Human Milk Samples Research Articles

Contaminant Exposure Profiles Demonstrate Similar Physiological Effects Across Environments Despite Unique Profile Composition in Formosa, Argentina, and Connecticut, USA.

Exposure to environmental contaminants is globally universal. However, communities vary in the specific combination of contaminants to which they are exposed, potentially contributing to variation in human health and creating "locally situated biologies." We investigated how environmental exposures differ across environments by comparing exposure profiles between two contexts that differ markedly across political, economic, and sociocultural factors-Namqom, Formosa, Argentina, and New Haven, Connecticut, United States. We collected infant urine, maternal urine, and human milk samples from mother-infant dyads in Formosa (n = 13) and New Haven (n = 21). We used untargeted liquid chromatography with high-resolution mass spectrometry (LC-HRMS) to annotate environmental contaminants and endogenous metabolites in these samples, and we analyzed the data using exposome-wide association studies (EWAS) followed by pathway enrichment. We found statistically significant differences between the chemical exposure profiles of the Argentinian and US mothers, mostly involving pesticides; however, we observed similarities in the infant urine and human milk environmental contaminant profiles, suggesting that the maternal body may buffer infant exposure through human milk. We also found that infants and mothers were exposed to contaminants that were associated with alterations in amino acid and carbohydrate metabolism. Infants additionally showed alterations in vitamin metabolism, including vitamins B1, B3, and B6. Differences in chemical exposure profiles may be related to structural factors. Despite variation in the composition of exposure profiles between the two study sites, environmental contaminant exposure was associated with similar patterns in human physiology when we considered contaminants comprehensively rather than individually, with implications for metabolic and cardiovascular disease risk as well as infant cognitive development.

Human milk oligosaccharide profiles remain unaffected by maternal pre-pregnancy body mass index in an observational study.

Human milk oligosaccharides (HMOs) are important carbohydrates in human milk that infants cannot digest, acting as prebiotics linked to infant health. The risk of childhood obesity increases with maternal obesity, potentially mediated through the gut microbiota affected by the available HMOs. Studies on whether maternal obesity affects HMO abundance, yield conflicting results. This study aimed to investigate the HMO profile and its association with maternal obesity measured by pre-pregnancy body mass index (BMI) and infant anthropometrics. The results were discussed in the context of existing literature. 90 human milk samples were collected at 3 months postpartum from mothers in three BMI-groups: 32 normal weight (BMI: 18.5-24.99 kg/m2), 34 overweight (BMI: 25-30 kg/m2), and 24 obese (BMI > 30 kg/m2). The samples were analyzed using nano liquid chromatography chip quadrupole time-of-flight mass spectrometry yielding 51 HMO structures and isomers. Their peak areas were integrated and normalized to determine relative abundances. Univariate and multivariate analysis showed associations between relative HMO abundance and donors' secretor status and specific infant anthropometric variables, but not with maternal pre-pregnancy BMI. This study does not support the hypothesis that maternal overweight influences the HMO profile and highlights the importance of reporting results despite absence of significant correlations.

Farm exposure is associated with human breast milk immune profile and microbiome.

Prenatal and early life farm exposure, and breastfeeding, are associated with protection from allergic diseases. We hypothesize that farm exposure influences the human breast milk microbiome and immune proteins. The immune protein profiles and microbial communities of 152 human breast milk samples were compared among three maternal farm exposure groups (traditional agrarian, farm, and non-farm) in rural Wisconsin to identify signatures associated with farm status and atopic disease. We found significant differences between farm groups for 23 immune proteins (p-adj<0.05), microbiome diversity (p=2.2E-05), and microbiome richness (p=8.0e-06). Traditional agrarian human breast milk had the highest immune protein levels and microbiome diversity and richness, followed by farm and non-farm human breast milk. Furthermore, Gram-positive bacterial species correlated with IL-23 mediated signaling events (p-adj<1.0E-05). These data suggest that increased farm exposures promotes human breast milk that is more microbially-diverse and rich in immune-associated proteins, ultimately influencing immune development in the infant.

