Influence of Camembert consumption on the composition and metabolism of intestinal microbiota: a study in human microbiota-associated rats

The objective of the present study was to evaluate the consequence of Camembert consumption on the composition and metabolism of human intestinal microbiota. Camembert cheese was compared with milk fermented by yoghurt starters andLactobacillus caseias a probiotic reference. The experimental model was the human microbiota-associated (HM) rat. HM rats were fed a basal diet (HMB group), a diet containing Camembert made from pasteurised milk (HMCp group) or a diet containing fermented milk (HMfm group). The level of micro-organisms from dairy products was measured in faeces using cultures on a specific medium and PCR–temporal temperature gradient gel electrophoresis. The metabolic characteristics of the caecal microbiota were also studied: SCFA, NH3, glycosidase and reductase activities, and bile acid degradations. The results showed that micro-organisms from cheese comprised 105–108bacteria/g faecal sample in the HMCp group.Lactobacillusspecies from fermented milk were detected in HMfm rats. Consumption of cheese and fermented milk led to similar changes in bacterial metabolism: a decrease in azoreductase activity and NH3concentration and an increase in mucolytic activities. However, specific changes were observed: in HMCp rats, the proportion of ursodeoxycholic resulting from chenodeoxycholic epimerisation was higher; in HMfm rats, α and β-galactosidases were higher than in other groups and both azoreductases and nitrate reductases were lower. The results show that, as for fermented milk, Camembert consumption did not greatly modify the microbiota profile or its major metabolic activities. Ingested micro-organisms were able to survive in part during intestinal transit. These dairy products exert a potentially beneficial influence on intestinal metabolism.

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Identification of the Bacterial Microflora in Dairy Products by Temporal Temperature Gradient Gel Electrophoresis

Applied and Environmental Microbiology ◽

10.1128/aem.68.8.3691-3701.2002 ◽

2002 ◽

Vol 68 (8) ◽

pp. 3691-3701 ◽

Cited By ~ 176

Author(s):

Jean-Claude Ogier ◽

Olivier Son ◽

Alexandra Gruss ◽

Patrick Tailliez ◽

Agnes Delacroix-Buchet

Keyword(s):

Temperature Gradient ◽

Gel Electrophoresis ◽

Dairy Products ◽

Bacterial Species ◽

Fermented Milk ◽

Multiple Control ◽

Gradient Gel Electrophoresis ◽

Temperature Gradient Gel Electrophoresis ◽

Yeasts And Molds

ABSTRACT Numerous microorganisms, including bacteria, yeasts, and molds, are present in cheeses, forming a complex ecosystem. Among these organisms, bacteria are responsible for most of the physicochemical and aromatic transformations that are intrinsic to the cheesemaking process. Identification of the bacteria that constitute the cheese ecosystem is essential for understanding their individual contributions to cheese production. We used temporal temperature gradient gel electrophoresis (TTGE) to identify different bacterial species present in several dairy products, including members of the genera Lactobacillus, Lactococcus, Leuconostoc, Enterococcus, Pediococcus, Streptococcus, and Staphylococcus. The TTGE technique is based on electrophoretic separation of 16S ribosomal DNA (rDNA) fragments by using a temperature gradient. It was optimized to reveal differences in the 16S rDNA V3 regions of bacteria with low-G+C-content genomes. Using multiple control strains, we first set up a species database in which each species (or group of species) was characterized by a specific TTGE fingerprint. TTGE was then applied to controlled dairy ecosystems with defined compositions, including liquid (starter), semisolid (home-made fermented milk), and solid (miniature cheese models) matrices. Finally, the potential of TTGE to describe the bacterial microflora of unknown ecosystems was tested with various commercial dairy products. Subspecies, species, or groups of species of lactic acid bacteria were distinguished in dairy samples. In conclusion, TTGE was shown to distinguish bacterial species in vitro, as well as in both liquid and solid dairy products.

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Composition and metabolism of the intestinal microbiota in consumers and non-consumers of yogurt

British Journal Of Nutrition ◽

10.1017/s0007114507243065 ◽

2007 ◽

Vol 97 (1) ◽

pp. 126-133 ◽

Cited By ~ 44

Author(s):

Elise Alvaro ◽

Claude Andrieux ◽

Violaine Rochet ◽

Lionel Rigottier-Gois ◽

Pascale Lepercq ◽

...

