Analysis of the intestinal microbiota of oligosaccharide fed mice exhibiting reduced resistance to Salmonella infection

2010 ◽  
Vol 1 (3) ◽  
pp. 271-281 ◽  
Author(s):  
A. Petersen ◽  
A. Bergström ◽  
J. Andersen ◽  
M. Hansen ◽  
S. Lahtinen ◽  
...  
Keyword(s):  
Denaturing Gradient Gel Electrophoresis ◽  
Short Chain Fatty Acids ◽  
Gut Health ◽  
Salmonella Infection ◽  
Faecal Samples ◽  
Before And After ◽  
Gradient Gel Electrophoresis ◽  
Bacteroidetes Phylum ◽  
Bifidogenic Effect ◽  
Stimulation Of

Certain indigestible carbohydrates, known as prebiotics, are claimed to be beneficial for gut health through a selective stimulation of certain gut microbes including bifidobacteria. However, stimulation of such microbes does not necessarily imply a preventive effect against pathogen infection. We recently demonstrated a reduced resistance to Salmonella infection in mice fed diets containing fructo-oligosaccharides (FOS) or xylo-oligosaccharides (XOS). In the present study, faecal and caecal samples from the same mice were analysed in order to study microbial changes potentially explaining the observed effects on the pathogenesis of Salmonella. Denaturing gradient gel electrophoresis revealed that the microbiota in faecal samples from mice fed FOS or XOS were different from faecal samples collected before the feeding trial as well as from faecal profiles generated from control animals. This difference was not seen for caecal profiles. Further analysis of faecal samples by real-time PCR demonstrated a significant increase in the Bacteroidetes phylum, the Bacteroides fragilis group and in Bifidobacterium spp. in mice fed FOS or XOS. The observed bifidogenic effect was more pronounced for XOS than for FOS. The Firmicutes phylum and the Clostridium coccoides group were reduced by both FOS and XOS. Surprisingly, no significant differences were detected between faecal samples collected before and after pathogen challenge in any of the groups. Furthermore, no effect of diets on caecal concentrations of short-chain fatty acids was recorded. In conclusion, diets supplemented with FOS or XOS induced a number of microbial changes in the faecal microbiota of mice. The observed effects of XOS were qualitatively similar to those of FOS, but the most prominent bifidogenic effect was seen for XOS. An increased level of bifidobacteria is thus not in itself preventive against Salmonella infection, since the same XOS or FOS-fed mice were previously reported to be more severely affected by Salmonella than control animals.

scholarly journals Investigations on the Effects of Dietary Essential Oils and Different Husbandry Conditions on the Gut Ecology in Piglets after Weaning

2009 ◽  
Vol 2009 ◽  
pp. 1-9 ◽  
Author(s):  
P. Janczyk ◽  
R. Pieper ◽  
V. Urubschurov ◽  
K. R. Wendler ◽  
W. B. Souffrant
Keyword(s):  
Essential Oils ◽  
Dry Matter ◽  
Distal Part ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Control Diet ◽  
Short Chain Fatty Acids ◽  
Growth Promoters ◽  
Gradient Gel Electrophoresis ◽  
Antibiotic Growth Promoters ◽  
Polymerase Chain

Essential oils (EO) are being considered as possible alternatives to in-feed antibiotic growth promoters in pig nutrition. The effects of an EO mixture consisting of limonene, eugenol and pinene (10.0, 2.0, and 4.8 mg/kg diet, resp.) on gut physiology and ecology were studied in piglets. The experiment was conducted at low (commercial farm) and high hygienic conditions (experimental farm), to elucidate interactions between EO supplementation and husbandry methods. Piglets were weaned at 28 days of age, when they were offered either a control diet (C) or C with EO. Four piglets were sacrificed in each group on day 29, 30, 33 and 39. Digesta from the third distal part of the small intestine and from the colon were sampled and analysed for pH, dry matter, lactic acid, short chain fatty acids and ammonia concentrations. Enterobacteria, enterococci, lactobacilli and yeast counts were obtained by plating. Genomic DNA was extracted from digesta and polymerase chain reaction—denaturing gradient gel electrophoresis was performed. Individual microbial communities were identified at each farm. Age affected the intestinal parameters. No effects of the EO with exception for a significant reduction in colon bacterial diversity at 39 days of age could be recorded at experimental farm.

