The objective of the study was to examine the effects of trans-9-octadecenoic acid (elaidic) and trans-II-octadecenoic acid (trans-vaccenic) on uptake and alteration of exogenous fatty acids by mouse mammary epithelial cells. Cells from a subclone of the COMMA-D cell line were plated on uncoated plastic petri dishes and grown to confluence. Supplemental fatty acids bound to bovine serum albumin were added to the medium applied to the confluent cell cultures. Treatments included 200 JlM octadecanoic acid (CI8:0) as a control and 100 JlM CI8:0 with one of the following JlM ratios of cis-octadecenoic acid (cis-CI8:I) to elaidic or trans-vaccenic: 100:0, 75:25, 50:50, 0:100. In addition, all treatments were conducted with or without lactogenic hormone supplementation. The cellular protein to DNA ratio and total amount of fatty acids per mg protein were decreased (P < .05) by addition of lactogenic hormones. In treatments without hormone supplementation, however, the total amount of cellular fatty acids per mg protein was decreased (P < .05) by addition of either trans-CI8:I isomer. Results indicated a significant (P < .05) relationship between the concentration oftrans-C18:1 in the media and uptake of trans-C 18: 1 isomers, and retroconversion of trans-C 18: 1 to trans-C 16: 1. The slopes of the lines for cellular C16:0, cis-C16:1, and cis-CI8:1 were less (P < .05) than zero as concentration oftrans-CI8:1 in the media increased. However, trans-CI8:1 isomers did not influence the proportion of polar and nonpolar lipids synthesized by the cells. It appears that trans fatty acids may depress milk fat output by decreasing de novo fatty acid synthesis and cis-C 18: I content. / Master of Science

Une étude comparative de la régulation nutritionnelle du métabolisme des lipides chez la vache et la chèvre laitières a été réalisée afin d’identifier les mécanismes et préciser les spécificités de ces 2 espèces en vue d’une meilleure maîtrise de la quantité et de la qualité de la matière grasse laitière (MGL). Les effets respectifs de régimes riches en concentrés non supplémenté (CTL) ou supplémentés en huile de maïs et en amidon (COS), ou en poudre d’algues (MAP) ou en huile de palme hydrogénée (HPO) sur la plasticité et la composition de la MGL, et sur des indicateurs des métabolismes ruminal, intermédiaire et mammaire ont été étudiés chez la vache et la chèvre (n=12 par espèce) conduites simultanément selon un carré latin 4X4. Les régimes n’ont pas eu d’effet sur la production laitière quelle que soit l’espèce. Cependant, une différence de réponse inter-espèces au régime COS a été observée avec une forte chute de la teneur en MGL (-45%) chez la vache mais pas chez la chèvre. Le régime MAP a conduit à une diminution de la teneur en MGL chez la vache (-22%) et, dans une moindre mesure, chez la chèvre (-15%), tandis que HPO l'a augmentée seulement chez la vache (+13%). Les différences majeures observées avec COS entre les 2 espèces sont attribuées 1/ à des différences de biohydrogénations ruminales (BHR) des AG polyinsaturés avec une plus grande stabilité des voies classiques de BHR chez la chèvre ; 2/ au métabolisme intermédiaire, avec une augmentation des lipides circulants chez la chèvre suggérant une plus grande disponibilité en AG longs pour la glande mammaire (GM). Les réponses à MAP ont été attribuées à des mécanismes similaires chez les 2 espèces mais différents en termes d’indicateurs des métabolismes ruminal et intermédiaire de ceux identifiés pour COS. Chez la vache HPO se distingue par une augmentation du C16:0 et C16:1 cis-9 du lait suggérant un transport et/ou un captage privilégié du C16:0 chez cette espèce. Quel que soit le régime, le métabolisme mammaire des lipides, étudié via l’abondance des ARNm de gènes de la lipogenèse, n’a pas été relié aux données de sécrétions des AG du lait. Nos résultats montrent que la plasticité de la MGL chez deux espèces de ruminants, à priori proches, est contrôlée par des mécanismes différents selon l’espèce et le régime. Nous avons construit une base de données phénotypiques sur 24 animaux de 2 espèces recevant 