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.     

Milk fat globule membrane proteins are involved in controlling the size of milk fat globules during conjugated linoleic acid–induced milk fat depression

Milk fat globule membrane (MFGM) proteins surround the triacylglycerol core comprising milk fat globules (MFG). We previously detected a decrease in the size of fat globules during conjugated linoleic acid (CLA)-induced milk fat depression (MFD), and other studies have reported that some MFGM proteins play a central role in regulating mammary cellular lipid droplet size. However, little is known about the relationship between MFD, MFG size, and MFGM proteins in bovine milk. The aim of this study was to investigate the profile of MFGM proteins during MFD induced by CLA. Sixteen mid-lactating Holstein cows (145 ± 24 d in milk) with similar body condition and parity were divided into control and CLA groups over a 10-d period. Cows were fed a basal diet (control, n = 8) or control plus 15 g/kg of dry matter (DM) CLA (n = 8) to induce MFD. Cow performance, milk composition, and MFG size were measured daily. On d 10, MFGM proteins were extracted and identified by quantitative proteomic analysis, and western blotting was used to verify a subset of the identified MFGM proteins. Compared with controls, supplemental CLA did not affect milk production, DM intake, or milk protein and lactose contents. However, CLA reduced milk fat content (3.73 g/100 mL vs. 2.47 g/100 mL) and the size parameters volume-related diameter D_[4,3] (3.72 μm vs. 3.35 μm) and surface area-related diameter D_[3,2] (3.13 μm vs. 2.80 μm), but increased specific surface area of MFG (1,905 m²/kg vs. 2,188 m²/kg). In total, 177 differentially expressed proteins were detected in milk from cows with CLA-induced MFD, 60 of which were upregulated and 117 downregulated. Correlation analysis showed that MFG size was negatively correlated with various proteins, including XDH and FABP3, and positively correlated with MFG-E8, RAB19, and APOA1. The results provide evidence for an important role of MFGM proteins in regulating MFG diameter, and they facilitate a mechanistic understanding of diet-induced MFD.     

Symposium review: Fat globules in milk and their structural modifications during gastrointestinal digestion

The fat globules in milk are unique oil droplets that are stabilized by a specific and structurally complex membrane, the milk fat globule membrane (MFGM). In the last decade, excellent progress has been made on studying the structure of the milk fat globules and the MFGM and how common processing treatments affect these structures to deliver dairy products with improved functional properties. Although the digestion of milk fat to deliver energy and lipid-soluble nutrients is essential for survival of the neonate, there is little understanding of the complex processes involved. The structural alterations to fat globules during gastrointestinal processing affect the way in which milk fat is digested, absorbed, and metabolized. The packaging of these globules within the MFGM or in other forms may affect the bioaccessibility of raw or processed milk fat globules; in turn, this may affect access of the gastrointestinal enzymes to the globules and, therefore, may influence the rate and extent of lipid digestion. This review focuses on recent advances in understanding milk fat globules during gastrointestinal digestion, including the effects of processing on their bioavailability and the kinetics of lipid digestion. Possible effects of the dairy matrix on lipid digestion and physiological responses are briefly described.     

乳脂肪球膜的特性、开发及在模拟母乳脂肪球结构中的应用

乳脂肪球膜（milk fat globule membrane,MFGM）是包裹在天然乳脂肪球外部的3 层膜状结构,然而牛乳基和大豆基婴儿配方奶粉缺少MFGM,脂肪球结构与母乳存在较大差异,因此添加外源MFGM以及制备与母乳脂肪球结构接近的婴儿配方奶粉成为了近期的研究焦点。本文综述了MFGM的相关特性和生产开发途径,以及牛乳MFGM在仿母乳脂肪球结构乳液和婴儿配方奶粉中的应用。体外模拟婴儿胃肠道消化实验以及啮齿动物体内实验结果表明,仿母乳脂肪球结构乳液和婴儿配方奶粉能够促进婴儿脂肪消化并且改善脂质代谢过程。     

Native vs. damaged milk fat globules: membrane properties affect the viscoelasticity of milk gels

The storage modulus G' of rennet and acid milk gels filled with milk fat globules was measured as a function of the fat globule surface composition (native milk fat globule membrane, caseins and whey proteins, or a mixture of the three due to mechanical treatments) and surface area (i.e., the fat globule size). By different technological procedures, it was possible to obtain fat globules of constant surface composition but various sizes, and vice-versa, which had never been done. For both rennet and acid gels, a critical fraction of the fat globule surface covered by caseins and whey proteins was identified (approximately 40%), beyond which G' increased. Below this threshold, the gel viscoelasticity was unaffected by mechanical treatments. When the diameter of native milk fat globules decreased, the G' of rennet gels increased slightly, whereas that of acid gels decreased sharply. For both types of gels, G' increased when the diameter of partially disrupted fat globules decreased. For recombined globules completely covered with caseins and few whey proteins, G' increased with fat globule surface area for rennet gels whereas it decreased for acid gels. With the help of confocal microscopy and in the light of general structural differences between rennet and acid gels, a mechanism is proposed for the effect of fat globule damage and diameter on G', depending on the interactions the globules can undergo with the casein network.     