Effects of gestational diabetes mellitus, parity and mode of delivery on polar lipids compositions of mature human milk

Polar lipids in human milk such as glycerophospholipids, sphingomyelin and plasmalogen play important role in growth and development of healthy infants. This work aims to investigate the effects of gestational diabetic mellitus (GDM), parity, delivery mode on the polar lipids’ compositions of human milk. A total of 56 mature human milk samples (1–6 months postpartum) were collected from breastfeeding mothers using electric breast pump from Ningbo, Zhejiang Province of China. Our results showed GDM and parity have limited effect on polar lipid content and composition. Delivery mode was found to significantly affect the polar lipids compositions in human milk with significantly higher total polar lipids in human milk of mothers who had cesarean delivery (938 nmol/mL) as compared to that of mothers who had vaginal delivery (740 nmol/mL). Discriminatory analysis shows human milk of mothers who have cesarean delivery demonstrated up-regulation of glycerophospholipids (GPL) and plasmalogen (PLG) esterified with unsaturated fatty acids (USFAs) [such as plasmenyl-ethanolamine 40:5 and 36:4].

Impact of Maternal Health and Stress on Steroid Hormone Profiles in Human Milk: Implications for Infant Development

Steroid hormones are biologically active factors in human milk (HM) that influence the physical and mental development of infants. Critically, maternal psychosocial stress has been associated with changes in HM steroid composition. This work aimed to characterize the steroid hormone profile of HM and pasteurized donor human milk (DHM) and assess the interplay between maternal physical and psychosocial status, the HM steroid profile, and infant outcomes.A targeted ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to quantify sixteen steroid hormones in HM samples. HM samples from mothers of term infants (N=42) and preterm infants (N=35) were collected at (i) recovery of birth weight or achievement of complete enteral nutrition, respectively, and (ii) 6 months later as well as DHM samples (N=19) from 11 donors. The physical and psychosocial status of mothers and infant neurodevelopment and temperament were assessed through structured interviews and validated questionnaires.Fourteen steroids were detected in HM/DHM samples, with cortisol, 20β-dihydrocortisol, dehydroepiandrosterone, pregnenolone, and cortisone being present in > 48% of samples. Pregnenolone, 17α-OH-progesterone, and dehydroepiandrosterone are reported for the first time in HM. Whereas milk cortisol levels were not directly related with maternal physical and psychosocial status nor with infant development, cortisone and pregnenolone correlated positively with maternal weight gain during pregnancy and were associated to maternal well-being and infant growth. The pasteurization process may have a detrimental effect on the steroid hormone levels in HM, what might influence the development of receptors.

P-phenylenediamines (PPDs) and PPD-quinones (PPD-Qs) in human urine and breast milk samples: Urgent need for focus on PPD-Qs and the establishment of health threshold criteria

PPDs and their oxidation products, PPD-Qs, are emerging environmental contaminants arising from the addition and oxidation of rubber products. Although numerous studies have been conducted to elucidate their risks, the primary focus has been on 6PPD and 6PPD-Q, with limited attention given to other PPDs and especially other PPD-Qs. This study comprehensively examines the occurrences of frequently used PPDs and their degradation products, PPD-Qs, in human urine and breast milk samples. The average concentrations of ΣPPDs and ΣPPD-Qs in urine were 27 ± 78 ng/mL and 16 ± 12 ng/mL, respectively. IPPD and DNPD were the predominant PPDs, while DPPD-Q, CPPD-Q, and IPPD-Q were the predominant PPD-Qs. Notably, the concentrations of 6PPD, CPPD, and DPPD were significantly lower than their oxidized quinone products. Weak or no correlations were observed between most PPDs and their corresponding PPD-Qs, suggesting that PPD-Qs in the human body are primarily derived from direct environmental intake rather than in vivo conversion of PPDs. PPDs and PPD-Qs were widely detected in breast milk, exhibiting concentrations and patterns similar to those found in urine, with comparable major pollutants. Estimated daily intakes of PPDs + PPD-Qs for infants were several μg/(kg·day), with the 95th percentile intake approaching 10 μg/(kg·day).