4 régimes dont l’analyse nous a permis de préciser les mécanismes de synthèse de la MGL. Ce projet de recherche pourrait permettre, in fine, de proposer des outils de monitoring via le phénotypage de la composition fine du lait et de proposer des stratégies d’élevage pour moduler les performances de l’animal. / A comparative study of the nutritional regulation of lipid metabolism in dairy cows and goats was performed to identify the mechanisms and clarify the specificities of these 2 ruminant species in order to better control milk fat yield and quality. The effects of diets containing no additional lipid (CTL) or supplemented with corn oil (5% dry matter intake (DMI)) and wheat starch (COS), marine algae powder (MAP) (1.5% DMI), or hydrogenated palm oil (HPO) (3% DMI), on milk fat plasticity and composition, and on indicators of ruminal, intermediary and mammary metabolisms were studied in cows and goats (n=12 per species) conducted simultaneously according to a 4x4 Latin square design. Dietary treatments had no significant effects on milk yield in both species. Conversely, species-specific response of milk fat content to dietary treatment were observed: in cows, milk fat content was lowered by COS (-45%) and MAP (-22%) and increased by HPO (+13%) compared with CTL, and in goats, only MAP had an effect compared with CTL by decreasing milk fat content by 15%. The major differences observed for COS among species were attributed 1/ at differences in the polyunsaturated fatty acids (FA) ruminal biohydrogenation (RBH) processes with a greater stability of the classical RBH pathways in goats; 2/ at the intermediary metabolism, with an increase in circulating lipids in goats suggesting a higher availability of long chain FA for mammary gland (MG). Responses on MAP treatment were attributed to similar mechanisms among species but different to those outlined for COS in terms of indicators of ruminal and intermediary metabolisms. In cows, HPO was characterized by an increased in milk 16:0 et cis-9 16:1 suggesting a favoured transport and/or uptake of 16:0 in this species. Whatever the dietary treatment the mammary lipid metabolism studied by the mRNA abundance of few lipogenic genes was not related with milk FA yields. Our results demonstrated that the milk fat plasticity in two closely related ruminant species is controlled by different mechanisms depending on species and dietary treatments. We produced a database on 24 animals of 2 species receiving 4 dietary treatments. The dataset analysis allowed us to enhance our knowledge on regulation mechanisms of milk fat synthesis. This research project will contribute for the development of monitoring tools based on milk composition phenotyping, and to propose husbandry strategies that modulate animal performance.

A procura dos consumidores por produtos mais saudáveis, combinada às descobertas científicas sobre os efeitos na saúde humana de diversos alimentos, gerou a busca por processos adequados à produção de alimentos. Neste contexto, óleos e gorduras modificados têm ganhado atenção especial. Uma opção industrial para modificar as propriedades dos óleos e gordura é por meio de reações de interesterificação catalisadas por agentes químicos ou bioquímicos. O presente trabalho teve como objetivo, verificar a influência do tipo de catalisador (metóxido de sódio e lipase de Rhizopus oryzae imobilizada em silica-PVA) nas propriedades sensoriais dos produtos obtidos por interesterificação da gordura do leite anidra e dois óleos vegetais (soja e canola). A avaliação sensorial feita através de um nariz eletrônico e um texturômetro, utilizando matérias-primas que estavam de acordo com as normas brasileiras e adequadas para as duas rotas de interesterificação. As reações de interesterificação química foram realizadas em rotaevaporador de baixa pressão contendo 320g da mistura (65% de gordura do leite e 35% de óleo vegetal) e 0,75% de metóxido de sódio a 60°C por 60min. As reações de interesterificação enzimática foram realizadas em reatores cilíndricos encamisados, carregados com 70g de meio (65% de gordura do leite