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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 detailConsumption patterns, bacteriological quality and risk factors for Salmonella contamination in meat-based meals consumed outside the home in Kigali, Rwanda
Niyonzima, Eugène; Ongol, Martin Patrick; Brostaux, Yves ULiege et al

in Food Control (2016)

Meat-based meals are consumed as a source of animal proteins and constitute one of the leading vehicles for food borne infections in humans. The main objective of this study was to determine the ... [more ▼]

Meat-based meals are consumed as a source of animal proteins and constitute one of the leading vehicles for food borne infections in humans. The main objective of this study was to determine the consumption pattern and the bacteriological quality of meat-based meals consumed outside households in Kigali. A survey on meat consumption patterns was carried out in 400 households by using a questionnaire, whereas different meat-based meals were sampled from 150 snack bars and restaurants. Enumeration of hygiene indicator bacteria (total mesophilic bacteria and Escherichia coli) and the qualitative detection of Salmonella were carried out by using conventional culture methods. The results indicated that goat was the type of meat that was consumed the most outside the home in Kigali and the meat intake varied significantly (p ≤0.05) with the social category of the household. The average levels of total aerobic bacteria and E. coli in meat-based meals were found to be 4.7 and 1.4 log cfu/g, respectively, whereas Salmonella was detected in 11.7% of all meat-based meals. Eight factors mostly linked to the cooking treatments and hygienic handling practices for cooked meals were found to be significantly (p ≤ 0.05) associated with the risk of Salmonella occurrence in meat-based meals consumed outside the home in Kigali. The findings from this study strongly suggest the need for proper cooking and/or improvements in hygiene in the establishments selling ready-to-eat meat-based meals in Kigali, particularly those located in rural localities. [less ▲]

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See detailComparative analysis of the respiratory microbiota of healthy dogs and dogs with canine idiopathic pulmonary fibrosis
Roels, Elodie ULiege; Taminiau, Bernard ULiege; Darnis, Elodie ULiege et al

in Journal of Veterinary Internal Medicine (2016, September)

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See detailStudy of the microbial diversity of microbial ring trials by metagenomic analysis : Quantification of alive bacteria by exclusion of dead bacteria
Fall, P.A.; Burteau, S.; Detry, E. et al

Poster (2016, July 19)

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See detailMetagenetics and predictive microbiology: a new tool to understand the kinetics of microbial subpopulations in Belgian white pudding
Cauchie, Emilie ULiege; Gand, Mathieu; Kergourlay, Gilles et al

Poster (2016, July 18)

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See detailChallenge testing with Brochothrix thermosphacta on minced pork meat shows interest to couple metagenetics to metabolomics to study food spoilage
Baré, Ghislain ULiege; Cauchie, Emilie ULiege; Leenders, Justine ULiege et al

Poster (2016, July)

The spoilage of perishable foods is mainly caused by bacterial activity. The risk of unwanted bacterial growth is particularly high in the minced pork meat. In this work, the natural microbial ... [more ▼]

The spoilage of perishable foods is mainly caused by bacterial activity. The risk of unwanted bacterial growth is particularly high in the minced pork meat. In this work, the natural microbial contaminants of the minced pork meat were followed by 16S ribosomal DNA deep sequencing (metagenetics) during aging tests at different temperatures. Brochothrix thermosphacta MM008 strain was selected as one of the main contaminants responsible for the spoilage of the meat. Minced pork meat previously sterilized by gamma irradiation was inoculated with B. thermosphacta MM008 for challenge tests measuring growth and then incubated at different temperatures. Minced meat samples were taken and analyzed by H-NMR 1D at time 0 and at final time (from 14 to 19 days, depending on the incubation temperature). Orthogonal partial least square discriminant analysis (OPLS-DA) showed that samples, regardless of the incubation temperature, could be splitted into 3 groups according to their spectral profile: 1) samples taken at time 0, 2) samples inoculated with B. thermosphacta and taken at final time, 3) samples uninoculated, taken at final time. From the analysis of the metabolomics data, higher concentrations of glycerol, glucose, taurine, lactate, carnitine, betaine and glycine were identified in the samples of uninoculated minced pork meat and an increased production of creatine, acetate and acetone was found in the samples of minced pork meat inoculated with B. thermosphacta MM008. These observations showed that -omics technologies (metagenetics and metabolomics) could be used conclusively to study microbial spoilage of minced pork meat. [less ▲]

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See detailEffects of protein source and cooking procedure on intestinal microbiota and on fermentation end-products in rats
POELAERT, Christine ULiege; Despret, Xavier; Portetelle, Daniel ULiege et al

Poster (2016, June)

Animal and plant proteins are major proteins sources in the human diet. After their enzymatic degradation in the upper gastro-intestinal tract, the undigested fraction of these proteins is available for ... [more ▼]

