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Quantitative real-time PCR assays to identify and quantify fecal Bifidobacterium species in infants receiving a prebiotic infant formula.

A healthy intestinal microbiota is considered to be important for priming of the infants' mucosal and systemic immunity. Breast-fed infants typically have an intestinal microbiota dominated by different Bifidobacterium species. It has been described that allergic infants have different levels of specific Bifidobacterium species than healthy infants. For the accurate quantification of Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium dentium, Bifidobacterium infantis, and Bifidobacterium longum in fecal samples, duplex 5' nuclease assays were developed. The assays, targeting rRNA gene intergenic spacer regions, were validated and compared with conventional PCR and fluorescent in situ hybridization methods. The 5' nuclease assays were subsequently used to determine the relative amounts of different Bifidobacterium species in fecal samples from infants receiving a standard formula or a standard formula supplemented with galacto- and fructo-oligosaccharides (OSF). A breast-fed group was studied in parallel as a reference. The results showed a significant increase in the total amount of fecal bifidobacteria (54.8% +/- 9.8% to 73.4% +/- 4.0%) in infants receiving the prebiotic formula (OSF), with a diversity of Bifidobacterium species similar to breast-fed infants. The intestinal microbiota of infants who received a standard formula seems to resemble a more adult-like distribution of bifidobacteria and contains relatively more B. catenulatum and B. adolescentis (2.71% +/- 1.92% and 8.11% +/- 4.12%, respectively, versus 0.15% +/- 0.11% and 1.38% +/- 0.98% for the OSF group). In conclusion, the specific prebiotic infant formula used induces a fecal microbiota that closely resembles the microbiota of breast-fed infants also at the level of the different Bifidobacterium species.

Bifidobacterium↗

Analysis of trace amounts of bovine beta-lactoglobulin in infant formulas by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection.

Bovine beta-lactoglobulin (betaLG) has been described by several authors as the main allergen present in cow's milk. It can induce allergic reactions even at the low concentration existing in hypoallergenic formulas based on hydrolyzed cow's milk proteins (generally lower than microM). In this paper, the usefulness of a capillary electrophoresis method with on-capillary derivatization and laser-induced fluorescence detection for the analysis of trace amounts of betaLG in a commercial hypoallergenic formula has been demonstrated. To confirm the identity of the peak of betaLG based on migration time, an immunorecognition step employing an anti-betaLG antibody was performed. BetaLG was quantitated in the whey and casein fractions of the hypoallergenic formula. The concentration of betaLG in the whey fraction of the formula was about 3 orders of magnitude lower than the average value present in cow's milk. In the casein fraction of the formula, the concentration of betaLG was about 1 order of magnitude lower than in the whey fraction. The method developed was also used for the quality control of three cereal-based infant foods formulated without milk to test the presence or absence of betaLG as an indicator of milk contamination during the fabrication process. BetaLG in a concentration of 10(-7) M or higher was not observed in any of the cereal-based infant formulas analyzed.

Animals↗

Interregional foodborne salmonellosis outbreak due to powdered infant formula contaminated with lactose-fermenting Salmonella virchow.

Spain's Salmonella surveillance system backed by regionally-based epidemiologists around the country made it possible to detect and halt the spread of a foodborne salmonellosis outbreak due to powdered infant formula contaminated with a lactose-fermenting strain of Salmonella virchow. Forty-eight cases involving children, mostly under 7 months old, were detected in 14 out of Spain's 17 Regions. The outbreak started in January and ended in June 1994. All cases were microbiologically confirmed. The implicated strain had a 3.6 kb plasmid, was susceptible to all antimicrobials tested except nitrofurantoin and was phagetype 4a. Isolates from 8 of 24 Brand "A" milk samples tested had the same characteristics as case isolates. All affected/suspect batches of Brand "A" milk were destroyed and the product withdrawn from sale, which led to the end of the outbreak. This incident underscores the importance of maintaining surveillance systems able to detect and prevent foodborne outbreaks and alert to the possibility of isolating unusual lactose-fermenting Salmonella serotypes in especially sensitive food products.

Animals↗

Graphite-furnace atomic absorption spectrometric determination of lead, cadmium, cobalt and nickel in infant formulas and evaporated milks after nitric-perchloric acid digestion and coprecipitation with ammonium pyrrolidine dithiocarbamate.

A graphite-furnace atomic absorption method, developed for lead and cadmium, was modified to enable simultaneous determination of lead, cadmium, cobalt and nickel in infant formulas and evaporated milks. The method was assessed on the basis of analytical quality assurance results during routine analysis of samples. Detection limits (ng g-1 based on 10 g sample size for ready-to-use formulas) were 0.04-0.21 for lead, 0.004-0.015 for cadmium, 0.04-0.2 for cobalt and 0.06-0.26 for nickel. Within-series repeatability and day-to-day reproducibility (among series) coefficients of variation (CVs) were, respectively, 11.7 and 16% for lead, 2.5 and 7.5% for cadmium, 7.2 and 18.8%for cobalt, and 8.1 and 8.6% for nickel at respective concentrations of 1.89, 0.40, 1.25 and 11.8 ng g-1.

