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Reducing agent can be omitted in the incubation medium of the batch in vitro fermentation model of the pig intestines

Published online by Cambridge University Press:  02 November 2017

C. Poelaert ,
G. Nollevaux ,
C. Boudry ,
B. Taminiau ,
C. Nezer ,
G. Daube ,
Y.-J. Schneider ,
D. Portetelle ,
A. Théwis  and
J. Bindelle
C. Poelaert
Affiliation:
Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium Microbiology and Genomics Unit, Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium
G. Nollevaux
Affiliation:
Laboratory of Cellular, Nutritional and Toxicological Biochemistry, Institute of Life Sciences, Université catholique de Louvain, 1348 Louvain-la-Neuve, Belgium
C. Boudry
Affiliation:
Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium
B. Taminiau
Affiliation:
Department of Food Science, Fundamental and Applied Research for Animal and Health (FARAH), Faculty of Veterinary Medicine, University of Liege, 4000 Liège, Belgium
C. Nezer
Affiliation:
Quality Partner S.A., 4040 Herstal, Belgium
G. Daube
Affiliation:
Department of Food Science, Fundamental and Applied Research for Animal and Health (FARAH), Faculty of Veterinary Medicine, University of Liege, 4000 Liège, Belgium
Y.-J. Schneider
Affiliation:
Laboratory of Cellular, Nutritional and Toxicological Biochemistry, Institute of Life Sciences, Université catholique de Louvain, 1348 Louvain-la-Neuve, Belgium
D. Portetelle
Affiliation:
Microbiology and Genomics Unit, Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium
A. Théwis
Affiliation:
Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium
J. Bindelle*
Affiliation:
Precision Livestock and Nutrition Unit, Gembloux Agro-Bio Tech, University of Liege, 5030 Gembloux, Belgium
*
†E-mail: [email protected]
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Abstract

Over the past decade, in vitro methods have been developed to study intestinal fermentation in pigs and its influence on the digestive physiology and health. In these methods, ingredients are fermented by a bacterial inoculum diluted in a mineral buffer solution. Generally, a reducing agent such as Na2S or cysteine-HCl generates the required anaerobic environment by releasing metabolites similar to those produced when protein is fermented, possibly inducing a dysbiosis. An experiment was conducted to study the impact of two reducing agents on results yielded by such in vitro fermentation models. Protein (soybean proteins, casein) and carbohydrate (potato starch, cellulose) ingredients were fermented in vitro by bacteria isolated from fresh feces obtained from three sows in three carbonate-based incubation media differing in reducing agent: (i) Na2S, (ii) cysteine-HCl and (iii) control with a mere saturation with CO2 and devoid of reducing agent. The gas production during fermentation was recorded over 72 h. Short-chain fatty acids (SCFA) production after 24 and 72 h and microbial composition of the fermentation broth after 24 h were compared between ingredients and between reducing agents. The fermentation residues after 24 h were also evaluated in terms of cytotoxicity using Caco-2 cell monolayers. Results showed that the effect of the ingredient induced higher differences than the reducing agent. Among the latter, cysteine-HCl induced the strongest differences compared with the control, whereas Na2S was similar to the control for most parameters. For all ingredients, final gas produced per g of substrate was similar (P>0.10) for the three reducing agents whereas the maximum rate of gas production (Rmax) was reduced (P<0.05) when carbohydrate ingredients were fermented with cysteine-HCl in comparison to Na2S and the control. For all ingredients, total SCFA production was similar (P>0.10) after 24 h of fermentation with Na2S and in the control without reducing agent. Molar ratios of branched chain-fatty acids were higher (P<0.05) for protein (36.5% and 9.7% for casein and soybean proteins, respectively) than for carbohydrate (<4%) ingredients. Only fermentation residues of casein showed a possible cytotoxic effect regardless of the reducing agent (P<0.05). Concerning the microbial composition of the fermentation broth, most significant differences in phyla and in genera ascribable to the reducing agent were found with potato starch and casein. In conclusion, saturating the incubation media with CO2 seems sufficient to generate a suitable anaerobic environment for intestinal microbes and the use of a reducing agent can be omitted.

Keywords

gut fermentationin vitro methodpigreducing agentpyrosequencing
Type
Research Article
Information
animal , Volume 12 , Issue 6 , June 2018 , pp. 1154 - 1164
Copyright
© The Animal Consortium 2017 

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