Concentrations and predictors of select nutrients in Canadian human milk samples from the Maternal-Infant Research on Environmental Chemicals pregnancy cohort

BackgroundHuman milk (HM) composition data are widely used in clinical, regulatory, and public health initiatives. The existing HM profiles in United States and Canadian nutrient databanks are outdated and now considered inappropriate to estimate current nutrient intakes. Recent reviews have underscored the limited North American data available to generate a new profile. ObjectivesTo describe concentrations and sources of variability of nutrients in HM from a large cohort collected in Canada. MethodsThe Maternal-Infant Research on Environmental Chemicals (MIREC) study recruited participants in the first trimester of pregnancy from 10 Canadian cities between 2008 and 2011. HM samples (n = 559–835, depending on nutrient) were collected 3–10 wk postpartum and analyzed for minerals (calcium, magnesium, phosphorus, potassium, sodium, manganese, molybdenum, zinc, copper, iodine, selenium), vitamin D [vitamin D3, 25-hydroxyvitamin D3], folate vitamers (folic acid, 5-methyltetrahydrofolate, total folates), and fatty acids (panel). We examined associations between participant characteristics and log-transformed nutrient concentrations using linear regression. ResultsConcentrations of HM components in MIREC samples were within the range observed in literature except for manganese, which was >100-fold lower than the value in the existing Canadian nutrient databank profile [2.43 (standard deviation 2.84) compared with 260 ng/g]. In multivariable models, concentrations of folate vitamers, vitamin D, and fatty acids demonstrated greater variability with maternal and sample characteristics than minerals. Factors such as relevant supplement use, body mass index, and for vitamin D, skin color and season, had a larger impact on nutrient concentrations than characteristics typically standardized in HM research, such as maternal or infant health, and method of collection. ConclusionsHM mineral concentrations from this study meet the methodological inclusion criteria for updating nutrient databank values and dietary reference intakes. Consideration of factors such as diet, skin color, and BMI will be important for selecting studies for developing representative reference values based on HM.

Maternal dietary protein and amino acid intake is not associated with the amino acid composition of human milk in an affluent environment.

Amino acids (AA) are essential nutrients in human milk (HM) and critical for infant growth and development. Several maternal lifestyle factors have been suggested to influence HM AA composition, with possible consequences for the breastfed infant. Whether maternal dietary protein and AA intake is associated with AA concentrations in HM is still largely unknown. Therefore, the aim of this study was to investigate the association between maternal dietary AA intake and AA concentrations in HM over the first month postpartum. Data from the observational longitudinal Amsterdam Mother's Milk study were used, consisting of 123 lactating women in their first postpartum month. HM samples were collected three times, on day 10, 17 and 24 postpartum. Maternal dietary protein and AA intake on these collection days was assessed using three 24-h recalls. HM protein-bound and free AA (BAA and FAA, respectively) were analysed by liquid chromatography. Associations between maternal AA intake and AA concentrations in HM were assessed using linear mixed models. Maternal intake was negatively associated with milk concentrations of free arginine (-0·0003; P = 0·01) and free lysine (-0·0004; P = 0·03) and was positively associated with free glutamine (0·002; P = 0·03) and free threonine (0·0008; P = 0·03). However, these associations were attenuated after correction for multiple testing. Both the quality and quantity of dietary protein intake in lactating women do not seem to influence the amino composition of their breast milk when living in an affluent environment.

Fat Content and Energy Calculation in Pasteurized Human Milk: Comparison Between Infrared Analysis and Creamatocrit Method.