Animals↗

Lipid peroxidation in human milk and infant formula: effect of storage, tube feeding and exposure to phototherapy.

Preformed lipid peroxidation products present in the feed may contribute to the total reactive oxygen radical load infants have to deal with and may play a role in the pathogenesis of necrotizing enterocolitis and bronchopulmonary dysplasia. In this study, the occurrence of lipid peroxidation in human milk and feeding formulas for preterm babies was evaluated in vitro. Free linoleic acid (18:2) and its hydroperoxide (18:2OOH) were measured by gas chromatography-mass spectrometry and the concentration of 18:2OOH and the 18:2OOH/18:2 ratio were used as indices of peroxidation. In all feeds peroxidation products were present, but the proportion of peroxidized 18:2 was greater in infant formula. Storage of human milk (+4 degrees C for four days) increased lipid peroxidation. Exposure to light during tube feeding increased peroxidation in infant formula but not in human milk. Different procedures for preparation, storage and feeding may decrease the concentration of these potentially toxic peroxidized lipids in human milk and infant formula.

Enteral Nutrition↗

Exposure of cadmium from infant formulas and weaning foods.

Cadmium levels were determined in 59 baby food samples, including milk-based, cereal and milk-based and soy-based formulas, recommended from 0 to 18 months of age. Determinations were performed by Zeeman graphite furnace atomic absorption spectrophotometry, after dry ashing, with parallel determinations of certified reference samples. Mean cadmium levels were found to range from 1.10 to 23.5 micrograms/kg fresh weight concentrated formulas. Levels were related to the composition of the diets. Formulas based on cow's milk had the lowest concentrations. Soy formulas contained approximately six times more cadmium than cow's milk formulas, and diets with a cereal content had 4-21 times higher mean levels. The mean weekly intakes of dietary cadmium were estimated to vary between 0.10 and 3.05 micrograms/kg body weight of the child, if the recommended amount of formula were to be consumed at the recommended age, and if the child were of average weight. This estimation however does not include the contribution of cadmium from drinking water. The highest intake on a body weight basis was found in 6-month-old children, consuming the recommended amount of wheat-, oat- and milk-based formulas. This intake is below the provisional tolerable weekly intake (PTWI) of 7 micrograms/kg body weight, established by a WHO/FAO expert group. However, the risk assessment is based on renal effects in adults. Furthermore, preparations of a formula by adding water with cadmium concentrations at the WHO guideline level could lead to intakes at the PTWI level. Compared to breast-fed children, the exposure of dietary cadmium from weaning diets can be up to 12 times higher in children fed infant formula.

Cadmium↗

Assessment of feeding different amounts of arachidonic and docosahexaenoic acids in preterm infant formulas on the fatty acid content of lipoprotein lipids.

This study evaluated preterm infants of less than 2.3 kg birth weight fed commercial formula (Preemie SMA) devoid of arachidonic acid (AA) and docosahexaenoic acid (DHA) and compared this control group with similar infant groups fed one of three formulas containing a range of 0.32-1.1% AA and 0.24-0.75% DHA in the fat component of the formula. An analogous group of infants fed on their mothers' breast milk and a breast milk fortifier was also studied. Individual lipoprotein fractions were isolated from blood samples collected at 12 d of age and after a further 4 wk of feeding. The fatty acid content of individual lipid components, isolated from each lipoprotein fraction was quantitatively determined in order to identify change in marker pools of essential fatty acid. The high density lipoproteins (HDL) and low density lipoproteins (LDL) phospholipid and cholesterol ester fractions contain most of the AA and DHA found in the lipoprotein fractions (total of 0.49% and 0.35%, respectively). Infants fed a formula without AA and DHA showed a reduction in AA level in the phospholipid fraction of all lipoproteins and in the HDL and LDL cholesterol ester fraction. A reduced level of DHA was also observed primarily in the lipoprotein phospholipid fraction in comparison with infants fed breast milk or infant formula containing AA and DHA. Supplementing infant formula with increasing levels of AA and DHA produced a clear dose response in the level of AA found in the HDL and LDL phospholipid fraction. From comparison of the fatty acid levels present in the lipoproteins it appears that a formula level of 0.49% AA and 0.35% DHA provides sufficient levels of these fatty acids to achieve a similar fatty acid content to that of infants fed breast milk for the major lipoprotein fractions examined.

Arachidonic Acid↗

Efficacy and safety of docosahexaenoic acid and arachidonic acid addition to infant formulas: can one buy better vision and intelligence?