Keyword(s):

Intestinal Microbiota ◽

Pcr Amplification ◽

Fermented Milk ◽

Lactobacillus Delbrueckii ◽

Galactosidase Activity ◽

Metabolic Characteristics ◽

Daily Food ◽

Regular Consumption ◽

Neutral Sterols ◽

The Impact

The objective of the present study was to evaluate the impact of a regular consumption of yogurt on the composition and metabolism of the human intestinal microbiota. Adult subjects were selected on the basis of daily food records and divided into two groups: yogurt consumers (at least 200 g yogurt consumed per d,n30); non-consumers (no yogurt,n21). Their faecal microbiota was analysed using molecular methods (in situhybridisation and PCR amplification combined with separation by denaturing gel electrophoresis) and its metabolic characteristics were assessed by measuring glycosidase, β-glucuronidase and reductase activities and profiling SCFA, neutral sterols and bile acids. The yogurt starterLactobacillus delbrueckiissp.bulgaricus(identity confirmed by 16S rRNA sequencing) was detected in 73 % of faecal samples from fermented milk consumersv. 28 % from non-consumers (P = 0·003). In yogurt consumers, the level of Enterobacteriaceae was significantly lower (P = 0·006) and β-galactosidase activity was significantly increased (P = 0·048). In addition, within this group, β-galactosidase activity and theBifidobacteriumpopulation were both positively correlated with the amount of fermented milk ingested (r0·66,P < 0·0001 andr0·43,P = 0·018, respectively). Apart from these effects, which can be considered beneficial to the host, no other major differences could be detected regarding the composition and metabolic activity of intestinal microbiota.

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Molecular Methods to Measure Intestinal Bacteria: A Review

Journal of AOAC International ◽

10.5740/jaoacint.sge_inglis ◽

2012 ◽

Vol 95 (1) ◽

pp. 5-23 ◽

Cited By ~ 34

Author(s):

G Douglas Inglis Agriculture ◽

Matthew C Thomas ◽

Dallas K Thomas ◽

Martin L Kalmokoff ◽

Stephen P J Brooks ◽

...

Keyword(s):

Intestinal Microbiota ◽

Gel Electrophoresis ◽

Denaturing Gradient Gel Electrophoresis ◽

Direct Sequencing ◽

Intestinal Bacteria ◽

Polymorphism Analysis ◽

Sample Collection ◽

Pcr Bias ◽

Gradient Gel Electrophoresis ◽

Temperature Gradient Gel Electrophoresis

Abstract The intestine is an exceptionally rich ecosystem encompassing a complex interaction among microorganisms, influenced by host factors, ingested food, and liquid. Characterizing the intestinal microbiota is currently an active area of research. Various molecular-based methods are available to characterize the intestinal microbiota, but all methods possess relative strengths, as well as salient weaknesses. It is important that researchers are cognizant of the limitations of these methods, and that they take the appropriate steps to mitigate weaknesses. Here, we discuss methodologies used to monitor intestinal bacteria including: (i) traditional clone libraries; (ii) direct sequencing using next-generation parallel sequencing technology; (iii) denaturing gradient gel electrophoresis and temperature gradient gel electrophoresis; (iv) terminal restriction fragment length polymorphism analysis; (v) fluorescent in situ hybridization; and (vi) quantitative PCR. In addition, we also discuss experimental design, sample collection and storage, DNA extraction, gene targets, PCR bias, and methods to reduce PCR bias.

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Effects of orally administeredLactobacillus caseiDN-114 001 on the composition or activities of the dominant faecal microbiota in healthy humans

British Journal Of Nutrition ◽

10.1079/bjn20051625 ◽

2006 ◽

Vol 95 (2) ◽

pp. 421-429 ◽

Cited By ~ 38

Author(s):

Violaine Rochet ◽

Lionel Rigottier-Gois ◽

Maléne Sutren ◽

Marie-Noëlle Krementscki ◽

Claude Andrieux ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Fold Increase ◽

Fermented Milk ◽

Milk Product ◽

Faecal Microbiota ◽

Specific Primers ◽

Healthy Humans ◽

Gradient Gel Electrophoresis ◽

Temperature Gradient Gel Electrophoresis

The composition and activities of the faecal microbiota in twelve healthy subjects analysed in a single open study were monitored before (1-week baseline step), during (10d supplementation step) and after (10d follow-up step) the ingestion of a fermented milk containingLactobacillus caseiDN-114001. Fluorescentin situhybridisation with group-specific DNA probes, real-time PCR usingL. paracaseigroup-specific primers and temporal temperature gradient gel electrophoresis (TTGE) using group-specific primers were carried out, together with bacterial enzyme activity and metabolite analyses to monitor the structure and activities of the faecal microbiota.L. caseiDNA was detected in the faeces of all of the subjects by TTGE after 10d supplementation. Its quantification by real-time PCR showed a 1000-fold increase during the test step compared with initial levels. No major modification in either the dominant members of the faecal microbiota or their activities was observed during the trial. In conclusion, the short-term consumption of a milk product containingL. caseiDN-114001 was accompanied by a high, transient increase in the quantity of this strain in the faeces of all of the subjects without markedly affecting biochemical or bacteriological factors.