Aims: This study aimed to analyze and compare the quantity of energy and fat using the infrared analysis and creamatocrit method in pasteurized human milk (HM) samples. Methods: This cross-sectional study analyzed 1,858 pasteurized human samples from 317 mothers at a single center. Infrared transmission spectrophotometry (Miris, Human Milk Analyser [HMA], Uppsala, Sweden) and the creamatocrit method were used to evaluate the quantity of energy and fat in pasteurized HM samples. Results: The average age of donor mothers was 29.7 ± 5.1 years, and the median duration of lactation was 22 days (interquartile range [IQ]: 7.7-59.2). Full-term births were observed in 196 (95.1%) of the women. The values of energy (difference: +8.96 kcal/dL, 95% CI: 8.52-9.44 kcal/dL; p < 0.001) and fat (difference: +0.40 g/dL, 95% CI: 0.35-0.45 g/dL; p < 0.001) in HM samples obtained by Miris were higher than those by the creamatocrit method. The energy calculated and the fat measured by Miris in the HM samples correlated moderately and directly with the obtained by creamatocrit (fat, r = 0.585; p < 0.001 and energy, r = 0.591; p < 0.01). The linear regression, adjusted for maternal age and lactation time, showed that the energy values calculated by creamatocrit were directly associated with those of Miris (energy kcal/dL = 38.43 + [0.516 × kcal/dL of creamatocrit]). Conclusion: The energy and fat quantity of pasteurized HM samples obtained by the creamatocrit and infrared methods were significantly correlated. However, the values calculated by the creamatocrit method were significantly lower than those by the infrared analyzer.

The effect of oral probiotics in the last trimester on the human milk and infant gut microbiotas at six months postpartum: A randomized controlled trial

ObjectiveThe main aim of this study was to evaluate the effect of oral probiotics on the human milk microbiota and determine whether that influenced infant microbiota development. MethodsA total of 27 pregnant women were recruited; 14 were assigned to the probiotic group, and the rest were assigned to the control group. Their infants were likewise assigned to the probiotic group or the control group. Pregnant women in the probiotic group received probiotic supplementation from 32 weeks of gestation until delivery. Human milk samples and infant fecal samples were collected at 6 months after delivery, and 16S rRNA sequencing was used to analyze the composition of the human milk and infant gut microbiota (NCT06241222). ResultsIn the control group, bacterial microbiota were detected in 8 out of 13 milk samples, whereas in the probiotic group, only 6 out of 14 milk samples contained bacterial microbiota. We examined the composition of the human milk and infant gut microbiota in both the control and probiotic groups. Spearman correlation analysis revealed that various genera in human milk were correlated with the infant gut microbiota. The Linear discriminant analysis effect size (LEfSe) showed that 6 bacteria in the human milk microbiota in the control group were significantly more abundant than those in the probiotic group. Nine bacteria were significantly more abundant in the human milk microbiota in the probiotic group than the control group. According to the LEfSe results, 11 bacteria in the infant gut microbiota in the control group were significantly more abundant than those in the probiotic group. Fourteen bacteria were significantly more abundant in the infant gut microbiota in the probiotic group than in the control group. ConclusionThe infant gut microbiota at 6 months has a complicated relationship with the maternal human milk microbiota. Oral probiotic supplementation can change the composition of the human milk microbiota and the infant gut microbiota.

Development and clinical implementation of an LC-HRMS method for ivacaftor, lumacaftor, tezacaftor and elexacaftor in human plasma and breast milk.

The four cystic fibrosis (CF) transmembrane conductance regulator (CFTR) modulators, ivacaftor, lumacaftor, tezacaftor, and elexacaftor, have revolutionised the treatment of CF by direct action on the protein target behind the disease's development. The aim was to develop and validate a quantification method for these CFTR modulators in plasma and breast milk to better understand inter-patient variability in pharmacokinetics and treatment outcome, including the risk of adverse drug reactions. The ability to monitor CFTR modulators in breast milk enables the estimation of the exposure of breastfed infant, with a potential concern for CFTR modulator-induced liver injury. The analysis was performed on a Thermo Vanquish Flex Binary UHPLC system coupled to a high-resolution mass spectrometer (HRMS), Thermo Q Exactive. The analytes were detected using positive electrospray ionisation in full scan mode. After sample preparation by protein precipitation, the supernatant was injected onto the LC system and the analytes were separated using a Zorbax SB-C18 Rapid Res HPLC column (3.5µm, 4.6 × 75mm). This is the first published method for CFTR modulators in breast milk. The validated quantification range for ivacaftor is 0.0050-10µg/mL with a coefficient of variation < 6% and a mean accuracy of 97-106%; for lumacaftor, tezacaftor, and elexacaftor, the validated quantification range is 0.050-100µg/mL with a coefficient of variation < 8% and a mean accuracy 93-106%. A simple and sensitive quantification method for CFTR modulators has been developed and used for routine analysis of human plasma and breast milk samples since 2022.

