Microstructure of fat globules in whole milk after thermosonication treatment

The structure of fat globules in whole milk was studied after heat and thermosonication treatments to observe what happens during these processes at the microscopic level using scanning electron microscopy. Raw whole milk was thermosonicated in an ultrasonic processor-Hielscher UP400S (400 W, 24 kHz, 120 microm amplitude), using a 22-mm probe at 63 degrees C for 30 min. Heat treatment involved heating the milk at 63 degrees C for 30 min. Color and fat content were measured to correlate the images with analytical measurements. The results showed that the surface of the fat globule was completely roughened after thermosonication. Ultrasound waves were responsible for disintegrating the milk fat globule membrane (MFGM) by releasing the triacylglycerols. Furthermore, the overall structure of milk after sonication showed smaller fat globules (smaller than 1 microm) and a granular surface. This was due to the interaction between the disrupted MFGM and some casein micelles. Minor changes in the aspect of the globules between thermal and raw milks were detected. Color measurements showed higher L* values for sonicated samples. Sonicated milk was whiter (92.37 +/- 0.20) and generally showed a better degree of luminosity and homogenization compared to thermal treated milk (88.25 +/- 0.67) and raw milk (87.82 +/- 0.18). Fat content analysis yielded a higher value after sonication (4.24%) compared to untreated raw milk (4.04%) because fat extraction is more efficient after sonication. The advantages of thermosonicated milk are that it can be pasteurized and homogenized in just 1 step, it can be produced with important cost savings, and it has better characteristics, making thermosonication a potential processing method for milk and most other dairy products.     

Analysis of major proteins and fat fractions associated with mare's milk fat globules

Although several studies have aimed to identify mare's milk proteins, only the major whey proteins and some caseins have yet been characterized. Incomplete sequencing of the equine genome and the difficulty of recovering highly hydrophobic proteins mean that little is known to date about the proteins associated with milk fat globules, which have been shown to play an important role in newborns' defense mechanisms. The fat fraction, in particular the distribution of unsaturated fatty acids, has been more extensively studied, but complex lipids are only partially elucidated. This study reports a 2-DE approach combined with a powerful method for de novo protein sequencing, and quali-quantitative data on complex lipid composition determined by high performance TLC (HPTLC) and GC. The presence in mare's milk of long-chain highly unsaturated fatty acids, and the evidence of close similarity between equine and human milk fat globule membrane proteins, support the use of mare's milk for human nutrition.     

Lipid rafts in the bovine milk fat globule membrane revealed by the lateral segregation of phospholipids and heterogeneous distribution of glycoproteins

This study reveals the lateral organisation of the milk fat globule membrane (MFGM). Using confocal laser scanning microscopy (CLSM) and a lipid soluble molecule, an exogenous phospholipid and two lectins as fluorescent probes we located triacylglycerols in the core of fat globules and investigated the organisation of the polar lipids and glycoproteins of the MFGM, in situ in milk. Lipid rafts corresponding to the lateral segregation of sphingolipids in liquid-ordered phases surrounded by liquid-disordered domains composed by the glycerophospholipids were observed in the MFGM. These lipid rafts which correspond to rigid sphingolipid-rich domains have a circular shape at room temperature. CLSM experiments revealed that glycoproteins and glycolipids are heterogeneously distributed around fat globules and that they are not located in the lipid rafts. The characterisations performed by in depth thin sectioning of fat globules and in dynamic as a function of time revealed chemical and structural heterogeneities in the MFGM. Schematic 3D and 2D representations of the MFGM are proposed and discussed. The physiological and nutritional consequences of the lateral organisation of polar lipids and glycoproteins in the MFGM are discussed but remain to be elucidated.     

