<i>In vitro</i>screening of mare's milk antimicrobial effect and antiproliverative activity

Guri, Anilda; Paligot, Michèle; Crèvecoeur, Sébastien; Piedboeuf, Benoît; claes, Jonathan; Daube, Georges; Corredig, Milena; Griffiths, Mansel W.; Delcenserie, Véronique

doi:10.1093/femsle/fnv234

Cited by 18 publications

(19 citation statements)

References 42 publications

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“…Typhimurium at 600 nm and after 4 h of exposure was 0.862 ± 0.078. From every tested housekeeping gene, gmk was the most stable and was chosen to normalize and adjust the results according to the efficiency of each pair of primers (virulence genes hilA, ssrB2 , and sopD ) monitored using qPCR (Guri et al, 2016 , Table 5 ). The CFSM of MRS2-3′SL medium fermented by B. bifidum induced a slight down-regulation of the ssrB2 gene (−2.1; P ≤ 0.05).…”

Section: Resultsmentioning

confidence: 99%

“…The method was adapted with some modifications from Tellez et al ( 2012 ), Mith et al ( 2014 ) and Guri et al ( 2016 ). After 4 h of incubation (Delcenserie et al, 2012 ), cells were collected by centrifugation at 5000 × g for 10 min at room temperature (Eppendorf Centrifuge 5804, Hamburg, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…The proteins produced by S . Typhimurium are controlled and activated by hilA and sopD genes and the ssrB2 gene is a major regulator of the T3SS (Guri et al, 2016 ). Regarding E. coli O157:H7, AE lesions are controlled by a pathogenicity island named locus of enterocyte effacement (LEE) through ler gene, which is regulated by qseA gene also implicated in quorum sensing (Medellin-Pena et al, 2007 ).…”

Section: Introductionmentioning

confidence: 99%

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Cell-Free Spent Media Obtained from Bifidobacterium bifidum and Bifidobacterium crudilactis Grown in Media Supplemented with 3′-Sialyllactose Modulate Virulence Gene Expression in Escherichia coli O157:H7 and Salmonella Typhimurium

Bondue¹,

Crèvecoeur²,

Brose³

et al. 2016

Front. Microbiol.

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Complex oligosaccharides from human milk (HMO) possess an antimicrobial activity and can promote the growth of bifidobacteria such as Bifidobacterium bifidum and Bifidobacterium longum subsp. infantis. In addition, fermentation of carbohydrates by bifidobacteria can result in the production of metabolites presenting an antivirulence effect on several pathogenic bacteria. Whey is rich in complex bovine milk oligosaccharides (BMO) structurally similar to HMO and B. crudilactis, a species of bovine origin, is able to metabolize some of those complex carbohydrates. This study focused on the ability of B. bifidum and B. crudilactis to grow in a culture medium supplemented in 3′-sialyllactose (3′SL) as the main source of carbon, a major BMO encountered in cow milk. Next, the effects of cell-free spent media (CFSM) were tested against virulence expression of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium. Both strains were able to grow in presence of 3′SL, but B. crudilactis showed the best growth (7.92 ± 0.3 log cfu/ml) compared to B. bifidum (6.84 ± 0.9 log cfu/ml). Then, CFSM were tested for their effects on virulence gene expression by ler and hilA promoter activity of luminescent mutants of E. coli and S. Typhimurium, respectively, and on wild type strains of E. coli O157:H7 and S. Typhimurium using RT-qPCR. All CFSM resulted in significant under expression of the ler and hilA genes for the luminescent mutants and ler (ratios of −15.4 and −8.1 respectively) and qseA (ratios of −2.1 and −3.1) for the wild type strain of E. coli O157:H7. The 3′SL, a major 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 overall phenotype of pathogenic agents and the exact nature of the active molecules.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cell-Free Spent Media Obtained from Bifidobacterium bifidum and Bifidobacterium crudilactis Grown in Media Supplemented with 3′-Sialyllactose Modulate Virulence Gene Expression in Escherichia coli O157:H7 and Salmonella Typhimurium

Bondue¹,

Crèvecoeur²,

Brose³

et al. 2016

Front. Microbiol.

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“…Mare's milk differs widely from the milk of other dairy animals in terms of its components, such as milk fat and protein concentration. A previous study by Guri et al (2015) showed that mare's milk has a lower casein concentration, but it contains twice whey than that of cow's milk. The antimicrobial activity of mare's milk is associated with whey fraction (Guri et al 2015).…”

Section: Introductionmentioning

confidence: 95%

Effect of mare’s milk prebiotic supplementation on the gut microbiome and the immune system following antibiotic therapy