Subcutaneous Semaglutide during Breastfeeding: Infant Safety Regarding Drug Transfer into Human Milk.

Postpartum mothers and their healthcare providers often face the challenge of limited data regarding the safety of drug therapies during lactation. Pregnancy can lead to sustained weight gain, and obesity can negatively impact both physical and psychological well-being. The introduction of GLP-1 agonists to augment weight loss has become a topic of interest for many postpartum mothers. Our study aims to investigate the transmission of semaglutide into human milk in the first steps to ensure the safety and health of both lactating mothers and their breastfed infants. Semaglutide quantification was performed using high-resolution liquid chromatography-mass spectrometry. InfantRisk Center Human Milk biorepository released milk samples from eight women collected at 0, 12 and 24 h post-semaglutide administration. Semaglutide was extracted using protein precipitation in methanol, followed by chromatographic separation. Linear calibration curves for the method ranged between 2.5-30 ng/mL, with a limit of detection of 1.7 ng/mL and a limit of quantification of 5.7 ng/mL (LLOQ). Semaglutide was not detected in any of the collected human milk samples. A worst-case scenario of the relative infant dose (RID) was calculated using the LLOQ as the drug concentration in milk when considering semaglutide's bioavailability and long-acting dose profile. The maximum RID projected was 1.26%, far below the standard 10% safety threshold. While questions about long-term infant outcomes, the safety of maternal nutrient intake, and the nutrient content of breast milk remain, our findings suggest that semaglutide concentrations in human milk are unlikely to pose clinical concerns for breastfed infants. These results support healthcare providers in making informed decisions regarding postpartum therapeutic interventions.

Extracting and Characterizing Lactic Acid Bacteria from Human Milk

The greatest diet for babies is breast milk since it meets all of their nutritional needs and promotes healthy growth and development. Human milk contains thousands of different bacteria, the most prevalent ones being lactobacillus and Bifido bacterium. Since the use of probiotics is growing daily, it's important to comprehend their properties and health advantages. The combination of protein, fats, carbs, lipids, minerals, and vitamins found in breast milk helps to nourish an infant's body. Additionally, the bacteria in breast milk offer defense against infections. We will learn about lactic acid bacteria from this study so that we can commercially make probiotics from strains of these bacteria without utilizing lactose, as certain people are lactose intolerant and do not consume any lactose-containing products. The purpose of this article is to identify and isolate lactic acid bacteria while also discussing the advantages of probiotics for health. To understand the role lactic acid bacteria (LAB) from human milk play in the development and health of neonates, LAB must be extracted and identified. This study's objective was to identify, characterize, and segregate LAB strains from human milk samples obtained from nursing mothers in good health. The first isolation was carried out using selective medium, and then morphological, biochemical, and molecular characterization were performed. Through 16S rRNA gene sequencing, the isolates were identified. All things considered, the development of the neonatal gut microbiota and the overall health of infants depend on lactic acid bacteria (LAB). This review's objective is to gather the most recent data on the identification and isolation of LAB from human milk, with an emphasis on the techniques employed, the types of LAB discovered, and any potential health risks. Human milk is an essential source of beneficial bacteria, including several species of lactobacilli (LAB), due to its complex and dynamic nature. Our ability to isolate and accurately identify these germs has improved due to technological advancements in the molecular and microbiological sciences. This paper provides an extensive overview of the methods utilized to extract LAB from human milk, the genera and species that are commonly detected, and the implications of these findings for the nutrition and health of neonates.