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Brussels sprouts, inulin and fermented milk alter the faecal microbiota of human microbiota-associated rats as shown by PCR-temporal temperature gradient gel electrophoresis using universal,Lactobacillus and Bifidobacterium16S rRNA gene primers

British Journal Of Nutrition ◽

10.1079/bjn20051372 ◽

2005 ◽

Vol 93 (5) ◽

pp. 677-684 ◽

Cited By ~ 20

Author(s):

Christèle Humblot ◽

Aurélia Bruneau ◽

Malène Sutren ◽

Evelyne F. Lhoste ◽

Joël Doré ◽

...

Keyword(s):

Control Diet ◽

Fermented Milk ◽

Initial Time ◽

Rrna Gene ◽

Final Time ◽

Brussels Sprouts ◽

Gradient Gel Electrophoresis ◽

The Mean ◽

Temperature Gradient Gel Electrophoresis ◽

Degree Of Similarity

We investigated the effect of Brussels sprouts, inulin and a fermented milk on the faecal microbiota diversity of human microbiota-associated (HMA) rats by PCR-temporal temperature gradient gel electrophoresis (PCR-TTGE) using universal and group-specific 16S rRNA gene primers. The HMA rats were submitted to a control diet for 10 d (initial time), then switched to the experimental diets for 4 weeks (final time). Using universal primers, the mean degree of similarity between all faecal samples at initial time was 80·8 %. In the group consuming the control diet throughout the experiment, the mean degree of similarity between the PCR-TTGE profiles at initialv.final time was 76·8 %, reflecting a spontaneous temporal variation. The mean degree of similarity between control and experimental groups at final time was lower, 72·4 %, 74·4 % and 75·6 % for inulin, Brussels sprouts and fermented milk, respectively, indicating a dietary effect on the predominant populations. Using specific primers, bifidobacteria could be detected only in those rats that had consumed inulin, showing a specific increasing effect of this dietary compound. TheLactobacilluspopulation was very heterogeneous at initial time but tended to homogenize within each dietary group. At final time, caecal contents were collected for analysis of SCFA and β-glucuronidase activity. Inulin and Brussels sprouts increased the butyrate and acetate proportion, respectively, while the fermented milk did not modify the caecal biochemistry. This experiment shows for the first time that cruciferous vegetables are able to alter the diversity and the metabolic activities of the digestive microbiota in HMA rats.

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Modulation of Gut Microbiota and Oxidative Status by β-Carotene in Late Pregnant Sows

Frontiers in Nutrition ◽

10.3389/fnut.2020.612875 ◽

2020 ◽

Vol 7 ◽

Author(s):

Xupeng Yuan ◽

Jiahao Yan ◽

Ruizhi Hu ◽

Yanli Li ◽

Ying Wang ◽

...

Keyword(s):

Gut Microbiota ◽

Dietary Supplementation ◽

Basal Diet ◽

Fecal Microbiota ◽

Control Group ◽

Metabolic Activities ◽

The Impact ◽

Positive Effect ◽

Β Carotene

Recent evidences suggest that gut microbiota plays an important role in regulating physiological and metabolic activities of pregnant sows, and β-carotene has a potentially positive effect on reproduction, but the impact of β-carotene on gut microbiota in pregnant sows remains unknown. This study aimed to explore the effect and mechanisms of β-carotene on the reproductive performance of sows from the aspect of gut microbiota. A total of 48 hybrid pregnant sows (Landrace × Yorkshire) with similar parity were randomly allocated into three groups (n = 16) and fed with a basal diet or a diet containing 30 or 90 mg/kg of β-carotene from day 90 of gestation until parturition. Dietary supplementation of 30 or 90 mg/kg β-carotene increased the number of live birth to 11.82 ± 1.54 and 12.29 ± 2.09, respectively, while the control group was 11.00 ± 1.41 (P = 0.201). Moreover, β-carotene increased significantly the serum nitric oxide (NO) level and glutathione peroxidase (GSH-Px) activity (P < 0.05). Characterization of fecal microbiota revealed that 90 mg/kg β-carotene increased the diversity of the gut flora (P < 0.05). In particular, β-carotene decreased the relative abundance of Firmicutes including Lachnospiraceae AC2044 group, Lachnospiraceae NK4B4 group and Ruminococcaceae UCG-008, but enriched Proteobacteria including Bilophila and Sutterella, and Actinobacteria including Corynebacterium and Corynebacterium 1 which are related to NO synthesis. These data demonstrated that dietary supplementation of β-carotene may increase antioxidant enzyme activity and NO, an important vasodilator to promote the neonatal blood circulation, through regulating gut microbiota in sows.

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