Milk Lipids Research Articles

Lipid profiling reveals the presence of unique lipid mediators in human milk from healthy and mastitic subjects

Milk lipids are an important energy source for infants, but the composition of milk lipids has not yet been clarified in detail. In this study, we analyzed free fatty acids and their metabolites in milk from humans and cows. In comparison to cow milk, human milk showed a higher content of free fatty acids including polyunsaturated fatty acids, especially ω-3 fatty acids and their metabolites. Polyunsaturated fatty acids were enriched at an early period of lactation, while saturated fatty acids did not change significantly over the period. Moreover, human milk contained high levels of ω-3 fatty acid metabolites, particularly 18-hydroxyeicosapentaenoic acid, an eicosapentaenoic acid-derived metabolite with anti-inflammatory activity. In comparison with human normal milk, thromboxane B2 and protectin D1 levels were significantly elevated in milk from individuals with mastitis, suggesting that these lipid mediators could be potential biomarkers of obstructive mastitis. Overall, the unique lipid profile of human milk supports the efficacy of breast-feeding for supply of more nutritional and bioactive lipids in comparison to artificial or cow milk to infants, in whom digestive and absorptive functions are still immature.

Effect of bypass fat on milk lipid quality and reproductive efficiency of dairy cows

Nariño (Colombia) has high dairy potential; however, their herds present difficulties in the nutritional component (energy imbalance) of the cows, affecting their productivity. This research evaluated the effect of a bypass fat supplement on lipid concentration in milk and the reproductive efficiency of dairy cows. The study was based on 21 Holstein x Simmental cows between first and third calving and the following was supplied: forage + concentrate (T1); forage + concentrate + 250 g/day of bypass fat (T2) and forage + concentrate + 250 g/day of bypass fat enriched with omega-three (T3). The experiment was conducted from day 15 pre-calving until day 105 of lactation. During this period, milk quantity, quality, composition, and reproductive behavior were recorded. For data analysis, a repeated measure mixed design with time-series data was used, where the fixed effects were the treatments, periods, and their interaction, the animals represented the random effect, and the covariable was the estimated milk yield during lactation. The results showed normal mobilization of adipose tissue; the compositional quality of the milk did not vary across treatments, although T2 presented a higher estimated production per lactation, and T3 presented a higher percentage of fatty acid C18:2. In the reproductive indicators, they were not influenced between the treatments; concluding that the base diet offered in the herd presents a nutritional balance appropriate to the requirements of the cows in production.

Microbial and metabolomic insights into the bovine lipometabolic responses of rumen and mammary gland to zymolytic small peptide supplementation.

Small peptides provide the easily utilized nitrogen for rumen microbial and promote acetate generation for milk fat synthesis. However, the impacts of peptide supplements on lipometabolic processes were still unclear. Therefore, a total of 800 multiparous dairy herds (with an average live weight of 667.6 ± 39.4 kg, an average lactation of 89.3 ± 18.8 days, and an average calving parity of 2.76 ± 0.47) were randomly allocated to the control (CON) and the small peptide (SP) supplement (100 g/day for each cow) treatments, respectively. A 35-day-long feeding procedure that includes a 7-day-long pretreatment test and a 28-day-long treatment test was followed for all cows. Dry matter intake (DMI) was recorded every day and calculated by the deviation between the supply and residue, while the daily milk production was automatically recorded through the rotary milking facilities. Milk samples were collected from each replicate on the last day, followed by the milk quality and milk lipid composition measurement. Rumen fluid samples were collected on the last day through esophageal tubing 3 h after morning feeding for the determination of the underlying mechanism of the small peptide on lipid metabolism through the measurement of rumen lipometabolic-related metabolites and rumen bacterial communities. Results indicated that dry matter intake showed an increasing trend, while milk production and the milk fat content remarkably increased after SP supplement (P < 0.05). Further detailed detection showed the mainly increased milk composition focused on monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA). Acetate-producing microbes, such as Acetitomaculum, Bifidobacterium, Succiniclasticum, and Succinivibrio, and butyrate-producing microbes, such as Shuttleworthia and Saccharofermentans, significantly proliferated, which causatively brought the increased ruminal content of acetate, isobutyrate, and butyrate after SP supplement (P < 0.05) compared with CON. Lipometabolic metabolites such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), triacylglycerol (TG), and Acetyl-CoA also significantly increased after SP supplement. In summary, SP supplements help to increase milk fat content through the proliferation of rumen bacterial communities, which provided more acetate and butyrate for milk fat synthesis combined with the promotion of ruminal lipometabolism.

