Crystallization behavior of milk fat obtained from linseed-fed cows

Milk with an increased content of unsaturated fatty acids was obtained by incorporating 60% of extruded linseed into the concentrate of cows. Two groups of Holstein cows (3 animals/group) were fed a concentrate (control or linseed enriched) together with the same roughage diet (ad libitum). After an adaptation period of 3 wk, evening and morning milk samples were collected every 7 d for 3 wk. Milk was decreamed and anhydrous milk fat (AMF) was isolated from the fat fraction by using the Bureau of Dairy Industries method. The objective of this study was to investigate if the crystallization mechanism of milk fat changed when the content of unsaturated fatty acids was increased. Therefore, the crystallization behavior of a milk fat enriched with unsaturated fatty acids was compared with that of a control milk fat. Nonisothermal crystallization was investigated with differential scanning calorimetry, and 1-step and 2-step isothermal crystallization behaviors were investigated using pulsed nuclear magnetic resonance, differential scanning calorimetry, and x-ray diffraction. A higher content of unsaturated fatty acids in AMF resulted in an increased proportion of low melting triglycerides. These triglycerides lowered the solid fat content profile, particularly at refrigerator temperatures. Furthermore, they induced some changes in the crystallization and melting behaviors of milk fat compared with a control AMF, although no fundamental changes in the crystallization mechanism could be revealed. Even though a lower melting point could be observed for milk fat with a higher content of unsaturated fatty acids, a similar degree of supercooling was needed to initiate crystallization, resulting in a shift in onset temperature of crystallization toward lower temperatures. In addition, slower crystallization kinetics were measured, such as a lower nucleation rate and longer induction times, although crystallization occurred in a similar polymorphic crystal lattice. During melting, a shift in offset temperature toward lower temperatures could be observed for the 3 melting fractions of AMF in addition to a higher proportion of low melting triglycerides. These results demonstrate that a higher content of unsaturated fatty acids has some effect on the crystallization behavior of milk fat. This knowledge could be used to produce dairy products of similar or superior quality compared with conventional products by intervening in the production process of dairy products.     

The potential of milk fat for the synthesis of valuable derivatives

The overall decline in milk fat consumption experienced in the last decades has promoted global research efforts seeking for alternate uses of this valuable natural fat. Milk fat possesses a pleasant flavor and a rich chemical composition, including a range of bioactive, health beneficial minor components. The main drawbacks of milk fat from the consumer point of view are its poor spreadability at refrigeration temperature and its high content in saturated fatty acids, which raises health concerns. However, the rich fatty acid composition of milk fat could be utilized for the production of a wide range of added-value derivatives in the food and cosmetic industries, including nutritionally enhanced modified fats, food emulsifiers, flavors, and tailor-made lipids. A promising strategy for the revalorization of milk fat encompasses the isolation and commercialization of the valuable minor components of milk fat, coupled with a broader utilization of physically or nutritionally improved milk fat fractions and derivative products.     

Distribution of cholesterol in milk fat fractions

Milk fat was fractionated into liquid (m.p. congruent to 12 degrees C), intermediate (m.p. congruent to 21 degrees C) and solid (m.p. congruent to 39 degrees C) fractions by three different processes--melt crystallization, short-path distillation and supercritical CO2 extraction--and the cholesterol content of these fractions determined. Cholesterol was enriched in the liquid fractions from all three processes, in particular about 80% of the cholesterol being found in the liquid fraction obtained by short-path distillation. The basis of migration of cholesterol into various milk fat fractions was explained by its affinity to various triglycerides (melt crystallization) and by vapour pressure and molecular weight (short-path distillation). It was more complex in the supercritical CO2 extraction process; the interplay of cholesterol affinity toward CO2 and its molar volume, and its vapour pressure enhancement under applied pressure play a role.     

