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Item The effect of microwave-assisted heating on bioactive and immunological compounds in donor human milk(Elsevier Ltd, 2022-05-01) Leite JAS; Robinson RC; Salcedo J; Ract JNR; Quintal VS; Tadini CC; Barile DLow-Temperature Long-Time pasteurization (LTLT) is normally applied in donor human milk from Human Milk Banks (HMBs) to guarantee microbiological safety; however, this treatment can modify the protein structures, decreasing their beneficial effects. Thus, this study aimed to determine the impact of microwave-assisted heating on the concentration of key biological compounds in donor human milk to verify whether a microwave heating technique can be used as an alternative to LTLT pasteurization in Human Milk Banks. The concentrations of oligosaccharides, immunoglobulins, lactoferrin and fatty acids were monitored in raw donor milk and after processing to assess the impact of the microwave and LTLT treatments. The concentration of oligosaccharides was determined by HPAEC-PAD, immunoglobulins and lactoferrin were quantified using ELISA kits and fatty acids were quantified by gas chromatography. Oligosaccharides and fatty acids were not significantly affected (p > 0.05) by LTLT and microwave processes; however, immunoglobulins and lactoferrin concentrations were better preserved when microwave-assisted heating was applied. For this reason, microwave-assisted heating can be considered a promising alternative to LTLT pasteurization of donor human milk in Human Milk Banks.Item Understanding the Effects of Lactose Hydrolysis Modeling on the Main Oligosaccharides in Goat Milk Whey Permeate(MDPI (Basel, Switzerland), 2019-09-10) Thum C; Weinborn V; Barile D; McNabb WC; Roy NC; Leite Nobrega de Moura Bell JM; Moreno DA; Villaño DEnzymatic hydrolysis of lactose is a crucial step to improve the efficiency and selectivity of membrane-based separations toward the recovery of milk oligosaccharides free from simple sugars. Response surface methodology was used to investigate the effects temperature (25.9 to 54.1 °C) and amount of enzyme (0.17 to 0.32% w/w) at 1, 2, and 4 h of reaction on the efficiency of lactose hydrolysis by Aspergillus oryzae β-galactosidase, preservation of major goat whey oligosaccharides, and on the de-novo formation of oligosaccharides. Lactose hydrolysis above 99% was achieved at 1, 2, and 4 h, not being significantly affected by temperature and amount of enzyme within the tested conditions. Formation of 4 Hexose (Hex) and 4 Hex 1 Hex and an increased de-novo formation of 2 Hex 1 N-Acetyl-Neuraminic Acid (NeuAc) and 2 Hex 1 N-Glycolylneuraminic acid (NeuGc) was observed in all treatments. Overall, processing conditions using temperatures ≤40 °C and enzyme concentration ≤0.25% resulted in higher preservation/formation of goat whey oligosaccharides.Item Infant Complementary Feeding of Prebiotics for the Microbiome and Immunity(MDPI (Basel, Switzerland), 2019-02-09) McKeen S; Young W; Mullaney J; Fraser K; McNabb WC; Roy NCComplementary feeding transitions infants from a milk-based diet to solid foods, providing essential nutrients to the infant and the developing gut microbiome while influencing immune development. Some of the earliest microbial colonisers readily ferment select oligosaccharides, influencing the ongoing establishment of the microbiome. Non-digestible oligosaccharides in prebiotic-supplemented formula and human milk oligosaccharides promote commensal immune-modulating bacteria such as Bifidobacterium, which decrease in abundance during weaning. Incorporating complex, bifidogenic, non-digestible carbohydrates during the transition to solid foods may present an opportunity to feed commensal bacteria and promote balanced concentrations of beneficial short chain fatty acid concentrations and vitamins that support gut barrier maturation and immunity throughout the complementary feeding window.Item Sequence-based genome-wide association study of individual milk mid-infrared wavenumbers in mixed-breed dairy cattle(BioMed Central Ltd, 2021-07-20) Tiplady KM; Lopdell TJ; Reynolds E; Sherlock RG; Keehan M; Johnson TJJ; Pryce JE; Davis SR; Spelman RJ; Harris BL; Garrick DJ; Littlejohn MDBACKGROUND: Fourier-transform mid-infrared (FT-MIR) spectroscopy provides a high-throughput and inexpensive method for predicting milk composition and other novel traits from milk samples. While there have been many genome-wide association studies (GWAS) conducted on FT-MIR predicted traits, there have been few GWAS for individual FT-MIR wavenumbers. Using imputed whole-genome sequence for 38,085 mixed-breed New Zealand dairy cattle, we conducted