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A medium for the presumptive detection of Enterobacter sakazakii in infant formula: interlaboratory study.

A standard method for the detection of Enterobacteriaceae was modified for the presumptive detection of Enterobacter sakazakii, and the modified method was validated in an interlaboratory trial with 16 laboratories from 8 European countries. The modification included a differential-elective medium for the isolation of E. sakazakii, consisting of nutrient agar (NA) supplemented with 4-methyl-umbelliferyl alpha-D-glucoside (alpha-MUG). A 25 g sample was added to 225 mL buffered peptone water. After incubation at 35 degrees or 37 degrees C for 16 or 20 h, 10 mL nonselective enrichment was transferred into 90 mL selective enrichment. The selective enrichment was streaked on violet-red bile glucose agar (VRBGA) and incubated at 37 degrees C for 24 h. It was streaked in parallel on NA plates supplemented with alpha-MUG at 50 mg/L and incubated at 25 degrees C for 16 h, and afterwards for an additional 24 h at room temperature in the dark. E. sakazakii appeared as vivid yellow colonies under normal light and showed blue/violet fluorescence under UV light on NA + alpha-MUG plates. Validation samples represented powdered infant formula without E. sakazakii (blanks) and with low (1-10 colony-forming units [CFU]/25 g) and medium (1-10 CFU/g) contamination levels. All samples contained Pseudomonas aeruginosa and Lactobacillus spp. as background flora. The specificity for blank samples was 100%. The sensitivity of the low contamination level was similar for VRBGA and NA + alpha-MUG, i.e., 66.7% (66.7% accordance, 53.9% concordance). For the medium level the sensitivities were 96.7% (93.3% accordance, 93.5% concordance) for VRBGA and 98.3% (96.9% accordance, 96.9% concordance) for NA + alpha-MUG.

Child↗

Iron bioavailability studied in infants: the influence of phytic acid and ascorbic acid in infant formulas based on soy isolate.

The influence of phytic acid and ascorbic acid content of soy formula on iron (Fe) bioavailability was investigated in infants by analysis of the incorporation of stable isotopes of Fe into red blood cells 14 d after administration using a double stable isotope technique. Paired comparisons were made with each infant acting as his or her own control. The geometric mean fractional Fe incorporation into red blood cells increased from 5.5 to 6.8% (p < 0.05) when soy formula with the native content of phytic acid was compared with a 83% dephytinized formula. A more pronounced effect was shown with soy formula containing no phytic acid; the mean fractional Fe incorporation increased from 3.9 (native phytic acid) to 8.7% (zero phytic acid; p < 0.001). A significant (p < 0.01) effect was also demonstrated when the Fe:ascorbic acid molar ratio in the native phytate-containing formula was increased from 1:2.1 to 1:4.2; mean fractional Fe incorporation increased from 5.9 to 9.6%. These results demonstrate that the Fe bioavailability from soy-based infant formulas can be similarly increased by either removing phytic acid or increasing the ascorbic acid content.

Ascorbic Acid↗

Iron retention from lactoferrin-supplemented formulas in infant rhesus monkeys.

Iron absorption from human milk and infant formula has received much attention, but experimental design problems have been common. In our study, iron retention from human milk, milk-based infant formula (IF) with and without supplemental ferrous sulfate, and IF supplemented with either human or bovine lactoferrin (Lf) was evaluated in infant rhesus monkeys. The exchange of 59Fe (III) Cl3 between the whey, casein, and fat fractions required up to 72 h to reach the same distribution as intrinsic iron, depending on the type of diet. Infant monkeys were intubated with labeled human milk or IF and counted in a whole body counter. Each infant received all five diets and was also intubated with a reference dose of 55Fe (II) ascorbate. There was no significant difference in iron retention (mean +/- SEM) from the experimental diets: human milk 32.5 +/- 5.1%; IF 32.1 +/- 8.0%; IF + Fe 23.0 +/- 3.9%; IF + human Lf 23.5 +/- 3.3%; IF + bovine Lf 22.7 +/- 4.9%. Therefore, infant monkeys absorb and retain iron similarly from human milk and infant formula. Supplementation of infant formula with human or bovine Lf resulted in similar iron retention to that of ferrous sulfate-supplemented infant formula.

