Macronutrient content of pasteurised donor human milk: Variability between batches from single-donor pools at an Australian milk bank.

AIM: This study aimed to characterise the between-batch variability of pasteurised donor human milk (PDHM) produced from single-donor pools at Australian Red Cross Lifeblood's milk bank and identify key donor characteristics that predict macronutrient content. METHODS: Macronutrient content from 200 batches of PDHM was measured using a mid-infrared human milk analyser (Miris, Uppsala, Sweden). Linear mixed models were used to study the impact of stage of lactation and gestational age on macronutrient content. Coefficients of determination (R2 ) were calculated to estimate the impact of the individual donor on overall variability. RESULTS: Macronutrient content of PDHM varied considerably, with between-batch variations of 2.8 and 6.4-fold for protein and fat content, respectively. Mean crude protein content was 1.16 g/100 mL, ranging from 0.7 to 1.96 g/100 mL. Mean fat content was 3.85 g/100 mL, ranging from 1.46 to 9.39 g/100 mL. Stage of lactation was identified as a predictor for protein content and gestational age at birth for fat content. Individual donor effect explained 55 and 35% of the variance for fat and protein content, respectively. CONCLUSIONS: This study highlights the variation in macronutrient content in PDHM at an Australian milk bank. Variability could be reduced through the implementation of targeted multiple-donor pooling using the key donor characteristics identified in this study along with the measurement of macronutrient content of individual donors at the time of first donation. However, the clinical benefit of a reduction in between-batch variation, achieved through multiple-donor pooling, would need to be assessed to justify additional efforts associated with PDHM processing changes.

The effect of physiological state, milk production traits and environmental conditions on milk fat globule size in cow's milk.

This research was carried out to quantify the effects of a range of variables on milk fat globule (MFG) size for a herd of Holstein-Friesian cows managed through an automatic milking system with year-round calving. We hypothesised that the overall variation in average MFG size observed between individual animals of the same herd cannot sufficiently be explained by the magnitude of the effects of variables that could be manipulated on-farm. Hence, we aimed to conduct an extensive analysis of possible determinants of MFG size, including physiological characteristics (parity, days in milk, days pregnant, weight, age, rumination minutes, somatic cell count) and milk production traits (number of milkings, milk yield, fat yield, protein and fat content, fat-protein ratio) on the individual animal level; and environmental conditions (diet, weather, season) for the whole herd. Our results show that when analysed in isolation, many of the studied variables have a detectable effect on MFG size. However, analysis of their additive effects identified days in milk, parity and milk yield as the most important variables. In accordance with our hypothesis, the estimated effects of these variables, calculated using a multiple variable linear mixed model, do not sufficiently explain the overall variation between cows, ranging from 2.70 to 5.69 µm in average MFG size. We further show that environmental variables, such as sampling day (across seasons) or the proportion of pasture and silage in the diet, have limited effects on MFG size and that physiological differences outweigh the effects of milk production traits and environmental conditions. This presents further evidence that the selection of individual animals is more important than the adjustment of on-farm variables to control MFG size.

Investigation on the suitability of milk-derived primary bovine mammary epithelial cells grown on permeable membrane supports as an in vitro model for lactation.

This study aimed to establish an in vitro model for lipid synthesis in primary bovine mammary epithelial cells (pbMECs) extracted from milk and cultured on Transwell permeable supports (TW culture). The suitability of these cells as a functional model for lactation was assessed by measuring κ-casein (CSN3) and diacylglycerol acyl transferase 1 (DGAT1) gene expression, the presence of intracellular lipid droplets, and the concentration of triacylglycerol in the cell lysates. The functionality of the milk-derived pbMECs cultured under lactogenic conditions, with and without oleic acid supplementation, was evaluated by comparing the cells grown on Transwell supports to cells grown on an extracellular matrix (ECM) gel (3D culture) or a plastic surface (2D culture). Furthermore, the functionality of milk-derived cells was compared to pbMECs obtained from bovine mammary tissue. Here, we show that in both tissue and milk-derived pbMECs, 3D culture offered the most suitable in vitro environment and led to increased levels of CSN3 and DGAT1 gene expression along with increased intracellular triacylglycerol content. The TW culture conditions also resulted in increased DGAT1 gene expression compared to the 2D conditions and milk-derived pbMECs cultured on TW inserts showed the highest viability compared to cells grown under 2D or 3D treatments. However, this was not observed for tissue-derived pbMECs, suggesting that TW culture may offer a beneficial environment specifically for milk-derived cells. We suggest that with further optimization of the culture conditions, TW culture may present a suitable model for the study of milk lipid synthesis in pbMECs.

Milk fat globule size development in the mammary epithelial cell: a potential role for ether phosphatidylethanolamine.

Milk fat globule (MFG) size is a milk production trait characteristic to the individual animal and has important effects on the functional and nutritional properties of milk. Although the regulation of MFG size in the mammary epithelial cell is not fully understood, lipid droplet (LD) fusion prior to secretion is believed to play a role. We selected cows that consistently produced milk with predominantly small or large MFGs to compare their lipidomic profiles, with focus on the polar lipid fraction. The polar lipid composition of the monolayer surrounding the LD is believed to either promote or prevent LD fusion. Using a targeted LC-MS/MS approach we studied the relative abundance of 301 detected species and found significant differences between the studied groups. Here we show that the lipidomic profile of milk from small MFG cows is characterised by higher phosphatidylcholine to phosphatidylethanolamine ratios. In contrast, the milk from large MFG cows contained more ether-phosphatidylethanolamine species. This is the first time that a potential role for ether-phosphatidylethanolamine in MFG size development has been suggested.