An Integrated Analysis of Lactation-Related miRNA and mRNA Expression Profiles in Donkey Mammary Glands.

Donkey milk is consumed by humans for its nutritional and therapeutic properties. MicroRNAs (miRNAs) and messenger RNAs (mRNAs) have been implicated in the regulation of milk component synthesis and mammary gland development. However, the regulatory profile of the miRNAs and mRNAs involved in lactation in donkeys is unclear. We performed mRNA-seq and miRNA-seq and constructed coexpression regulatory networks for the mammary glands during the lactating and nonlactating period of jennies. We identified 3144 differentially expressed (DE) mRNAs (987 upregulated mRNAs and 2157 downregulated mRNAs) and 293 DE miRNAs (231 upregulated miRNAs and 62 downregulated miRNAs) in the lactating group compared to the nonlactating group. The DE miRNA target mRNA were significantly associated with pathways related to RNA polymerase, glycosphingolipid biosynthesis, mRNA surveillance, ribosome biogenesis in eukaryotes, glycerophospholipid metabolism, Ras signaling, and the fly hippo signaling pathway. The mRNA–miRNA coregulation analysis showed that novel-m0032-3p, miR-195, miR-26-5p, miR-23-3p, miR-674-3p, and miR-874-3p are key miRNAs that target mRNAs involved in immunity and milk lipid, protein, and vitamin metabolism in the jenny mammary gland. Our results improve the current knowledge of the molecular mechanisms regulating bioactive milk component metabolism in the mammary glands and could be used to improve milk production in donkeys.

Human Milk Lipid Profiles around the World: A Systematic Review and Meta-Analysis.

Reported breast milk lipid concentrations may vary with geographical region, postnatal age, and year of sample collection. In this review, we summarized data on the concentrations of total fat, total phospholipids, cholesterol, and fatty acids in human milk worldwide and their variation according to lactation stage, study area, and sample collection year. A systematic literature search was performed using the PubMed, Embase, Web of Science, and Medline databases for English-language papers and Wanfang and China National Knowledge Infrastructure databases for Chinese-language papers. A total of 186 studies evaluating the human milk lipid profiles were included. According to random-effects models based on worldwide data, the summarized means (95% CIs) as percentages of total fat were 42.2% (41.1%, 43.3%) for SFAs, 36.6% (35.6%, 37.5%) for MUFAs, and 21.0% (19.3%, 22.7%) for PUFAs. However, the study heterogeneity was high for most types of fatty acids (I2>99%). Human milk from Western countries had higher concentrations of MUFAs and 18:1n-9 (ω-9), but lower concentrations of PUFAs, 18:2n-6, 20:4n-6, 18:3n-3, 20:5n-3, 22:6n-3, and total n-6 PUFA compared with those from non-Western countries (P < 0.001-0.011). Significant lactation stage differences were observed for total fat and some individual fatty acids. The concentrations of SFAs and 16:0 were significantly negatively correlated with sampling year (P<0.001-0.028). In contrast, a significant positive correlation between the concentrations of 18:2n-6 and 18:3n-3 and sampling year was observed (P<0.001-0.035). Our results suggest that the pooling of data on human milk lipid profiles in different studies should be done with caution due to the high between-study heterogeneity. The concentration of lipids, including total fat, cholesterol, and specific fatty acids, differs in human milk according to lactation stage, geographical region, and year of sample collection.

Neolithic culinary traditions revealed by cereal, milk and meat lipids in pottery from Scottish crannogs

Cereal cultivation in Britain dates back to ca. 4000 BCE, probably introduced by migrant farmers from continental Europe. Widespread evidence for livestock appears in the archaeozoological record, also reflected by ubiquitous dairy lipids in pottery organic residues. However, despite archaeobotanical evidence for domesticated plants (such as cereals), organic residue evidence has been near-absent. Our approach, targeting low-abundance cereal-specific markers, has now revealed evidence for cereals (indicating wheat) in Neolithic pottery from Scottish ‘crannogs’, dating to ca. 3600 – 3300 BCE. Their association with dairy products suggests cereals may have been regularly prepared together as a milk-based gruel. We also observed a strong association between the occurrence of dairy products and smaller-mouthed vessels. Here, we demonstrate that cereal-specific markers can survive in cooking pots for millennia, revealing the consumption of specific cereals (wheat) that are virtually absent from the archaeobotanical record for this region and illuminating culinary traditions among early farming communities.

