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Human milk is the sole and recommended nutrition for the newborn infant and contains one of the largest constituents of diverse oligosaccharides, dubbed human milk oligosaccharides (HMOs). Preclinical and clinical association studies indicate that HMOs have multiple physiological functions largely mediated through the establishment of the gut microbiome. Until recently, HMOs were not available to investigate their role in randomized controlled intervention trials. To our knowledge, this is the first report on the effects of 2 HMOs on establishing microbiota in newborn infants. We provide a detailed description of the microbiota changes observed upon feeding a formula with 2 HMOs in comparison to breastfed reference infants' microbiota. Then, we associate the microbiota to long-term health as assessed by prescribed antibiotic use. Human milk oligosaccharides (HMOs) may provide health benefits to infants partly by shaping the development of the early-life intestinal microbiota. In a randomized double-blinded controlled multicentric clinical trial, healthy term infants received either infant formula (control) or the same formula with two HMOs (2′-fucosyllactose and lacto-N- neo tetraose; test) from enrollment (0 to 14 days) to 6 months. Then, all infants received the same follow-up formula without HMOs until 12 months of age. Breastfed infants (BF) served as a reference group. Stool microbiota at 3 and 12 months, analyzed by 16S rRNA gene sequencing, clustered into seven fecal community types (FCTs) with marked differences in total microbial abundances. Three of the four 12-month FCTs were likely precursors of the adult enterotypes. At 3 months, microbiota composition in the test group ( n  = 58) appeared closer to that of BF ( n  = 35) than control ( n  = 63) by microbiota alpha (within group) and beta (between groups) diversity analyses and distribution of FCTs. While bifidobacteriaceae dominated two FCTs, its abundance was significantly higher in one (FCT BiH for Bifidobacteriaceae at high abundance) than in the other (FCT Bi for Bifidobacteriaceae ). HMO supplementation increased the number of infants with FCT BiH (predominant in BF) at the expense of FCT Bi (predominant in control). We explored the association of the FCTs with reported morbidities and medication use up to 12 months. Formula-fed infants with FCT BiH at 3 months were significantly less likely to require antibiotics during the first year than those with FCT Bi. Previously reported lower rates of infection-related medication use with HMOs may therefore be linked to gut microbiota community types. (This study has been registered at ClinicalTrials.gov under registration number NCT01715246.) IMPORTANCE Human milk is the sole and recommended nutrition for the newborn infant and contains one of the largest constituents of diverse oligosaccharides, dubbed human milk oligosaccharides (HMOs). Preclinical and clinical association studies indicate that HMOs have multiple physiological functions largely mediated through the establishment of the gut microbiome. Until recently, HMOs were not available to investigate their role in randomized controlled intervention trials. To our knowledge, this is the first report on the effects of 2 HMOs on establishing microbiota in newborn infants. We provide a detailed description of the microbiota changes observed upon feeding a formula with 2 HMOs in comparison to breastfed reference infants' microbiota. Then, we associate the microbiota to long-term health as assessed by prescribed antibiotic use.

Levels of harmful nitrogen-containing substances have increased in many foods and drinks around the world. The impacts of dietary nitrate and nitrite on human health have been controversial topics for many years. The present study aimed to quantify the levels of nitrate and nitrite in infant formulas and baby foods sold in Iran and to estimate non-carcinogenic human health risk from exposure to these substances. The samples were gathered randomly and subsequently analyzed to detect nitrate and nitrite via Spectrophotometry. The ranges of nitrate and nitrite in the infant formulas were 0.221–1.347 (mean 0.645) mg/kg and 0.045–0.263 (mean 0.151) mg/kg, respectively. For baby foods, the ranges of nitrate and nitrite were 0.24–1.93 (mean 0.99) mg/kg and 0.04– 1.45 (0.36) mg/kg. Estimated Daily Intake (EDI) values of nitrate and nitrite in all the samples were below the acceptable daily intake (nitrate ADI = 3.7 mg/kg bw/day and nitrite ADI = 0.07 mg/kg bw/day established by FAO/WHO), showing that levels of these contaminants in infant formulas and baby foods may not cause toxicity in the infant and baby population. None of the infant formulas and baby foods in this study showed evidences of a non-cancer risk to the consumers. However, it is suggested that levels of nitrate and nitrite in these products and their related health risks be constantly monitored to prevent significant health hazards in the future.

BACKGROUNDOsteopontin (OPN), a multifunctional protein, is present abundantly in human milk, but not in bovine milk and infant formulas. A recent randomized clinical trial showed that supplementing infant formula with bovine milk OPN (bOPN) resulted in better immune outcomes.METHODSHuman milk OPN (hOPN) concentrations were analyzed by ELISA. Plasma samples were obtained from infants receiving one of four treatments: breast milk (BF), unsupplemented formula (F0), formula supplemented with 65 mg/L bOPN (F65), or with 130 mg/L bOPN (F130). Plasma samples were analyzed for hOPN and bOPN by ELISA.RESULTSThe hOPN concentration was high in early lactation (D1 to D8), decreased gradually after D9, and deceased significantly after 1 month. At 4 and 6 months, higher levels of hOPN were found in plasma samples from the BF, F65, and F130 groups than in samples from the F0 group; the plasma bOPN concentration in the F130 group was greater than that in the F65 group.CONCLUSIONDynamic changes in the concentration of milk OPN may reflect infant needs for different amounts of milk OPN for various functions at different developmental stages. Supplemental bOPN in infant formula may exert its beneficial effects by increasing endogenous OPN in plasma.

