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In an attempt to develop strategies for enhancing the nutritional value of sheep milk fat, dairy ewe diet was supplemented with 3 incremental levels of marine algae (MA), in combination with sunflower oil, to evaluate the effects of these marine lipids on milk fatty acid (FA) profile and animal performance. Fifty Assaf ewes in mid lactation were distributed in 10 lots of 5 animals each and allocated to 5 treatments (2 lots per treatment): no lipid supplementation (control) or supplementation with 25 g of sunflower oil/kg of DM plus 0 (SO), 8 (SOMA(1)), 16 (SOMA(2)), or 24 (SOMA(3)) g of MA (56.7% ether extract)/kg of DM. Milk production and composition, including FA profile, were analyzed on d 0, 3, 7, 14, 21, and 28 of treatment. Neither intake nor milk yield were significantly affected by lipid addition, but all MA supplements decreased milk fat content from d 14 onward, reaching a 30% reduction after 28 d on SOMA(3). This milk fat depression might be related not only to the joint action of some putative fat synthesis inhibitors, such as trans-9,cis-11 C18:2 and probably trans-10 C18:1, but also to the limited ability of the mammary gland to maintain a desirable milk fat fluidity, that would have been caused by the noticeable increase in trans-C18:1 together with the lowered availability of stearic acid for oleic acid synthesis through Delta(9)-desaturase. Furthermore, all lipid supplements, and mainly MA, reduced the secretion of de novo FA (C6:0-C14:0) without increasing the yield of preformed FA (>C16). Supplementation with sunflower oil plus MA resulted in larger increases in cis-9,trans-11 C18:2 than those observed with sunflower oil alone, achieving a mean content as high as 3.22% of total FA and representing a more than 7-fold increase compared with the control. Vaccenic acid (trans-11 C18:1) was also significantly enhanced (on average +794% in SOMA treatments), as was C22:6 n-3 (DHA) content, although the transfer efficiency of the latter, from the diets to the milk, was very low (5%). However, the highest levels of MA inclusion (SOMA(2) and SOMA(3)) reduced the milk n-6:n-3 ratio, but MA supplements caused an important increase in trans-10 C18:1, which would rule out the possibility that this milk has a healthier fat profile before determining the specific role of each individual FA and ensuring that this trans-FA is at least innocuous in relation to cardiovascular disease risk.

17Background: Milk fat intake is often associated with a high risk of suffering from 18 cardiovascular disease (CVD) due to its high saturated fat content. However, not all 19 saturated fatty acids (SFA) are equal and they present structural differences that 20 promote distinct effects on the biological processes. In addition, there is a growing 21 scientific consensus pointing to dairy fat as a natural source of bioactive components. 22 23 Scope and Approach: The present review provides the most recent knowledge on the 24 bioactive properties of fatty acids detected in dairy products and their potential effects 25 on consumer health. The metabolic processes that involve these fatty acids and serious 26 chronic diseases such as CVD, obesity, diabetes or cancer are explained and discussed 27 throughout the text based on in vitro, animal and human studies. Moreover, information 28 gaps are highlighted to inspire further research in the field. 29 30 Key Findings and Conclusions: Recent investigations support that milk SFA should no 31 longer be considered as a single group in terms of metabolism or negative effects in 32 case of excess. Even they suggest that individual SFA possess specific properties 33 associated with important physiological functions. Whole dairy products would also 34 promote human health due to the presence of certain bioactive fatty acids. Among them, 35 it is worth mentioning the maintenance of gut microbiota and weight control from short 36 and medium-chain SFA, the essential role of branched-chain SFA in gut health at birth 37 and the prevention of chronic inflammatory diseases by vaccenic and rumenic acids.38 39 Keywords: bioactive lipids, dairy fat, human health, trans fat, conjugated linoleic acid 40 41 42 48 total fat, while the remaining 2% comprises diacylglycerides, monoacylglycerides, free 49 fatty acids (FFA), phospholipids, sterols and hydrocarbons. TAG composition is 50 extremely complex as more than 400 different fatty acids (FA) can be esterified in the 51 three positions (sn-1, sn-2 and sn-3) of the glycerol backbone at different 52 concentrations, which mainly depend on ruminant diet and its lactation stage (Jensen, 53

The objective of this study was to assess the effects of dietary supplementation of extruded linseed on animal performance and fatty acid (FA) profile of ewe milk for the production of n-3 FA- and conjugated linoleic acid-enriched cheeses. A Manchega ewe flock (300 animals) receiving a 60:40 forage:concentrate diet was divided into 3 groups supplemented with 0, 6, and 12 g of extruded linseed/100 g of dry matter for the control, low, and high extruded linseed diets, respectively. Bulk and individual milk samples from 5 dairy ewes per group were monitored at 7, 14, 28, 45, and 60 d following supplementation. Manchego cheeses were made with bulk milk from the 3 treatment groups. Milk yield increased in dairy ewes receiving extruded linseed. Milk protein, fat, and total solids contents were not affected by linseed supplementation. Milk contents of alpha-linolenic acid increased from 0.36 with the control diet to 1.91% total FA with the high extruded linseed diet. Similarly, cis-9 trans-11 C18:2 rose from 0.73 to 2.33% and its precursor in the mammary gland, trans-11 C18:1, increased from 1.55 to 5.76% of total FA. This pattern occurred with no significant modification of the levels of trans-10 C18:1 and trans-10 cis-12 C18:2 FA. Furthermore, the high extruded linseed diet reduced C12:0 (-30%), C14:0 (-15%) and C16:0 (-28%), thus significantly diminishing the atherogenicity index of milk. The response to linseed supplementation was persistently maintained during the entire study. Acceptability attributes of n-3-enriched versus control cheeses ripened for 3 mo were not affected. Therefore, extruded linseed supplementation seems a plausible strategy to improve animal performance and nutritional quality of dairy lipids in milk and cheese from ewes.