Animals↗

Evaluation of powdered infant formula milk as chelating agent for copper under simulated gastric conditions of a baby's stomach.

The objective of this study was to determine the complexing capacity of four types of powdered commercial milks with copper(II) using square wave adsorptive stripping voltammetry. Two were types of cow milk adapted for babies under one year (A and B), one was soymilk (C) and the other was normal milk (D). Milk solutions were prepared following the instructions shown on the milk container, and they were mixed with a pepsin solution simulating a baby's stomach conditions (pepsin and salts concentration, pH and temperature). Complexing capacity was determined by titrating milk samples with aliquots of a standard copper solution until the peak current due to solvated or free copper ions was increasing. Assuming a 1:1 copper-milk complex, the apparent stability constant was found using the pseudopolarogram method. The log K'(Cu-milk) values were 4.9, 5.0, 3.0 and 5.1 for A, B, C and D types of milk, respectively. Voltammograms of the milk solution as a function of copper added show that the binding properties of the four types of powdered milk studied were different and that saturation of the four types, occurs at different copper concentrations. Concentrations obtained were: 4.9, 5.8, 1.1 and 10.1 mM for A, B, C and D types, respectively. The best complexing agent was the solution prepared with powdered milk D and the worst was that of C. This is important for the bioavailability of this element as a micronutrient.

Animals↗

Potential aluminium toxicity in infants fed special infant formula.

Aluminium was measured in samples of plasma and samples of feed obtained from 74 infants with normal renal function established on various feeds (breast, whey-based, fortified whey-based, preterm, soy, and casein hydrolysate). All infants were bolus fed, and blood samples were collected midway between feeds. Aluminium was measured using electrothermal atomization and atomic absorption spectrometry. Mean aluminium concentrations in milks were as follows: breast, 9.2 micrograms/L [95% confidence interval (CI), 5.6-12.7]; whey-based, 165 micrograms/L (95% CI, 151-180); fortified, 161 micrograms/L (95% CI, 143-180); preterm, 300 micrograms/L (95% CI, 272-328); soy, 534 micrograms/L (95% CI, 470-598); casein hydrolysate, 773 micrograms/L (95% CI, 632-914). Mean plasma aluminium concentrations in infants receiving different milks were as follows: breast, 8.6 micrograms/L (95% CI, 5.6-10.6); whey-based, 9.2 micrograms/L (95% CI, 7.4-11.0); fortified, 10.3 micrograms/L (95% CI, 8.3-12.3); preterm, 9.7 micrograms/L (95% CI, 5.3-17.1); soy, 12.5 micrograms/L (95% CI, 5.0-20.0); casein hydrolysate, 15.2 micrograms/L (95% CI, 10.7-19.8). Mean plasma aluminium concentration was significantly different in infants fed casein hydrolysate formulae than in those fed breast milk (difference, 6.7 micrograms/L; 95% CI, 2.8-10.5; p = 0.028). We conclude that infants may be at risk from aluminium toxicity when consuming formula containing > 300 micrograms/L--in particular, casein hydrolysate formulae. We speculate that the aluminium compounds found in breast milk are more bioavailable than those found in other milks and that some constituents of infant formula affect aluminium absorption from the gut lumen.

Aluminum↗

Growth and development of premature infants fed predominantly human milk, predominantly premature infant formula, or a combination of human milk and premature formula.