Buffalo vs. cow milk fat globules: Size distribution,zeta-potential,compositions in total fatty acids and in polar lipids from the milk fat globule membrane

Although buffalo milk is the second most produced milk in the world, and of primary nutritional importance in various parts of the world, few studies have focused on the physicochemical properties of buffalo milk fat globules. This study is a comparative analysis of buffalo and cow milk fat globules. The larger size of buffalo fat globules, 5 vs. 3.5 μm, was related to the higher amount of fat in the buffalo milks: 73.4 ± 9.9 vs. 41.3 ± 3.7 g/kg for cow milk. Buffalo milks contained significantly lower amount of polar lipids expressed per gram of lipids (0.26% vs. 0.36%), but significantly higher amount of polar lipids per litre of milk (+26%). Buffalo and cow milk fat globule membranes contain the same classes of polar lipids; phosphatidylethanolamine, sphingomyelin (SM) and phosphatidylcholine (PC) being the main constituents. A significant higher percentage of PC and lower percentage of SM were found for buffalo milks. The fatty acid analysis revealed that saturated fatty acids, mainly palmitic acid, trans fatty acids, linolenic acid (ω3) and conjugated linolenic acid were higher in buffalo milk than in cow milk. Such results will contribute to the improvement of the quality of buffalo milk-based dairy products.     

Transfer of dietary zinc and fat to milk--evaluation of milk fat quality, milk fat precursors, and mastitis indicators

The present study demonstrated that the zinc concentration in bovine milk and blood plasma is significantly affected by the intake of saturated fat supplements. Sixteen Holstein cows were used in a 4 × 4 Latin square design with 4 periods of 12 d, and 4 dietary treatments were conducted. A total mixed ration based on corn silage, grass-clover silages, and pelleted sugar beet pulp was used on all treatments. A high de novo milk fat diet was formulated by adding rapeseed meal and molasses in the total mixed ration [39 mg of Zn/kg of dry matter (DM)], and a low de novo diet by adding saturated fat, fat-rich rapeseed cake, and corn (34 mg of Zn/kg of DM). Dietary Zn levels were increased by addition of ZnO to 83 and 80 mg of Zn/kg of DM. Treatments did not affect daily DM intake, or yield of energy-corrected milk, milk fat, or milk protein. The high de novo diet significantly increased milk fat percentage and milk content of fatty acids with chain length from C6 to C16, and decreased content of C18 and C18:1. Treatments did not influence milk free fatty acids at 4°C at 0 or 28 h after milking. The average diameter of milk fat globules was significantly greater in milk from cows offered low de novo diets. Furthermore, the low de novo diet significantly increased the concentration of nonesterified fatty acids and d-β-hydroxybutyrate in blood plasma, the latter was also increased in milk. Treatments did not affect the enzyme activity of lactate dehydrogenase and N-acetyl-β-d-glucosaminidase in milk or the activity of isocitrate dehydrogenase and malate dehydrogenase in blood plasma. The low de novo diet significantly increased plasma Zn and milk Zn content, whereas dietary Zn level did not in itself influence these parameters. This indicates that the transfer of fat from diet to milk might facilitate transfer of Zn from diet to milk.     

Size distribution of fat globules in human colostrum, breast milk, and infant formula

Only a few results are available on the size of human milk fat globules (MFG), despite its significance regarding fat digestion in the infant, and no data are available at <24 h postpartum (PP). We measured the MFG size distribution in colostrum and transitional human milk in comparison with fat globules of mature milk and infant formula. Colostrum and transitional milk samples from 18 mothers were collected regularly during 4 d PP and compared with mature milk samples of 17 different mothers and 4 infant formulas. The size distribution was measured by laser light scattering. For further characterization, the ζ-potential of some mature MFG was measured by laser Doppler electrophoresis. The MFG diameter decreased sigmoidally in the first days. At <12 h PP, the mode diameter was 8.9 ± 1.0 μm vs 2.8 ±0.3 μm at 96 h PP. Thus, the surface area of MFG increased from 1.1 ±0.3 to 5.4 ±0.7 m²/g between colostrum and transitional milk. In mature milk, the MFG diameter was 4 μm on average and increased with advancing lactation, whereas the droplets in infant formula measured 0.4 μm. The ζ potential of mature MFG was −7.8 ± 0.1 mV. The fat globules are larger in early colostrum than in transitional and mature human milk and in contrast with the small-sized fat droplets in infant formula. Human MFG also have a low negative surface charge compared with bovine globules. These structural differences can be of nutritional significance for the infant.     

