Maternal dietary intervention during lactation impacts the maternal faecal and human milk microbiota.

AIMS: To determine the effect of a two-week reduced fat and sugar and increased fibre maternal dietary intervention on the maternal faecal and human milk (HM) microbiomes. METHODS AND RESULTS: Faecal swabs and HM samples were collected from mothers (n = 11) immediately pre-intervention, immediately post-intervention, and 4 and 8 weeks post-intervention, and were analysed using full-length 16S rRNA gene sequencing. Maternal macronutrient intake was assessed at baseline and during the intervention. Maternal fat and sugar intake during the intervention were significantly lower than pre-intervention (P = <0.001, 0.005, respectively). Significant changes in the bacterial composition of maternal faeces were detected after the dietary intervention, with decreases in the relative abundance of Bacteroides caccae (P = <0.001) and increases in the relative abundance of Faecalibacillus intestinalis (P = 0.006). In HM, the diet resulted in a significant increase in Cutibacterium acnes (P = 0.001) and a decrease in Haemophilus parainfluenzae (P = <0.001). The effect of the diet continued after the intervention, with faecal swabs and HM samples taken 4 and 8 weeks after the diet showing significant differences compared to baseline. CONCLUSION: This pilot study demonstrates that short-term changes in maternal diet during lactation can alter the bacterial composition of the maternal faeces and HM.

Acute changes to breast milk composition following consumption of high-fat and high-sugar meals.

Breast milk composition is influenced by habitual diet, yet little is known about the short-term effects of changes in maternal diet on breast milk macronutrient concentrations. Our aim was to determine the acute effect of increased consumption of sugar/fat on breast milk protein, lactose and lipids. Exclusively breastfeeding women (n = 9) were provided with a control, higher fat (+28 g fat) and higher sugar (+66 g sugar) diet over three separate days at least 1 week apart. Hourly breast milk samples were collected concurrently for the analysis of triglycerides, cholesterol, protein, and lactose concentrations. Breast milk triglycerides increased significantly following both the higher fat and sugar diet with a greater response to the higher sugar compared to control diet (mean differences of 3.05 g/dL ± 0.39 and 13.8 g/dL ± 0.39 in higher fat and sugar diets, respectively [P < 0.001]). Breast milk cholesterol concentrations increased most in response to the higher sugar diet (0.07 g/dL ± 0.005) compared to the control (0.04 g/dL) and the higher fat diet (0.05 g/dL) P < 0.005. Breast milk triglyceride and lactose concentrations increased (P < 0.001, P = 0.006), whereas protein decreased (p = 0.05) in response to the higher fat diet compared to the control. Independent of diet, there were significant variations in breast milk composition over the day; triglycerides and cholesterol concentrations were higher at end of day (P < 0.001), whereas protein and lactose concentrations peaked at Hour 10 (of 12) (P < 0.001). In conclusion, controlled short-term feeding to increase daily sugar/fat consumption altered breast milk triglycerides, cholesterol, protein and lactose. The variations observed in breast milk protein and lactose across the 12 h period is suggestive of a circadian rhythm.

A Systematic Review of Collection and Analysis of Human Milk for Macronutrient Composition.

BACKGROUND: As human milk (HM) composition varies by time and across even a single feed, methods of sample collection can significantly affect the results of compositional analyses and complicate comparisons between studies. OBJECTIVE: The aim was to compare the results obtained for HM macronutrient composition between studies utilizing different sampling methodologies. The results will be used as a basis to identify the most reliable HM sampling approach. METHODS: EMBASE, MEDLINE/PubMed, Cochrane Library, Scopus, Web of Science, and ProQuest databases were searched for relevant articles. Observational and interventional studies were included, and at least 2 authors screened studies and undertook data extraction. Quality assessment was conducted using the Newcastle-Ottawa scale and previously published pragmatic score. RESULTS: A total of 5301 publications were identified from our search, of which 101 studies were included (n = 5049 breastfeeding women). Methods used for HM collection were divided into 3 categories: collection of milk from all feeds over 24 h (32 studies, n = 1309 participants), collection at one time point (62 studies, n = 3432 participants), and "other methods" (7 studies, n = 308 participants). Fat and protein concentrations varied between collection methods within lactation stage, but there were no obvious differences in lactose concentrations. There was substantial variability between studies in other factors potentially impacting HM composition, including stage of lactation, gestational age, and analytical method, which complicated direct comparison of methods. CONCLUSIONS: This review describes the first systematic evaluation of sampling methodologies used in studies reporting HM composition and highlights the wide range of collection methods applied in the field. This information provides an important basis for developing recommendations for best practices for HM collection for compositional analysis, which will ultimately allow combination of information from different studies and thus strengthen the body of evidence relating to contemporary HM composition. This trial was registered at PROSPERO as CRD42017072563, https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42017072563.