BACKGROUND: In a recent meta-analysis, human milk feeding of low birth-weight (LBW) infants was associated with a 5.2 point improvement in IQ tests. However, in the studies in this meta-analysis, feeding regimens were used (unfortified human milk, term formula) that no longer represent recommended practice. OBJECTIVE: To compare the growth, in-hospital feeding tolerance, morbidity, and development (cognitive, motor, visual, and language) of LBW infants fed different amounts of human milk until term chronologic age (CA) with those of LBW infants fed nutrient-enriched formulas from first enteral feeding. METHODS: The data in this study were collected in a previous randomized controlled trial assessing the benefit of supplementing nutrient-enriched formulas for LBW infants with arachidonic acid and docosahexaenoic acid. Infants (n = 463, birth weight, 750-1,800 g) were enrolled from nurseries located in Chile, the United Kingdom, and the United States. If human milk was fed before hospital discharge, it was fortified (3,050-3,300 kJ/L, 22-24 kcal/oz). As infants were weaned from human milk, they were fed nutrient-enriched formula with or without arachidonic and docosahexaenoic acids (3,300 kJ/L before term, 3,050 kJ/L thereafter) until 12 months CA. Formula fed infants were given nutrient-enriched formula with or without added arachidonic and docosahexaenoic acids (3,300 kJ/L to term, 3,050 kJ/L thereafter) until 12 months CA. For the purposes of this evaluation, infants were categorized into four mutually exclusive feeding groups: 1) predominantly human milk fed until term CA (PHM-T, n = 43); 2) >/= 50% energy from human milk before hospital discharge (>/= 50% HM, n = 98); 3) < 50% of energy from human milk before hospital discharge (< 50% HM, n = 203); or 4) predominantly formula fed until term CA (PFF-T, n = 119). RESULTS: PFF-T infants weighed approximately 500 g more at term CA than did PHM-T infants. This absolute difference persisted until 6 months CA. PFF-T infants were also longer (1.0-1.5 cm) and had larger head circumferences (0.3-1.1 cm) than both PHM-T and >/= 50% HM infants at term CA. There was a positive association between duration of human milk feeding and the Bayley Mental Index at 12 months CA (P = 0.032 full and P = 0.073 reduced, statistical models) after controlling for the confounding variables of home environment and maternal intelligence. Infants with chronic lung disease fed >/= 50% HM until term CA (n = 22) had a mean Bayley Motor Index about 11 points higher at 12 months CA compared with infants PFF-T (n = 24, P = 0.033 full model). CONCLUSION: Our data suggest that, despite a slower early growth rate, human milk fed LBW infants have development at least comparable to that of infants fed nutrient-enriched formula. Exploratory analysis suggests that some subgroups of human milk fed LBW infants may have enhanced development, although this needs to be confirmed in future studies.

Adult↗

Prebiotics in infant formulas.

BACKGROUND: The intestinal flora of breast-fed infants is an important physiologic factor in the function of the gut and in the development of the immune system. The current research is part of a group of studies performed to answer the question whether a bovine milk formula supplemented with a prebiotic mixture from galactooligosaccharides and fructooligosaccharides can stimulate an intestinal flora similar to that of breast-fed infants. METHODS: The prebiotic effect of the oligosaccharide mixture was tested in preterm and term infants by measuring fecal flora using plating as well as fluorescent in situ hybridization techniques. The effect of the oligosaccharides on the bacterial metabolism was studied by measuring short-chain fatty acid production in vitro and the short-chain fatty acid pattern in the stools of a group of term infants. RESULTS: The oligosaccharide mixture increases significantly the number of bifidobacteria and reduces the number of pathogens in term as well as in preterm infants when compared with a group of infants fed an unsupplemented formula. Using a concentration of 0.8 g oligosacchrides/100 mL formula, the amount of bifidobacteria is similar to that typical of breast-fed infants. In vitro, the short-chain fatty acids produced by the mixture of oligosaccharides under study were similar to those produced by the human milk oligosaccharides fraction. In clinical trials the pattern of fecal short-chain fatty acids in infants fed the oligosaccharide mixture was similar to that of breast-fed infants but was significantly different from that of a group of infants fed with an unsupplemented formula. Additionally, the fecal pH was significantly higher in the group fed an unsupplemented formula than in the groups fed either breast milk or a supplemented formula. CONCLUSION: The data obtained indicate that the prebiotic mixture under study is able to stimulate the development of a microbial flora similar to that of breast-fed infants. Several biota, whose growth is enhanced by this prebiotic mixture, represent important factors in the postnatal development of the immune system. On this evidence it can be suggested that prebiotics may play a role as modulators of the postnatal development of the immune system.