Identification of ABCA1 and ABCG1 in milk fat globules and mammary cells--implications for milk cholesterol secretion

The ATP-binding cassette (ABC) transporters ABCA1 and ABCG1 play an important role in cellular cholesterol homeostasis, but their function in mammary gland (MG) tissue remains elusive. A bovine MG model that allows repeated MG sampling in identical animals at different functional stages was used to test whether 1) ABCA1 and ABCG1 protein expression and subcellular localization in mammary epithelial cells (MEC) change during the pregnancy-lactation cycle, and 2) these 2 proteins were present in milk fat globules (MFG). Expression and localization in MEC were investigated in bovine MG tissues at the end of lactation, during the dry period (DP), and early lactation using immunohistochemical and immunofluorescence approaches. The presence of ABCA1 and ABCG1 in MFG isolated from fresh milk was determined by immunofluorescence. The ABCA1 protein expression in MEC, expressed as arbitrary units, was higher during the end of lactation (12.2 ± 0.24) and the DP (12.5 ± 0.22) as compared with during early lactation (10.2 ± 0.65). In contrast, no significant change in ABCG1 expression existed between the stages. Throughout the cycle, ABCA1 and ABCG1 were detected in the apical (41.9 ± 24.8 and 49.0 ± 4.96% of cows, respectively), basal (56.2 ± 28.1 and 54.6 ± 7.78% of cows, respectively), or entire cytoplasm (56.8 ± 13.4 and 61.6 ± 14.4% of cows, respectively) of MEC, or showed combined localization. Unlike ABCG1, ABCA1 was absent at the apical aspect of MEC during early lactation. Immunolabeling experiments revealed the presence of ABCA1 and ABCG1 in MFG membranes. Findings suggest a differential, functional stage-dependent role of ABCA1 and ABCG1 in cholesterol homeostasis of the MG epithelium. The presence of ABCA1 and ABCG1 in MFG membranes suggests that these proteins are involved in cholesterol exchange between MEC and alveolar milk.     

Free oil and rheology of Cheddar cheese containing fat globules stabilized with different proteins

Cheddar cheese was manufactured from recombined milk containing fat globules coated with alpha(s1)-CN (casein), alpha(s2)-CN, beta-CN, kappa-CN, alpha-lactalbumin, or beta-lactoglobulin. The effect of the coating on fat globule structure, free oil formation, and cheese rheology was investigated to determine if globule coating affected the physical structure of cheese. Fat globule size and shape were determined in cheese using confocal laser scanning microscopy, and the rheological properties measured by uniaxial compression after maturation for 35 and 70 d. Fat globules were elongated and clustered in the control cheese coated with native membrane material and in cheese where the globules were coated with alpha(s2)-CN, but were more circular and distinct than all others. Cheese containing globules coated with alpha(s2)-CN fractured at a lower strain and with a lower stress than other experimental cheeses. Free oil decreased in cheese as the stress at fracture of the cheese protein matrix increased. Strain at fracture increased as pH increased from 4.7 to 5.3. There was no correlation between free oil and fat globule circularity. Cheddar cheese aroma was not evident in experimental cheeses.     