Animal and plant proteins are major proteins sources in the human diet. After their enzymatic degradation in the upper gastro-intestinal tract, the undigested fraction of these proteins is available for fermentation by the microbiota of the large intestine leading to the production of short-chain fatty acids (SCFA), branched-chain fatty acids (BCFA), ammonia, biogenic amines, sulphur metabolites, phenols and indoles. As some of these compounds have genotoxic and cytotoxic effects, protein fermentation is considered as detrimental to the host’s epithelial health. BCFA are usually used as a marker of intestinal protein fermentation. We studied in vivo the impact of proteins from animal and plant origin, raw or after a cooking procedure, on the composition of gut microbiota and on fermentation end-products. Weanling rats were used as models of the human gut microbiota. Eight experimental diets were formulated with beef meat (Longissimus dorsi), chicken meat (Pectoralis major), white pea beans (Phaseolus vulgaris), soybeans (Glycine max), used raw and cooked, as sole source of protein in the diet. One casein diet was used as control. All diets, formulated to contain 15% of raw protein, were given to seven rats for four weeks. After euthanasia, caecal contents were collected. Pyrosequencing analyses (Roche 454 GS Junior Genome Sequencer) were performed to study the microbial composition. SCFA and BCFA were measured using HPLC (Waters 2690). Microbial composition in the caecum is associated to the type of dietary protein and to the cooking procedure applied. The proportion of BCFA in the caecal content is mainly affected by the type of protein. So BCFA represent respectively 04-06% and 35-44% of total SCFA with diets based on plant and on animal proteins. In conclusion, both the type of protein and the cooking procedure could impact the gut microbiota in terms of composition and of fermentative capacity. [less ▲]

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See detailLaboratory identification of anaerobic bacteria isolated on Clostridium difficile selective medium
Rodriguez Diaz, Cristina ULiege; Warszawski, Nathalie; Korsak Koulagenko, Nicolas ULiege et al

in Acta Microbiologica et Immunologica Hungarica (2016)

Despite increasing interest in the bacterium, the methodology for Clostridium difficile recovery has not yet been standardised. Cycloserine cefoxitin fructose taurocholate (CCFT) has historically been the ... [more ▼]

Despite increasing interest in the bacterium, the methodology for Clostridium difficile recovery has not yet been standardised. Cycloserine cefoxitin fructose taurocholate (CCFT) has historically been the most used medium for C. difficile isolation from human, animal, environmental and food samples, and presumptive identification is usually based on colony morphologies. However, CCFT is not totally selective. This study describes the recovery of 24 bacteria species belonging to 10 different genera other than C. difficile, present in the environment and foods of a retirement establishment that were not inhibited in the C. difficile selective medium. These findings provide insight for further environmental and food studies as well as for isolation of C. difficile on supplemented CCFT. [less ▲]

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See detailAssociation of Targeted metagenomic analysis and classical microbiology for Clostridium difficile detection and microbial ecosystem mapping of surfaces hands and foodstuffs in a meat processing plant
Rodriguez Diaz, Cristina ULiege; Oukbir, Meriem; Taminiau, Bernard ULiege et al

Poster (2016, May 11)

INTRODUCTION Zoonoses are infectious that can be transmitted between animals and humans through direct contact, close proximity or the environment. Since domestic and food animals frequently test positive ... [more ▼]

INTRODUCTION Zoonoses are infectious that can be transmitted between animals and humans through direct contact, close proximity or the environment. Since domestic and food animals frequently test positive for the bacterium, it seems plausible that C. difficile could be zoonotic. PURPOSE This study aimed to determine the prevalence and the epidemiology of C. difficile in cattle farms and the possible spread of the bacterium among animals and farmers. METHODS A total of 176 faecal samples of cattle were collected from 5 different Belgian farms. A stool sample of each farmer was also requested. Detection of C. difficile was performed by classical culture on C. difficile selective medium (cycloserine cefoxitin fructose cholate). Isolates were characterised by PCR-ribotyping and Genotype Cdiff test (Hain Lifescience), which allows the detection of all toxin genes, mutations in gyrA gene and the deletion in the regulator gene tcdC. RESULTS C. difficile was detected in 14/178 (7.9%) animal samples. Isolates were grouped into five different types, including PCR-ribotype 015. All of them were identified as toxigenic. In contrast, none of the 5 farmers studied were positive for the bacterium. SIGNIFICANCE Results obtained indicate that PCR-ribotypes commonly isolated from hospitalised patients are also present in cattle, indicating an animal reservoir. However, a zoonotic transmission could be not demonstrated in the farms studied. [less ▲]

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See detailDaily intake and bacteriological quality of meat consumed in the households of Kigali, Rwanda
Niyonzima, Eugene ULiege; Ongol, Martin Patrick; Brostaux, Yves ULiege et al

in Food Control (2016), 69

Meat is consumed worldwide as a source of animal proteins, but it is recognized as one of the most important vehicles for food borne infections in humans. This study was conducted to determine the daily ... [more ▼]