e 35% de óleo vegetal) incubadas com derivado imobilizado na proporção fixa de 500 unidades de atividade por grama de meio (500U/g). As reações foram conduzidas por 6 horas, com agitação magnética, em atmosfera inerte (N2), ao abrigo da luz, a 45°C. Os resultados mostraram que os sensores do nariz eletrônico discriminaram as amostras satisfatoriamente e que o tipo de catalisador influenciou de forma marcante no aroma dos produtos interesterificados, independentemente do tipo de óleo usado. Este fato pode ser atribuído à especificidade do catalisador enzimático (enzima 1,3 específica) que atua nos carbonos 1,3 do glicerol enquanto na interesterificação química a distribuição dos triacilgliceróis ocorre ao acaso, imprimindo no painel dos componentes principais do aroma posições opostas. Especificamente, verificou-se que os aromas dos produtos interesterificados pela rota enzimática ficaram localizados em posição mais próxima à dos óleos vegetais, enquanto pela rota química os produtos obtidos ficaram mais próximos aos componentes dos aromas da gordura de leite e das misturas gordura e óleos vegetais. Apesar desta metodologia não permitir identificar que tipo de aroma poderia ser mais agradável ao consumidor, é possível afirmar que o catalisador tem uma influência importante no aroma de produtos obtidos por interesterificação. Com relação à textura, ambas as rotas geraram produtos com consistências adequadas ao critério estabelecido por Haigton. / Consumers\' demand for healthier products, combined with scientific discoveries about the effects of different foods on human health, has generated the search for suitable processes for the production of food. In this context, oils and modified fats have gained special attention. Reactions catalysed by chemical or biochemical agents are an industrial option to modify the properties of the oils and fat by means of its interesterification. This study aimed to verify the influence of the type of catalyst (sodium methylate and Rhizopus oryzae imobilizada on silica PVA) on the sensory properties of the products obtained by interesterification anhydrous fat milk and two vegetable oils (soybean and canola). Sensory evaluation was made through an electronic nose and texturometer, using raw materials that were in accordance with Brazilian rules and appropriate standards for the two routes of interesterification. The chemical interesterification reactions were performed at low pressure rotaevaporator, the mixture containing 320g (65% milk fat and 35% vegetable oil) and 0.75% sodium methoxide at 60° C for 60min. The enzymatic interesterification reactions were carried out in cylindrical jacketed reactors loaded with 70g of médium (65% milk fat and 35% vegetable oil) incubated with immobilized derivative in a fixed proportion of 500 activity units per gram of reaction medium (500U / g). The reactions were conducted for 6 hours, with magnetic stirring under inert atmosphere (N2), protected from light, at 45 ° C. The results showed that the sensors of the electronic nose discriminated samples satisfactorily and that the type of catalyst influenced markedly in the aroma of interesterified products regardless of the type of oil used. This fact can be attributed to the specificity of the enzyme catalyst (1.3-specific enzyme) that acts on the 1,3 carbons of the glycerol as chemical interesterification of triacylglycerols distribution occurs randomly, printing on the panel of the components of the aroma opposite positions. Specifically, it was found that the scent of interesterified product by enzymatic route were located in the nearest position to the vegetable oil, while by the chemical route the products obtained were closest to the components of milk fat flavors and mixtures fat and vegetable oils . Although this methodology cannot identify what kind of aroma could be more pleasant to the consumer, it is clear that the catalyst has an important influence on product aroma obtained by interesterification. Regarding the texture, both routes generated products with suitable consistency to the criteria established by Haigton.