The short-term impact of dietary fat and sugar intake on breast milk composition: A clinical trial protocol.

BACKGROUND: Breast milk is uniquely designed for the infant and contains the key nutrients and bioactive factors required to support optimal infant health and development. While previous studies have reported that maternal obesity can influence milk composition, whether this relationship is driven by maternal or dietary factors remains unclear. AIM: The aim of this study is to assess the impact of test meals varying in fat and sugar content on post-prandial concentrations of macronutrients and metabolic hormones in the breast milk. METHODS: This open label crossover study will include 25 lactating women. On the three days of the intervention, women will be randomized to receive a breakfast meal with a fat and sugar content consistent with the Australian Guide to Healthy Eating (9 g fat, 25 g of sugar) or a breakfast meal containing higher levels of fat (28 g fat, 18 g of sugar) or sugar (5 g fat, 56 g of sugar). All breakfast meals will be similar in composition (cereal, milk, yogurt, toast and spread) and matched for total energy content. This study will measure breast milk concentrations of metabolic hormones (leptin, insulin, adiponectin, ghrelin and glucagon-like peptide-1) and macronutrients in the following 12 hours. RESULTS AND CONCLUSION: The results of this study will provide novel direct evidence of the impact of variations in dietary fat and sugar content to alter the macronutrient and/or metabolic hormone concentrations in breast milk. Data on the effect of maternal diet on milk composition is critical given the established importance of nutritional exposures in early infancy for an individual's life-long health outcomes.

Human milk lactoferrin and lysozyme concentrations vary in response to a dietary intervention.

It is known that human milk (HM)1 antimicrobial protein composition varies during lactation. However, the impact of maternal diet on these antimicrobial proteins, particularly lactoferrin and lysozyme remains unknown. In addition, it is unclear whether daily, circadian, and between breast variations exist for lactoferrin and lysozyme concentrations. We investigated the impact of a low sugar, low fat, high fibre dietary intervention on HM lysozyme and lactoferrin concentrations. HM was sampled across a 3-week period; daily, at different times of day, and from both breasts to measure the level of intraindividual variation. The intervention significantly reduced maternal sugar, total fat, and saturated fat intake. HM lactoferrin concentration declined significantly over the course of the intervention however the effect size was relatively small. In addition, lactoferrin and lysozyme concentrations were variable over time, and differed significantly within and across the day but not between breasts.

Effect of a reduced fat and sugar maternal dietary intervention during lactation on the infant gut microbiome.