GWAS on 895 individual FT-MIR wavenumber phenotypes, and assessed the value of these direct phenotypes for identifying candidate causal genes and variants, and improving our understanding of the physico-chemical properties of milk. RESULTS: Separate GWAS conducted for each of 895 individual FT-MIR wavenumber phenotypes, identified 450 1-Mbp genomic regions with significant FT-MIR wavenumber QTL, compared to 246 1-Mbp genomic regions with QTL identified for FT-MIR predicted milk composition traits. Use of mammary RNA-seq data and gene annotation information identified 38 co-localized and co-segregating expression QTL (eQTL), and 31 protein-sequence mutations for FT-MIR wavenumber phenotypes, the latter including a null mutation in the ABO gene that has a potential role in changing milk oligosaccharide profiles. For the candidate causative genes implicated in these analyses, we examined the strength of association between relevant loci and each wavenumber across the mid-infrared spectrum. This revealed shared association patterns for groups of genomically-distant loci, highlighting clusters of loci linked through their biological roles in lactation and their presumed impacts on the chemical composition of milk. CONCLUSIONS: This study demonstrates the utility of FT-MIR wavenumber phenotypes for improving our understanding of milk composition, presenting a larger number of QTL and putative causative genes and variants than found from FT-MIR predicted composition traits. Examining patterns of significance across the mid-infrared spectrum for loci of interest further highlighted commonalities of association, which likely reflects the physico-chemical properties of milk constituents.Item Extraction of milk oligosaccharides from lactose mother liquor : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Bioprocess Engineering at Massey University, Manawatū, New Zealand.(Massey University, 2016) Peacey, Malcolm"Oligosaccharides are a diverse group of bioactive sugars that are a critical part of a new-born’s nutrition. In particular, sialylated oligosaccharides have been shown to be beneficial in stimulating brain development and providing resistance to infection in the first months of life, among many other benefits. Breast milk is naturally rich in these molecules and is the best possible diet for an infant, whereas infant formula based on cows’ milk is much lower in oligosaccharides. Because of the large throughput of the dairy industry, there is an opportunity to separate and enrich the small fraction of oligosaccharides to create an ingredient with a higher oligosaccharide content that could be added to infant formula to provide formula-fed babies with a level of nutrition closer to breast milk. Lactose mother liquor is a low value by-product of lactose manufacture and has been identified to have one of the highest oligosaccharide contents of any dairy stream, including a significant portion of sialylated oligosaccharides. The objective of this work was to develop a food grade, industrial scale process for enriching oligosaccharides from mother liquor to at least 4% on a dry basis. While the laboratory characterisation and high purity isolation of milk oligosaccharide species has developed significantly in the past 20 years, there is comparatively little understanding of high volume separations from process sources. This presents an opportunity to research the behaviour of oligosaccharides and other components of lactose mother liquor in different separation systems. Technologies were initially evaluated by mass balances and economic criteria, with reference to final oligosaccharide purity and yield, likely production cost, and food safety considerations. Nanofiltration and size exclusion chromatography had distinct advantages in these areas, while other technologies were likely to either give poor purity for their cost or product with nonfood grade chemicals. A process was selected that centres on simulated moving bed chromatography, a continuous separation of the smaller lactose, monosaccharide, and mineral components of ultra-filtered mother liquor which are more strongly retained on a size exclusion resin from the larger oligosaccharides. A four-column laboratory scale system was built and used in trials to determine its suitability, with results showing that a raffinate containing up to 35% oligosaccharides on a dry basis could be obtained at a flow rate of 145 mL/hour (1.03 column volumes/hour). Nanofiltration was also trialled for the separation of oligosaccharides from ultra-filtered mother liquor, but the size difference between oligosaccharides and lactose is too small for current membrane