Physical properties of palm kernel olein-anhydrous milk fat mixtures transesterified using mycelium-bound lipase from Rhizomucor miehei

The transesterification activity of mycelium-bound lipase from Rhizomucor miehei on palm kernel olein:anhydrous milk fat (PKO:AMF) blends was investigated. Commercial immobilised R. miehei lipase preparation, Lipozyme IM60 (Novo Nordisk), was used as a comparison. Mixtures of PKO:AMF, at ratios of 100:0, 70:30, 60:40, 50:50 and 0:100, were transesterified using either enzyme in a solvent-free system. The triglyceride (TG) profile, slip melting point, solid fat content, melting thermogram and the polymorphic form of the unreacted and transesterified mixtures were evaluated. Results indicated that transesterification by either enzyme was able to produce an oil mixture with new TG profiles, generally lower slip melting points and solid fat contents. The melting thermograms from differential scanning calorimetry analysis indicated changes in the triglyceride's crystalline composition and an overall shift to lower melting TG. Although the catalytic activities were similar for both lipases, Lipozyme-catalysed mixtures produced higher degrees of transesterification (43–51%) than mycelium-bound lipase-catalysed (22–34%) mixtures. This study also demonstrated that the transesterified PKO:AMF mixture at 70:30 ratio completely melted at 25C, and this meets the melting criteria for fat used in ice cream formulation.     

Performance of selected milk fat fractions in cold-spreadable butter.

Based on solid fat content profiles, milk fat fractions produced by fractional crystallization procedures employing melted milk fat and milk fat dissolved in acetone were selected for incorporation into soft butter samples. Butter samples made from low melting liquid fractions or a combination of primarily low melting liquid fractions and a small amount of high melting solid fractions exhibited good spreadability at refrigerator temperature (4 degrees C) but were almost melted at room temperature (21 degrees C). Butters made with a high proportion of low melting liquid fraction, a small proportion of high melting solid, and a small proportion of very high melting solid fractions were still spreadable at refrigerator temperature and maintained their physical form at room temperature. Very high melting fractions, which provided key structural functionality in spreadable butter, were obtained from acetone fractionation. Because the use of acetone in processing may hinder or prevent commercialization of these fractions, other means to obtain very high melting fractions from milk fat should be pursued.     

Fractionation of milk fat by short-path distillation

Fractionation of milk fat by short-path distillation changes the chemical composition and physical properties of the resulting fractions. Increases in distillation temperature from 125 to 250 degrees C increased distillate yield from 0.3 to 42.7% (wt/wt). The distillate was enriched in short- and medium-chain fatty acids and low molecular weight acylglycerols, while the retentate was enriched in long-chain saturated and unsaturated fatty acids as well as high molecular weight acylglyerols. As distillation temperature increased, dropping points of the distillate increased. Relative to native milk fat, the solid fat content (SFC) vs. temperature melting profile of the distillate was depressed and that of the retentate was augmented, which correlated with the saturated long-chain fatty acid content in the fractions. Retentate crystallization parameters obtained by fitting the Avrami model to SFC-time data, did not change as a function of distillation temperature, but varied as a function of the degree of undercooling. Changes in microstructure observed by polarized light microscopy also appeared to be solely a function of the degree of undercooling, with no observable differences between retentates obtained at the different distillation temperatures. In addition, no changes in the retentate's free energy of nucleation (deltaGc) as a function of distillation temperature were found. The compressive storage modulus of the crystallized retentate increased as a function of increasing distillation temperature.     

