Food-based strategies to modulate the composition of the intestinal microbiota and their associated health effects.

The most well known food-based strategies to modulate the composition of the intestinal microbiota are the dietary use of prebiotics, probiotics and their combination, synbiotics. Currently established prebiotic compounds are mainly targeting the bifidobacteria population of the colon microbiota. A good illustration of the importance of high colonic bifidobacteria levels is the observation that breast milk creates an environment in the colon (because of its high amount in galacto-oligosaccharides with prebiotic activity) favouring the development of a simple flora, dominated by bifidobacteria to which various health benefits have been ascribed. Currently, high colonic bifidobacteria levels has been considered favourably at all ages and strategies to augment their presence have been demonstrated in placebo-controlled intervention studies; e.g. in toddlers to reduce sickness events, in adults to reduce the risk for developing gastrointestinal diseases and in the elderly to re-enhance their declining immune activity. The intestinal microbiota can be considered as a metabolically adaptable and rapidly renewable organ of the body. However, unbalances in its microbial community and activities are found to be implicated in disease initiation and progression, such as chronic inflammatory bowel diseases and colonic cancers. Restoration of this balance by increasing bifidobacteria levels has demonstrated to reduce disease severity of patients and to improve well-being in healtly volunteers. New emerging evidence on the difference in the composition of the colonic microbiota between obese and lean volunteers has opened new areas for pre-, pro- and synbiotic research. Additionally, as knowledge will increase about the microbial bio-conversion of polyphenolic compounds into bioactive metabolites in the colon and whether food-based strategies can augment such bioconversion into more potent compounds with anti-oxidant and/or anti-inflammatory activity new areas of research will be discovered. This paper provides an up-to-date review of the health benefits associated to the induction of high bifidobacteria levels in the colon by the use of prebiotics (inulin and oligofructose). New areas of emerging science will be discussed as well.

Inulin and oligofructose as prebiotics in the prevention of intestinal infections and diseases.

Health and wellbeing are challenged constantly by pathogens. A number of defence mechanisms exist to protect the body from pathogen colonisation and invasion, with an important role to play for the natural intestinal bacterial flora (mainly by bifidobacteria and lactobacilli). The present paper reviews the evidence on the effects of inulin and oligofructose on colonisation and translocation of pathogens and the prevention of intestinal diseases. In vitro experiments have shown that lactic acid-producing bacteria have antagonistic (antibacterial) activity against pathogens partly because of the production of organic acids which are the endproducts of inulin and oligofructose fermentation. In addition, studies with epithelial layers have shown that inulin and oligofructose inhibit pathogen colonisation and that endproducts of their fermentation have the ability to support barrier function. Furthermore, studies in various animal models have shown that inulin and oligofructose accelerate the recovery of beneficial bacteria, slow down pathogen growth, decreasing pathogen colonisation and systemic translocation. Finally, data from human intervention trials either in patients with intestinal disorders or disease, or prone to critical illness, found that inulin and oligofructose restore the balance when the gut microbial community is altered, inhibit the progression of disease or prevent it from relapsing and/or developing. To conclude, the dietary use of inulin and oligofructose offers a promising approach to restore microbial communities and to support barrier function of the epithelia by their prebiotic action. This may offer the host protection against invasion and translocation of pathogens (endogenous and/or exogenous) and in the prevention of gastrointestinal diseases.

Do thickening properties of locust bean gum affect the amount of calcium, iron and zinc available for absorption from infant formula? In vitro studies.

