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.

Anaerobic batch culture fermenters were used for a preliminary screening of the in vitro utilization by human gut microflora of dextran and novel oligodextrans (I, II and III) produced in the University of Reading (UK). Glucose and fructooligosaccharides (FOS) were used as reference carbohydrates. As expected, FOS acted as a good prebiotic in that it selectively increased numbers of bifidobacteria in the early stages of the fermentation. Dextran and oligodextrans each resulted in an enrichment of bifidobacteria in the batch cultures, with high levels of persistence up to 48 h. They also produced elevated levels of butyrate ranging from 5 to 14·85 mmol/l. To more effectively simulate conditions that prevail in different regions of the large intestine, a three-stage continuous culture cascade system was used to study further the fermentation of dextran, a low-molecular-mass oligodextran (IV) and maltodextrin. Oligodextran IV was shown to be the best substrate for bifidobacteria and lactobacilli with steady-state populations of bifidobacteria and lactobacilli being higher in all three vessels of the gut model than the respective populations resulting from dextran and maltodextrin. A maximum difference of 1·9 log was observed in vessel 1 for both bifidobacteria and lactobacilli in the case of dextran fermentation, while 1·4 log and 0·8 log in vessel 3 were the maximum differences for bifidobacteria and lactobacilli when maltodextrin was used as the carbohydrate source. Moreover, dextran and oligodextran appeared to stimulate butyrate production, with a maximum production up to 25·39 mmol/l in vessel 3 when fermenting dextran, followed by 21·70 mmol/l in the case of oligodextran IV and only 12·64 mmol/l in the case of maltodextrin.

The Maillard reaction produces coloured, macromolecular materials (melanoidins) in a variety of foods, on heating. Significant quantities may enter the human gut on a daily basis, but there is little information on their metabolism in the human colon. As the large bowel contains a diverse population of bacteria involved in normal bowel function, it is possible that melanoidins are metabolized therein. Depending on the bacteria involved, there may be disease or health implications. The aim of the present study was to use in vitro models to determine the digestibility of melanoidins and the effect of melanoidins on colonic bacteria in the gastrointestinal tract. Melanoidins were prepared and the effects of simulated upper-gut secretions on their stability determined in a model system. The effects of faecal bacteria were also determined, in batch culture, with a combination of phenotypic and genotypic (probes) criteria being used to identify the microbial diversity involved. Simulation of peptic and pancreatic digestion showed that the melanoidins did not produce detectable amounts of low-molecular-mass degradation products. However, melanoidins affected the growth of gut bacteria during mixed culture growth. The effect was to cause a non-specific increase in the anaerobic bacteria enumerated. This in vitro study indicates that melanoidins can affect the growth of human large-bowel bacteria and serves to demonstrate possible effects that may occur in vivo. Given the large and varied number of food items that contain Maillard reaction products, this may have relevance for lower-gut health.