Role of lipidomics in assessing the functional lipid composition in breast milk.

Breast milk is the ideal source of nutrients for infants in early life. Lipids represent 2–5% of the total breast milk composition and are a major energy source providing 50% of an infant’s energy intake. Functional lipids are an emerging class of lipids in breast milk mediating several different biological functions, health, and developmental outcome. Lipidomics is an emerging field that studies the structure and function of lipidome. It provides the ability to identify new signaling molecules, mechanisms underlying physiological activities, and possible biomarkers for early diagnosis and prognosis of diseases, thus laying the foundation for individualized, targeted, and precise nutritional management strategies. This emerging technique can be useful to study the major role of functional lipids in breast milk in several dimensions. Functional lipids are consumed with daily food intake; however, they have physiological benefits reported to reduce the risk of disease. Functional lipids are a new area of interest in lipidomics, but very little is known of the functional lipidome in human breast milk. In this review, we focus on the role of lipidomics in assessing functional lipid composition in breast milk and how lipid bioinformatics, a newly emerging branch in this field, can help to determine the mechanisms by which breast milk affects newborn health.

Profiles of total and sn-2 fatty acid of human mature milk and their correlated factors: A cross-sectional study in China.

Fatty acid (FA) in breast milk is beneficial to the growth and neurodevelopment of infants. However, the structure profiles of breast milk FAs and the influencing factors which are crucial for normal function have not been fully elucidated. This study aimed to characterize the profiles of total and sn-2 FAs in human mature milk based on two representative urban areas in China and explore potential sociodemographic determinants. Mothers (n = 70) at 40–100 d postpartum from Beijing and Danyang were recruited according to unified inclusion and exclusion criteria. Total and sn-2 FA compositions were examined by gas chromatography and quantified. Using the Spearman correlation and multiple regression model, we found that the location and maternal education level were the most conspicuous correlated factor. The milk of mothers from Beijing had higher levels of the n-6 series of long-chain polyunsaturated fatty acids (LCPUFA) (C20:2, C20:3n-6, C20:4n-6, n-6PUFA/n-3PUFA, LA/ALA, and ARA/DHA) than that of Danyang, while the opposite was observed in the n-3 series of LCPUFA (C18:3n-3 and Total n-3PUFA). Compared to the milk of mothers with a high school degree or below, those with a bachelor's degree or above had lower SFAs (C10:0, C12:0, C14:0, and Total SFA), n-3 series of LCPUFA (C18:3n-3 and Total n-3PUFA), C18:1n-9t, and higher n-6 series of LCPUFA (C18:2n-6c, C20:2, C20:4n-6, Total n-6PUFA, and n-6PUFA/n-3PUFA). Maternal age, infant gender, pre-conception body mass index (BMI), parity, delivery mode, and gestational weight gain were also associated with total FAs. However, fewer associations were found between the above factors and sn-2 FAs. This study will promote an understanding of human breast milk's lipid profile and help develop a formula more suitable for infants.

Effect of Delta-9-tetrahydrocannabinol and cannabidiol on milk proteins and lipid levels in HC11 cells.

Pregnant and lactating women have been discouraged from using cannabis by Health Canada. However, the increasing rate of cannabis use among pregnant women has presented an urgent need to investigate its physiological effects during the perinatal period. During pregnancy, the mammary gland (MG) undergoes remodeling, which involves alveolar differentiation of mammary epithelial cells (MECs), which is essential for breast milk production and secretion. Limited evidence has been reported on the impact of cannabis or its components, delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), on MG development or MEC differentiation. In this study, we investigated the effects of THC and CBD on the differentiation of MECs by assessing changes in cellular viability, lipid accumulation, and gene and protein expression of major milk protein and lipid synthesizing markers. using the HC11 cells as a model. We hypothesized that THC and CBD will negatively impact the synthesis of milk proteins and lipids, as well as lipid markers in HC11 cells. Our results demonstrated that THC and CBD reduced cellular viability at concentrations above 30μM and 20μM, respectively. Relative to control, 10μM THC and 10μM CBD reduced mRNA levels of milk proteins (CSN2 and WAP), lipid synthesizing and glucose transport markers (GLUT 1, HK2, FASN, FABP4, PLIN2 and LPL), as well as whey acidic protein and lipid levels. In addition, co-treatment of a CB2 antagonist with THC, and a CB2 agonist with CBD, reversed the impact of THC and CBD on the mRNA levels of key markers, respectively. In conclusion, 10μM THC and CBD altered the differentiation of HC11 cells, in part via the CB2 receptor.