Locust bean gum acts as a milk thickener in infant formula because of its high apparent viscosity. The effects of such thickening agents on metabolic and physiologic responses during infancy have not been clarified sufficiently. Due to the increased volume of the digest and the bulking and trapping effects, digestion and absorption of nutrients may be influenced in presence of locust bean gum. The central question addressed in this paper is whether the thickening properties of locust bean gum affect the availability of calcium, iron, and zinc. Increasing amounts of powdered locust bean gum were homogenised with infant formula and samples were diluted to 0.14, 0.27, 0.42 and 0.71 g/100 ml. Viscosity of the samples was measured by a Carrie-Med CSL100 rheometer. Available amounts of calcium, iron, and zinc were evaluated using a continuous-flow dialysis model with preliminary digestion. Elemental contents of samples and dialysates were analysed with atomic absorption spectrometry. The first set of experiments showed that addition of locust bean gum to infant formulas increased the viscosity of the luminal contents. Correlations between the locust bean gum concentration and the viscosity of the samples before and after gastric or intestinal digestion were highly significant (0.97). In the second set of experiments, the correlations between the locust bean gum concentration and the amounts of calcium trapped by the locust bean gum fraction also showed high significance (0.93). In the third experimental design, again strong correlations were found between the viscosity of the intestinal digest and the amounts of calcium trapped by the fibre fraction (0.90). For iron and zinc, no such relationships were found. From this experimental set-up it appears that locust bean gum influences calcium availability in infant formulas by means of its physical properties to act as thickening agent, rather than its chemical ability to form complexes as demonstrated earlier with respect to iron and zinc.

Calcium, iron and zinc availability from weaning meals.

UNLABELLED: During the weaning period, solid foods are introduced in the diet to complement human milk or infant formula. Consequently, marginal deficiencies may easily develop because of inappropriate feeding and low bioavailability of nutrients. The objective of this study was to determine calcium, iron and zinc availability from weaning foods that contained a variety of ingredients but almost the same dietary fibre content, and to describe dietary factors that influence availability in a linear matter. Availability measurements from eight meals were made in vitro by a continuous-flow dialysis system with preliminary intraluminal digestion. Calcium, iron and zinc were analysed by flame atomic absorption spectrometry; ascorbic acid content was taken from food-composition tables; and macronutrient and mineral contents were obtained from the manufacturer. Calcium availability was higher in fruits (mean +/- SD, 39.2 +/- 4.1%) than vegetables (31.7 +/- 5.8%) (p < or = 0.001). Iron availability was marginally significant higher in vegetables (13.0 +/- 4.7%) than fruits (10.2 +/- 1.6%) (p = 0.027). Zinc availability showed higher availability in fruits (52.3 +/- 22.3%) than vegetables (22.1 +/- 5.6%) (p < or = 0.001). CONCLUSION: A major portion (47.8%) of the variance in calcium availability, as determined by multiple regression, could be explained by protein (p = 0.058, beta = -0.691). Major predictors of iron availability among the nutrients were iron content of the meal (p = 0.014, beta = -0.838), carbohydrates (p = 0.044, beta = -0.610) and dietary fibre (p = 0.235, beta = -0.285). Zinc availability was merely influenced by the protein (p = 0.010, beta = -0.876) and fat content of the meals (p = 0.295, beta = 0.255) in relation to other nutrients.

In vitro availability of zinc from infant foods with increasing phytic acid contents.

An in vitro method was used to determine the availability of Zn from infant foods containing increasing amounts of phytate, and to quantify the effect of the phytate:Zn molar ratio on the availability. During the in vitro assay, digestive conditions of infants, younger and older than 4 months of age, were carefully simulated since the solubility of phytate-Zn complexes during digestion is pH dependent. Availability was measured with a continuous flow dialysis in vitro procedure with previous intralumen digestive stage. Zn concentrations were determined with flame atomic absorption spectrometry. Phytic acid content was measured with HPLC. Adding phytate to infant formula lowered Zn availability to 2.84 (sd 0.17) % when the phytate:Zn molar ratio increased to 2.2 as compared with cows' milk-based formula (6.65 (sd 0.55) %). Availability from vegetables (23.83 (sd 2.17) %) significantly decreased at a ratio > 7.9 (15.12 (sd 1.63) %). Zn availability from soyabean-based formula (2.26 (sd 0.36) %) was lower compared with cows' milk-based formula (6.65 (sd 0.55) %). Availability between soyabean- and cows' milk-based formula was similar when a phytate:Zn ratio of 2.2 (2.84 (sd 0.17) %) was obtained in the cows' milk formula. The negative effect of phytic acid on Zn availability was dependent on the type of the food and the phytate content, and should be considered when using soyabean-based formulas during early infancy.

Binding of mineral elements to locust bean gum influences availability in vitro.