Hydrolysis of bovine milk fat globules by lipoprotein lipase: inhibition by proteins extracted from milk fat globule membrane

Extraction of membrane proteins from milk fat globules by GuHCl or by MgCl2 made the lipids more accessible to lipolysis by added lipoprotein lipase. The increase in lipolysis paralleled the loss of membrane proteins and was continuous up to 2.5 M GuHCl, which was the highest concentration used. About twice as much protein was extracted with 2.5 M GuHCl as with buffer only. The amount of protein lost was about 50% of total milk fat globule protein. Lipolysis of milk fat globules was inhibited by addition of the extracted protein. The extracted proteins also reduced lipolysis when added to whole milk. More protein was needed to inhibit lipolysis of milk fat globules treated with GuHCl compared with globules treated with buffer only. The inhibition by a given amount of protein decreased if more milk fat globules were used. Protein extracted with MgCl2 had similar effects as those extracted with GuHCl. The major components extracted with MgCl2 migrated in the 40 to 50-kdalton region on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. By gel filtration chromatography, two protein fractions were obtained, which inhibited lipolysis more efficiently than the total extract. As has previously been found for inhibition of lipolysis by skim milk, the amount of extracted protein needed to inhibit lipolysis varied between preparations of milk fat globules. Milk with propensity to cold-induced ("spontaneous") lipolysis was normalized by addition of extracted proteins.     

Effect of feed on the composition of milk fat.

Researchers attending the Wisconsin Milk Board 1988 Milk Fat Roundtable indicated that the ideal nutritional milk fat would contain 10% polyunsaturated fatty acids, 8% saturated fatty acids, and 82% monounsaturated fatty acids. This cannot be accomplished by modifying diets of lactating cows. Monounsaturated fatty acid (C18:1) content can be increased by 50 to 80% and may approach 50% of milk fatty acids by feeding lipids rich in 18-carbon fatty acids. Because of ruminal hydrogenation and intestinal and mammary desaturase activity, degree of unsaturation of dietary 18-carbon fatty acids is not critical in influencing milk fat C18:1. Feeding low roughage diets increases the proportion of C18:1 in milk fat, and effects of feeding low roughage diets and lipid may be additive. Palmitic acid (C16:0) content of milk fat can be reduced by 20 to 40% unless the supplemented lipid is rich in C16:0. Milk fat alteration is dependent on the level of lipid supplementation. Limited evidence indicates frequency of lipid feeding and physical form of oil (free oil vs. oilseed), and heat treatment of oilseeds has relatively little influence on modification of milk fat. Significant changes in milk fat composition can be achieved on farm via nutritional modifications.     

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.     

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。     

Buffalo milk fat globules and their biological membrane: in situ structural investigations

Milk fat globules and their surrounding biological membrane (the MFGM) are not well understood despite the importance of these milk components in human nutrition and the role of fat globules in determining the properties of dairy products. The objectives of this study were to investigate these unique colloidal assemblies and the microstructure of the MFGM in buffalo milk, which is the second largest global source of dairy products. In-situ structural investigations were performed at room temperature using confocal microscopy with multiple fluorescent probes (Nile Red, Rh-DOPE, the lectin WGA-488). Microscopic observations showed cytoplasmic crescents around fat globules and the heterogeneous distribution of glycosylated molecules and polar lipids with the occurrence of lipid domains. The lipid domains in the buffalo MFGM appear to form by the segregation of lipids with a high phase transition temperature (e.g. sphingomyelin and saturated phosphatidylcholine molecular species) and cholesterol resulting in a gel phase or a Lo phase forming circular domains. The structure of the buffalo MFGM results from a non-random mixing of components, consistent with observations for other species. Structural heterogeneities of the MFGM could affect the processability of buffalo fat globules and the bioavailability of milk lipids.     

Compositional effects on milk fat crystallization.

Seasonal and regional variability in milk fat composition causes differences in crystallization behavior, which can, for example, result in variability in fractionation efficiency and physical properties of butter. However, the specific compositional causes of variability in milk fat crystallization behavior are still only poorly understood. In this work, the seasonal and regional variations in composition of anhydrous milk fat were analyzed and related to crystallization behavior. Although there were no clear-cut trends in chemical composition (triacylglycerol, fatty acid, and minor lipid profiles) among the milk fats, significant differences in crystallization behavior were observed. For anhydrous milk fat samples made from sweet cream and obtained from the same milk supply through a period of 14 mo, no significant trends in either composition or crystallization behavior were observed. This suggests that seasonal variability of milk fat may be reduced by current feeding strategies in the United States. More detailed examination of the triacylglycerol profiles led to the conclusion that the relative contents of certain triacylglycerol combinations correlated well with crystallization behavior. In particular, the ratio of higher-melting to lower-melting triacylglycerols could be used to predict crystallization behavior. Higher ratios of higher-melting triacylglycerols led to higher crystallization rate.     