Analysis of polycyclic aromatic hydrocarbons in Brazilian human milk: A simple and effective approach

Human milk is the best source of nutrition for infants due to its essential biologically active components. However, it can contain potentially harmful chemicals, such as polycyclic aromatic hydrocarbons, originating from contaminated air and food, which can cause adverse effects to infants. Using a Plackett-Burman design, a QuEChERS protocol was optimized to determine 13 polycyclic aromatic hydrocarbons in 79 Brazilian human milk samples by gas chromatography high-resolution mass spectrometry. Limits of quantification below 1 μg/kg were achieved for all compounds, with recoveries ranging between 61 % and 126 %. Six PAHs, including benzo(g,h,i)perylene, dibenzo(a,h)anthracene, fluoranthene, indeno(1,2,3-cd)pyrene, phenanthrene, and pyrene were detected in 40 samples. Mature milk exhibited the highest incidence of PAHs, with approximately 44 % of the positive samples, compared to 27 % for both colostrum and transitional milk. The concentrations detected varied between < LOQ and 1.14 μg/kg, with pyrene being the most frequently found PAH in the human milk samples. The estimated daily intake and margin of exposure of PAHs showed the highest values for babies aged 0–15 days for all PAHs evaluated. Despite the low levels of PAHs found in the analyzed samples, discontinuing breastfeeding is not recommended. This represents the first study on the incidence of PAHs in Brazilian human milk.

Presepsin in Human Milk Is Delivery Mode and Gender Dependent.

Breast milk (BM) is a unique food due to its nutritional composition and anti-inflammatory characteristics. Evidence has emerged on the role of Presepsin (PSEP) as a reliable marker of early sepsis diagnosis. In the present study, we aimed to investigate the measurability of PSEP in BM according to different maturation stages (colostrum, C; transition, Tr; and mature milks, Mt) and corrected for delivery mode and gender. We conducted a multicenter prospective case-control study in women who had delivered 22 term (T) and 22 preterm (PT) infants. A total of 44 human milk samples were collected and stored at -80 °C. BM PSEP (pg/mL) levels were measured by using a rapid chemiluminescent enzyme immunoassay. PSEP was detected in all samples analyzed. Higher (p < 0.05) BM PSEP concentrations were observed in the PT compared to the T infants. According to the grade of maturation, higher (p < 0.05) levels of PSEP in C compared to Tr and Mt milks were observed in the whole study population. The BM subtypes' degrees of maturation were delivery mode and gender dependent. We found that PSEP at high concentrations supports its antimicrobial action both in PT and T infants. These results open the door to further studies investigating the role of PSEP.

Association Between Prenatal Exposure to Per- and Poly-Fluoroalkyl Substances From Electronic Waste Disassembly Areas and Steroid Hormones in Human Milk Samples.

Per- and poly-fluoroalkyl substances (PFAS), which are long-lasting environmental contaminants that are released into the environment during the e-waste disassembly process, pose a threat to human health. Human milk is a complex and dynamic mixture of endogenous and exogenous substances, including steroid hormones and PFAS. Therefore, in this study, we aimed to investigate the association between PFAS and steroid hormones in human milk from women living close to an e-waste disassembly area. In 2021, we collected milk samples from 150 mothers within 4weeks of delivery and analyzed them via liquid chromatography-tandem mass spectrometry to determine the levels of 21 perfluorinated compounds and five steroid hormones (estrone, estriol, testosterone, progesterone, and androstenedione [A-dione]). We also performed multiple linear regression analysis to clarify the association between maternal PFAS exposure and steroid hormone concentrations. Our results indicated that PFOA and PFOS were positively associated with estrone (β, 0.23; 95% CI, 0.08-0.39) and A-dione (β, 0.186; 95% CI, 0.016-0.357) concentrations in human milk, respectively. Further, the average estimated daily intake of PFOA and PFOS were 36.5ng/kgbw/day (range, 0.52-291.7ng/kgbw/day) and 5.21ng/kgbw/day (range, 0.26-32.3ng/kgbw/day), respectively. Of concern, the PFAS intake of breastfeeding infants in the study area was higher than the recommended threshold. These findings suggested that prenatal exposure to PFAS from the e-waste disassembly process can influence steroid hormones levels in human milk. Increased efforts to mitigate mother and infant exposure to environmental pollutants are also required.