Experiments were conducted to evaluate the extent to which element binding of locust bean gum (LBG) affects the availability of calcium, iron, and zinc in the gut. Infant formula was supplemented with increasing amounts of LBG and subjected to an intraluminal digestion procedure. Element binding was measured by eliminating the complexes by twofold centrifugation. Availability of the elements was determined using a validated continuous-flow dialysis technique. Elemental content of the samples, supematants, and dialysates was analyzed with validated atomic absorption spectrometry. LBG provided small amounts of intrinsic calcium (1.13+/-0.02 mg/g) and trace amounts of iron (0.02+/-0.00 mg/g) and zinc (0.01+/-0.00 mg/g), which were strongly bound to the LBG molecule (respectively: 76.6+/-3.3%, 83.4+/-1.2%, 96.7+/-6.6%). Correlation analysis, between percent element bound by LBG after centrifugation and percent trapped after dialysis, yielded significant correlation only for the data of zinc (r = 0.93). For calcium and iron, no correlation could be demonstrated; however, for iron a similar trend was observed. These findings suggest that element binding of LBG has a major influence on the availability of zinc and maybe of iron. For calcium, other factors might also be involved, affecting availability.

Effect of thickening agents, based on soluble dietary fiber, on the availability of calcium, iron, and zinc from infant formulas.

Although it is accepted that dietary fiber probably is not needed in the diets of infants younger than 1 y, babies suffering from regurgitation are often fed with infant formulas thickened with considerable amounts of fiber. The effect of increasing amounts of alginic acid, locust-bean gum, and guar gum was studied from casein and whey-based infant formulas. A dialysis in vitro method with preliminary intraluminal digestion, adapted to the conditions of infants younger than 6 mo, was used. Human milk was used as the reference standard. Elemental contents of samples and dialysates were determined by atomic absorption spectrometry. Soluble dietary fiber inhibited mineral availability more in casein than in whey-based formulas. Mineral availabilities from casein- and whey-based formulas supplemented with 0.42 g of locust-bean gum/100 mL were 9.4% (0.7) and 10.4% (0.6) for calcium (P < 0.05), 0.32% (0.08) and 1.45% (0.17) for iron (P < 0.05), and 3.2% (0.2) and 5.6% (0.5) for zinc (P < 0.05), respectively. Calcium availability from the whey formula decreased in the presence of each fiber source, especially guar gum and alginic acid. Supplementing 2 g of alginic acid-based agents per 100 mL depressed calcium availability from 13.3% (1.2) to 5.3% (0.3; P < 0.05). With respect to iron and zinc, availabilities increased from 1.28% (0.28) to 6.05% (0.96; P < 0.05) and from 6.7% (0.6) to 10.2% (1.0), respectively, with the addition of 2 g of alginic acid (P < 0.05). Both gums lowered iron and zinc availabilities, and guar gum affected iron availability more severely than locust-bean gum did. Iron availabilities were 1.45% (0.17) from formula thickened with locust-bean gum (0.42 g/100 mL) and 0.92% (0.15) from formula thickened with guar gum (P < 0.05). Adding thickening agents based on soluble dietary fiber to traditional infant formulas probably affects calcium, iron, and zinc availability in various ways.

A method for in vitro determination of calcium, iron and zinc availability from first-age infant formula and human milk.

A method for in vitro determination of available calcium, iron and zinc content from infant food after digestion was evaluated. This method introduced an intraluminal digestive phase, adapted to the gastrointestinal conditions of infants younger than 6 months of age, prior to continuous flow dialysis of the resultant gastric digest. Precautions handling the method were discussed and enzymatic parameters were defined. Ruggedness of the method was determined from the availability of calcium, iron and zinc at different gastrointestinal conditions. Availability of all three elements was higher at gastric pH of 2 (20.0 +/- 1.1% for calcium, 4.06 +/- 0.66% for iron and 17.5 +/- 1.3% for zinc), than from the normal procedure (pH 4) (15.6 +/- 1.2% for calcium, 1.18 +/- 0.26% for iron and 8.2 +/- 0.9% for zinc). At pH 5, however, calcium availability appeared to be lower (11.7 +/- 1.0%) (P < 0.05). The intestinal pH also had a major influence on the availability. At low intestinal pH (5.5), availability was 40.5 +/- 2.3% for calcium, 3.01 +/- 0.58% for iron and 26.8 +/- 1.8% for zinc, which was higher compared with the normal procedure (P < 0.05). Moreover, other factors, such as digestion time, mixing and filtration pressure, also affected the availability. Recovery tests yielded mean values of 94 +/- 3% for calcium, 109 +/- 9% for iron and 106 +/- 4% for zinc. Mean intra- and inter-batch precision of the availability procedure was 4.1 CV% and 6.6 CV% for calcium, 14.5 CV% and 19.2 CV% for iron, and 4.0 CV% and 13.6 CV% for zinc. The method provides adequate accuracy, acceptable precision and good recovery. It offers the advantage of being simple, rapid and inexpensive, since it takes only 1 day to run the whole availability procedure (including four replicates per sample), and the low costs of the dialysis equipment. It can therefore be considered as suitable for predicting the availability of essential elements from foods used during the first months of infancy.

