Profiling of aminoxyTMT-labeled bovine milk oligosaccharides reveals substantial variation in oligosaccharide abundance between dairy cattle breeds.

Free milk oligosaccharides are bioactive molecules that function as prebiotics and prevent infections that commonly afflict developing infants. To date, few publications have examined the factors affecting bovine milk oligosaccharide production among cattle in the dairy industry. Here we have applied a high-throughput isobaric labeling technique to measure oligosaccharide abundances in milk collected from Danish Holstein-Friesian and Jersey dairy cattle by liquid chromatography-mass spectrometry. With a total of 634 milk samples, this collection represents the largest sample set used for milk oligosaccharide profiling in the current literature. This study is also the first to use isobaric labeling for the purpose of measuring free oligosaccharides in a real sample set. We have identified 13 oligosaccharides that vary significantly by breed, with most structures being more abundant in the milk of Jersey cattle. The abundances of several oligosaccharides were increased in second-parity cows, and correlations between the abundances of oligosaccharide pairs were identified, potentially indicating similarities in their synthetic pathways. Fucosylated oligosaccharide structures were widely identified among both breeds. Improving our understanding of oligosaccharide production will aid in developing strategies to recover these compounds from processing streams and may enable their use as a functional ingredient in foods for infants and adults

uc_Milk: An ontology for scientifically-based unambiguous characterization of mammalian milks, their composition and the biological processes giving rise to their creation

Abstract -Recent efforts in biological ontology go to great lengths to unambiguously categorize biological entities and phenomena of the natural world, as well as their relationships with each other. This paper illustrates the importance of unambiguously characterizing mammalian milk because milk is a complex mixture of many chemical components and thus represents a key role in infant nourishment and development. In addition to the build of a computable knowledge base around mammalian milk, ontological modeling of this aspect of biology and chemistry enable increased understanding of mammalian milk composition and the biological structures and biochemical processes giving rise to their creation. Utilizing unambiguous vocabularies to compare human milk with other mammalian milks relative to the biological and behavioral survival challenges facing varied mammalian organisms and the phenotypic qualities each milk confers, is a fundamental goal of this project. Keywords -mammalian milk composition; ontological modeling; biological processes One foundational design pattern for creating uc_Milk is an ontology for unambiguous characterization of mammalian milks, their composition and the biological processes giving rise to their creation. Available online at the GitHub website github.com/IC-FOODS/uc_Milk, uc_Milk is part of a larger multi-ontology framework [1] currently being housed within the International Center for Food Ontology Operability, Data, and Semantics. IC-FOODS at UC Davis We used the Basic Formal Ontology as an upper ontology because of its ubiquity in biological and biomedical ontologies, for facilitating integration by way of meta-concepts. In addition to characterizing the biological and biochemical structures and processes related to mammalian milks, since milk is a commodity as well as an essential biological fluid, uc_Milk ontology considers social, commercial, and environmental entities related to milk consumption and production (both as a food and as an ingredient in foods). These additional classifications hold promise for uc_Milk ontology usability relative to traceability and trust in commercial milk and all milk products, and provide a platform for improvement of processing and production techniques, information about milk distribution, availability and choice, as well as increased health and wellbeing of consumers. uc_Milk maps directly to entities from several other ontologies. Yet because milk is often at the "leaf" end of existing ontologies as in the "Environment Ontology". The "Uber Anatomy Ontology (Uberon)" and the "Gene Ontology" and "Vertebrate Taxonomy Ontology" respectively provide structures for mammary gland structures and development, the lactogenesis biological process, and formal mammalian phylogeny. uc_Milk also maps to the "Cell Ontology (CL)" for the intracellular anatomical structures and processes. uc_Milk is currently expanding feeding behaviors by mapping to the burgeoning uc_Eating ontology as well as to anatomical entities and the roles in the development of neural infant tissues in the NeuroBehavior Ontology. uc_Milk increases availability of structured knowledge relative to milk. Lactation is the hallmark underlying biological force driving the mother-infant dyad in Mammalia. The significant energetic and metabolic costs of lactation imposed to the mother suggests that milk's role in infant health extend well beyond simple nutrition. Millennia of selective pressures on the mother-infant dyad, have conferred in milk a dual purpose: an optimal source of nutrients, and a delivery vehicle for bioactive agents. Milk is known as nature's most complete food because of its complex mixture of bioactive components and essential nutrients such as protein, fat, carbohydrate, minerals, vitamins, and physiologically active substances. Aside from nutritional values of milk, biologically active compounds such as casein and whey proteins have been found to be increasingly important for physiological and biochemical functions that have crucial impacts on human metabolism and health. Accordingly, the infant gut and its associated microbiota are adapted not only for the utilization and absorption of milk macronutrients, but also to respond to non-nutritive, yet bioactive molecular entities. Important non-nutritive, bioactive agents in human milk include an astonishing high number of complex oligosaccharides, and a plethora of glycoconjugated proteins and lipids. Complex prebiotic oligosaccharides are known to be present in domestic animal milks, yet are in low abundance relative to human milk,

An improved method for the purification of milk oligosaccharides by graphitised carbon-solid phase extraction

Milk oligosaccharides (OS) are bioactive molecules that impart a variety of health benefits to the consumer. Techniques commonly used to analyse and quantify OS require optimised extraction methods to separate the OS from more abundant milk components. Solid phase extraction (SPE) is frequently used to isolate milk OS from lactose; however, the literature contains no formal studies on its efficacy in this application. In this study, established SPE conditions were modified to improve the technique's effectiveness in purifying OS from lactose. Low concentrations of acetonitrile (ACN) and trifluoroacetic acid (TFA) were tested for solid phase washing. Lactose removal and retention of many OS were significantly improved when using 4% ACN/0.1% TFA compared with the more common water washing technique. Different behaviours between acidic and neutral OS were evident. The new SPE technique improves extraction efficiency for bovine milk OS in applications that do not require prior lactose hydrolysis

Profiling of aminoxyTMT-labeled bovine milk oligosaccharides reveals substantial variation in oligosaccharide abundance between dairy cattle breeds.

Free milk oligosaccharides are bioactive molecules that function as prebiotics and prevent infections that commonly afflict developing infants. To date, few publications have examined the factors affecting bovine milk oligosaccharide production among cattle in the dairy industry. Here we have applied a high-throughput isobaric labeling technique to measure oligosaccharide abundances in milk collected from Danish Holstein-Friesian and Jersey dairy cattle by liquid chromatography-mass spectrometry. With a total of 634 milk samples, this collection represents the largest sample set used for milk oligosaccharide profiling in the current literature. This study is also the first to use isobaric labeling for the purpose of measuring free oligosaccharides in a real sample set. We have identified 13 oligosaccharides that vary significantly by breed, with most structures being more abundant in the milk of Jersey cattle. The abundances of several oligosaccharides were increased in second-parity cows, and correlations between the abundances of oligosaccharide pairs were identified, potentially indicating similarities in their synthetic pathways. Fucosylated oligosaccharide structures were widely identified among both breeds. Improving our understanding of oligosaccharide production will aid in developing strategies to recover these compounds from processing streams and may enable their use as a functional ingredient in foods for infants and adults