Objective: A growing body of literature has shown that maternal diet during pregnancy is associated with infant gut bacterial composition. However, whether maternal diet during lactation affects the exclusively breastfed infant gut microbiome remains understudied. This study sets out to determine whether a two-week of a reduced fat and sugar maternal dietary intervention during lactation is associated with changes in the infant gut microbiome composition and function. Design: Stool samples were collected from four female and six male (n = 10) infants immediately before and after the intervention. Maternal baseline diet from healthy mothers aged 22-37 was assessed using 24-h dietary recall. During the 2-week dietary intervention, mothers were provided with meals and their dietary intake was calculated using FoodWorks 10 Software. Shotgun metagenomic sequencing was used to characterize the infant gut microbiome composition and function. Results: In all but one participant, maternal fat and sugar intake during the intervention were significantly lower than at baseline. The functional capacity of the infant gut microbiome was significantly altered by the intervention, with increased levels of genes associated with 28 bacterial metabolic pathways involved in biosynthesis of vitamins (p = 0.003), amino acids (p = 0.005), carbohydrates (p = 0.01), and fatty acids and lipids (p = 0.01). Although the dietary intervention did not affect the bacterial composition of the infant gut microbiome, relative difference in maternal fiber intake was positively associated with increased abundance of genes involved in biosynthesis of storage compounds (p = 0.016), such as cyanophycin. Relative difference in maternal protein intake was negatively associated with Veillonella parvula (p = 0.006), while positively associated with Klebsiella michiganensis (p = 0.047). Relative difference in maternal sugar intake was positively associated with Lactobacillus paracasei (p = 0.022). Relative difference in maternal fat intake was positively associated with genes involved in the biosynthesis of storage compounds (p = 0.015), fatty acid and lipid (p = 0.039), and metabolic regulator (p = 0.038) metabolic pathways. Conclusion: This pilot study demonstrates that a short-term maternal dietary intervention during lactation can significantly alter the functional potential, but not bacterial taxonomy, of the breastfed infant gut microbiome. While the overall diet itself was not able to change the composition of the infant gut microbiome, changes in intakes of maternal protein and sugar during lactation were correlated with changes in the relative abundances of certain bacterial species.Clinical trial registration: Australian New Zealand Clinical Trials Registry (ACTRN12619000606189).

Daily variation of macronutrient concentrations in mature human milk over 3 weeks.

Human milk (HM) composition is known to be highly variable, both between individuals and across the duration of lactation. It is less clear, however, to what extent fat, lactose and protein concentrations in HM change daily over shorter time periods in mature HM, and no studies have evaluated this to date. The aim of this study was to systematically assess and compare HM macronutrient concentrations in samples collected at different times of day, from left and right breasts and daily across a 3-week period in the same woman. Fifteen lactating women (1.6-4.9 months postpartum) collected daily pre-feed HM samples from both breasts each morning for 21 consecutive days and completed intensive sampling once a week (morning, afternoon and evening samples) during this period. Concentrations of fat, protein and lactose in HM did not differ according to time of day, day of week or breast used for collection. The results of this study suggest that pre-feed samples collected at any point across a 3-week period and from either the left or right breast provide comparable measures of fat, protein and lactose concentrations in mature HM, in pragmatic studies where women are collecting their own HM samples.Clinical trial registration: Australian New Zealand Clinical Trials Registry (ACTRN12619000606189).

Reduction in Maternal Energy Intake during Lactation Decreased Maternal Body Weight and Concentrations of Leptin, Insulin and Adiponectin in Human Milk without Affecting Milk Production, Milk Macronutrient Composition or Infant Growth.

Maternal diet has the potential to affect human milk (HM) composition, but very few studies have directly assessed the effect of maternal diets on HM composition. The primary aim of this study was to assess the effect of improving dietary quality in lactating women over 2 weeks on the concentrations of macronutrients and metabolic hormones in HM. The secondary aims were to assess the impact of the dietary intervention on 24 h milk production, maternal body composition and infant growth. Fifteen women completed a 1-week baseline period followed by a 2-week dietary intervention phase targeted towards reducing fat and sugar intake. Maternal anthropometric and body composition and infant growth measurements were performed weekly. Total 24 h milk production was measured before and after the dietary intervention, and HM samples were collected daily. Maternal intakes of energy (-33%), carbohydrate (-22%), sugar (-29%), fat (-54%) and saturated fat (-63%) were significantly reduced during the dietary intervention. HM insulin, leptin and adiponectin concentrations were 10-25% lower at the end of the dietary intervention, but HM concentrations of macronutrients were unaffected. Maternal body weight (-1.8%) and fat mass (-6.3%) were significantly reduced at the end of the dietary intervention, but there were no effects on 24 h milk production or infant growth. These results suggest that reducing maternal energy, carbohydrate, fat and sugar intake over a 2-week period is associated with significant reductions in HM insulin, leptin and adiponectin concentrations. These changes may be secondary to decreases in maternal weight and fat mass. The limited studies to date that have investigated the association between metabolic hormone concentrations in HM and infant growth raise the possibility that the changes in HM composition observed in the current study could impact infant growth and adiposity, but further studies are required to confirm this hypothesis.

The impact of maternal obesity on human milk macronutrient composition: A systematic review and meta-analysis.

