expression of E. coli O157:H7. However, this trend has to be validated with the further replicates on the SHIME® system. [less ▲]

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See detailImpact of 3’-­‐sialyllactose and Bifidobacterium crudilactis on infant feces microbial composition and virulence modulation of Escherichia coli O157:H7, using the SHIME® gastrointes6nal model
Bondue, Pauline ULiege; Lebrun, Sarah ULiege; Crevecoeur, Sébastien ULiege et al

Poster (2017, June)

Bifidobacterium crudilactis, a bacterial species from bovine origin, growths on bovine milk oligosaccharides (BMO). Cell free spent media (CFSM) from B. crudilactis and 3’-sialyllactose (3’SL), a major ... [more ▼]

Bifidobacterium crudilactis, a bacterial species from bovine origin, growths on bovine milk oligosaccharides (BMO). Cell free spent media (CFSM) from B. crudilactis and 3’-sialyllactose (3’SL), a major BMO, modulated Escherichia coli O157:H7 virulence gene expressiona. In this study, the SHIME® gastrointestinal model was inoculated with 4 different treatments: 3’SL (ttm1), B. crudilactis (ttm2), 3’SL and B. crudilactis (ttm3) and CFSM from 3’SL and B. crudilactis culture (ttm4). In each section of the colon, samples were collected and analysed for short chain fatty acid (SCFA) concentration using HPLC, microbial populations using 16S rDNA metagenetic analysis and correlated with E. coli O157:H7 virulence gene expression. The results showed that SCFA levels were stable during the experiments. Metagenetic analysis showed a microbial diversity in transverse (TC) and descending colon (DC) close to feces, dominated by Bacteroides, Prevotella and Fusobacterium, while the ascending colon (AC) showed a different microbial diversity dominated by Veillonella. Ttm4 and ttm2 induced mainly a down-regulation of virulence genes: fliC in DC with ttm4, and luxS, stx1, qseA in AC, DC or TC with ttm2. Ttm1 also showed a down-regulation of fliC in DC, similar to the one observed with ttm4, but this was associated with an up-regulation of fliC and stx1 in AC or TC. Finally, ttm3 showed slight upregulation of ler, fliC and qseA in AC. These results show that ttm4 and ttm2 might have a positive effect against virulence expression of E. coli O157:H7. However, this trend has to be validated with the further replicates on the SHIME® system. [less ▲]

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See detailMetabolites from media supplemented with 3’-sialyllactose and fermented by bifidobacteria have an antivirulent effect against intestinal pathogens
Bondue, Pauline ULiege; Daube, Georges ULiege; Delcenserie, Véronique ULiege

Poster (2016, October 21)

Complex oligosaccharides from human milk (HMO) promote growth of Bifidobacterium bifidum. Oligosaccharides from cow milk (BMO), similar to HMO, are mainly represented in colostrum by 3’-sialyllactose ... [more ▼]

Complex oligosaccharides from human milk (HMO) promote growth of Bifidobacterium bifidum. Oligosaccharides from cow milk (BMO), similar to HMO, are mainly represented in colostrum by 3’-sialyllactose (3’SL). Bifidobacterium crudilactis, a species from bovine origin and encoding for β galactosidases and α-glucosidases, could be able to metabolise them. Also, fermentation products could have antivirulent activity against intestinal pathogens. This study focused on capacity of bifidobacteria to metabolise 3’SL and on potential antivirulent effect of cell-free spent media (CFSM) against pathogenic bacteria. B. bifidum BBA1 and B. crudilactis FR/62/B/3 isolated respectively from breastfed children feces and cow raw milk cheese were grown on media supplemented with 3’SL as sole source of carbon. Next, CFSM 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. Both strains were able to grow in presence of 3’SL. CFSM 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 with S. Typhimurium. Little is known about CFSM metabolites and they have to be isolated and identified. The potential synbiotic effect between 3’SL and bifidobacteria will be tested using the Shime®, a human gastrointestinal model. [less ▲]

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See detailMetabolites from media supplemented with 3’-sialyllactose and fermented by bifidobacteria have an antivirulent effect against intestinal pathogens
Bondue, Pauline ULiege

Poster (2016, September 16)

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 ... [more ▼]

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 1. Garrido D. et al. (2013). Microbiology 159: 649-664. 2. Urashima T. et al. (2013). Biosci Biotechnol Biochem 77: 455-466. 3. Sela D. A. (2011). Int J Food Microbiol 149: 58-64. 4. Milani C. et al. (2014). Appl Environ Microbiol 80: 6290-6302. 5. Bondue P. & Delcenserie V. (2015). Korean J Food Sci Anim Resour 35: 1-9. 6. Medellin-Pena M. J. et al. (2007). Appl Environ Microbiol 73: 4259-4267. 7. Bayoumi M. A. & Griffiths M. W. (2012). Int J Food Microbiol 156: 255-263. 8. Champagne C. P. et al. (2014). Can J Microbiol 60: 287-295. [less ▲]