A simple step to improve fat and energy delivery of human milk delivered via bottle-feeding pump: An experimental study.

Enteral feeding pump systems deliver decreased amounts of macronutrients in human milk to neonates. This study determined the macronutrient loss associated with a bottle-feeding pump system and the effect of manually mixing the human milk during extended feeds. Macronutrient content from samples of donor human milk was analyzed after simulated extended feeds with a bottle-feeding pump system, using a human milk analyzer. Simulations were repeated using manual mixing of the bottle every 30 min during feeding. The percentage of the baseline was calculated, and one-sample t tests and analysis of variance compared the effect of manual mixing and the duration of feeding on macronutrient delivery. The delivery of fat and energy was lower over time, but manual mixing considerably improved retention. The length of feeding impacted fat delivery, with less fat delivered over time (P < 0.001). Manually mixing significantly increased fat delivery (P < 0.001). Similar results were found for energy, with a significant reduction in energy delivery over time (P < 0.001) and significantly more energy delivered with mixing (P < 0.001). Mixing and the duration of feeding had minimal effect on protein or carbohydrate delivery. Bottle-feeding pump systems are associated with a significant reduction in the delivery of fat and energy of donor human milk. The manual mixing of donor human milk during prolonged feeds is a simple way to improve fat and energy delivery to the neonate.

Consumption of different combinations of human milk oligosaccharides in the first 6 mo of infancy is positively associated with early cognition at 2 y of age in a longitudinal cohort of Latino children

BackgroundLactation has been widely associated with optimal neurocognitive development, but the underlying mechanism remains unknown. Human milk oligosaccharides (HMOs) are complex sugars that support brain development, but previous studies examining their associations with cognition have yielded inconsistent findings. ObjectivesThis study aimed to provide a broader understanding of how HMOs jointly influence cognition. MethodsWe used data from an ongoing longitudinal cohort of Latino mother–infant dyads. Human milk samples from 1 mo (n = 157) and 6 mo (n = 107) postpartum were assessed for the 19 most abundant HMOs. Cognitive performance was assessed at 2 y using the Bayley Scale of Infant and Toddler Development. A partial least squares model identified HMO combinations predictive of cognitive scores. ResultsAt 1 mo, the combination of higher concentrations of lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), lacto-N-fucopentaose (LNFP)-III, 6′-sialyllactose, and 2′-fucosyllactose (FL) with lower concentrations of sialyllacto-N-tetraose (LST) b, LNFP-II, fucodisialyllacto-N-hexaose, and 3-FL significantly predicted higher cognitive scores (β: 0.61; 95% confidence interval [CI]: 0.30, 0.92), explaining an additional 8% of the variance over a model with only nuisance covariates (11%). Additional analyses revealed that the combination of higher LNFP-III and lower LSTb alone explained 5% more of the variation in cognitive scores (β: 0.66; 95% CI: 0.24, 1.09). At 6 mo (n = 107), higher LNnT, LNT, and LNFP-III and lower 3FL and LSTb concentrations explained an extra 6% of the variance in cognitive scores (β: 0.43; 95% CI: 0.12, 0.75). ConclusionsThis study highlights specific HMO combinations in early life influencing cognitive performance at 2 y.

