Impact of milk secretor status on the fecal metabolome and microbiota of breastfed infants.

Maternal secretor status has been shown to be associated with the presence of specific fucosylated human milk oligosaccharides (HMOs), and the impact of maternal secretor status on infant gut microbiota measured through 16s sequencing has previously been reported. None of those studies have confirmed exclusive breastfeeding nor investigated the impact of maternal secretor status on gut microbial fermentation products. The present study focused on exclusively breastfed (EBF) Indonesian infants, with exclusive breastfeeding validated through the stable isotope deuterium oxide dose-to-mother (DTM) technique, and the impact of maternal secretor status on the infant fecal microbiome and metabolome. Maternal secretor status did not alter the within-community (alpha) diversity, between-community (beta) diversity, or the relative abundance of bacterial taxa at the genus level. However, infants fed milk from secretor (Se+) mothers exhibited a lower level of fecal succinate, amino acids and their derivatives, and a higher level of 1,2-propanediol when compared to infants fed milk from non-secretor (Se-) mothers. Interestingly, for infants consuming milk from Se+ mothers, there was a correlation between the relative abundance of Bifidobacterium and Streptococcus, and between each of these genera and fecal metabolites that was not observed in infants receiving milk from Se- mothers. Our findings indicate that the secretor status of the mother impacts the gut microbiome of the exclusively breastfed infant.

Chicken liver and eggshell crackers as a safe and affordable animal source food for overcoming micronutrient deficits during pregnancy and lactation in Indonesia: a double-blind, randomised placebo-controlled trial (SISTIK Growth Study).

Background: Indonesia ranks fifth in terms of the number of stunted children and there has been little change in the stunting prevalence in the last decade. In earlier observational studies conducted in 2014-2015, we identified several key underlying problems with the potential to impact stunting in Sumedang district, West Java, Indonesia. Deficits in intakes of growth-limiting micronutrients were observed, most notably calcium, iron, zinc, and vitamin A, emphasizing the need for a food-based intervention to overcome these micronutrient deficits in the diets of mothers and their infants. Methods: A double-blind placebo-controlled cluster randomised trial comparing the effect of daily consumption of 75 grams of locally produced micronutrient-enriched crackers (MEC) (intervention group) compared to placebo crackers (control group) by mothers at two-time intervals: (i) from the 8-14 weeks of pregnancy to delivery (i.e., 28-34 weeks of consumption of MEC) on birth length, and (ii) from the 8-14 weeks of pregnancy to 5 months post-partum on attained linear growth and linear growth velocity of breast-fed infants. A total of 324 pregnant women from 28 clusters (villages) located in 3 sub-districts in Sumedang district, West Java, Indonesia, will be randomly assigned to either intervention (n=14 villages) or control (n=14 villages). Discussi on: This will be the first study in Indonesia to use crackers based on powdered eggshells and chicken liver, in a form which is acceptable, safe, and has a long shelf life. If daily consumption of MEC for 6 months during pregnancy can enhance birth length, or their continued daily consumption for 5 months postpartum improves both attained and incremental linear growth at 5 months of age, then scaling-up in Indonesia may be considered. Trial Registration: https://clinicaltrials.gov/ct2/show/NCT04564222 ; 25 th September 2020.

Determination of modifiable risk factors for length-for-age z-scores among resource-poor Indonesian infants.

To reduce the burden of early-life linear growth faltering in low- and middle-income countries, interventions have focused on nutrition strategies, sometimes combined with water quality, sanitation, and hygiene (WASH). However, even when combined, their effects on linear growth have been inconsistent. Here, we investigate potential predictors of length-for-age z-scores (LAZ) in a cohort of resource-poor rural Indonesian infants to inform the optimal strategies to reduce linear growth faltering. Apparently healthy rural breastfed Indonesian infants were randomly selected from birth registries at age 6 months (n = 230) and followed up at 9 (n = 202) and 12 (n = 190) months. Using maximum likelihood estimation, we examined longitudinal relationships among socio-demographic status, maternal height, infant sex, age, water source, sanitation facility, energy, protein, micronutrient intakes and biomarkers (serum ferritin, zinc, retinol binding protein (RBP), selenium-adjusted for inflammation), and α-1-acid glycoprotein (AGP) and C-reactive protein (CRP) (systemic inflammation biomarkers) at age 6 and 9 months on LAZ at age 9 and 12 months. Stunting (LAZ <-2) at 6, 9, and 12 months was 15.7%, 19.3%, and 22.6%, respectively. In the full model, the predictor variable at age 6 months that was most strongly associated with infant LAZ at 9 months was maternal height (0.18 (95% CI 0.03, 0.32) SD). At age 9 months, the strongest predictors of LAZ at 12 months were improved drinking water source (-0.40 (95% CI -0.65, -0.14) vs. not improved), elevated AGP compared to not elevated (0.26 (95%CI -0.06, 0.58), maternal height (0.16 (95% CI 0.02, 0.31) SD), sex (0.22 (95% CI -0.02,0.45) female vs. male), serum RBP (0.12 (95% CI -0.01, 0.25) SD), and protein intake (0.17 (95% CI -0.01, 0.35) SD). Health promotion that includes exclusive breastfeeding up to the first six months and follows microbial water quality guidelines to ensure water intake is always safe should be considered.