A procura dos consumidores por produtos mais saudáveis, combinada às descobertas científicas sobre os efeitos na saúde humana de diversos alimentos, gerou a busca por processos adequados à produção de alimentos. Neste contexto, óleos e gorduras modificados têm ganhado atenção especial. Uma opção industrial para modificar as propriedades dos óleos e gordura é por meio de reações de interesterificação catalisadas por agentes químicos ou bioquímicos. O presente trabalho teve como objetivo, verificar a influência do tipo de catalisador (metóxido de sódio e lipase de Rhizopus oryzae imobilizada em silica-PVA) nas propriedades sensoriais dos produtos obtidos por interesterificação da gordura do leite anidra e dois óleos vegetais (soja e canola). A avaliação sensorial feita através de um nariz eletrônico e um texturômetro, utilizando matérias-primas que estavam de acordo com as normas brasileiras e adequadas para as duas rotas de interesterificação. As reações de interesterificação química foram realizadas em rotaevaporador de baixa pressão contendo 320g da mistura (65% de gordura do leite e 35% de óleo vegetal) e 0,75% de metóxido de sódio a 60°C por 60min. As reações de interesterificação enzimática foram realizadas em reatores cilíndricos encamisados, carregados com 70g de meio (65% de gordura do leite e 35% de óleo vegetal) incubadas com derivado imobilizado na proporção fixa de 500 unidades de atividade por grama de meio (500U/g). As reações foram conduzidas por 6 horas, com agitação magnética, em atmosfera inerte (N2), ao abrigo da luz, a 45°C. Os resultados mostraram que os sensores do nariz eletrônico discriminaram as amostras satisfatoriamente e que o tipo de catalisador influenciou de forma marcante no aroma dos produtos interesterificados, independentemente do tipo de óleo usado. Este fato pode ser atribuído à especificidade do catalisador enzimático (enzima 1,3 específica) que atua nos carbonos 1,3 do glicerol enquanto na interesterificação química a distribuição dos triacilgliceróis ocorre ao acaso, imprimindo no painel dos componentes principais do aroma posições opostas. Especificamente, verificou-se que os aromas dos produtos interesterificados pela rota enzimática ficaram localizados em posição mais próxima à dos óleos vegetais, enquanto pela rota química os produtos obtidos ficaram mais próximos aos componentes dos aromas da gordura de leite e das misturas gordura e óleos vegetais. Apesar desta metodologia não permitir identificar que tipo de aroma poderia ser mais agradável ao consumidor, é possível afirmar que o catalisador tem uma influência importante no aroma de produtos obtidos por interesterificação. Com relação à textura, ambas as rotas geraram produtos com consistências adequadas ao critério estabelecido por Haigton. / Consumers\' demand for healthier products, combined with scientific discoveries about the effects of different foods on human health, has generated the search for suitable processes for the production of food. In this context, oils and modified fats have gained special attention. Reactions catalysed by chemical or biochemical agents are an industrial option to modify the properties of the oils and fat by means of its interesterification. This study aimed to verify the influence of the type of catalyst (sodium methylate and Rhizopus oryzae imobilizada on silica PVA) on the sensory properties of the products obtained by interesterification anhydrous fat milk and two vegetable oils (soybean and canola). Sensory evaluation was made through an electronic nose and texturometer, using raw materials that were in accordance with Brazilian rules and appropriate standards for the two routes of interesterification. The chemical interesterification reactions were performed at low pressure rotaevaporator, the mixture containing 320g (65% milk fat and 35% vegetable oil) and 0.75% sodium methoxide at 60° C for 60min. The enzymatic interesterification reactions were carried out in cylindrical jacketed reactors loaded with 70g of médium (65% milk fat and 35% vegetable oil) incubated with immobilized derivative in a fixed proportion of 500 activity units per gram of reaction medium (500U / g). The reactions were conducted for 6 hours, with magnetic stirring under inert atmosphere (N2), protected from light, at 45 ° C. The results showed that the sensors of the electronic nose discriminated samples satisfactorily and that the type of catalyst influenced markedly in the aroma of interesterified products regardless of the type of oil used. This fact can be attributed to the specificity of the enzyme catalyst (1.3-specific enzyme) that acts on the 1,3 carbons of the glycerol as chemical interesterification of triacylglycerols distribution occurs randomly, printing on the panel of the components of the aroma opposite positions. Specifically, it was found that the scent of interesterified product by enzymatic route were located in the nearest position to the vegetable oil, while by the chemical route the products obtained were closest to the components of milk fat flavors and mixtures fat and vegetable oils . Although this methodology cannot identify what kind of aroma could be more pleasant to the consumer, it is clear that the catalyst has an important influence on product aroma obtained by interesterification. Regarding the texture, both routes generated products with suitable consistency to the criteria established by Haigton.

