Daily and within-feed variation of macro- and trace-element concentrations in human milk and implications for sampling.

BACKGROUND: Daily variations of macro- and trace-elements in human milk (HM) are not well characterised and sampling protocols are highly variable between studies. OBJECTIVES: To investigate diurnal and within-feed variation of HM macro- and trace-elements using pre- and post- feed concentrations and to compare infant intake estimates using limited samples with measured 24-hour intake. METHODS: HM Samples were collected pre- and post- every feed in a 24-hour period from 11 mother-infant dyads. Test-weighing was used to determine the volume of HM consumed in each feed. For macro- and trace-elements within-feed and daily variation was measured. Intake estimated from a morning pre-feed sample was compared to the measured milk intake calculated from every feed over 24-hours. Macro- and trace-elements concentrations were measured using ICP-MS. Linear mixed modelling was used for statistical analysis. RESULTS: Average intake of HM was 737 ± 63 mL for infants aged 1-6 months and 508 ± 50 for infants aged 6-12 months. Pre- and post-feed HM variation was found for phosphorus, calcium, manganese, iron, copper, zinc, selenium, molybdenum, and iodine (p < 0.05). Variation across 24 h was found for magnesium, phosphorus, potassium, manganese, iron, and selenium (p < 0.05). Estimated intake using morning, pre-feed samples resulted in significantly lower intake when compared to measured milk intake for iron, phosphorus, selenium, and manganese (p < 0.05). CONCLUSION: Standardised sampling protocols using large sample volumes and multiple collections over 24-hours provide a calculated intake that is more reflective of actual infant HM macro- and trace-elements intake.

Reproducibility of the creamatocrit technique for the measurement of fat content in human milk.

Quantification of human milk (HM) fat is important for determining the energy intake of infants. The simplest and most rapid method is the creamatocrit method. However, the reliability of the creamatocrit has not been comprehensively investigated. The aims of this study were to test the inter- and-intra-rater reliability of: 1) HM sampling after hand- or-machine mixing methods and 2) HM fat measurement by the creamatocrit method. Inter-and-intra rater HM sampling after hand- or-machine mixing methods had high intraclass correlation coefficient (>0.91). Inter-rater reliability of measurement of HM with low fat (<2%) resulted in high variability (median coefficient of variations (CVs) > 15%). Intra- and inter-rater reliability of measurement of HM with higher fat (>3.5%) had low variability (median CVs < 10%). As the greatest variation in the creamatocrit method occurred during the measurement of HM samples with low fat, duplicate readings are necessary to reduce discrepancies in every HM fat determination.

Mother's Own Milk Biomarkers Predict Coming to Volume in Pump-Dependent Mothers of Preterm Infants.

OBJECTIVE: To assess serial secretory activation biomarker concentrations (sodium [Na], potassium [K], Na:K, protein, lactose, and citrate) in mother's own milk (MOM) from breast pump-dependent mothers of preterm infants to determine associations with coming to volume (CTV), defined as producing at least 500 mL/day MOM by day 14 postpartum. STUDY DESIGN: We collected serial MOM samples and pumped MOM volume data for 14 days postpartum in mothers who delivered at <33 weeks of gestation. Regression models and the Mann-Whitney U test were used to evaluate associations. RESULTS: Among 40 mothers, 39 (mean gestational age, 28.8 weeks; 67% overweight/obese; 59% nonwhite) had paired MOM volume and biomarker data; 33% achieved CTV between postpartum days 6 and 14. In univariate models, MOM Na on postpartum day 5 and Na:K on days 3 and 5 were associated with CTV. Mothers achieving CTV were more likely to have postpartum Na:K ≤1 on day 3 (75% vs 25%; P = .06) and ≤0.8 on day 5 (69% vs 10%; P < .01). In a multivariable regression model, day 5 Na:K (1 unit decrease in Na:K: OR, 18.7; 95% CI, 1.13-311.41; P = .049) and maternal prepregnancy body mass index (BMI) (1 unit increase in BMI: OR, 0.88; 95% CI, 0.78-0.99; P = .04) were associated with CTV between postpartum days 6 and 14. CONCLUSIONS: Secretory activation and CTV were compromised in breast pump-dependent mothers with preterm delivery. CTV was predicted by MOM Na level and Na:K. These biomarkers have potential as objective point-of-care measures to detect potentially modifiable lactation problems in a high-risk population.

Maternal and infant infections stimulate a rapid leukocyte response in breastmilk.

Breastmilk protects infants against infections; however, specific responses of breastmilk immune factors to different infections of either the mother or the infant are not well understood. Here, we examined the baseline range of breastmilk leukocytes and immunomodulatory biomolecules in healthy mother/infant dyads and how they are influenced by infections of the dyad. Consistent with a greater immunological need in the early postpartum period, colostrum contained considerable numbers of leukocytes (13-70% out of total cells) and high levels of immunoglobulins and lactoferrin. Within the first 1-2 weeks postpartum, leukocyte numbers decreased significantly to a low baseline level in mature breastmilk (0-2%) (P<0.001). This baseline level was maintained throughout lactation unless the mother and/or her infant became infected, when leukocyte numbers significantly increased up to 94% leukocytes out of total cells (P<0.001). Upon recovery from the infection, baseline values were restored. The strong leukocyte response to infection was accompanied by a more variable humoral immune response. Exclusive breastfeeding was associated with a greater baseline level of leukocytes in mature breastmilk. Collectively, our results suggest a strong association between the health status of the mother/infant dyad and breastmilk leukocyte levels. This could be used as a diagnostic tool for assessment of the health status of the lactating breast as well as the breastfeeding mother and infant.