The milk fat globule size governs a physiological switch for biofilm formation by Bacillus subtilis.

Milk lipids are organized in the form of milk fat globules (MFG), ranging in size from 0. 1 to 15 μm. The MFG size is closely associated with the composition of fatty acids, polar lipids, sphingolipids, cholesterol and the content of the MFG membrane (MFGM). Also, the MFGM integral proteins and glycoconjugates differ in composition and structure between different MFG size groups. These compositional differences may modulate the functionality of the MFG and its interaction with microbial cells. We report that small (2.3 μm) MFG facilitates the growth of the Gram-positive bacterium Bacillus subtilis whereas induction of biofilm formation was found in the presence of large (7.0 μm) MFG. Attempting to distinguish between the role played by the size from that played by the composition of the MFG, we compared phospholipid composition between treatments. We found that adjusting the phosphatidylethanolamine (PE) level to the concentration found in the small MFG, increased growth but suppressed biofilm formation in the presence of large MFG. The same normalization protocol for phosphatidylinositol (PI) or sphingomyeline (SM) did not exert a similar effect, suggesting a specific role for PE in regulating bacteria proliferation. We suggest that the content of MFGM, affected by MFG size, governs the ability of B. subtilis to utilize lipids from milk fat. This process might affect the bacterial decision-making toward biofilm formation or growth.

Influence of gestational diabetes mellitus on lipid signatures in breast milk and association with fetal physical development.

Gestational diabetes mellitus (GDM) commonly leads to adverse pregnancy outcomes and long-term metabolic complications in offspring. Breastfeeding has been shown to rewrite the fetal “metabolic programming” resulting from maternal diabetes and finally lead to a lower risk of future metabolic disease. Lipids in breast milk act like hormones to promote infant growth and development, but there is minimal information invested thus far in constitution changes of lipids in breast milk, especially in the context of GDM. In the present study, we performed a lipidomics analysis to compare the lipid composition in breast milk collected from women with or without GDM. We further revealed the correlations of dysregulated lipids in breast milk with maternal glucose and infant physical development. A total of 833 lipid species from 15 classes were identified, 60 of which were found to be significantly altered in response to the high glucose, suggesting a remarkable lipid profiling change in breast milk induced by GDM. Our results showed significant associations between dysregulated lipids (e.g., neutral lipids, phospholipids, sphingolipids) and maternal glucose. Furthermore, correction analysis demonstrated that GDM related lipids were also associated with indicators of infant physical development, including body weight, length, and head circumference. These findings may help to understand the protective effects of breastfeeding especially during GDM pregnancy.

Lipidomic profiling using GC and LC-MS/MS revealed the improved milk quality and lipid composition in dairy cows supplemented with citrus peel extract

Eight lactating cows were used to determine the effects of citrus peel extract (CPE) on milk performance, antioxidant properties, and milk lipids composition. CPE supplementation up to 150 g/d (CPE150) increased milk yield and the proportions of unsaturated fatty acids of conjugated linoleic acid. CPE with abundant polyphenol and flavonoids can transfer these bioactive substances to mammary gland and improve the antioxidant properties of milk obtained from cows. Lipidomics revealed that 56 lipid species were altered between CON vs CPE150, and there were five key differential metabolic pathways. In particular, milk phosphatidylethanolamine and phosphatidylcholine were significantly increased with dietary CPE supplementation. In summary, our results provide insights into the modifications in the milk components and milk quality of dairy cows received CPE. The inclusion of CPE in the diet of dairy cows may be an effective and natural way to increase the antioxidant amounts and beneficial lipids in milk.

Comparative Lipidomics Analysis of Human and Ruminant Milk Reveals Variation in Composition and Structural Characteristics.

In the present study, the different lipidomes between human milk and ruminant milk were compared. The 471, 376, 467, and 87 differential lipids were identified in human versus cow, goat, sheep, and camel groups, respectively. According to multivariate statistical analysis, lipids in human and camel milk were closer but differed from other milk. The distributions of long-chain and polyunsaturated fatty acids of triglycerides (TGs), the proportions of functional TGs (OPO, OPL, and PPO), and many kinds of phospholipids (PLs) (PS, PI, GD, GM3, and Cer) in human milk were similar to those in camel milk. The similar structure of TGs and proportion of PLs in human milk to camel milk might contribute to their similar digestion and bioactivity properties. Camel milk could be considered as a new resource of lipid base for infant formula. Minor PLs should also be considered for designing formula. Our results provide a new sight for humanized lipids in infant formula.