Safety and persistence of orally administered human Lactobacillus sp. strains in healthy adults

2011 ◽  
Vol 2 (1) ◽  
pp. 79-90 ◽  
Author(s):  
P. Hütt ◽  
P. Kõll ◽  
J. Stsepetova ◽  
B. Alvarez ◽  
R. Mändar ◽  
...  
Keyword(s):  
Denaturing Gradient Gel Electrophoresis ◽  
Blood Parameters ◽  
Second Phase ◽  
Biotechnological Applications ◽  
Gut Health ◽  
Blood Indices ◽  
Day Range ◽  
Gradient Gel Electrophoresis ◽  
Liver And Kidney ◽  
High Doses

The aim of the study was to evaluate the safety and persistence of selected Lactobacillus strains in the gastrointestinal tract (GIT) of healthy adult volunteers after oral consumption of high doses of lactobacilli to identify potential candidates for probiotic and biotechnological applications. In the first phase of the study, nine individuals consumed capsules containing Lactobacillus gasseri 177 and E16B7, Lactobacillus acidophilus 821-3, Lactobacillus paracasei 317 and Lactobacillus fermentum 338-1-1 (each daily dose 1×1010 cfu) for 5 consecutive days. Data on gut health, blood parameters, and liver and kidney function were collected. The persistence of Lactobacillus strains was assessed by culturing combined with arbitrarily primed polymerase chain reaction (AP-PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) on days 0, 5, 8, 10 and 20 from faecal samples. All strains survived gastrointestinal passage and were detected on the 5th day. L. acidophilus 821-3 was detected in four volunteers on the 8th day (4.3 to 7.0 log10 cfu/g) and in two on the 10th day (8.3 and 3.9 log10 cfu/g, respectively). In the second phase of the study, five additional volunteers consumed L. acidophilus 821-3 (daily 1×1010 cfu) for 5 consecutive days. The strain was subsequently detected in faeces of all individuals using real-time PCR on the 10th day (range 4.6-6.7; median 6.0 log10 cell/g) in both phases of the study for at least 5 days after discontinuation of consumption. The administration of high doses of different Lactobacillus strains did not result in any severe adverse effects in GIT and/or abnormal values of blood indices. Thus, the strain L. acidophilus 821-3 is a promising candidate for probiotic and biotechnological applications. Further studies will be performed to confirm the strain persistence and safety in a larger number of individuals.

scholarly journals Differences between human subjects in the composition of the faecal bacterial community and faecal metabolism of linoleic acid

Microbiology ◽  
2009 ◽  
Vol 155 (2) ◽  
pp. 513-520 ◽  
Author(s):  
Estelle Devillard ◽  
Freda M. McIntosh ◽  
Delphine Paillard ◽  
Nadine A. Thomas ◽  
Kevin J. Shingfield ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Vaccenic Acid ◽  
Human Subjects ◽  
Microbial Community Composition ◽  
Intestinal Bacteria ◽  
Short Chain Fatty Acids ◽  
Gut Health ◽  
Metabolic Characteristics ◽  
Faecal Samples ◽  
Cla Isomers

Conjugated linoleic acid (CLA) is formed from linoleic acid (LA; cis-9,cis-12-18 : 2) by intestinal bacteria. Different CLA isomers have different implications for human health. The aim of this study was to investigate LA metabolism and the CLA isomers formed in two individuals (V1 and V2) with different faecal metabolic characteristics, and to compare fatty acid metabolism with the microbial community composition. LA incubated with faecal samples was metabolized at similar rates with both subjects, but the products were different. LA was metabolized extensively to stearic acid (SA; 18 : 0) in V1, with minor accumulation of CLA and more rapid accumulation of vaccenic acid (VA; trans-11-18 : 1). CLA accumulation at 4 h was almost tenfold higher with V2, and little SA was formed. At least 12 different isomers of CLA were produced from LA by the colonic bacteria from the two individuals. The predominant (>75 %) CLA isomer in V1 was rumenic acid (RA; cis-9,trans-11-18 : 2), whereas the concentrations of RA and trans-10,cis-12-18 : 2 were similar with V2. Propionate and butyrate proportions in short-chain fatty acids were higher in V1. A 16S rRNA clone library from V1 contained mainly Bacteroidetes (54 % of clones), whereas Firmicutes (66 % of clones) predominated in V2. Both samples were devoid of bacteria related to Clostridium proteoclasticum, the only gut bacterium known to metabolize VA to SA. Thus, the CLA formed in the intestine of different individuals may differ according to their resident microbiota, with possibly important implications with respect to gut health.

scholarly journals Temperature and Denaturing Gradient Gel Electrophoresis Analysis of 16S rRNA from Human Faecal Samples

2000 ◽  
Vol 19 (2) ◽  
pp. 93-98 ◽  
Author(s):  
Antoon D.L. AKKERMANS ◽  
Erwin G. ZOETENDAL ◽  
Christine F. FAVIER ◽  
Hans G.H.J. HEILIG ◽  
Wilma M. AKKERMANS-VAN VLIET ◽  
...  
Keyword(s):  
16S Rrna ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Faecal Samples ◽  
Gradient Gel Electrophoresis

scholarly journals Impact of Grain Sorghum Polyphenols on Microbiota of Normal Weight and Overweight/Obese Subjects during In Vitro Fecal Fermentation