Differences in Micronutrient Intakes of Exclusive and Partially Breastfed Indonesian Infants from Resource-Poor Households are Not Accompanied by Differences in Micronutrient Status, Morbidity, or Growth.

BACKGROUND: When maternal micronutrient intakes and statuses are compromised, reductions in micronutrient concentrations in neonatal stores and human milk may result in suboptimal micronutrient intakes, statuses, and functional outcomes of breastfed infants during the critical first 6-month period. OBJECTIVES: We compared the adequacy of micronutrient intakes and statuses at 2 and/or 5 months and morbidity and growth faltering at 2, 5, and 12 months in a cohort of exclusively breastfed (EBF) and partially breastfed (PBF) infants from low-resource Indonesian households. METHODS: At 2 and 5 months, the breastfeeding status and human milk intake of 212 infants were determined using the deuterium oxide dose-to-mother technique, and intakes were calculated from milk micronutrient concentrations and 3-d weighed food intakes. At 5 months, five infant micronutrient biomarkers, hemoglobin, C-reactive protein, and α-1-acid-glycoprotein were measured. Infant morbidity, weight, and length were measured at 2, 5, and 12 months. Means, medians, or proportions were reported for each group and differences between groups were statistically determined. RESULTS: Median intakes of iron, thiamin, niacin, and vitamin B-12 were higher in PBF than EBF infants at 5 months (all P values < 0.05), but intakes in all infants were below adequate intakes. At 5 months, anemia was <20% in both groups, although fewer PBF versus EBF infants had vitamin B-12 deficiency (11.5% vs. 28.6%, respectively; P = 0.011). The mean ± SD length-for-age z-scores for EBF versus PBF infants at 2 months were 0.7 ± 0.9 versus -0.5 ± 1.1, respectively  (P = 0.158), declining to -1.4 ± 0.9 versus -1.1 ± 1.2, respectively, at 12 months (P = 0.059). Reported morbidity rates were generally low, with no evidence of a difference between infant groups (all P values > 0.126). CONCLUSIONS: Irrespective of exclusive or partial breastfeeding status, micronutrient intakes of infants were low, statuses were compromised, and growth faltering during the critical 6 months period of early infancy was present. The findings highlight the importance of improving maternal nutritional statuses and evaluating their impacts on infant outcomes.

Association of maternal diet, micronutrient status, and milk volume with milk micronutrient concentrations in Indonesian mothers at 2 and 5 months postpartum.

BACKGROUND: Maternal micronutrient deficits during preconception and pregnancy may persist during lactation and compromise human milk composition. OBJECTIVE: We measured micronutrient concentrations in human milk and investigated their association with maternal micronutrient intakes, status, and milk volume. METHODS: Infant milk intake (measured via a deuterium dose-to-mother technique), milk micronutrient and fat concentrations, and maternal micronutrient intakes were assessed at 2 and 5 mo postpartum in 212 Indonesian lactating mother-infant pairs. Maternal hemoglobin, ferritin, transferrin receptors, retinol binding protein (RBP), zinc, selenium, and vitamin B-12 were measured at 5 mo (n = 163). Multivariate or mixed effects regression examined associations of milk micronutrient concentrations with maternal micronutrient intakes, status, and milk volume. RESULTS: Prevalence of anemia (15%), and iron (15% based on body iron), selenium (2.5%), and vitamin B-12 deficiency (0%) were low compared with deficiencies of zinc (60%) and vitamin A (34%). The prevalence of inadequate intakes was >50% for 7 micronutrients at 2 and 5 mo. Median milk concentrations for most micronutrients were below reference values, and nearly all declined between 2 and 5 mo postpartum and were not associated substantially with milk volume (except for ß-carotene, α-carotene, and ß-cryptoxanthin). At 5 mo postpartum, associations between maternal micronutrient status and corresponding milk concentrations reported as mean percentage difference in human milk concentration for each unit higher maternal biomarker were significant for hemoglobin (1.9%), iron biomarkers (ranging from 0.4 to 7%), RBP (35%), selenium (70%), and vitamin B-12 (0.1%), yet for maternal intakes only a positive association with ß-carotene existed. CONCLUSIONS: Most milk micronutrient concentrations declined during lactation, independent of changes in human milk production, and few were associated with maternal micronutrient intakes. The significant associations between maternal biomarkers and milk micronutrient concentrations at 5 mo warrant further study to investigate whether the declines in milk micronutrients are linked to shifts in maternal status.