婴儿配方乳粉脂质母乳化的研究进展

人乳脂肪是最复杂的天然脂质混合物之一,具有独特的脂肪酸组成、分布和许多复杂脂质。乳脂肪几乎提供着新生婴儿所需的一半能量,同时还会影响婴儿日后的代谢方式和整体发育。因此,随着对母乳脂肪及婴儿生长需求研究的深入,以母乳脂质组成为模板,对婴儿配方乳粉脂质进行母乳化模拟,可以缩小母乳与婴儿配方乳粉喂养婴儿之间的差异。目前,婴儿配方乳粉脂质母乳化主要集中在脂肪酸组成模拟、sn-2棕榈酸结构脂的应用以及乳脂肪球膜成分的添加方面。另外,利用乳脂肪球膜包裹脂滴实现脂肪球结构模拟的相关研究也在逐步开展。本文着重对这几个方面的研究进展进行总结,在关注婴儿营养需求的同时,为婴儿配方乳粉的研发提供理论依据。     

Composition and Structure of the Bovine Milk Fat Globule Membrane—Some Nutritional and Technological Implications

Milk fat is dispersed in milk as small, spherical globules, stabilized in the form of emulsion by its surrounding membrane. This membrane, called the milk fat globule membrane (MFGM), is created in the secretory cells of the mammary gland, and represents an ordered and unique biophysical system. This review characterizes the main milk fat globule components, their structure, and intracellular origin. The milk fat globule membrane has many potentially bioactive components. These are discussed in terms of their health effects for the native and processed globules. Because of their functional and nutritional properties, MFGM components can be used as valuable ingredients in the manufacture of new functional foods.     

乳脂肪球膜的组成、营养及制备研究进展

乳中的脂肪以乳化的脂肪球形态分散于水相中,脂肪球被三层脂肪球膜包裹,近年来的研究表明脂肪球膜中的成分对婴幼儿的生长发育有至关重要的作用。本文介绍了乳脂肪球膜特别是其磷脂的组成及含量,添加入婴幼儿配方奶粉中乳脂肪球膜的营养作用,以及目前市售的富含脂肪球膜的产品及制备方式,为缩小配方奶与婴幼儿营养需求的差异提供科学依据。     

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.     

Proteolysis of milk fat globule membrane proteins during in vitro gastric digestion of milk

The influence of gastric proteolysis on the physicochemical characteristics of milk fat globules and the proteins of the milk fat globule membrane (MFGM) in raw milk and cream was examined in vitro in simulated gastric fluid (SGF) containing various pepsin concentrations at pH 1.6 for up to 2 h. Apparent flocculation of the milk fat globules occurred in raw milk samples incubated in SGF containing pepsin, but no coalescence was observed in either raw milk samples or cream samples. The changes in the particle size of the fat globules as a result of the flocculation were dependent on the pepsin concentration. Correspondingly, the physical characteristics of the fat globules and the composition of the MFGM proteins in raw milk changed during incubation in SGF containing pepsin. The major MFGM proteins were hydrolyzed at different rates by the pepsin in the SGF; butyrophilin was more resistant than xanthine oxidase, PAS 6, or PAS 7. Peptides with various molecular weights, which altered with the time of incubation and the pepsin concentration, were present at the surfaces of the fat globules.     

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.     

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.     

牛乳脂肪球膜组分分离鉴定及其功能性研究进展

乳脂肪球膜（milk fat globule membrane,MFGM）是牛乳中脂肪球外表面包裹着的一层膜,在牛乳中的含量非常微小。但由于其具有良好的生理活性以及潜在的食品加工应用潜力,关于其展开的基础研究和应用基础研究受到广泛关注。本综述介绍MFGM的分离制备技术、结构、组分鉴定分析、生理活性及其应用特性的研究进展,以期为MFGM在食品工业尤其是在功能性食品中的应用提供一定的理论支持。     

乳脂肪球膜功能特性的研究进展

乳脂肪以脂肪球的形式存在,乳脂肪球膜是包裹在乳脂肪球周围的三层生物薄膜,具有很高的营养价值。随着食品科学研究的深入和分离技术的发展,乳脂肪球膜中的活性成分及其功能作用正在逐渐被揭示。乳脂肪球膜是含有蛋白质、磷脂、鞘脂、神经节苷脂、胆碱、唾液酸和胆固醇的混合物,这些成分是具有重要功能的食品成分,应用于配方食品生产。本文综述了乳脂肪球膜中常见的蛋白质、脂质及其生物活性,综述了近年来乳脂肪球膜及其成分在改善肠道健康、改善大脑发育、改善肥胖及相关并发症、改善老年人虚弱、抗癌、抗氧化和缓解疲劳等方面的体内研究和临床研究进展,并讨论了其可能的作用机制,以期为乳脂肪球膜配料的研发及其在配方食品中的应用提供借鉴和参考。     