Long chain polyunsaturated fatty acids (LCPUFA) namely arachidonic acid (ARA, 20:4n-6) and docosahexaenoic acid (DHA, 22:6n-3) are highly concentrated in the phospholipid bilayer of biologically active brain and retinal neural membranes and are important in phototransduction and neuronal function. The rationale for adding these LCPUFA to infant formula (IF) was primarily because of their presence in large quantities in the retina and brain and in human milk. In addition, infants fed IF containing LCPUFA and breastfed infants have comparable ARA and DHA levels in red cell and plasma, in contrast to the lower ARA and DHA levels in those fed IF containing only the essential fatty acids: linoleic (LA, 18:2n-6) and linolenic (LNA, 18:3n-3), the precursors to ARA and DHA, respectively. However, functional benefits in particular visual or neural development from IF containing LCPUFA remains controversial. Potential for excessive and/or imbalanced intake of n-6 and n-3 fatty acids exists with increasing fortification of LCPUFA to infant foods other than IF.

Arachidonic Acid↗

Detection of counterfeit and relabeled infant formulas by high-pH anion-exchange chromatography-pulsed amperometric detection for the determination of sugar profiles.

High pH anion-exchange separation with pulsed amperometric detection (HPAE-PAD) is used to characterize various milk-based, soy-based, and protein hydrolysate infant formulas based on carbohydrate profiles. Counterfeit and relabeled formulas are compared to authentics. Figures of merit are shown for glucose, fructose, lactose, sucrose, and maltose.

Animals↗

Content of thyroid hormones in human and animal milk and in cow milk based infant formulas.

The concentration of thyroxine (T4), triiodothyronine (T3), thyroxine binding globulin (TBG) as well as thyroxine binding to proteins (TBP) and free thyroxine index (FTI) were measured in human milk. In addition, the concentration of T4 in milk of rats, rabbits and cows and in cow milk based infant formulas was measured. It was found that the concentration of T4 and T3 in human milk or that of T4 in rat and rabbit milk is much higher than in corresponding colostrum. The concentrations of TGB and also TBP and FTI in human milk were much less than in blood serum. In cow milk and in cow milk based products used for infant nutrition the concentration of T4 was either undetectable or only traces were found. The data suggest that the hormonal interrelation between the mother and the fetus should operate even in the postnatal period via the excretion of some hormones by milk.

Animals↗

Choline, phosphatidylcholine and sphingomyelin in human and bovine milk and infant formulas.

Choline is a precursor for the biosynthesis of phosphatidylcholine (lecithin), sphingomyelin, and choline plasmalogens--all essential constituents of membranes. Choline is also needed to make acetylcholine, a major neurotransmitter. The major choline-containing compounds of human milk (unesterified choline, phosphatidylcholine, sphingomyelin) were measured in samples obtained from mothers of full-term infants. Unesterified choline concentrations were highest (greater than 600 nmol/ml) during the first week of lactation, but thereafter remained relatively constant at 70-200 nmol/ml. There was no difference among foremilk, middle milk and hind milk, nor was there a diurnal pattern of variation in unesterified choline concentrations. Milk phosphatidylcholine and sphingomyelin concentrations remained relatively constant throughout lactation (100-200 nmol/ml). Hind milk always contained more of these phospholipids than did foremilk or middle milk. There was no consistent diurnal pattern of variation in milk concentrations of phosphatidylcholine or sphingomyelin. Milk contained no phospholipase activity capable of forming free choline from phosphatidylcholine or sphingomyelin. Bovine milk contained approximately the same concentrations of choline, phosphatidylcholine and sphingomyelin as did human milk from mothers more than 15 d postpartum. The same was true of "humanized" infant formulas made from cow's milk. Soy protein-based formulas had much more unesterified choline (up to 650 nmol/ml) and much less sphingomyelin than did mature human milk.

Adult↗

[New infant formula based on soy milk and corn flour].

In recent years the use of infant feeding formulas based on soy bean has been increasing. We report a trial of a new, soy bean-based formula to which starch was added. A control group was given a cow milk-based formula widely used in this country ("Materna" with cornflour, Ma'abaroth Products Ltd.). Healthy infants, between 3-6 months of age and of normal length and weight were enrolled at random at Infant Welfare Centers in Jerusalem. 25 received the soy formula, and 21 the cow milk-based formula. During the 2-month trial the infants did not receive any other foods. They were examined by the senior author every 2 weeks and weight, length and head circumference were measured at their Infant Welfare Clinic visits. The mothers filled questionnaires concerning the amount of formula ingested, the nature and number of stools, and complaints, such as crying. Both groups gained weight at the normal rate during the trial. In each group, the mothers were satisfied with the product and wanted to continue with it after the trial. We conclude that a soy-based formula containing starch is comparable to a cow milk formula containing star.

Body Height↗