Background: Post-natal gut maturation in infants interrelates maturation of the morphology, digestive, and immunological functions and gut microbiota development. Here, we explored both microbiota development and markers of gut barrier and maturation in healthy term infants during their early life to assess the interconnection of gut functions during different infant formulae regimes. Methods: A total of 203 infants were enrolled in this randomized double-blind controlled trial including a breastfed reference group. Infants were fed starter formulae for the first four weeks of life, supplemented with different combination of nutrients (lactoferrin, probiotics (Bifidobacterium animal subsp. Lactis) and prebiotics (Bovine Milk-derived Oligosaccharides—BMOS)) and subsequently fed the control formula up to eight weeks of life. Stool microbiota profiles and biomarkers of early gut maturation, calprotectin (primary outcome), elastase, α-1 antitrypsin (AAT) and neopterin were measured in feces at one, two, four, and eight weeks. Results: Infants fed formula containing BMOS had lower mean calprotectin levels over the first two to four weeks compared to the other formula groups. Elastase and AAT levels were closer to levels observed in breastfed infants. No differences were observed for neopterin. Global differences between the bacterial communities of all groups were assessed by constrained multivariate analysis with hypothesis testing. The canonical correspondence analysis (CCA) at genus level showed overlap between microbiota profiles at one and four weeks of age in the BMOS supplemented formula group with the breastfed reference, dominated by bifidobacteria. Microbiota profiles of all groups at four weeks were significantly associated with the calprotectin levels at 4 (CCA, p = 0.018) and eight weeks of age (CCA, p = 0.026). Conclusion: A meaningful correlation was observed between changes in microbiota composition and gut maturation marker calprotectin. The supplementation with BMOS seems to favor gut maturation closer to that of breastfed infants.

BackgroundExcessive exposure to toxic trace elements through food ingestion can adversely affect infant health. This study assesses the carcinogenic and non-carcinogenic risks associated with trace elements (iron, selenium, zinc, chromium, mercury, cadmium, aluminium, barium, and strontium) in infant formula and complementary foods.MethodsA total of 80 formula milks and 27 baby food samples from 6 commercial brands have been collected from June 2020 to June 2021 from the Iranian market. The potential lifetime health risk posed to infants and toddlers is measured based on daily intake of elements, non-cancer hazard indices (HIs), hazard quotient (HQ), and cancer risks (CR).ResultsThe average concentrations of aluminium, cobalt, chromium, copper, iron, and zinc in infant formula and complementary foods significantly exceeded FAO/WHO standards across all seasons (P < 0.05). All HI values exceeded the safety threshold (HI > 1) for all age groups. Cancer risks ranged from 9.55 × 10⁻⁵ to 3.57 × 10⁻⁴, indicating potential carcinogenic and non-carcinogenic risks for infants and children.ConclusionsTherefore, authoritative action limits should be set for baby food manufacturers to evaluate both their finished products for toxic trace elements and phase out products that have high toxic trace elements.

The relationships between nutrition, metabolic response, early growth and later body weight have been investigated in human studies. The aim of this follow-up study was to assess the long-term effect of infant feeding on growth and to study whether the infant metabolome at the age of 4 months might predict anthropometry at 4 years of age. The Belgrade-Munich infant milk trial (BeMIM) was a randomized controlled trial in which healthy term infants received either a protein-reduced infant formula (1.89 g protein/100 kcal) containing alpha-lactalbumin enriched whey and long-chain polyunsaturated fatty acids (LC-PUFA), or a standard formula (2.2 g protein/100 kcal) without LC-PUFA, focusing on safety and suitability. Non-randomized breastfed infants were used as a reference group. Of the 259 infants that completed the BeMIM study at the age of 4 months (anthropometry assessment and blood sampling), 187 children participated in a follow-up visit at 4 years of age. Anthropometry including weight, standing height, head circumference, and percent body fat was determined using skinfolds (triceps, subscapular) and bioelectrical impedance analysis. Plasma metabolite concentration, collected in samples at the age of 4 months, was measured using flow-injection tandem mass spectrometry. A linear regression model was applied to estimate the associations between each metabolite and growth with metabolites as an independent variable. At 4 years of age, there were no significant group differences in anthropometry and body composition between formula groups. Six metabolites (Asn, Lys, Met, Phe, Trp, Tyr) measured at 4 months of age were significantly associated with changes in weight-for-age z-score between 1 to 4 months of age and BMI-for-age z-score (Tyr only), after adjustment for feeding group. No correlation was found between measured metabolites and long-term growth (up to 4 years of age). No long-term effects of early growth patterns were shown on anthropometry at 4 years of age. The composition of infant formula influences the metabolic profile and early growth, while long-term programming effects were not observed in this study.