Examination of Carbohydrate Products in Feces Reveals Potential Biomarkers Distinguishing Exclusive and Nonexclusive Breastfeeding Practices in Infants.

BACKGROUND: The stable isotope deuterium dose-to-mother (DTM) technique to estimate nonbreast milk water intake demonstrates that maternal self-report methods of infant feeding overestimate the true prevalence of exclusively breastfeeding practices. OBJECTIVE: We aimed to determine potential monosaccharide and oligosaccharide markers that distinguish between exclusively breastfed (EBF) versus nonexclusively breastfed (non-EBF) infants utilizing LC-MS-based methods. METHODS: Data for the analysis were collected as part of a larger, longitudinal study of 192 breastfed Indonesian infants aged 2 mo and followed up at 5 mo. Feces samples were collected from infants aged 2 mo (n = 188) and 5 mo (n = 184). EBF and non-EBF strata at each time point were determined via the DTM technique. Feces samples were analyzed to determine monosaccharide content using ultra-high-performance LC-triple quadrupole MS (UHPLC-QqQ MS). Relative abundances of fecal oligosaccharides were determined using nano-LC-Chip-quadrupole time-of-flight MS (nano-LC-Chip-Q-ToF MS). RESULTS: At age 2 mo, monosaccharide analysis showed the abundance of fructose and mannose were significantly higher (+377% and +388%, respectively) in non-EBF compared with EBF infants (P <0.0001). Fructose and mannose also showed good discrimination with areas under the curve (AUC) of 0.86 and 0.82, respectively. Oligosaccharide analysis showed that a 6-hexose (Hex6) isomer had good discrimination (AUC = 0.80) between EBF and non-EBF groups at 5 mo. CONCLUSION: Carbohydrate products, particularly fecal mono- and oligosaccharides, differed between EBF and non-EBF infants aged under 6 mo and can be used as potential biomarkers to distinguish EBF versus non-EBF feeding practices.

Poor dietary diversity and low adequacy of micronutrient intakes among rural Indonesian lactating women from Sumedang district, West Java.

Information on micronutrient adequacy of diets of rural Indonesian lactating women is lacking, despite their high nutrient requirements. This is of concern because deficits in micronutrient intakes may compromise the health of both mothers and infants. This study aimed to assess micronutrient adequacy and dietary diversity (DD) among rural lactating women and explore relationships between micronutrient adequacy, DD, and intakes of energy and food groups consumed. We measured in-home 12-h weighed food records and 12-h recalls over three non-consecutive days from 121 exclusively breastfeeding women at 2-5 months postpartum. Next, we calculated intakes of energy and 11 micronutrients and estimated probability of adequacy (PA) for usual intakes of 11 micronutrients for each women taking into account national fortification of wheat flour with thiamin, riboflavin, folate, zinc, and iron. We assessed DD from nine food groups consumed. Energy and macronutrient balance were within recommended ranges, yet population prevalence of adequacy was less than 50% for niacin, vitamins B6 and C, and less than 60% for calcium, vitamin B12 and vitamin A, all micronutrients not targeted by the national wheat flour fortification program. In contrast, population prevalence of adequacy for the fortified micronutrients was at least 60%, with iron and zinc attaining 79% and 97%, respectively. Overall mean population prevalence of micronutrient adequacy was 57% and mean (±SD) DD score was 4.3±1.2. Mean PAs, a composite measure based on individual PAs over 11 micronutrients, were strongly correlated with energy intakes and with DD scores. In the multivariate models with maternal education and wealth index as covariates, organ meats were the most important determinant of mean PA after controlling for energy intake. In conclusion, despite wheat flour fortification, lactating mothers remained at risk of multiple micronutrient inadequacies. Increasing intakes of animal source foods including organ meats, and fruits and vegetables should be considered.

Micronutrient intakes of lactating mothers and their association with breast milk concentrations and micronutrient adequacy of exclusively breastfed Indonesian infants.

