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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.