ABSTRACT Effects of Milk Processing on the Milk Fat Globule Membrane Constituents Xiomara E. Elías-Argote The milk fat globule membrane (MFGM) is avidly studied by many groups of scientists around the world due to its unprecedented nutritional and functional properties; however, limited research has been performed on the effects of milk processing on the chemical changes of the MFGM. Thus, this study highlights the changes that lipids and proteins undergo from the time milk leaves cow’s udders. Cooling (4 °C) was included along with subsequent pasteurization by different traditional thermal processes and cold pasteurization by pulse light ultra violet treatment. Cooling milk to 4 °C had a measureable effect on the MFGM composition, resulting in protein alterations, particularly to butyrophilin and adipophilin. Thermal treatments disturbed the native structures of molecules and increased the adsorption of milk and whey proteins on the globules, especially a-lactalbumin and b-lactoglobulin. As the heat intensity increased, the aggregation of fat globules became more pronounced due to protein interactions. Intrinsic MFGM proteins also varied in relative abundance during the processing steps. The concentrations of polar lipids did not change during processing, with the exception of phosphatidylserine, which decreased during the cooling and thermal treatments. Cold pasteurization (UV treatment) had a minimal effect on fat globules and MFGM proteins. Since the MFGM promises to deliver nutritional effects and more when included in food products, currently HTST pasteurization was shown to be the best method to process milk and obtain MFGM isolates for further supplementation.

Milk fat globule membrane surrounds the fat droplets of milk. It is a biopolymer containing primarily membrane glycoproteins and polar lipids which contribute to its properties as a possible neutraceutical. The aims of the studies were to determine if dietary milk fat globule membrane: (1) confers protection against colon carcinogenesis; and (2) promotes gut mucosal integrity while decreasing inflammation compared to diets containing corn oil or anhydrous milk fat. Aim 1. Three dietary treatments differing only in the fat source were formulated: (1) AIN-76A, corn oil; (2) AIN-76A, anhydrous milk fat; and (3) AIN-76A, 50% milk fat globule membrane, 50% anhydrous milk fat. Each diet was formulated to contain 50 g/kg diet of fat and to be identical in macro and micro nutrient content. To assess protection against colon carcinogenesis, male, weanling Fischer-344 rats were randomly assigned to one of the three dietary treatments. Animals were injected with 1,2-dimethylhydrazine once per week at weeks 3 and 4. After 13 weeks animals were sacrificed, colons were removed, and aberrant crypt foci were counted by microscopy. Rats fed the milk fat globule membrane diet (n = 16) had significantly fewer aberrant crypt foci (20.9 ± 5.7) compared to rats fed corn oil (n = 17) or anhydrous milk fat (n = 16) diets (31.3 ± 9.5 and 29.8 ± 11.4 respectively; P < 0.05). Aim 2. Male BALB/c mice were randomly assigned to one of two diets: AIN- 76A, corn oil or AIN-76A, 50% milk fat globule membrane, 50% anhydrous milk fat. After 5 weeks mice were injected with saline vehicle control or lipopolysaccharide and gavaged with dextran-FITC. To assess gut mucosal integrity and inflammation, serum samples were assayed for dextran-FITC 24 and 48 hours after gavage, and a panel of 16 cytokine concentrations was analyzed. Serum concentrations of IL-6, IL-10, IL-17, MCP-1, IFNγ, and TNFα decreased and gut permeability decreased 45% in lipopolysaccharide challenged mice fed milk fat globule membrane diet compared to control diet at 24 hours (P < 0.05). Overall, the results of these aims suggest that diets containing milk fat globule membrane are protective against colon carcinogenesis, inhibit the inflammatory response, and protect against gastrointestinal stress.