Nutrients ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 217 ◽  
Author(s):  
Danielle Ashley ◽  
Daya Marasini ◽  
Cindi Brownmiller ◽  
Jung Lee ◽  
Franck Carbonero ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Batch Fermentation ◽  
Short Chain Fatty Acids ◽  
Negative Control ◽  
Normal Weight ◽  
Weight Class ◽  
Gut Health ◽  
Sorghum Bran ◽  
Before And After

The human gut microbiota is considered as a crucial mediator between diet and gut homeostasis and body weight. The unique polyphenolic profile of sorghum bran may promote gastrointestinal health by modulating the microbiota. This study evaluated gut microbiota and modulation of short-chain fatty acids (SCFA) by sorghum bran polyphenols in in vitro batch fermentation derived from normal weight (NW, n = 11) and overweight/obese (OO, n = 11) subjects’ fecal samples. Six separate treatments were applied on each batch fermentation: negative control (NC), fructooligosaccharides (FOS), black sorghum bran extract (BSE), sumac sorghum bran extract (SSE), FOS + BSE, or FOS + SSE; and samples were collected before and after 24 h. No significant differences in total and individual SCFA production were observed between NW and OO subjects. Differential responses to treatment according to weight class were observed in both phyla and genera. Sorghum bran polyphenols worked with FOS to enhance Bifidobacterium and Lactobacillus, and independently stimulated Roseburia and Prevotella (p < 0.05). Our results indicate that sorghum bran polyphenols have differential effects on gut health and may positively impact gut ecology, with responses varying depending on weight class.

scholarly journals The Effects of Non-Nutritive Artificial Sweeteners, Aspartame and Sucralose, on the Gut Microbiome in Healthy Adults: Secondary Outcomes of a Randomized Double-Blinded Crossover Clinical Trial

Nutrients ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3408
Author(s):  
Samar Y. Ahmad ◽  
James Friel ◽  
Dylan Mackay
Keyword(s):  
Gut Microbiota ◽  
Day Treatment ◽  
Daily Intake ◽  
Short Chain Fatty Acids ◽  
Artificial Sweeteners ◽  
Microbiota Composition ◽  
Faecal Samples ◽  
Before And After ◽  
Double Blinded ◽  
Gut Microbiota Composition

Non-nutritive artificial sweeteners (NNSs) may have the ability to change the gut microbiota, which could potentially alter glucose metabolism. This study aimed to determine the effect of sucralose and aspartame consumption on gut microbiota composition using realistic doses of NNSs. Seventeen healthy participants between the ages of 18 and 45 years who had a body mass index (BMI) of 20–25 were selected. They undertook two 14-day treatment periods separated by a four-week washout period. The sweeteners consumed by each participant consisted of a standardized dose of 14% (0.425 g) of the acceptable daily intake (ADI) for aspartame and 20% (0.136 g) of the ADI for sucralose. Faecal samples collected before and after treatments were analysed for microbiome and short-chain fatty acids (SCFAs). There were no differences in the median relative proportions of the most abundant bacterial taxa (family and genus) before and after treatments with both NNSs. The microbiota community structure also did not show any obvious differences. There were no differences in faecal SCFAs following the consumption of the NNSs. These findings suggest that daily repeated consumption of pure aspartame or sucralose in doses reflective of typical high consumption have minimal effect on gut microbiota composition or SCFA production.

Changes in Fecal and Colonic Mucosal Microbiota of Patients with Refractory Constipation after a Subtotal Colectomy

2015 ◽  
Vol 81 (2) ◽  
pp. 198-206 ◽  
Author(s):  
Xiaobo Feng ◽  
Yong Su ◽  
Jun Jiang ◽  
Ning Li ◽  
Weiwei Ding ◽  
...  
Keyword(s):  
Subtotal Colectomy ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Intestinal Flora ◽  
Diversity Indices ◽  
Control Subjects ◽  
Before And After ◽  
Gradient Gel Electrophoresis ◽  
Healthy Control ◽  
Polymerase Chain ◽  
Bacterial Groups

The purpose of this study was to investigate the changes in gut microbiota of patients with refractory constipation 6 months after a subtotal colectomy. Feces and mucosal samples of five healthy volunteers and 17 patients with refractory constipation before and six months after subtotal colectomy were collected. Denaturing gradient gel electrophoresis (DGGE) and polymerase chain reaction techniques were used for quantitative analysis of main bacterial groups and archeal methanogens. No significant differences were found in the DGGE profiles among the three groups. After subtotal colectomy, a significantly decreased similarity coefficient was observed in the fecal. The Shannon diversity indices had no significant differences among the three groups. The numbers of predominant bacteria (Bacteriodetes, Clostridium coccoides group, and Clostridium leptum group) did not significantly change in patients before and after surgery compared with healthy control subjects, and the number of total bacteria, Firmicutes, Bacteriodetes, and Clostridium leptum group bacteria in the feces decreased after surgery. However, the numbers of these bacteria remained the same in mucosa from postoperative patients. The numbers of Bifidobacteria and Lactobacilli in feces and the number of Bifidobacteria in mucosa were significantly lower in preoperative compared with healthy control subjects and increased after the surgery. The number of methanogens in the mucosa was decreased in preoperative patients but returned to normal levels postoperatively. In conclusion, although there was no difference in the structure of the predominant bacteria between refractory constipated patients and healthy control subjects, the number of probiotics ( Bifidobacteria and Lactobacilli) was significantly lower in refractory constipated patients. However, subtotal colectomy can significantly normalize the number of intestinal flora.