technology to separate completely and a maximum enrichment of 2.5% (dry basis) oligosaccharides was reached. This is lower than the 4% minimum oligosaccharide requirement which is needed to meet the minimum oligosaccharide to lactose ratio. Attempts to optimise nanofiltration using lactose crystallisation and multiple nanofiltration stages gave small improvements but still fell short of the 4% target. A sequence consisting of decanting, ultrafiltration, simulated moving bed chromatography, and evaporation was chosen as the recommended process for oligosaccharide enrichment"--SummaryItem Effects of dietary caprine milk oligosaccharides enriched fraction on maternal large intestine and the consequences for the development of the offspring : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy, Massey University, Palmerston North, New Zealand(Massey University, 2015) Thum, CarolineThe colonisation of the neonate gastrointestinal tract by health-promoting microbiota is likely to improve the overall health of the infant and may also have health benefits in later life. Initial development and maturation of the foetal/neonatal gastrointestinal tract is heavily influenced by the in utero environment which itself, may be altered by the maternal diet and gastrointestinal tract microbiota composition. The maternal gastrointestinal tract microbiota can be altered by supplementation with synthetic oligosaccharides; however, positive effects on the health and well-being of the offspring have not been adequately established. Human milk contains natural oligosaccharides known to improve the gastrointestinal tract colonisation and the development and maturation of the infant gastrointestinal tract. Among domestic farm animals, caprine milk has oligosaccharides structurally similar to human milk and potentially similar beneficial effects for the infant. We hypothesised that feeding caprine milk oligosaccharide enriched product to pregnant and lactating mice would induce changes in the maternal large intestine microbiota and milk composition, accelerating the development and maturation of the offspring’s large intestine tissue and altering the gastrointestinal tract microbiota composition. The aim of this project was to obtain bifidobacteria from the faeces of breast-fed human infants and determine which were of capable fermenting caprine milk oligosaccharide enriched product. Subsequently, the effects of the best strains on the morphology and metabolic pathways of the colonic mucosa of germ-free and conventionally raised mice, supplemented with dietary caprine milk oligosaccharide enriched product. The present study is the first to report New Zealand Saanen caprine colostrum, milk and whey. An enrichment method previously described was used to produce a caprine milk oligosaccharide enriched product for in vitro and in vivo assessment of its health effects. Caprine milk oligosaccharide enriched product was shown to differentially stimulate the growth of bifidobacteria, commonly found in the gastrointestinal tract of breast-fed infants. Among the bifidobacterial species tested, Bifidobacterium bifidum utilised caprine milk oligosaccharide enriched product most efficiently when compared to Bifidobacterium breve and Bifidobacterium longum subsp. longum. B. bifidum (AGR2166) was shown to ferment the sialyloligosaccharides, 3’- and 6’-sialyl-lactose present in caprine milk oligosaccharide enriched product through cell-associated sialidase expression. Augmented microbial biomass associated with enhanced growth and in vitro fermentation of caprine milk oligosaccharide enriched product, increased the production of microbial fermentation end products such as acetate and lactate. These findings indicate that in vivo caprine milk oligosaccharide enriched product may stimulate the growth and fermentation of bifidobacteria within the gastrointestinal tract. Germ-free mice or mice mono-associated with B. bifidum (AGR2166) were used to test the in vivo effects of maternal caprine milk oligosaccharide enriched product consumption during pregnancy and the effects on the foetus. Caprine milk oligosaccharide enriched product diet showed no effects on maternal gastrointestinal tract or foetal growth regardless of microbial status. Mice inoculated with B. bifidum (AGR2166) and fed caprine milk oligosaccharide enriched product diet, however, showed an increased bacterial translocation from maternal gastrointestinal tract to organs and placenta (inferred by the presence of the bifidobacteria 16S rRNA gene in the maternal organs). Increased translocation of commensal bacteria from maternal gastrointestinal tract to the foetus may have