Effects of milk powders in milk chocolate

The physical characteristics of milk powders used in chocolate can have significant impact on the processing conditions needed to make that chocolate and the physical and organoleptic properties of the finished product. Four milk powders with different particle characteristics (size, shape, density) and "free" milk fat levels (easily extracted with organic solvent) were evaluated for their effect on the processing conditions and characteristics of chocolates in which they were used. Many aspects of chocolate manufacture and storage (tempering conditions, melt rheology, hardness, bloom stability) were dependent on the level of free milk fat in the milk powder. However, particle characteristics of the milk powder also influenced the physical and sensory properties of the final products.     

Fractionation and characterization of edible sheep tail fat

Edible sheep tail fat was effectively fractionated by an acetone crystallization. Each of the fractions and filtrates were analyzed for melting point, refractive index, iodine value, fatty acid composition, and characterized by differential scanning calorimetry. Fatty acid analysis indicated that as the fractionation temperature decreased, the concentration of unsaturated fatty acids in the fractions increased. The liquid fraction had a differential scanning calorimetry melting curve similar to commercial salad oil and the curve of one of the filtrates resembled that of cocoa butter.     

Short communication: Effect of antioxidant supplementation on milk production,milk fat synthesis,and milk fatty acids in dairy cows when fed a diet designed to cause milk fat depression

This study evaluated the effect of a blend of synthetic antioxidants on the yield of milk and milk components and milk fatty acid composition in dairy cows fed a diet designed to cause milk fat depression (MFD). We hypothesized that supplementing a synthetic antioxidant to diets with a high rumen unsaturated fatty acid load (RUFAL) would decrease the severity of MFD. Sixteen lactating Holstein cows (163 ± 47 d in milk), in a crossover design with two 21-d periods, were fed a corn silage and grass silage-based diet containing 15% distillers grains. The diet contained 34% neutral detergent fiber, 18% crude protein, 26% starch, and 4.3% total fatty acids (dry matter basis). Cows were fed the diet without supplementation (control; CON) or supplemented with 0.02% (dry matter basis) of a synthetic antioxidant (AOX; Agrado Plus, Novus International Inc., St. Charles, MO). Dry matter intake and milk yields were recorded daily. Milk samples were collected at the start of the study for baseline values and the end of each period (d 20–21) and analyzed for milk components and fatty acid composition. Dry matter intake and milk yield were unaffected by treatment and averaged 25.9 and 50.2 kg/d, respectively. Similarly, we observed no effect of treatment on yields of fat, protein, lactose, 3.5% fat-corrected milk, energy-corrected milk, feed efficiency, body weight, or body condition score. Milk fat concentration and yield were both reduced by the high RUFAL diets. We observed a tendency for AOX to increase the concentration of milk fat and decrease the concentration of milk protein. Yields of de novo and preformed fatty acids were not affected by treatment, although we detected a trend for a slight increase in the yield of 16-carbon fatty acid for AOX compared with CON. Treatment had only minor effects on individual milk fatty acids, except for the concentration and yield of linoleic acid, which were over 90% higher for AOX compared with CON. In conclusion, milk fat concentration and yield were reduced by a high RUFAL diet containing 15% distillers grains; however, supplementation with AOX did not overcome the MFD induced by this diet.     

