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Effect of dietary grape seed extract on chicken intestinal ecosystem studied by RFLP.

INTRODUCTION

Previous studies have demonstrated that grape phenolic compounds have antimicrobial properties. Because a large fraction of the intestinal community is difficult or not posible to culture, molecular approaches such as Restriction Fragment Length Polymorphism (RFLP), has been shown to be an useful tool for the study of diet-associated changes in gut microbial communities.

An experiment was conducted to examine changes on chicken gut microbial ecosystem in response to the addition of a grape seed extract (GSE) in the diet.

MATERIAL AND METHODS

Two similar diets (Control and GSE diets) differing in the addition of GSE (0 and 7.2 g/kg DM) were given to birds (12/diet) for 18 days. GSE contained more than 80% of total polyphenols of which 60% were procyanidols and 0.75% of anthocyanidins.

Caecum content from 2 birds were pooled (6 replicate/diet). Cecal DNA was extracted, purified and subjected to a restriction with HhAI. Plausible community composition was studied using the Microbial Community Analysis (MiCA) web-based tool, to obtain:

Biodiversity degree: Microbial richness, the number of terminal restriction fragments (TRF)

Frequency of detection: % of animals that present a TRF compatible with the presence of a bacterium

RESULTS

Effect of inclusion of grape seed extract on biodiversity degree and frequency of detection (%) of identified, uncultured and unidentified bacteria at caecum in 18 d old chickens.
                                              Control   GSE    p (1)

BIODIVERSITY DEGREE, no TRF                   35.6      43.8   0.03

FREQUENCY OF DETECTION, %

Bacterial group   Genus

Actinobacteria    Nocardiodes                 0.00      100    0.001
                  Bifidobacterium             16.7      33.3   NS
Bacteroidetes     Flavobacterium              50.0      50.0   NS
Firmicutes        Lactobacillus               50.0      83.3   NS
                  Bacillus/Paenibacillus      16.7      83.3   0.02
                  Enterococcus                16.7      33.3   NS
                  Clostridium                 66.7      100    NS
                  Desulfitobacterium          33.3      66.7   NS
Proteobacteria    Sphingomonas                50.0      33.3   NS
                  Flexibacter                 33.3      16.7   NS
                  Thiomonas                   33.3      33.3   NS
                  Pseudomonas Acinetobacter   16.7      100    0.003
                  Enterobacter, Citrobacter   33.3      66.7   NS
                  Desulfovibrio               33.3      50.0   NS

(1) P<0.05: Significant differences

                  TRF, bp(2)                  Control   GSE    p

Uncultured        78                          50.0      50.0   NS
                  80                          0.00      66.7   0.01
                  109                         16.7      83.3   0.02
                  126                         33.3      100    0.01
                  137                         0.00      50.0   0.05
                  148                         50.0      0.00   0.05
                  265                         50.0      83.3   NS
                  500                         33.3      50.0   NS
                  539                         0.00      50.0   0.05
Unidentified      77                          83.3      16.7   0.02
                  121                         50.0      33.3   NS
                  128                         0.00      83.3   0.003
                  133                         33.3      0.00   NS
                  247                         0.00      50.0   0.05
                  258                         50.0      0.00   0.05
                  275                         16.7      83.3   0.02
                  494                         33.3      83.3   0.08
                  498                         0.00      66.7   0.01

(2)TRF, bp: size (base pairs) of terminal restriction fragment


CONCLUSIONS

Dietary addition of GSE promoted changes on caecal microbial ecosystem:

increasing the biodiversity degree

increasing the frequency of detection of several potential phenol degrading bacteria.

affected the presence of uncultured and unidentified bacteria.

S. CHAMORRO (1), A. VIVEROS (2), A. BRENES (1)

(1) Dpto. Metabolismo y Nutricion, ICTAN-CSIC, Madrid, Spain CSIC

(2) Facultad de Veterinaria, Universidad Complutense Madrid, Spain
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Article Details
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Title Annotation:articulo en ingles
Author:Chamorro, S.; Viveros, A.; Brenes, A.
Publication:Revista Complutense de Ciencias Veterinarias
Date:Jan 1, 2012
Words:538
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