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See detailOptmization of culture media for Bifidobacterium bifidum and Bifidobacterium crudilactis and study of the antimicrobial effect of culture supernatants
Bondue, Pauline ULiege; Delcenserie, Véronique ULiege; Crevecoeur, Sébastien ULiege et al

Poster (2015, October)

Complex oligosaccharides from human milk (HMO) contribute to infant health. Bifidobacteriumbifidum mainly found in breast-fed infant microbiota has all the enzymatic machinery for degradation of HMO. On ... [more ▼]

Complex oligosaccharides from human milk (HMO) contribute to infant health. Bifidobacteriumbifidum mainly found in breast-fed infant microbiota has all the enzymatic machinery for degradation of HMO. On the other hand, whey is rich in complex bovin milk oligosaccharides (BMO) very similar to HMO, including 3’-sialyllactose (3’SL). They are very likely to be metabolised by B. bifidum too, but also by B. crudilactis, a bovine origin strain. Fermentation of HMO or BMO by bifidobacteria can result in production of metabolites modulating virulence expression of several pathogenic bacteria. Two strains of bifidobacteria were used in this study: B. bifidum, isolated from breastfed infant feces and B. crudilactis, isolated from bovine raw milk. The ability of those strains to metabolise culture media enriched in glucose, whey and 3’SL has been assessed. Then, the obtained culture supernatant has been tested against virulence genes expression of E. coli O157:H7.  Both strains were able to grow in presence of BMO and 3’SL. B. crudilactis presented the best growth on all media. All culture supernatants obtained after supplementation with 3’SL resulted in significant under-expression of genes ler and qseA. The trend of genes stxB2 and luxS was also toward a down-regulation. BMO combined to some bovine or human origin bifidobacteria strains could be interesting synbiotics to maintain or restore the intestinal health of young children. These effects observed in vitro require further investigations to ensure repeatability in humans and to identify the exact nature of molecules obtained from fermentation media by B. bifidum and B. crudilactis. [less ▲]

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See detailOptimisation de milieux de culture pour Bifidobacterium bifidum et Bifidobacterium crudilactis et étude de l’effet antimicrobien des surnageants de culture
Bondue, Pauline ULiege; Delcenserie, Véronique ULiege; Crevecoeur, Sébastien ULiege et al

Poster (2015, June)

Modulé entre autre par notre alimentation, le microbiote intestinal influence notre santé. Un enfant allaité sera en meilleure santé qu’un enfant nourri avec des formulations commerciales. Ceci est ... [more ▼]

Modulé entre autre par notre alimentation, le microbiote intestinal influence notre santé. Un enfant allaité sera en meilleure santé qu’un enfant nourri avec des formulations commerciales. Ceci est notamment dû à la présence d’oligosaccharides complexes dans le lait maternel (HMO). Les oligosaccharides ajoutés au lait maternisé sont d’origine végétale et leur structure est très éloignée de celle des HMO. Parce que les HMO ressemblent aux glycans de la paroi épithéliale, les pathogènes intestinaux infantiles s’y fixent et sont expulsés naturellement. Le lait de vache possède des oligosaccharides complexes (BMO), dont la structure est très similaire à celle des HMO. Les bifidobactéries rencontrées majoritairement dans les matières fécales d’un nourrisson devraient pouvoir métaboliser les BMO, tout comme celles isolées dans le lait de vache. Les objectifs de cette étude étaient d’étudier le potentiel de croissance de bifidobactéries d’origine bovine ou humaine sur des milieux de culture enrichis en lactosérum et BMO. Le deuxième objectif était de vérifier si le catabolisme de ces sucres complexes induisaient une synthèse de métabolites influençant l’expression de virulence de certains pathogènes tels qu’Escherichia coli O157 :H7. Une souche de Bifidobacterium bifidum, isolée à partir des matières fécales d’un nourrisson exclusivement allaité et une souche de Bifidobacterium crudilactis, isolée à partir de fromage au lait cru, ont été mises en culture dans des milieux contenant différentes sources d’hydrates de carbone (glucose, lactosérum naturellement riche en lactose et BMO, et 3’-syalillactose (3’SL)). Le 3’SL est un oligosaccharide complexe majoritaire parmi les BMO. Les surnageants des différents milieux de culture ont été prélevés et concentrés par lyophilisation puis mis en contact avec E. coli O157 :H7. L’expression relative de différents gènes de virulence d’E. coli O157:H7, fliC, ler, stx2b et luxS a été étudiée. Dans le futur, les BMO et certaines souches d’origine bovine ou humaine pourraient s’avérer être des compléments alimentaires intéressants pour maintenir ou rétablir la santé intestinale des jeunes enfants. [less ▲]

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See detailGenome of Bifidobacteria and Carbohydrate Metabolism
Bondue, Pauline ULiege; Delcenserie, Véronique ULiege

in Korean Journal for Food Science of Animal Resources (2015)

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