Composition of milk fat from cows selected for milk fat globule size and offered either fresh pasture or a corn silage-based diet

The objective of this study was to examine the synthesis and composition of milk produced by dairy cows that secrete either small milk fat globules (SMFG) or large milk fat globules (LMFG), and to study their response to diets known to alter milk composition. Four groups of 3 multiparous dairy cows were assigned to 2 isoenergetic feeding treatments: a corn silage treatment supplemented with soybean meal, and fresh pasture supplemented with cereal concentrate. The 4 groups comprised 2 groups of 3 dairy cows that produced SMFG (3.44 μm) and 2 groups of 3 dairy cows that produced LMFG (4.53 μm). The SMFG dairy cows produced higher yields of milk, protein, and calcium. Nevertheless, their milk had lower fat and protein contents. Both SMFG and LMFG cows secreted similar amounts of milk fat; therefore, higher globule membrane contents in milk fat were observed in SMFG cows. Higher calcium mineralization of the casein micelles in SMFG cows suggests that it may be possible to improve cheese-making properties even if the lower protein content may lead to lower cheese yields. The SMFG cows secrete milk fat with a higher concentration of monounsaturated fatty acids and a lower concentration of short-chain fatty acids. They also have a higher C18:1/C18:0 ratio than LMFG cows. This suggests that SMFG cows have more significant fatty acid elongation and desaturation. The pasture treatment led to an increase in milk and protein yields because of increased energy intake. It also resulted in lower milk fat yield and fat and protein contents. The pasture treatment led to a decrease in milk fat globule size and, as expected, an increase in monounsaturated and polyunsaturated fatty acid contents. However, it induced a decrease in the protein content, and in calcium mineralization of casein micelles, which suggests that this type of milk would be less suitable for making cheese. This study also shows that there is no correlation between the cows, based on milk fat globule size and diet. These results open up possibilities for improving milk fat quality based on milk fat globule size, and composition. The mechanisms involved in milk fat globule secretion are still to be determined.     

Removal of milk fat globules from whey protein concentrate 34% to prepare clear and heat-stable protein dispersions

Whey protein concentrates (WPC) are low-cost protein ingredients, but their application in transparent ready-to-drink beverages is limited due to turbidity caused by fat globules and heat instability. In this work, fat globules were removed from WPC 34% (WPC-34) to prepare heat-stable ingredients via the Maillard reaction. The removal of fat globules by acid precipitation and centrifugation was observed to be the most complete at pH 4.0, and the loss of protein was caused by micrometer-sized fat globules and protein aggregates. Spray-dried powder prepared from the transparent supernatant was glycated at 130°C for 20 and 30 min or 60°C for 24 and 48 h. The 2 groups of samples had comparable heat stability and degree of glycation, evaluated by free amino content and analytical ultracentrifugation, but high-temperature, short-time treatment reduced the color formation during glycation. Therefore, WPC-34 can be processed for application in transparent beverages.     

Impact of milking frequencies on the level of free fatty acids in milk, fat globule size, and fatty acid composition

The aim of the present study was to study the effect of milking cows 4 times daily on free fatty acids (FFA) in the milk compared with milking twice daily. An experiment was performed during 2 wk in which half udders in 11 cows were milked 2 or 4 times daily. Milk yield was measured, and milk was analyzed for fat content, FFA, fatty acid composition, fat globule size, and activity of γ-glutamyl transpeptidase. Concentration of FFA was greater (1.49 mEq/100 g of fat) in milk from half udders milked 4 times daily than in milk from the half udders milked twice daily (1.14 mEq/100 g of fat). Further, it was noted that milk from the half udder milked 4 times daily contained milk fat globules with larger average diameters. Increased milking frequency increased milk yield by 9% compared with the udder half milked twice daily, but fat content and fat yield were not affected. The results are of importance for further understanding the mechanisms behind the increased content of FFA that is frequently observed in automatic milking systems.     