Factors affecting the concentration of sphingomyelin in bovine milk

Sphingomyelin is a phospholipid located in the outer leaflet of the plasma membrane of most cells and is a component of the milk fat globule membrane. Sphingomyelin and its digestion products participate in several antiproliferative pathways that may suppress oncogenesis. Although milk and dairy products are important sources of sphingomyelin in the human diet, little is known about factors that influence sphingomyelin concentrations in milk fat or whether concentrations can be modified via the nutrition of cows. Sphingomyelin concentrations were determined in milk from Holstein and Jersey cows matched for parity and stage of lactation. Sphingomyelin was more concentrated in milk fat from Holstein cows than in milk fat from Jersey cows (1,044 vs. 839 μg/g of fat). Concentrations in whole milk did not differ because of greater milk fat content for milk from Jerseys. Differences between breeds may be related to the greater fat globule size in milk from Jerseys. Sphingomyelin content in whole milk increased with increasing days in milk because of associated increases in milk fat content. Regardless of breed, primiparous cows had greater amounts of stearic acid and less palmitic acid in sphingomyelin than did older cows. The sphingomyelin concentration in milk fat of cows in a commercial Jersey herd was lower for cows in their fourth or greater parity. Sphingomyelin content in whole milk was greater for cows in late lactation because of greater milk fat content. Feed restriction of multiparous Holstein cows to 37% of ad libitum dry matter intake increased milk fat content but did not affect milk sphingomyelin content or milk fat globule size. Supplementation of the diet with 4% soybean oil did not affect milk composition, sphingomyelin content, or milk fat globule size. Milk was sampled seasonally from 7 herds throughout Illinois during a 2-yr period. Sphingomyelin concentration in milk fat was greatest during summer and least during winter, but whole milk concentrations did not vary across seasons. We conclude that 1) sphingomyelin content of milk fat is greater in milk from Holsteins than that from Jerseys, 2) sphingomyelin content in whole milk increases with stage of lactation, and 3) sphingomyelin content of milk fat is greater during summer. However, efforts to produce milk with a greater sphingomyelin content through altering management and nutrition are unlikely to be successful.     

Physical,textural, and rheological properties of whipped cream affected by milk fat globule membrane protein

This work aims at improving the textural and whipping properties of whipped cream by the addition of milk fat globule membrane protein. The determination of particle size distribution and average diameter of whipped cream showed that the small particle size was shifted to a larger range after milk fat globule membrane protein was added. The average particle size (d_3,2) of whipped cream reached a maximum value of 5.05 µm at 1% milk fat globule membrane protein, while slowly decreased with increasing milk fat globule membrane protein levels from 2% to 5%. In addition, the partial coalescence of fat increased with the increase of milk fat globule membrane protein levels, and the correlation between the whipping time and the overrun of whipped cream was positive. The addition of milk fat globule membrane protein also altered the rheological behaviour of whipped cream, resulting in the increase of modulus G′ and the loss modulus G″. The results also indicated that higher milk fat globule membrane protein level decreased the serum loss of whipped cream while improved its stability. While milk fat globule membrane protein levels had no significant effect on viscosity, its increasing levels effectively improved the hardness, consistency, and viscosity of whipped cream.     

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.     

The effect of dietary forage to concentrate ratio and forage type on milk fatty acid composition and milk fat globule size of lactating cows

We examined the effects of 2 grass silage-based diets differing in forage:concentrate (FC) ratio and those of a red clover silage-based diet on intake, milk production, ruminal fatty acid (FA) biohydrogenation, milk FA composition, and milk fat globule (MFG) size distribution. Ten multiparous Nordic Red cows received the following treatments: grass silage-based diets containing high (70:30, HG) or low (30:70, LG) FC ratio or a red clover silage-based diet with an FC ratio of 50:50 (RC) on a dry matter basis. Determinations of MFG were performed from fresh milk samples without addition of EDTA so the results of fat globules >1 µm in diameter are emphasized instead of the entire globule population. Lower FC ratio in grass silage-based diets increased milk production with no effect on daily fat yield, leading to 13% lower milk fat concentration. The effect of FC ratio on MFG size was moderate. It did not affect the volume-weighted diameter in grass silage-based diets, although LG lowered the volume-surface diameter of MFG in the size class >1 µm compared with HG. Compared with HG, feeding LG moderately decreased the biohydrogenation of 18:2n-6, leading to a higher level of polyunsaturated fatty acids in milk fat. Feeding RC lowered milk fat concentration and daily milk fat yield compared with grass silage-based diets. The volume-weighted diameter of MFG in the size class >1 µm was smaller in RC milk compared with grass silage-based diets. Feeding RC increased the flow of 18:3n-3 at the omasum by 2.4-fold and decreased the apparent ruminal 18:3n-3 biohydrogenation compared with grass silage-based diets despite similar intake of 18:3n-3. It also resulted in the lowest amount of saturated FA and the highest amounts of cis-9 18:1, 18:3n-3, and polyunsaturated FA in milk. In conclusion, LG decreased milk fat content and induced minor changes in MFG size distribution compared with HG, whereas RC lowered milk fat production, altered milk FA composition to nutritionally more beneficial direction, and led to smaller MFG compared with grass silage-based diets.     

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.