Short communication: Eicosatrienoic acid and docosatrienoic acid do not promote vaccenic acid accumulation in mixed ruminal cultures

Previous research found that docosahexaenoic acid (C22:6n-3) was a component of fish oil that promotes trans-C18:1 accumulation in ruminal cultures when incubated with linoleic acid. The objective of this study was to determine if eicosatrienoic acid (C20:3n-3) and docosatrienoic acid (C22:3n-3), n-3 fatty acids in fish oil, promote accumulation of trans-C18:1, vaccenic acid (VA) in particular, using cultures of mixed ruminal microorganisms. Treatments consisted of control, control plus 5 mg of C20:3n-3 (ETA), control plus 5 mg of C22:3n-3 (DTA), control plus 15 mg of linoleic acid (LA), control plus 5 mg of C20:3n-3 and 15 mg of linoleic acid (ETALA), and control plus 5 mg of C22:3n-3 and 15 mg of linoleic acid (DTALA). Treatments were incubated in triplicate in 125-mL flasks, and 5 mL of culture contents was taken at 0 and 24 h for fatty acid analysis by gas-liquid chromatography. After 24 h of incubation, the concentrations of trans-C18:1 (0.87, 0.88, and 0.99 mg/culture), and VA (0.52, 0.56, and 0.62 mg/culture) were similar for the control, ETA, and DTA cultures, respectively. The concentrations of trans-C18:1 (5.51, 5.41, and 5.36 mg/culture), and VA (4.78, 4.62, and 4.59 mg/culture) were also similar between LA, ETALA, and DTALA cultures, respectively. These data suggest that C20:3n-3 and C22:3n-3 are not the active components in fish oil that promote VA accumulation when incubated with linoleic acid.     

Effect of supplementation with fish oil or microalgae on fatty acid composition of milk from cows managed in confinement or pasture systems

The objective of this study was to examine the interaction between lipid supplement (LS) and management system (MS) on fatty acid (FA) composition of milk that could affect its healthfulness as a human food. Forty-eight prepartal Holstein cows were blocked by parity and predicted calving date and deployed across pasture (PAS; n = 23) or confinement (CONF; n = 25) systems. Cows within each system were assigned randomly to a control (no marine oil supplement) or to 1 of 2 isolipidic (200 g/d) marine oil supplements: fish oil (FO) or microalgae (MA) for 125 ± 5 d starting 30 d precalving. The experiment was conducted as a split-plot design, with MS being the whole-plot treatment and LS as the subplot treatment. Cows were housed in a tie-stall barn from −30 until 28 ± 10 d in milk (DIM) and were fed total mixed rations with similar formulations. The PAS group was then adapted to pasture and rotationally grazed on a perennial sward until the end of the experiment (95 ± 5 DIM). Milk samples were collected at 60 and 90 DIM for major components and FA analyses. Milk yield (kg/d) was lower in PAS (34.0) compared with CONF (40.1) cows. Milk fat percentage was reduced with MA compared with FO (3.00 vs. 3.40) and the control (3.56) cows. However, milk fat yield (kg/d) was not affected by lipid supplements. Compared with CONF, PAS cows produced milk fat with a lower content of 12:0 (−38%), 14:0 (−28%), and 16:0 (−17%), and more cis-9 18:1 (+32%), 18:3 n-3 (+30%), conjugated linoleic acid (CLA; +70%) and trans 18:1 (+34%). Both supplements, regardless of MS, reduced similarly the milk fat content of 16:0 (−12%) and increased CLA (+28%) and n-3 long-chain polyunsaturated FA (n-3 LC-PUFA; +150%). Milk fat content of trans 18:1 (trans-6 to trans-16) was increased with FO or MA, although the effect was greater with MA (+81%) than with FO (+42%). The interaction between MS and LS was significant only for trans-11 18:1 (vaccenic acid, VA) and cis-9,trans-11 CLA (rumenic acid). In contrast to CONF, feeding FO or MA to PAS cows did not increase milk fat content of VA and rumenic acid. We concluded that compared with CONF, milk from PAS cows had a more healthful FA composition. Feeding either FO or MA improved n-3 long-chain polyunsaturated FA and reduced levels of 16:0 in milk fat, regardless of MS, but concurrently increased the trans 18:1 isomers other than VA, at the expense of VA, particularly in grazing cows.     