et al. 2020

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Abstract. Nurgaziyev M, Atenov Y, Khassenbekova Z, Akpanova S, Rysbekov K, Kozhakhmetov S, Nurgozhina A, Sergazy S, Chulenbayeva L, Ospanova Z, Tuyakova A, Mukhambetganov N, Sattybayeva R, Urazova S, Galymgozhina N, Zhumadilova A, Gulyaev A, Kushugulova A. 2020. Effect of mare’s milk prebiotic supplementation on the gut microbiome and the immune system following antibiotic therapy. Biodiversitas 21: 5065-5071. Antibiotic treatment can severely affect the gut microbiome for short-term and long-term consequences. Probiotic and prebiotic supplements are widely prescribed to modulate the composition and function of the human gut microbiome. The current study aims to determine the impacts of mare’s milk prebiotics on the diversity of gut bacterial communities and the local immune system when administered during and after a course of antibiotic therapy. Six children aged 4 to 5 years diagnosed with bilateral bronchopneumonia were prescribed cephalosporin (cefuroxime) antibiotics. During the 60 days of the study, three children consumed mare’s milk prebiotics, while the other three did not. Fecal samples were collected daily during antibiotic therapy and every five days after the last day of antibiotic treatment. Total DNA was isolated, and the taxonomic composition of the gut microbiome was analyzed by sequencing the 16S rRNA gene (V1-V3 region). The MULTIPLEX MAP platform was used to evaluate the local immune status. The relative abundance of 11 genera was reduced and did not recover until the last day of the study. The abundance of Bacteroides was not significantly altered in either group. Christensenella, Rothia, Abiotrophia, Acinetobacter, Anaerotruncus, Holdemania, and Turicibacter numbers were significantly increased on day five and remained at the same level during the study period. Cephalosporin administration also reduced the levels of pro-inflammatory and anti-inflammatory cytokines/chemokines (MIP1α, TNFα, GMCSF, GCSF, sCD40L, FGF2, TGFα, IL1α, and IP10).

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“…The RNA was then subjected to reverse transcription polymerase chainTo highlight the effects of filtrated supernatants on virulence gene expressions of E. coli O157:H7 ATCC 43890, the expression of ler, fliC, stx1, luxS, and qseA genes was determined using qPCR with gnd as a reference housekeeping gene[51][52]. For S. Typhimurium ATCC 14028, the virulence expressions of hilA, ssrB2 and sopD genes were assessed using gmk as a reference housekeeping gene[53]. Quantitative PCR amplification was performed using the GoTaq ® qPCR Master Mix (Promega, Leiden, Netherlands) and using the Light Cycler 480 (Roche Diagnostics, Mannheim, Germany).…”

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confidence: 99%

Bifidobacterium mongoliense genome is particularly adapted to milk oligosaccharides digestion leading to production of antivirulent metabolites

Bondue

Milani

Arnould

et al. 2019

Preprint

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Human milk oligosaccharides (HMO) could promote the growth of bifidobacteria, improving young children’s health. In addition, fermentation of carbohydrates by bifidobacteria can result in the production of metabolites presenting an antivirulent activity against intestinal pathogens. Bovine milk oligosaccharides (BMO), structurally similar to HMO, are found at high concentration in cow whey. This is particularly observed for 3’-sialyllactose (3’SL). This study focused on enzymes and transport systems involved in HMO/BMO metabolism contained in B. crudilactis and B. mongoliense genomes, two species from bovine milk origin. The ability of B. mongoliense to grow in media supplemented with whey or 3’SL was assessed. Next, the effects of cell-free spent media (CFSM) were tested against the virulence expression of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium. Due to the presence of genes encoding β-galactosidases, β-hexosaminidases, α-sialidases and α-fucosidases, B. mongoliense presents a genome more sophisticated and more adapted to the digestion of BMO/HMO than B. crudilactis (which contains only β-galactosidases). In addition, HMO/BMO digestion involves genes encoding oligosaccharide transport systems found in B. mongoliense but not in B. crudilactis. B. mongoliense seemed able to grow on media supplemented with whey or 3’SL as main source of carbon (8.3±1.0 and 6.7±0.3 log cfu/mL, respectively). CFSM obtained from whey resulted in a significant under-expression of ler, fliC, luxS, stx1 and qseA genes (-2.2, -5.3, -2.4, -2.5 and -4.8, respectively) of E. coli O157:H7. CFSM from 3’SL resulted in a significant up-regulation of luxS (2.0) gene and a down-regulation of fliC (-5.0) gene. CFSM obtained from whey resulted in significant up-regulations of sopD and hil genes (2.9 and 3.5, respectively) of S. Typhimurium, while CFSM obtained from 3’SL fermentation down-regulated hil and sopD genes (-2.7 and -4.2, respectively). From enzymes and transporters highlighted in the genome of B. mongoliense and its potential ability to metabolise 3’SL and whey, B. mongoliense seems well able to digest HMO/BMO. The exact nature of the metabolites contained in CFSM has to be identified still. These results suggest that BMO associated with B. mongoliense could be an interesting synbiotic formulation to maintain or restore intestinal health of young children.

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In vitroscreening of mare's milk antimicrobial effect and antiproliverative activity

Cited by 18 publications

References 42 publications

Cell-Free Spent Media Obtained from Bifidobacterium bifidum and Bifidobacterium crudilactis Grown in Media Supplemented with 3′-Sialyllactose Modulate Virulence Gene Expression in Escherichia coli O157:H7 and Salmonella Typhimurium

Cell-Free Spent Media Obtained from Bifidobacterium bifidum and Bifidobacterium crudilactis Grown in Media Supplemented with 3′-Sialyllactose Modulate Virulence Gene Expression in Escherichia coli O157:H7 and Salmonella Typhimurium

Effect of mare’s milk prebiotic supplementation on the gut microbiome and the immune system following antibiotic therapy

Bifidobacterium mongoliense genome is particularly adapted to milk oligosaccharides digestion leading to production of antivirulent metabolites

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