Reverting to a Healthy Diet during Lactation Normalizes Maternal Milk Lipid Content of Diet-Induced Obese Rats and Prevents Early Alterations in the Plasma Lipidome of the Offspring.

ScopeThis study aims to assess in rats whether normalizing maternal diet during lactation prevents the harmful effects of western diet (WD) consumption during the whole perinatal period on the lipidomic profile in maternal milk and offspring plasma.Methods and ResultsControl dams (CON‐dams), fed with standard diet (SD); WD‐dams, fed with WD prior and during gestation and lactation; and reversion dams (REV‐dams), fed as WD‐dams but moved to SD during lactation are followed. Lipidomic analysis is performed in milk and plasma samples from pups. Milk of WD‐dams presents a different triacylglycerol composition and free fatty acid (FA) profile compared to CON‐dams, including an increased ratio of pro‐inflammatory to anti‐inflammatory long‐chain polyunsaturated FA. Such alterations, which are also present in the plasma of their offspring, are widely reversed in the milk of REV‐dams and the plasma of their pups. This is related with the recovery of control adiponectin expression levels in the mammary gland, and the presence of decreased expression of pro‐inflammatory factors.ConclusionImplementing a healthy diet during lactation prevents early alterations in the plasma lipidome of pups associated to the maternal intake of an obesogenic diet, which may be related to the normalization of milk lipid content and the inflammatory state in the mammary gland.

The Role of Human Milk Lipids and Lipid Metabolites in Protecting the Infant against Non-Communicable Disease.

Non-communicable diseases continue to increase globally and have their origins early in life. Early life obesity tracks from childhood to adulthood, is associated with obesity, inflammation, and metabolic dysfunction, and predicts non-communicable disease risk in later life. There is mounting evidence that these factors are more prevalent in infants who are formula-fed compared to those who are breastfed. Human milk provides the infant with a complex formulation of lipids, many of which are not present in infant formula, or are present in markedly different concentrations, and the plasma lipidome of breastfed infants differs significantly from that of formula-fed infants. With this knowledge, and the knowledge that lipids have critical implications in human health, the lipid composition of human milk is a promising approach to understanding how breastfeeding protects against obesity, inflammation, and subsequent cardiovascular disease risk. Here we review bioactive human milk lipids and lipid metabolites that may play a protective role against obesity and inflammation in later life. We identify key knowledge gaps and highlight priorities for future research.

Development and Application of Feature-Based Molecular Networking for Phospholipidomics Analysis.

Phospholipids are small but critical lipids in milk. Conventional lipidomics is a powerful method for the analysis of lipids in milk. Although the number of lipidomics software has drastically increased over the past five years, reducing false positives and obtaining structurally accurate annotations of phospholipids remain a significant challenge. In this study, we developed a rapid and accurate method for measuring a wide spectrum of phospholipids in milk. The developed approach that employed information-dependent acquisition (IDA) mode and feature-based molecular networking has exhibited better performance on data processing and lipid annotation when compared with sequential window acquisition of all theoretical mass spectra (SWATH) and MS-DIAL. This validated method was further evaluated using three kinds of sheep milk. A total of 150 phospholipids were identified, including rarely reported phospholipids containing ethers or vinyl ethers. The result indicated that phospholipids could be used as potential markers to distinguish sheep milk from different varieties and origins. The experimental and computational methods provide a rapid and reliable method for the investigation of phospholipids in milk.

Babies, Bugs, and Barriers: Dietary Modulation of Intestinal Barrier Function in Early Life.

The intestinal barrier is essential in early life to prevent infection, inflammation, and food allergies. It consists of microbiota, a mucus layer, an epithelial layer, and the immune system. Microbial metabolites, the mucus, antimicrobial peptides, and secretory immunoglobulin A (sIgA) protect the intestinal mucosa against infection. The complex interplay between these functionalities of the intestinal barrier is crucial in early life by supporting homeostasis, development of the intestinal immune system, and long-term gut health. Exclusive breastfeeding is highly recommended during the first 6 months. When breastfeeding is not possible, milk-based infant formulas are a safe alternative. Breast milk contains many bioactive components that help to establish the intestinal microbiota and influence the development of the intestinal epithelium and the immune system. Importantly, breastfeeding lowers the risk for intestinal and respiratory tract infections. Here we review all aspects of intestinal barrier function and the nutritional components that impact its functionality in early life, such asmicronutrients, bioactive milk proteins, milk lipids, and human milk oligosaccharides. These components are present in breast milk and can be added to milk-based infant formulas to support gut health and immunity.