Human milk macro- and trace-elements: Simultaneous analysis in sub-milliliter amounts by ICP-MS and application to assessing acute supplementation effects

Adequate concentrations of human milk (HM) nutrients, including macro- and trace-elements, are essential for healthy growth and development of exclusively breastfed (EBF) infants. To monitor potential risk of deficiencies, and evaluate the effects of interventions like supplementation, accurate analysis is crucial. Even recent methods reporting on HM macro- and/or trace-elements describe multiple methodological approaches and the need for several milliliters. We optimized and validated a comprehensive method for simultaneous analysis of 13 macro- and trace-elements for simultaneous analysis by inductively-coupled plasma-mass spectrometry. 100–600 μL HM were microwave digested with ≤1.5 mL HNO3 (70 %). The digest was diluted to 5 % final acid concentration. He-Kinetic Energy Discrimination (KED; Na, K, P, Ca, Mg, Fe, Cu, Zn, Cr, Mo) and O2-Dynamic Reaction Cell (DRC; As, Mn, Se) modes minimized remaining interferences. Accuracy (NIST SRM 1869 infant formula; n = 15, 4 weeks) varied from 93.2 to 103 % (CV: 2.8–8.5 %) with trueness ranging from 93.9 to 104 %. Inter-day variation of a HM-pool (n = 20, 3 weeks) varied between 4.1 and 8.5 % for most elements; Cr, Mo, Mn (all<5 μg L−1) had higher variation, up to 25 %. Analyzing HM from 18 Bangladeshi mothers (2–4 months postpartum; day 1 = baseline, n = 17; day 2/3 = supplementation, n = 21 each) revealed higher concentrations for P, Ca, and Zn post-supplementation (p < 0.05, Friedman's Chi-Square Test). Na, Mg, Zn, and Se had the highest number of samples (>80 %) with concentrations below the Adequate Intake. Our method allows for simultaneous and reproducible analysis of macro- and trace-elements with concentrations ranging over 6 orders of magnitude, without the need for separate analytics and sample preparations, and requiring only sub-milliliter amounts of HM. Additional elements may be included after optimization and validation. The results from Bangladeshi HM samples indicate selective supplementation effects and concerningly low concentrations for some elements, which could adversely affect the EBF infant.

Analysis of Human Milk Microbiota in Northern Greece by Comparative 16S rRNA Sequencing vs. Local Dairy Animals.

Milk is a biological fluid with a dynamic composition of micronutrients and bioactive molecules that serves as a vital nutrient source for infants. Milk composition is affected by multiple factors, including genetics, geographical location, environmental conditions, lactation phase, and maternal nutrition, and plays a key role in dictating its microbiome. This study addresses a less-explored aspect, comparing the microbial communities in human breast milk with those in mature milk from species that are used for milk consumption. Since mature animal milk is used as a supplement for both the infant (formula) and the child/adolescent, our main aim was to identify shared microbial communities in colostrum and mature human milk. Using 16S rRNA metagenomic sequencing, we focused on characterizing the milk microbiota in the Northern Greek population by identifying shared microbial communities across samples and comparing the relative abundance of prevalent genera. We analyzed ten human milk samples (from five mothers), with five collected three days postpartum (colostrum) and five collected thirty to forty days postpartum (mature milk) from corresponding mothers. To perform an interspecies comparison of human milk microbiota, we analyzed five goat and five bovine milk samples from a local dairy industry, collected fifty to seventy days after birth. Alpha diversity analysis indicated moderate diversity and stability in bovine milk, high richness in goat milk, and constrained diversity in breast milk. Beta diversity analysis revealed significant distinctions among mammalian species, emphasizing both presence/absence and abundance-based clustering. Despite noticeable differences, shared microbial components underscore fundamental aspects across all mammalian species, highlighting the presence of a core microbiota predominantly comprising the Proteobacteria, Firmicutes, and Actinobacteriota phyla. At the genus level, Acinetobacter, Gemella, and Sphingobium exhibit significant higher abundance in human milk compared to bovine and goat milk, while Pseudomonas and Atopostipes are more prevalent in animal milk. Our comparative analysis revealed differences and commonalities in the microbial communities of various mammalian milks and unraveled the existence of a common fundamental milk core microbiome. We thus revealed both species-specific and conserved microbial communities in human, bovine, and goat milk. The existence of a common core microbiome with conserved differences between colostrum and mature human milk underscores fundamental similarities in the microbiota of milk across mammalian species, which could offer valuable implications for optimizing the nutritional quality and safety of dairy products as well as supplements for infant health.