In vitro availability of calcium, iron, and zinc from first-age infant formulae and human milk.

BACKGROUND: Variation in the bioavailability of calcium (Ca), iron (Fe), and zinc (Zn) occurs because of interactions of food components in the gastrointestinal microenvironment. Bioavailability is preferably determined by in vivo tests, but these are expensive, labor-intensive, time consuming, and often unethical. As an alternative, in vitro methods can be used to predict bioavailability of nutrients from foodstuffs. METHODS: A continuous-flow dialysis model with preliminary intraluminal digestive phase, adapted to the gastrointestinal conditions of infants younger than 6 months, was used. Human milk was the reference standard. Ca, Fe, and Zn content of samples and dialysates after digestion were analyzed by atomic absorption spectrometry. RESULTS: Ca availability is similar in human milk (13.1%+/-0.8%), whey (13.3%+/-1.2%), and soy-based formulae (13.0%+/-1.2%; P > 0.05), and higher in casein-predominant formula (21.2%+/-0.6%; P < 0.05). Availability of Fe is highest in human milk (8.12%+/-0.27%: P < 0.05). Fe availability in whey (1.28%+/-0.28%) and soy formulae (1.48%+/-0.28%) is similar (P > 0.05), but availability is lower in casein-predominant formula (0.48%+/-0.22%; P < 0.05). Zn availability is also highest in human milk (13.1%+/-0.7%; P < 0.05). However, Zn availability is similar in whey (6.7%+/-0.6%) and casein formulae (8.5%+/-1.6%; P > 0.05), but lower in soy formula (2.3%+/-0.4%; P < 0.05). CONCLUSIONS: Our observations are in agreement with previous data from in vivo studies in term infants. This in vitro procedure is an inexpensive, simple, rapid, and reliable method that predicts the bioavailability of Ca, Fe, and Zn in foods.

Thickening infant formula with digestible and indigestible carbohydrate: availability of calcium, iron, and zinc in vitro.

BACKGROUND: Regurgitation is common in infants and is usually due to gastroesophageal reflux. Often parental reassurance and dietary management by feeding thickened formulas are the only therapeutic steps necessary. Adding fibers may interfere with the absorption of micronutrients. METHODS: A continuous-flow dialysis in vitro method with a preliminary intraluminal digestive phase, modified to simulate the conditions of infants less than 6 months of age and children from 6 months of age on, was used to study the availability of calcium, iron, and zinc from thickened and nonthickened first-and second-age infant formulas. Pooled mature human milk was used as the reference standard. The elemental content of the samples and dialysate fractions of calcium, iron, and zinc after digestion was determined by atomic absorption spectrometry. RESULTS: In human milk, calcium, iron, and zinc were highly available for absorptive processes. Availability of calcium, iron, and zinc from nonthickened first- and second-age infant formulas tends to be significantly better than in the corresponding products thickened with locust bean gum. Thickening infant formulas with pregelatinized rice starch, however, does not affect the availability of calcium, iron, and zinc. CONCLUSIONS: It appears that human milk provides optimal conditions for the availability of calcium, iron, and zinc. Availability of calcium, iron, and zinc seems to lower when infant formulas are thickened with indigestible carbohydrates, whereas it does not by adding digestible carbohydrates.