BACKGROUND: Breast milk is the sole source of nutrition for exclusively breastfed infants in the first 6 mo of life, yet few studies have measured micronutrient concentrations in breast milk in light of maternal diet and subsequent infant micronutrient intakes. OBJECTIVES: We evaluated the adequacy of micronutrient intakes of exclusively breastfed Indonesian infants by measuring milk volume and micronutrient concentrations and assessed maternal micronutrient intakes and their relationship with milk concentrations. METHODS: Mother-infant (2-5.3 mo) dyads (n = 113) were recruited for this cross-sectional study. Volume of breast-milk intake via the deuterium dose-to-mother technique over 14 d and analyzed micronutrient concentrations were used to calculate micronutrient intakes of exclusively breastfed infants. Maternal 3-d weighed food records were collected to assess median (IQR) micronutrient intakes. Multivariate regression analyses examined the association of usual maternal micronutrient intakes with milk micronutrient concentrations after adjustment for confounding variables. RESULTS: Mean ± SD intake of breast-milk volume was 787 ± 148 mL/d. Median daily infant intakes of iron, zinc, selenium, magnesium, sodium, and B-vitamins (thiamin, riboflavin, niacin, pantothenic acid, B-6, and B-12) were below their respective Adequate Intakes. Inadequacies in maternal intakes (as % < estimated average requirements) were >40% for calcium, niacin, and vitamins A, B-6, and B-12. Significant positive associations existed between maternal usual intakes of vitamin A, niacin and riboflavin and milk retinol, nicotinamide, and free riboflavin concentrations in both unadjusted and adjusted (for infant age, milk volume, and parity) analyses (all P < 0.05). CONCLUSIONS: The majority of micronutrient intakes for these exclusively breastfed infants and their mothers fell below recommendations, with associations between maternal intakes and breast-milk concentrations for 3 nutrients. Data on nutrient requirements of exclusively breastfed infants are limited, and a better understanding of the influence of maternal nutritional status on milk nutrient concentrations and its impact on the breastfed infant is needed.

Multimicronutrient Biomarkers Are Related to Anemia during Infancy in Indonesia: A Repeated Cross-Sectional Study.

BACKGROUND: Anemia during infancy in Indonesia is common, with iron deficiency (ID) assumed to be the major cause. Other micronutrients besides iron may have a role in determining hemoglobin (Hb) but have not yet been explored in Indonesia. OBJECTIVE: We investigated 7 micronutrient biomarkers and selected nonnutritional factors as potential predictors of Hb and anemia at ages 6, 9, and 12 mo in a cohort of Indonesian infants at risk of coexisting micronutrient deficiencies. METHODS: Apparently healthy breastfed infants were randomly selected from birth registries at 6 mo (n = 230) and followed-up at 9 mo (n = 202) and 12 mo (n = 190). Hb, serum micronutrient biomarkers-iron [as ferritin and serum soluble transferrin receptor (sTfR)], zinc, selenium, folate, vitamin A [as retinol-binding protein (RBP)], vitamin B-12, and vitamin D (as 25-hydroxyvitamin D) (adjusted for inflammation, where appropriate)-and maternal sociodemographic status, health, BMI, heminthiasis, and selected Hb genetic disorders were measured. Multivariate analysis examined relations between micronutrient biomarkers and nonnutritional factors (except helminthiasis and genetic Hb disorders) with Hb and anemia at 6 and 12 mo. RESULTS: ID (based on ferritin) was a predictor of lower Hb and anemia at both 6 and 12 mo of age (P < 0.02). Additional predictors at 6 mo were tertiary education and higher maternal Hb for higher Hb, sex (being male) and inflammation (P < 0.05) for both lower Hb and anemia, and greater maternal height (P = 0.036) for anemia only. At 12 mo, a significant biomarker predictor besides ID was RBP (P = 0.035) for Hb. CONCLUSION: ID was a major contributor to lower Hb and anemia, although RBP was also associated.

An Acceptability Trial of Desiccated Beef Liver and Meat Powder as Potential Fortifiers of Complementary Diets of Young Children in Indonesia.

The addition of desiccated beef liver to infant and young child complementary foods can be used to overcome nutrient deficits, however its acceptability is unknown. We conducted a series of studies to test the acceptability of complementary foods fortified with either powdered beef liver, beef meat, beef liver + meat or placebo among 96 Indonesian children aged 12 to 23 mo. This was achieved by determining liking of a single test food with added study powder, followed by a 2-wk home trial and focus group discussions to assess liking during repeated consumption of the study powders added to daily meals. The test food with added beef powders were well liked by mothers, with liking scores never falling below neutral on a 7-point scale. After home use, mothers reported that their children moderately liked their meals with added powder, with scores ranging between 3.3 and 3.5 on a 5-point scale. With the exception of lower liking for the combination beef liver + meat powder, there were no detectable differences in mothers' overall perception of child's liking between the placebo and any of the study powders. The low disappearance rate of the study powders during the home trial was a concern, with mothers reporting a strong smell and fishy odor as the major reason why children did not like their meals. Nonetheless, mothers declared they would continue using the powder on account of the nutritional value and perceived health benefits. Strategies are underway to minimize the level of fishy odor in the beef liver powder.