scholarly journals Dietary resistant and butyrylated starches have different effects on the faecal bacterial flora of azoxymethane-treated rats

2011 ◽  
Vol 105 (10) ◽  
pp. 1480-1485 ◽  
Author(s):  
Guy C. J. Abell ◽  
Claus T. Christophersen ◽  
Alexandra L. McOrist ◽  
Julie M. Clarke
Keyword(s):  
Denaturing Gradient Gel Electrophoresis ◽  
Bacterial Flora ◽  
Epidemiological Studies ◽  
Rrna Gene ◽  
Maize Starch ◽  
Faecal Microbiota ◽  
Faecal Samples ◽  
Gradient Gel Electrophoresis ◽  
High Amylose ◽  
Bowel Health

Epidemiological studies have suggested that dietary fibre lowers the risk of colorectal cancer, which may be due to increased butyrate production from colonic fermentation of a type of fibre, resistant starch (RS). The present study investigated the effects of dietary RS and butyrylated RS on the faecal microbiota of rats treated with azoxymethane. A total of four groups of nine rats were fed diets containing either standard maize starch (low-amylose maize starch (LAMS), low RS), LAMS with 3 % tributyrin (LAMST), cooked 10 % high-amylose maize starch (HAMS, high RS) or cooked 10 % butyrylated HAMS (HAMSB). Faecal samples were examined by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments. Multivariate analysis demonstrated no differences between faecal microbiota before treatment but revealed differences in DGGE patterns between diet groups, with the exception of the two low-RS groups (LAMS and LAMST). Subsequent analysis identified eleven DGGE bands contributing significantly to the differentiation between diets. These phylotypes belonged to Clostridiales (five),Lactobacillus(one) and Bacteroidetes (five) lineages. Rats fed HAMS had increased concentration of propionate in their distal colonic digesta and developed faecal populations containingRuminococcus bromii-like bacteria. HAMSB increased propionate and butyrate concentrations in distal colonic digesta and was associated with the appearance of two non-butyrate-producing bacteria,Lactobacillus gasseriandParabacteroides distasonis.In conclusion, supplementation with specific dietary RS leads to changes in faecal microbiota profiles that may be associated with improved bowel health.

Structure of bacterial communities in diverse freshwater habitats

2012 ◽  
Vol 58 (3) ◽  
pp. 326-335 ◽  
Author(s):  
Yana Aizenberg-Gershtein ◽  
Dalit Vaizel-Ohayon ◽  
Malka Halpern
Keyword(s):  
Drinking Water ◽  
Water Samples ◽  
Bacterial Communities ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Rrna Gene ◽  
Source Water ◽  
Before And After ◽  
Different Seasons ◽  
Gradient Gel Electrophoresis

The structures and dynamics of bacterial communities from raw source water, groundwater, and drinking water before and after filtration were studied in four seasons of a year, with culture-independent methods. Genomic DNA from water samples was analyzed by the polymerase chain reaction – denaturing gradient gel electrophoresis system and by cloning of the 16S rRNA gene. Water samples exhibited complex denaturing gradient gel electrophoresis genetic profiles composed of many bands, corresponding to a great variety of bacterial taxa. The bacterial communities of different seasons from the four sampling sites clustered into two major groups: (i) water before and after filtration, and (ii) source water and groundwater. Phylogenetic analyses of the clones from the autumn sampling revealed 13 phyla, 19 classes, and 155 operational taxonomic units. Of the clones, 66% showed less than 97% similarities to known bacterial species. Representatives of the phyla Proteobacteria, Bacteroidetes, and Actinobacteria were found at all four sampling sites. Species belonging to the phylum Firmicutes were an important component of the microbial community in filtered water. Representatives of Enterobacteriaceae were not detected, indicating the absence of fecal pollution in the drinking water. Differences were found in the bacterial populations that were sampled from the same sites in different seasons. Each water habitat had a unique bacterial profile. Drinking water harbors diverse and dynamic microbial communities, part of which may be active and resilient to chlorine disinfection. This study provides, for the first time, basic data for uncultivable drinking water bacteria in Israel.

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