Meat is consumed worldwide as a source of animal proteins, but it is recognized as one of the most important vehicles for food borne infections in humans. This study was conducted to determine the daily intake; the levels of hygiene indicator bacteria, namely the total mesophilic bacteria (TMC) and Escherichia coli counts (ECC); and the prevalence of Salmonella in meat consumed within the households of Kigali (Rwanda). The survey on meat consumption was carried out in 400 households by using a questionnaire, whereas the bacteriological analyses of meat samples were performed by using conventional culture methods. The results from the survey indicated that beef was the type of meat mostly consumed in Kigali city households, and the daily meat intake significantly varied with the social category of the household. No significant difference was observed between daily meat intakes in different age classes of household members. In the samples where microorganisms were detected, the average levels of TMCs and ECCs in raw meat were found to be 5.4 and 1.6 log cfu/g, respectively, whereas in cooked meat they were significantly reduced to 3.1 and 1.1 log cfu/g, respectively. The prevalence of Salmonella was reduced from 21.4% in raw meat to 3.4% in ready-to-eat cooked meat. Salmonella was not detected in cooked meat consumed in high-income households. The results from this study highlight the need for hygiene improvements in meat shops as well as in the households of Kigali, particularly those with low and medium incomes. [less ▲]

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See detailOral administration of a Spirulina extract protects old mice against hepatic “ammaging”
Neyrinck, A.; Taminiau, Bernard ULiege; Daube, Georges ULiege et al

in Nutrition Clinique et Metabolisme (2016), 30(1), 64

Background & aims: Ageing predisposes to hepatic dysfunction and inflammation that can evolve to non-alcoholic fatty liver disease. Spirulina, a cyanobacterium used as a food additive or food supplement ... [more ▼]

Background & aims: Ageing predisposes to hepatic dysfunction and inflammation that can evolve to non-alcoholic fatty liver disease. Spirulina, a cyanobacterium used as a food additive or food supplement, has been shown to impact immune functions and to improve non-alcoholic steatohepatitis in obese mice. The aim of the present study was to test the potential hepatoprotective effects of Spirulina extract supplementation in aged mice and to determine whether these effects can be related to a modulation of the gut microbiota. Methods: Old mice of 24 months were fed a control diet supplemented with or without 5% Spirulina extract (Biores, Liège, Belgium) and were compared to young mice of 3 months during 6 weeks. Results: Combination of pyrosequencing and qPCR analyses of the 16S rRNA gene revealed a decrease in total bacteria and -among specific changes of gut microbiota composition,- an increase in Allobaculum, Blautia, Roseburia, and Lactobacillus populations by Spirulina treatment. Interestingly, parameters related to the innate immunity, especially T Regulatory cells (FoxP3), CD11b-dendritic cells, cytokines (IL6, IFN MCP-1) and antimicrobial peptides (Pla2g2, Reg3) were upregulated in the small intestine of Spirulina-treated mice. Aged mice exhibited inflammation and oxidative stress in the liver tissue as compared to young mice. The supplementation with Spirulina extract reduced several hepatic inflammatory and oxidative stress markers in old mice. Conclusions: Our study shows for the first time that the oral administration of a Spirulina is able to modulate the gut microbiota, to activate immune system in the gut, thereby improving hepatic inflammation in aged mice. Those data allow to envisagesuggest a new therapeutic tools in the management of immune and metabolic alterations in ageing, based on gut microbes-host interactions. [less ▲]

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See detailOral administration of a spirulina extract protects old mice against hepatic "inflammaging"
Neyrinck, A.; Taminiau, Bernard ULiege; Daube, Georges ULiege et al

Poster (2016, February 18)

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See detailNo favorable effect of reduced tillage on microbial community diversity in a silty loam soil (Belgium)
Degrune, Florine ULiege; Theodorakopoulos, Nicolas ULiege; Dufrêne, Marc ULiege et al

in Agriculture, Ecosystems & Environment (2016), 224

Among the soil management practices used to promote sustainable agriculture, reduced tillage and retention of residues from the previous crop are reported to enhance significantly both soil fertility and ... [more ▼]

Among the soil management practices used to promote sustainable agriculture, reduced tillage and retention of residues from the previous crop are reported to enhance significantly both soil fertility and crop productivity. Here, high-throughput sequencing (454 technology) was used to see how the tillage regime (conventional vs. reduced tillage) and the fate of crop residues (retention or removal) affect microbial communities at two sampling depths (top soil: 0–5 cm and deeper soil: 15–20 cm) in a fertile silty loam soil in Belgium. All combinations of these three factors were studied. After 6 years of conversion from conventional to reduced tillage, depth emerged as the main factor responsible for variation in microbial diversity, tillage regime ranked second, and finally, crop residue fate had no influence on microbial diversity. For both bacteria and fungi, the diversity appeared higher in the top soil than in the deeper soil, and surprisingly, higher under conventional than under reduced tillage. These differences are explained by changes in community composition due to taxon loss rather than taxon replacement. The specific local set of environmental conditions (a loess-derived soil and an oceanic temperate climate) may explain these results. These observations raise the question: does impoverishment in indicator taxa influence soil processes, and thus crop production? To answer this question, we discuss how the presence of certain indicator taxa liable to play an ecological role might relate to crop productivity. [less ▲]

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