HPLC-MS, GC and NMR Profiling of Bioactive Lipids of Human Milk and Milk of Dairy Animals (Cow, Sheep, Goat, Buffalo, Camel, Red Deer)

For non-bovine milks, information regarding bioactive lipids is fragmented, unreliable or unavailable. The purpose of the current study was to analyse bioactive lipids in the milk of dairy animals using modern analytical methods to achieve the most reliable results. Bioactive lipids in human milk were also analysed and used as a reference. A suite of modern analytical methods was employed, namely High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS), Gas Chromatography (GC) and Nuclear Magnetic Resonance (NMR). The total lipid content was determined, and phospholipid, fatty acid, neutral glycosphingolipids and ganglioside (GM3 and GD3) levels were measured. Lipid classes in selected milks were reliably characterised for the first time, including gangliosides in deer, camel and sheep; cerebrosides in deer, camel and buffalo; plasmalogens in deer, buffalo and goat and phospholipids in deer. Our study demonstrated the advantage of utilising a range of analytical techniques in order to characterise a diverse set of bioactive lipids.

Complete Genome Sequence of Chryseobacterium mulctrae KACC 21234T: A Potential Proteolytic and Lipolytic Bacteria Isolated from Bovine Raw Milk

Chryseobacterium mulctrae KACC 21234T is a novel species isolated from raw bovine milk. Psychrotrophic bacteria are considered contaminants and are hypothesized to originate from the environment. In this investigation, the C. mulctrae KACC 21234T genome was determined to be 4,868,651 bp long and assembled into four contigs with a G+C ratio of 33.8%. In silico genomic analyses revealed the presence of genes encoding proteases (endopeptidase Clp, oligopeptidase b, carboxypeptidase) and lipases (phospholipase A(2), phospholipase C, acylglycerol lipase) that can catalyze the degradation of the proteins and lipids in milk, causing its quality to deteriorate. Additionally, antimicrobial resistance and putative bacteriocin genes were detected, potentially intensifying the pathogenicity of the strain. The genomic evidence presented highlights the need for improved screening protocols to minimize the potential contamination of milk by proteolytic and lipolytic psychrotrophic bacteria.

Dietary Fiber to Starch Ratio Affects Bovine Milk Oligosaccharide Profiles

ABSTRACTBackgroundBovine milk oligosaccharides (BMOs) have several demonstrated and hypothesized benefits including roles in cognitive development and antipathogenic activities, making them promising ingredients for infant formulas and nutraceutical applications. BMO extraction from bovine milk is challenged by low concentrations relative to nonbioactive simple sugars like lactose. BMO abundances are known to vary with a cow's lactation stage, breed, and parity, but these characteristics are difficult to modify in existing dairy herds. In contrast, diet modification is an accessible target, and is already known to influence milk yield, lipid content, protein levels, and monosaccharide compositions.ObjectivesTo determine the impact of a low starch high fiber versus a high starch low fiber diet on overall BMO profiles and individual BMO abundances in Holstein dairy cattle.MethodsMilk samples were collected from 59 midlactation Holsteins in a crossover study featuring dietary modification with either a low starch high fiber or high starch low fiber feed. BMO profiles were evaluated by nano-LC quadrupole time-of-flight tandem MS, and differences in BMO abundances between diets were evaluated using linear mixed effects modeling.ResultsA total of 19 BMOs were identified across the sample set, including 4 large fucosylated compounds. Seven BMOs were found to have significantly more positive percent changes in yield-adjusted abundance from the pre-experiment baseline period for milk samples collected during feeding with the low starch high fiber diet compared with the high starch low fiber diet.ConclusionsConsuming the low starch high fiber diet promoted greater overall BMO production than the high starch low fiber diet in a population of midlactation Holsteins. Additionally, this study afforded the opportunity to investigate the impact of other factors potentially influencing BMO abundances, furthering understanding of how dairy herd management practices can positively impact milk composition and support the potential use of BMOs as functional ingredients.