Ruminant diet supplementation with sunflower oil (SO) and fish oil (FO) has been reported as a good strategy for enhancing some milk fat compounds such as conjugated linoleic acid (CLA) and n-3 polyunsaturated fatty acids in dairy cows, but no information is available regarding dairy sheep. In this work, ewe diet was supplemented with FO, alone or in combination with SO, with the aim of improving milk nutritional value and evaluating its effect on animal performance. Sixty-four Assaf ewes in mid lactation, fed a high-concentrate diet, were distributed in 8 lots of 8 animals each and assigned to 4 treatments (2 lots/treatment): no lipid supplementation (control) or supplementation with 20 g of SO/kg (SO), 10 g of FO/kg (FO), or 20 g of SO plus 10 g of FO/kg (SOFO). Milk production and composition, including a complete fatty acid profile, were analyzed on d 0, 3, 7, 14, 21, and 28 of treatments. Supplementation with FO tended to reduce dry matter intake compared with the control treatment (-15%), and its use in combination with SO (SOFO) resulted in a significant decrease in milk yield as well (-13%). All lipid supplements reduced milk protein content, and FO also reduced milk fat content by up to 21% alone (FO) and 27% in combination with SO (SOFO). Although the mechanisms involved in FO-induced milk fat depression are not yet well established, the observed increase in some milk trans-FA that are putative inhibitors of milk fat synthesis, such as trans-9,cis-11 CLA, and the 63% decrease in C18:0 (consistent with the theory of reduced milk fat fluidity) may be involved. When compared with the control, lipid supplementation remarkably improved the milk content of rumenic acid (cis-9,trans-11 CLA; up to 4-fold increases with SO and SOFO diets), whereas FO-containing diets also increased milk n-3 polyunsaturated fatty acids, mainly docosahexaenoic acid (with mean contents of 0.29 and 0.38% of total fatty acids for SOFO and FO, respectively), and reduced the n-6:n-3 FA ratio to approximately half the control value. All lipid supplements resulted in high levels of some trans-FA, mainly trans-11 C18:1 (vaccenic acid) but also trans-10 C18:1.

Feeding vegetable oils rich in linoleic acid has been demonstrated to be an effective strategy to enrich milk with conjugated linoleic acid (CLA). However, high amounts of vegetable oil in the diet in free form could adversely affect animal performance, mainly in sheep. The aim of this work was to improve the ewe milk fatty acid profile by increasing potentially healthy acids such as CLA without any detrimental effects on milk production and ruminal fermentation with soybean oil (SBO) diet supplementation. Twenty-four ewes were assigned to 2 treatments and fed 2 diets (control or supplemented with 6% of SBO; 2 lots of 6 animals per treatment) and fed ad libitum for 4 wk. The forage:concentrate ratio was 20:80. Batch cultures of rumen microorganisms were used to study in vitro rumen fermentation. Changes in fatty acid profile were characterized as a reduction in C6:0 to C16:0 at the expense of an increase in C18:0, C18:1 isomers, and CLA concentrations. Proportions of milk CLA and trans-11 C18:1 (vaccenic acid) went from 1.04 to 3.44 and 2.08 to 6.20 g/100 g of total fatty acids, respectively. However, the SBO diet also increased trans-10 C18:1 and other trans C18:1 content. No significant decreases were found in the treatments for dry matter intake and milk production. The notable increases in trans-10, cis-12 and trans-9, cis-11 were not accompanied by fat level decreases in ewe milk. Concerning in vitro ruminal fermentation, no significant differences were found in the extent and rate of gas production, effective degradability, in vitro true digestibility, and volatile fatty acid production. The results demonstrate that dairy sheep milk CLA content can be substantially increased (more than 3-fold) by adding high levels of SBO in the diet as free oil, without any negative effects on animal performance.

An improved rapid method for separating lipids from milk to determine the fatty acid composition using 2 centrifugations at room temperature (20 degrees C) was compared with the ISO-IDF reference procedure based on solvent extraction. The new method is useful for research and routine quality control and has a number of advantages over the reference procedure--mainly no solvents are required and it saves time. Applicability of the rapid separation method was confirmed in fats with different physical characteristics from ewe and goat milk samples. Minor differences were found in the proportions of some fatty acids in the reference and centrifugation methods. Milk fat separated by centrifugation at room temperature did not differ in fatty acid composition from milk centrifuged at 4 degrees C.

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