important effects on foetal immunological programming. The consumption of caprine milk oligosaccharide enriched product, during gestation and lactation were also tested in conventional rodents and it had no effects on maternal gastrointestinal tract microbiota and morphology. Changes on maternal lipid metabolism and increased maternal milk protein, however, were observed. These modifications may have positively affected the development of the pups, relative abundance of gastrointestinal tract bifidobacteria and butyric acid production at weaning. Important changes in the plasma and urine metabolites involved in bile acid and fatty acid metabolism were also observed in the pups as a consequence of maternal caprine milk oligosaccharide-enriched diet. The effects of maternal caprine milk oligosaccharide enriched product diet on pups, were no longer apparent after 30 days of consuming a control diet post-weaning, however, detrimental physiological characteristics such as an increased body fat were observed. Further studies, are needed to understand the physiological effects of caprine milk oligosaccharides on the maternal/infant pair.Item Strategies for the removal of raffinose family oligosaccharides from navy bean flour : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Bioprocess Engineering at Massey University, Palmerston North, New Zealand(Massey University, 2013) Brain, Jarod ChristopherNavy beans are legumes with highly nutritious qualities. However they are underutilised in the processed food industry due to the undesirable bloating, abdominal discomfort and excessive flatulence associated with ingestion of the raffinose family oligosaccharides (RFOs) present in them. If a suitable technology were available to sufficiently reduce the concentration of RFOs then navy bean flour could find application in many food production processes. The traditional method for decreasing RFO content in navy beans and other similar legumes has been through soaking in large quantities of water, thereby leaching the RFOs from the bean. However this is a slow process and results in the depletion of all water soluble solids (up to 25% of the dry weight of the bean). The leaching process can be sped up dramatically through the decrease in particle size achieved by milling the beans into navy bean flour prior to the addition of water. However this process makes dewatering of the flour difficult. Rates of moisture uptake and RFO leaching were characterised for navy bean flour and dehulled navy bean cotyledons demonstrating that RFO leaching is slower but of a similar magnitude as moisture uptake and that these rates are dramatically increased with decrease in particle size. The addition of α-galactosidase to the leaching water enables the rapid removal of RFOs from navy bean flour without the need to separate the flour from the leaching water because the RFOs leached out of the flour are hydrolysed into simple sugars. Galactose is a product of the hydrolysis of RFOs and its presence at high concentrations can result in the inhibition of α-galactosidase. However at the concentrations likely to be experienced during RFO reduction in bean flour the inhibition effect is minimal. Rates of hydrolysis were studied for raffinose and stachyose hydrolysis by α-galactosidase in the concentration range expected during processing of navy bean flour. Enzymatic processing using very small amounts of moisture addition was investigated demonstrating partial RFO removal at moisture contents as low as 28% (wet basis). Reduction in RFO content to a level at which the flatus response is negligible can be achieved at moisture contents as low as 38% (wet basis) which significantly minimises the drying required to produce a dry navy bean ingredient or could allow direct application in an extrusion process. The application of α-galactosidase at low moisture content is a novel process for the depletion of RFOs in navy bean flour. The mechanism for this process is explained by preliminary modelling of moisture and RFO diffusion which demonstrates how partial RFO removal is possible. From these mechanisms a series of commercially practical methods for RFO depletion of navy bean flour were explored for several targeted product applications. This process also has potential applications for low moisture enzyme processing in a range of food and other biological systems. The simple processes developed in this work open up the commercial use of navy bean flour for the food industry to develop products that take advantage of its functional and nutritional properties without the negative nutritional problems usually associated with this material.