Animals↗

Formula tolerance in postbreastfed and exclusively formula-fed infants.

OBJECTIVE: Perceived intolerance to infant formula is a frequently reported reason for formula switching. Formula intolerance may be related to perceived symptoms of constipation, fussiness, abdominal cramps, and excessive spit-up or vomit. Commercially available formulas differ from each other in processing and in sources and levels of protein, lipids, and micronutrients. These differences may affect tolerance. The objective of this article was to compare the tolerance of two commercially available powder infant formulas that differ in composition. Measures of tolerance in exclusively breastfed infants weaned to an infant formula and exclusively formula-fed infants were evaluated. METHODS: Two clinical studies were conducted. In study 1, 82 healthy, full-term infants who were exclusively breastfed at the time of enrollment were randomized at weaning to formula A (commercially available Similac With Iron Powder) or formula B (previously available Enfamil With Iron Powder). Parents completed daily records of tolerance during exclusive breast milk feeding, during the weaning period, and for a 2-week exclusive formula-feeding period. In study 2, 87 healthy, full-term infants who were exclusively formula-fed at the time of study enrollment (by 2 weeks of age) were fed a standard cow milk-based formula (previously commercially available Similac With Iron Powder) and then randomized to receive formula A or B for a 2-week period. Parents completed daily records of tolerance throughout the study. Formula A was a cow milk-based formula with a whey:casein ratio of 48:52 and a fat blend of 42% high-oleic safflower, 30% coconut, and 28% soy oils. Formula B was a cow milk-based formula with a whey:casein ratio of 60:40 and a fat blend of 45% palm olein, 20% soy, 20% coconut, and 15% high-oleic sunflower oils. Both formulas had lactose as the source of carbohydrate and contained 12 mg of iron per liter. Only formula A contained nucleotides at the time of the study. Measures of tolerance included volume of each formula feeding, occurrences of spit-up and/or vomit, and the color (yellow, green, brown, or black) and consistency (water, loose/mushy, soft, formed, or hard) of each stool. RESULTS: In both studies, volume of formula intake, weight gain, and incidence of spit-up or vomit did not differ between feeding groups. In study 1, stool frequency decreased significantly from the exclusive breast milk period to weaning. Stools also became firmer as infants moved from breast milk to weaning and to exclusive formula feeding. When formula was introduced into the diet, stools became less yellow and more green. Infants weaned to formula B had less frequent stools, fewer brown stools, and more yellow stools than did infants fed formula A. In both studies, infants fed formula B experienced significantly firmer stools than did those fed formula A. CONCLUSIONS: The present clinical studies indicate that the composition and/or processing of milk-based powder iron-fortified infant formulas affect stool characteristics experienced by infants. The inclusion of palm olein oil in formula B may be the reason for the observed differences in stool characteristics. Palm olein is used in infant formulas to provide palmitic acid at a level similar to that found in breast milk. However, palmitic acid from palm olein is arranged differently from that in breast milk triglyceride and is poorly absorbed. Unabsorbed palmitic acid tends to react with calcium to form insoluble soaps, and the level of these soaps is correlated with stool hardness. The pattern of softer stools and greater frequency of stooling associated with formula A is similar to the stool pattern in the exclusively breastfed infant. Thus, the use of formula A may ease the transition from breast milk to formula feeding and ameliorate parents' perception that constipation is associated with iron-fortified formula.

Animals↗

Extensively and partially hydrolysed infant formulas for allergy prophylaxis.