Elevated concentrate-to-forage ratio in dairy cow rations is associated with a shift in the diameter of milk fat globules and remodeling of their membranes

We examined the effects of concentrate-to-forage ratio in dairy cow rations on milk-fat composition, with a specific focus on the structure of milk fat globules (MFG). Twenty-four Holstein cows, 153 d in milk, were assigned to 2 dietary treatments in a crossover design study. Treatments were (1) high-concentrate (65%), low-forage (35%; HCLF) diet and (2) low-concentrate (35%), high-forage (65%; LCHF) diet. The mean diameter of the MFG; plasma concentrations of insulin, glucose, and nonesterified fatty acids (FA); and the composition and concentrations of milk FA and polar lipids were determined. Concentrations of insulin were 56% higher, and those of nonesterified FA 46% lower, in the HCLF than in the LCHF diet. The milk yield was 8.5 kg/d higher and yields of fat, protein, and lactose were 180, 350, and 403 g/d higher, respectively, in the HCLF versus LCHF diet. Milk FA composition differed between treatments, with 1.5 and 1.0 percentage units higher saturated and polyunsaturated FA concentrations, respectively, in the HCLF versus LCHF diet. Mean MFG diameter tended to be smaller (0.2 μm) in the HCLF diet than in the LCHF diet, associated with increased daily phospholipids yield (34%), lower phosphatidylserine and higher phosphatidylcholine concentrations. In conclusion, the decreased milk and fat yields in the LCHF diet were associated with remodeling of the MFG membrane and with the secretion of larger MFG. Membrane remodeling of the mammary epithelium membranes seems to play a role in regulating MFG size.     

Invited review: Anti-adhesive properties of bovine oligosaccharides and bovine milk fat globule membrane-associated glycoconjugates against bacterial food enteropathogens

The prevalence of the main raw milk and raw milk–derived dairy product enteropathogens (Campylobacter, Shiga toxin-producing Escherichia coli, Listeria, and Salmonella) is higher than the number of epidemiological cases related to ingesting these foodstuffs. Bovine milk oligosaccharides and milk fat globule membrane (MFGM)-linked glycoconjugates interact with foodborne enteropathogens to inhibit their adhesion to intestinal cells and tissues. This review examines the main mechanisms and strategies used by enteropathogens to adhere to their target, details the anti-adhesive properties of MFGM against enteropathogens and enterotoxins, assesses the integrity of bacteria–MFGM complexes during dairy product manufacture and digestion, and discusses the potential for using these macromolecules and glycoconjugates in foods for public health.     

Comparison and characterisation of fat and protein composition for camel milk from eight Jordanian locations

Camel milk was collected from eight locations in Jordan followed by studying molecular characteristics and fatty acids contents. Lactoferrin content ranged between non-detectable amount (Al Hamra and Al Jweideh) and 0.43% of total protein (Al Hazeem). Serum albumin ranged between non-detectable amount (Al Hazeem) and 0.34% of total protein (Wadi Araba). Lactophorin variants A and B were in non-detectable amount in milk collected from Al Khalidyah and Al Hazeem, whereas the highest content of lactophorin was in milk collected from Al Qatrana. Saturated fatty acids content ranged between 50.15% (Al Umari) and 64.12% (Al Qatrana) of total fatty acids. Unsaturated fatty acids content ranged between 35.89% (Al Qatrana) and 49.85% (Al Umari) of total fatty acids. unsaturated fatty acids (C 18:1–C 18:3) varied significantly and ranged between 17.13% (Al Qatrana) and 38.85% (Al Umari) of total fatty acids.     

淡水鱼油性质测定及其在人乳替代脂市场的应用研究

对巴沙鲶鱼油等市场上常见的10种淡水鱼油脂肪酸、sn-2脂肪酸组成进行了测定与分析。结果表明:大部分淡水鱼油中棕榈酸含量在15%~25%、油酸含量在25%~45%,和人乳脂肪较为接近;巴沙鲶鱼油、黑鱼油、鲈鱼油、鲤鱼油、白鲢鱼油sn-2棕榈酸含量分别为44.27%、40.91%、31.31%、30.02%和31.02%,且sn-2棕榈酸分布均高于50%;巴沙鲶鱼油、黑鱼油的sn-1,3油酸含量在50%左右,并通过UPLC-MS/MS验证了OPO的存在,推测适合作为人乳替代脂的基料油;鲤鱼油中中碳链脂肪酸C10∶0和C12∶0总含量超过10%,易于婴幼儿吸收;鲈鱼油中DHA含量为12.73%,对于婴幼儿智力发育有着关键作用,鲤鱼油和鲈鱼油适合作为人乳替代脂的配料油。     