Effects of protein and fat levels in milk on cheese and whey compositions

Bulk tank milk was standardised to six levels of fat (3·0, 3·2, 3·4, 3·6, 3·8, 4·0%) and similarly to six levels of protein, thus giving a total of 36 combinations in composition. Milk was analyzed for total solids, fat, protein, casein, lactose and somatic cell count and was used to make laboratory-scale cheese. Cheese samples from each batch were assayed for total solids, fat, protein and salt. Losses of milk components in the whey were also determined. Least squares analysis of data indicated that higher protein level in milk was associated with higher protein and lower fat contents in cheese. This was accompanied by lower total solids (higher moisture) in cheese. Inversely, higher fat level in milk gave higher fat and lower protein and moisture contents in cheese. Higher fat level in milk resulted in lower retention of fat in cheese and more fat losses in the whey. Higher protein level in milk gave higher fat retention in cheese and less fat losses in the whey. Regression analysis showed that cheese fat increased by 4·22%, while cheese protein decreased by 2·61% for every percentage increase in milk fat. Cheese protein increased by 2·35%, while cheese fat decreased by 6·14% per percentage increase in milk protein. Milk with protein to fat ratio close to 0·9 would produce a minimum of 50% fat in the dry matter of cheese.     

Conjugated linoleic acid and other anticarcinogenic agents of bovine milk fat.

Prevention is an important strategy for conquering cancer. Milk fat contains a number of components, such as conjugated linoleic acid, sphingomyelin, butyric acid, ether lipids, beta-carotene, and vitamins A and D that have anticancer potential. Conjugated linoleic acid inhibits the growth of a number of human cancer cell lines and suppresses chemically-induced tumor development at a number of sites in animal models. As little as 0.1% of dietary conjugated linoleic acid inhibits the development of rat mammary tumors, independent of the amount and type of fat in the diet. Sphingomyelin, through its metabolites ceramide and sphingosine, participates in multiple antiproliferative pathways associated with suppression of carcinogenesis. Dietary sphingomyelin inhibits murine colon tumor development. Butyric acid, uniquely present in ruminant milk, is a potent antineoplastic agent and may ameliorate its potency through synergy with other milk fat components. Dietary butyric acid inhibits mammary carcinoma development in rats. In humans, ether lipids, beta-carotene, and vitamins A and D are associated with anticancer effects. Cows have the ability to extract anticarcinogenic components from pasture and feed and transfer them to milk. Use of genetic engineering and other techniques to increase the range and level of anticarcinogens in pasture and supplements may increase the anticancer potential of milk.     

Functional properties of cream from dairy cows with experimentally altered milk fat composition

Modification of milk fat composition might be a desirable method to alter manufacturing characteristics or produce dairy products low in saturated fat that more closely meet consumer dietary preferences. The aim of this research was to evaluate functional properties of cream obtained from milks with fat composition modified by altering the profile of long-chain fatty acids (FA) absorbed from the intestine. A control and 5 mixtures of long-chain free FA were infused into the abomasum of lactating dairy cows in a 6 × 6 Latin square design with 21-d periods. Treatments were as follows: (1) control (no FA infused), (2) mostly saturated FA (C16:C18 = 0.74), (3) low linoleic palm FA (C16:C18 = 0.73), (4) palm FA (C16:C18 = 0.73), (5) soy FA (C16:C18 = 0.10), and (6) high palmitic soy FA (C16:C18 = 0.73). All treatments included meat solubles and Tween 80 as emulsifiers. Viscosity, overrun, whipping time, foam firmness, and foam stability were evaluated in creams (33% fat). Cream from cows infused with soy FA (treatment 5) had the longest whipping time and lowest overrun, foam stability, viscosity, melting point, firmness, and solid fat content at 5 and 20°C because the fat had the highest unsaturated FA content. Increasing palmitic acid content of soy FA (treatment 6) improved functional variables in cream relative to soy FA alone. Differences among treatments 1 to 4 were less pronounced because of the effect of C18:1 trans in treatments 3 and 4 on milk fat yield and composition. Milk fat from cows infused with palm FA (treatment 4) exhibited comparable or better functionality than control cream. Increased polyunsaturated FA in milk fat resulted in increased amounts of triglyceride (TG) fractions with 28, 30, 38, and 40 carbon numbers, increased oleic acid resulted in increased 50-carbon TG, and higher palmitic and myristic acids resulted in greater 44, 46, and 48 carbon number TG. These TG groups consistently correlated with functional properties of creams from different treatments. Our results indicated that optimal functionality of cream is dependent more on its content of palmitic acid than on unsaturated FA. However, an optimal composition of milk fat for cream functional characteristics might be obtained through nutritional manipulation of diets for dairy cows to deliver an optimal profile of FA.     