Video files: Some images may require stereoscopic glasses / Fat in milk and cream is present as tiny droplets, which are each enveloped in a thin membrane, called the milk fat globule membrane (MFGM). The MFGM can easily be damaged by factors such as pumping the milk and applying other forms of agitation. MFGM damage is believed to reduce processing efficiency and compromise the quality of manufactured products. A comprehensive review of the literature showed that our understanding of changes occurring in the MFGM post secretion of the fat globule by the mammary secretory cell is still rudimentary. Furthermore, it was found that a fundamental understanding of MFGM damage in raw milk is lacking. Hence, this study sought to develop analytical techniques for studying the MFGM. Fluorescent probes were identified that associated with the MFGM (bovine, ovine, human) in one of two ways: either by embedding in the phospholipid bilayer (lipophilic probe) or by binding to carbohydrate moieties of glycosylated chains in the glycocalyx (lectin probes). The use of these probes, in combination with either conventional fluorescence microscopy or confocal laser scanning microscopy, allowed 2-D images and 3-D images of fat globules to be made. Application of water-soluble lipophilic probes and the lectin wheat germ agglutinin (WGA) directly to milk allowed the staining of the MFGM in its native environment. Variable distribution patterns of the probes in the MFGM were observed, which suggests that the MFGM of fat globules in harvested milk is structurally and chemically heterogeneous both within and among globules from the same species and between species, and even among fat globules within the milk of an individual animal. Furthermore, the binding behaviour of WGA to the MFGM of native fat globules (in bovine milk) and washed fat globules (in model systems) following heat treatment implicated β-lactoglobulin, α-lactalbumin, immunoglobulin M and/or the glycosylated proteins Periodic acid Schiff 6/7 in the disappearance of fat globule aggregation upon elevated heat treatment of milk. The results of the current study showed that the use of membrane-specific fluorescent probes, particularly in combination with confocal laser scanning microscopy, has significant potential for providing real time structural and chemical information about the MFGM in matrices such as harvested milk and milk products. In addition to the fluorescence microscopy techniques, development of other techniques was also conducted. Flow cytometry was shown to have significant potential for the quantitative determination of various properties of fat globules and their membranes. Although no suitable sample preparation technique could be developed in this study, atomic force microscopy is believed to have significant potential for studying structural and physical properties of the MFGM. Selective harvesting of individual fat globules was shown to be possible by using a micromanipulator. In future work, this technique is expected to be used in combination with fluorescence microscopy, or atomic force microscopy. The present study has shown that the development and application of novel analytical techniques has advanced, and in the future will further advance, understanding of the MFGM.

This thesis deals with the composition of milk fat and the proportion of fatty acids in bovine milk fat. Then we evaluated factors that affect the fatty acids in milk fat - in particular the effect of diet. In results is as well mentioned one of the biological factors - the effect of the breed.

Changes occur in milk during the technological processing of raw milk, which also affect the properties of milk fat. Dairy products may have different properties compared to the raw milk. Chemical and microbial processes can alter the presence and distribution of individual fatty acids and thus influence the nutritional value of the product. The aim of this thesis was to assess the presence of fatty acids in milk fat of goat milk and to evaluate differences in the spectrum of fatty acids in goat milk and yoghurt produced from it.

Global studies show the effect of polymorphism of selected genes on dairy production and fatty acid spectrum. The aim of the diploma thesis was genotyping of candidate FASN locus with a focus on milk yield and fatty acid spectrum depending on genotype. The thesis describes the characteristics of cow's milk, milk fat and fatty acids in milk fat. Furthermore, the thesis deals with the genome of cattle and the potential influence of polymorphism of candidate genes affecting fatty acids of milk fat. Genotypes for FASN were determined by the PCR-RFLP method, the milk yield of selected dairy cattle was statistically evaluated according to the milk production indices for 1st lactation in individual breeds, and the determination of fatty acids was performed by spectrophotometry followed by statistical evaluation. The resulting genotypes in selected breeds were only two, the GG genotype with a higher relative frequency than the genotype AG. There is no statistically significant difference between FASN genotypes depending on the milk yield and the fatty acids spectrum.