The effect of milk fat globules on adherence and internalization of Salmonella Enteritidis to HT-29 cells

Milk fat globules were extracted from bovine and goat milk and incubated with HT-29 human adenocarcinoma cells to assess the attachment and internalization of Salmonella Enteritidis. Because the expression of bacterial adhesins is highly affected by the presence of antibiotic, the attachment was studied with and without antibiotic in the cell growth medium. Although no inhibitory effect of the fat globules was observed in the presence of the antibiotic, milk fat globules significantly inhibited the binding and internalization of Salmonella in medium free of antibiotic. The fat globules from both bovine and goat milk markedly reduced bacterial binding and invasion compared with controls, and the cells treated with goat milk-derived fat globules demonstrated greater protective properties than those derived from bovine milk. The effect of heat treatment on bovine fat globules was also investigated, and it was shown that the fat globules from heated milk had a higher degree of inhibition than those from unheated milk.     

Effect of safflower oil, flaxseed oil, monensin, and vitamin E on concentration of conjugated linoleic acid in bovine milk fat

Conjugated linoleic acid (CLA) refers to a mixture of conjugated octadecadienoic acids of predominantly ruminant origin. The main isomer in bovine milk fat is the cis-9, trans-11 CLA. Interest in CLA increased after the discovery of its health-promoting properties, including potent anticarcinogenic activity. Two experiments were conducted to evaluate dietary strategies aimed at increasing the concentration of CLA in bovine milk fat. Both experiments were organized as a randomized complete block design with a repeated measures treatment structure. In Experiment 1, 28 Holstein cows received either a control diet or one of 3 treatments for a period of 2 wk. The control diet consisted of 60% forage (barley silage, alfalfa silage, and alfalfa hay) and 40% concentrate on a dry matter (DM) basis, fed as a total mixed ration (TMR). The concentrate was partially replaced in the treatment groups with 24 ppm of monensin (MON), 6% of DM safflower oil (SAFF), or 6% of DM safflower oil plus 24 ppm of monensin (SAFF/M). Average cis-9, trans-11 CLA levels in milk fat after 2 wk of feeding were 0.45, 0.52, 3.36, and 5.15% of total fatty acids for control, MON, SAFF, and SAFF/M, respectively. In Experiment 2, 62 Holstein cows received either a control diet or one of 5 treatment diets for a period of 9 wk. The control diet consisted of 60% forage (barley silage, alfalfa silage, and alfalfa hay) and 40% concentrate on a DM basis, fed as a TMR. The concentrate was partially replaced in the treatment groups with 6% of DM safflower oil (SAFF), 6% of DM safflower oil plus 150 IU of vitamin E/kg of DM (SAFF/E), 6% of DM safflower oil plus 24 ppm of monensin (SAFF/M), 6% of DM safflower oil plus 24 ppm of monensin plus 150 IU of vitamin E/kg of DM (SAFF/ME), or 6% of DM flaxseed oil plus 150 IU of vitamin E/kg of DM (FLAX/E). Average cis-9, trans-11 CLA levels during the treatment period were 0.68, 4.12, 3.48, 4.55, 4.75, and 2.80% of total fatty acids for control, SAFF, SAFF/E, SAFF/M, SAFF/ME, and FLAX/E, respectively. The combination of safflower oil with monensin was particularly effective at increasing milk fat CLA. The addition of vitamin E to the diet partially prevented the depression in milk fat associated with oilseed feeding, but had no significant effect on the concentration of CLA in milk.     