Maternal obesity has been associated with changes in the macronutrient concentration of human milk (HM), which have the potential to promote weight gain and increase the long-term risk of obesity in the infant. This article aimed to provide a synthesis of studies evaluating the effects of maternal overweight and obesity on the concentrations of macronutrients in HM. EMBASE, MEDLINE/PubMed, Cochrane Library, Scopus, Web of Science, and ProQuest databases were searched for relevant articles. Two authors conducted screening, data extraction, and quality assessment independently. A total of 31 studies (5078 lactating women) were included in the qualitative synthesis and nine studies (872 lactating women) in the quantitative synthesis. Overall, maternal body mass index (BMI) and adiposity measurements were associated with higher HM fat and lactose concentrations at different stages of lactation, whereas protein concentration in HM did not appear to differ between overweight and/or obese and normal weight women. However, given the considerable variability in the results between studies and low quality of many of the included studies, further research is needed to establish the impact of maternal overweight and obesity on HM composition. This is particularly relevant considering potential implications of higher HM fat concentration on both growth and fat deposition during the first few months of infancy and long-term risk of obesity.

A methodological approach to identify the most reliable human milk collection method for compositional analysis: a systematic review protocol.

BACKGROUND: Breast milk composition has been reported to vary significantly between individual women and between different populations. However, the composition is also known to vary within the same woman between different days, within the same day, and even across the same feed. Therefore, it is unclear to what extent variations in composition are due to variations in sampling methodology between studies. The purpose of this systematic review is to compare the results obtained for breast milk macronutrient composition between studies utilizing different sampling methodologies and to use this as a basis to determine the most robust and consistent sampling approach as an alternative to full expression (gold standard). METHODS: The EMBASE, MEDLINE/PubMed, Cochrane Library, Scopus, Web of Science, and ProQuest Dissertations and Theses Global databases will be searched for relevant articles. Observational studies, including cross-sectional, comparative cohort, and longitudinal cohort studies which involve lactating women who are breastfeeding (exclusively or not) or expressing (manually or using a breast pump) at any lactation stage will be included. This review will compare different methods of breast milk collection used in research studies which report macronutrient levels (protein, fat, lactose). Two review authors will independently screen titles and abstracts of studies identified by the literature search to determine articles for the full text screening. Quality assessment of included articles will be conducted independently by two review authors using the Newcastle-Ottawa scale. DISCUSSION: It is important to identify the most reliable and practical method of human milk collection which best represents the average composition of the milk that is being consumed by the infant. This systematic review will be critical for ensuring that we determine a robust and consistent sampling approach to use in future studies of evaluating breast milk composition in a larger population. Identifying a recommended standard collection protocol will also provide more opportunities for sharing and combining data from different research groups, thus enhancing replicability and knowledge in the field. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42017072563.

Zinc is a critical regulator of placental morphogenesis and maternal hemodynamics during pregnancy in mice.

Zinc is an essential micronutrient in pregnancy and zinc deficiency impairs fetal growth. We used a mouse model of moderate zinc deficiency to investigate the physiological mechanisms by which zinc is important to placental morphogenesis and the maternal blood pressure changes during pregnancy. A 26% reduction in circulating zinc (P = 0.005) was exhibited in mice fed a moderately zinc-deficient diet. Zinc deficiency in pregnancy resulted in an 8% reduction in both near term fetal and placental weights (both P < 0.0001) indicative of disrupted placental development and function. Detailed morphological analysis confirmed changes to the placental labyrinth microstructure. Continuous monitoring of maternal mean arterial pressure (MAP) revealed a late gestation decrease in the zinc-deficient dams. Differential expression of a number of regulatory genes within maternal kidneys supported observations on MAP changes in gestation. Increased MAP late in gestation is required to maintain perfusion of multiple placentas within rodent pregnancies. Decreased MAP within the zinc-deficient dams implies reduced blood flow and nutrient delivery to the placenta. These findings show that adequate zinc status is required for correct placental morphogenesis and appropriate maternal blood pressure adaptations to pregnancy. We conclude that insufficient maternal zinc intake from before and during pregnancy is likely to impact in utero programming of offspring growth and development largely through effects to the placenta and maternal cardiovascular system.