The allergy preventive effect of extensively (N) and partially (PH) hydrolysed cows' milk formulas compared with a regular formula (RM) was assessed in 155 infants with a family history of allergy. No cows' milk was given during the first nine months of life and no egg and fish up to 12 months of age. Breast feeding mothers avoided the same foods. At weaning the infants were randomised to one of the formula groups. The cumulative incidence of atopic symptoms at 18 months was 51, 64, and 84% in the N, PH, and RM groups, respectively. From 6 to 18 months there were significantly less cumulative atopic symptoms in the N group compared with the RM group, and significantly less than the PH group up to 6 (N = 25%; PH = 46%) and 9 months (N = 34%, PH = 58%). At 9 months significantly fewer infants in the N group (10%) than in the PH group (33%) had a positive skin prick test to eggs. The findings support an allergy preventive effect of an extensively hydrolysed formula, but not of a partially hydrolysed formula, during the first 18 months of life of high risk infants.

Animals↗

Quantitative and qualitative study of gastric lipolysis in premature infants: do MCT-enriched infant formulas improve fat digestion?

Intragastric fat digestion was investigated by analyzing the products of lipolysis and the gastric lipase (HGL) levels of premature infants fed with a formula enriched with medium chain triglycerides (MCT) and those of infants fed with human milk. Infants were fed using a gastric tube and the gastric contents were aspirated twice a day for 5 d, before and at various times after gavage feeding. HGL levels were measured using the pHstat technique. After extraction, lipids were separated and quantified using thin-layer chromatography coupled to a flame ionization detector. Fatty acid methyl esters were analyzed by gas chromatography. HGL concentration increased during digestion, reaching 77.4 +/- 43.1 microg/mL (around 75% of those recorded in adults). Mean HGL output was 115 +/- 43 microg for 3 h and the overall intragastric lipolysis was 6.1 +/- 2.6%. Although the formula was enriched with octanoic and decanoic acid, the main fatty acids released in the stomach were palmitic (C16:0, 17.03 +/- 0.23% wt/wt) and oleic (C18:1 n-9, 28.23 +/- 1.26% wt/wt) acid. Similar results were obtained with infants fed with human milk. MCT supplementation has no quantitative or qualitative effects on the intragastric lipolysis, which is not higher in premature infant than in adults.

Gastric Mucosa↗

Considerations and approaches in determining the protein and energy composition of preterm infant formulas.

Optimal early nutritional support is considered a crucial issue in the care of the preterm infant, particularly of those with very low- or extremely low-brithweight. Unfortunately, this goal is seldom satisfactorily attained. Several conditions such as hypoxia, acidosis, patent ductus arteriosus, drug therapy, reduced intestinal motility may interfere with an adequate nutritional delivery in the early neonatal period. Moreover, there is still concern about metabolic and intestinal tolerance of the currently suggested intakes and a lack of uniformity in the nutritional program among different NICUs. Finally, the vast majority of the available preterm formulas are not fully adequate to the real nutritional needs of these infants. Inadequate protein content and inappropriate protein energy ratio of most preterm formulas represent a matter of major concern, since there is a strict relationship between formula composition and the quantity and quality of weight gain. As a consequence there is a need of at least two milk formulas for different preterm infants: one for LBW and VLBW infants, the other for preterm infants weighing >1500 g.

Energy Intake↗

Copper absorption from human milk, cow's milk, and infant formulas using a suckling rat model.

Since copper deficiency is known to occur during infancy, it becomes important to assess copper uptake from various infant diets. We have investigated the uptake of copper from human milk, cow's milk, cow's milk formulas, cereal/milk formula and soy formula, compensating for the decay of 64Cu and using the suckling rat as a model. Radiocopper was added to the diet in trace amounts. Ultracentrifugation, ultrafiltration, and gel filtration were used to show that the added 64Cu bound to milk fractions and individual binding compounds in a manner analogous to the distribution of native copper, thus validating the use of extrinsically labeled diets. Labeled diets were intubated into 14-day-old suckling rats. Animals were killed after 6 h and tissues removed and counted. Liver copper uptake was 25% from human milk, 23% from cow's milk formula, 18% from cow's milk, 17% from premature (cow's milk based) infant formula, 17% from cereal/milk formula and 10% from soy formula. These results show that the rat pup model may provide a rapid, inexpensive, and sensitive method to assay bioavailability of copper from infant foods.

Animals↗