Effects of canola oil dilution on anhydrous milk fat crystallization and fractionation behavior

Blends of anhydrous milk fat (AMF) and canola oil (CO) were cooled from 35 to 5°C at 0.1°C/min, held for 24 h, and centrifuged to separate the liquid and crystalline fractions. The blends’ crystallization behaviors and microstructures depended on the level of CO present. Onset and half times of crystallization reflected a slower crystallization mechanism at higher levels of CO dilution. These differences were accompanied by a change in microstructure from large spherulites to smaller particles. The biggest change occurred between the 1:4 and 1:5 blends. Canola oil dilution also influenced the polymorphism of milk fat. Whereas only the β′ polymorph was observed in the crystallized 1:2 blend, the β polymorph predominated in the 1:8 blend. Some solubilization of AMF solids into CO was observed. This increased gradually with increasing CO concentration. Compositional analysis revealed the exchange of AMF and CO species between the liquid and crystalline fractions. The crystalline fractions were slightly enriched in AMF triacylglycerols, particularly with the more dilute blends (1:7 and 1:8). Large amounts of oil were trapped in the crystalline fractions, particularly for the concentrated AMF:CO blends where the β′ crystals and spherulitic microstructures were observed. Although the solid fat content profiles of the fractionated blends were marginally higher than those of the starting blends, the samples were very soft and oily. This strategy of using CO to fractionate milk fat was limited by the poor separation of solids and liquid during centrifugation.     

Detection of adulteration by caprine body fat and mixtures of caprine body fat and groundnut oil in bovine and buffalo ghee using differential scanning calorimetry

Differential scanning calorimetry was used to detect adulteration of pure ghee with caprine body fat when added singly (at 5, 10 and 15%) and in combination with groundnut oil (GNO) (at 5, 10 and 15%). Samples were analysed for transition behaviour in terms of crystallising and melting curves. When compared to pure ghee, adulterated ghee samples showed a shift in the midrange temperature of thermal curves, indicating the presence of foreign fats. The results revealed that the detection of adulteration was possible at the lowest level of the study (5%), irrespective of the nature of the adulterants.     

Efficient in vitro digestion of lipids and proteins in bovine milk fat globule membrane ingredient (MFGMi) and whey-casein infant formula with added MFGMi

The relative immaturity of the infant digestive system has the potential to affect the bioavailability of dietary lipids, proteins, and their digested products. We performed a lipidomic analysis of a commercial bovine milk fat globule membrane ingredient (MFGMi) and determined the profile of lipids and proteins in the bioaccessible fraction after in vitro digestion of both the ingredient and whey-casein-based infant formula without and with MFGMi. Test materials were digested using a static 2-phase in vitro model, with conditions simulating those in the infant gut. The extent of digestion and the bioaccessibility of various classes of neutral and polar lipids were monitored by measuring a wide targeted lipid profile using direct infusion–mass spectrometry. Digestion of abundant proteins in the ingredient and whey–casein infant formula containing the ingredient was determined by denaturing PAGE with imaging of Coomassie Brilliant Blue stained bands. Cholesterol esters, diacylglycerides, triacylglycerides, phosphatidylcholines, and phosphatidylethanolamines in MFGMi were hydrolyzed readily during in vitro digestion, which resulted in marked increases in the amounts of free fatty acids and lyso-phospholipids in the bioaccessible fraction. In contrast, sphingomyelins, ceramides, and gangliosides were largely resistant to simulated digestion. Proteins in MFGMi and the infant formulas also were hydrolyzed efficiently. The results suggest that neutral lipids, cholesterol esters, phospholipids, and proteins in MFGMi are digested efficiently during conditions that simulate the prandial lumen of the stomach and small intestine of infants. Also, supplementation of whey-casein-based infant formula with MFGMi did not appear to alter the profiles of lipids and proteins in the bioaccessible fraction after digestion.