The effect of butter grains on physical properties of butter-like emulsions

Milk fat exists as globules in its natural state in milk. The potential of using globular fat to modulate the rheological properties and crystallization behavior in butter-like emulsions was studied in the present work. We conducted a comparative study of butter-like emulsions, with a fat phase consisting of 0, 10, 25, 50, or 100% anhydrous milk fat (AMF), the remaining fat being butter grains, and all samples containing 20% water, to obtain systematic variation in the ratio of globular fat. All emulsions were studied over 4 wk of storage at 5°C. By combining small and large deformation rheology, we conducted a detailed characterization of the rheological behavior of butter-like emulsions. We applied differential scanning calorimetry to monitor thermal behavior, confocal laser scanning microscopy for microstructural analysis, and low-field pulsed nuclear magnetic resonance spectrometry to measure solid fat content. By combining these techniques, we determined that increasing the fraction of globular fat (by mixing with butter grains) decreases the hardness of butter-like emulsions up to an order of magnitude at d 1. However, no difference was observed in thermal behavior as a function of butter grain content, as all emulsions containing butter grains revealed 2 endothermal peaks corresponding to the high (32.7°C ± 0.6) and medium (14.6°C ± 0.1) melting fractions of fatty acids. In terms of microstructure, decreasing the amount of butter grains in the emulsions resulted in formation of a denser fat crystal network, corresponding to increased hardness. Moreover, microstructural analysis revealed that the presence of butter grains resulted in faster formation of a continuous fat crystal network compared with the 100% AMF sample, which was dominated by crystal clusters surrounded by liquid oil. During storage, hardness remained stable and no changes in thermal behavior were observed, despite an increase in solid fat content of up to 5%. After 28 d of storage, we observed no difference in either microstructural or rheological properties, indicating that formation of primary bonds occurs primarily within the first day of storage. The rheological behavior of butter-like emulsions is not determined solely by hardness, but also by stiffness related to secondary bonds within the fat crystal network. The complex rheological behavior of milk fat-based emulsions is better characterized using multiple parameters.     

Bovine milk fat globule membrane proteome

Milk fat globule membranes (MFGM) were isolated from the milk of mid-lactation Holstein cows. The purified MFGM were fractionated using 1-dimensional SDS gels. Tryptic peptides from gel slices were further fractionated on a micro-capillary high performance liquid chromatograph connected to a nanospray-tandem mass spectrometer. Analysis of the data resulted in 120 proteins being identified by two or more unique peptide sequences. Of these 120 proteins, 71% are membrane associated proteins with the remainder being cytoplasmic or secreted proteins. Only 15 of the proteins identified in the cow MFGM were the same as proteins identified in previous mouse or human MFGM proteomic studies. Thus, the bulk of the proteins identified are new for bovine MFGM proteomics. The proteins identified were associated with membrane/protein trafficking (23%), cell signalling (23%), unknown functions (21%), fat transport/metabolism (11%), transport (9%), protein synthesis/folding (7%), immune proteins (4%) and milk proteins (2%). The proteins associated with cell signalling or membrane/protein trafficking may provide insights into MFGM secretion mechanisms. The finding of CD14, toll like receptor (TLR2), and TLR4 on MFGM suggests a direct role for the mammary gland in detecting an infection.