Effect of linoleic acid and dietary vitamin E supplementation on sustained conjugated linoleic acid production in milk fat from dairy cows

Conjugated linoleic acid (CLA; cis-9,trans-11 18:2), a bioactive fatty acid (FA) found in milk and dairy products, has potential human health benefits due to its anticarcinogenic and antiatherogenic properties. Conjugated linoleic acid concentrations in milk fat can be markedly increased by dietary manipulation; however, high levels of CLA are difficult to sustain as rumen biohydrogenation shifts and milk fat depression (MFD) is often induced. Our objective was to feed a typical Northeastern corn-based diet and investigate whether vitamin E and soybean oil supplementation would sustain an enhanced milk fat CLA content while avoiding MFD. Holstein cows (n = 48) were assigned to a completely randomized block design with repeated measures for 28 d and received 1 of 4 dietary treatments: (1) control (CON), (2) 10,000 IU of vitamin E/d (VE), (3) 2.5% soybean oil (SO), and (4) 2.5% soybean oil plus 10,000 IU of vitamin E/d (SO-VE). A 2-wk pretreatment control diet served as the covariate. Milk fat percentage was reduced by both high-oil diets (3.53, 3.56, 2.94, and 2.92% for CON, VE, SO, and SO-VE), whereas milk yield increased significantly for the SO-VE diet only, thus partially mitigating MFD by oil feeding. Milk protein percentage was higher for cows fed the SO diet (3.04, 3.05, 3.28, and 3.03% for CON, VE, SO, and SO-VE), implying that nutrient partitioning or ruminal supply of microbial protein was altered in response to the reduction in milk fat. Milk fat concentration of CLA more than doubled in cows fed the diets supplemented with soybean oil, with concurrent increases in trans-10 18:1 and trans-11 18:1 FA. Moreover, milk fat from cows fed the 2 soybean oil diets had 39.1% less de novo synthesized FA and 33.8% more long-chain preformed FA, and vitamin E had no effect on milk fat composition. Overall, dietary supplements of soybean oil caused a reduction in milk fat percentage and a shift in FA composition characteristic of MFD. Supplementing diets with vitamin E did not overcome the oil-induced reduction in milk fat percentage or changes in FA profile, but partially mitigated the reduction in fat yield by increasing milk yield.     

Milk production and composition from cows fed small amounts of fish oil with extruded soybeans

Eight Holstein (189 ± 57 DIM) and 4 Brown Swiss (126 ± 49 DIM) multiparous cows were used in a replicated 4 × 4 Latin square with 28-d periods to determine the minimal dietary concentration of fish oil necessary to maximize milk conjugated linoleic acid (CLA) and vaccenic acid (VA). Treatments consisted of a control diet with a 50:50 ratio of forage to concentrate (dry matter basis), and 3 diets with 2% added fat consisting of 0.33% fish oil, 0.67% fish oil, and 1% fish oil with extruded soybeans providing the balance of added fat. Dry matter intake (23.1, 22.6, 22.8, and 22.9 kg/d, for control, low, medium, and high fish oil diets, respectively) was similar for all diets. Milk production (21.5, 23.7, 22.7, and 24.2 kg/d) was higher for cows fed the fat-supplemented diets vs. the control. Milk fat (4.42, 3.81, 3.80, and 4.03%) and true protein (3.71, 3.58, 3.54, and 3.55%) concentrations decreased when cows were fed diets containing supplemental fat. Concentration of milk cis-9,trans-11 CLA (0.55, 1.17, 1.03, and 1.19 g/100 g of fatty acids) was increased similarly by all diets containing supplemental fat. Milk VA (1.12, 2.47, 2.13, and 2.63 g/100 g of fatty acids) was increased most in milk from cows fed the low and high fish oil diets. Milk total n-3 fatty acids were increased (0.82, 0.96, 0.92, and 1.01 g/100 g of fatty acids) by all fat-supplemented diets. The low fish oil diet was as effective at increasing VA and CLA in milk as the high fish oil diet, showing that only low concentrations of dietary fish oil are necessary for increasing concentrations of VA and CLA in milk.     

Milk fatty acid composition of grazing dairy cows when supplemented with linseed oil

The effects of varying amounts of linseed oil (LSO) in grazing dairy cows’ diet on milk conjugated linoleic acid (cis-9, trans-11 CLA) were investigated in this study. Twelve Holstein cows in midlactation (150 ± 19 DIM) were placed on alfalfa-based pasture and assigned to 4 treatments using a 4 × 4 Latin square design with 3-wk periods. Treatments were: 1) control grain supplement; 2) control grain supplement containing 170 g of LSO (LSO1); 3) control grain supplement containing 340 g of LSO (LSO2); and 4) control grain supplement containing 510 g of LSO (LSO3). Grain supplements were offered at 7 kg/d. Additional 100 g/d of algae, divided evenly between the 2 feeding times, were added to every treatment diet. Milk samples were collected during the last 3 d of each period and analyzed for chemical and fatty acid composition. Treatments had no effect on milk production (18.9, 18.5, 19.6, and 19.1 kg/d for treatments 1 to 4, respectively). Linseed oil supplementation caused a quadratic increase in milk fat (3.23, 3.44, 3.35, and 3.27% for treatments 1 to 4, respectively) and protein (3.03, 3.19, 3.12, and 3.08%) contents. Concentrations (g/100 g of fatty acids) of milk cis-9, trans-11 CLA (1.12, 1.18, 1.39, and 1.65 for treatments 1 to 4, respectively) and VA (3.39, 3.62, 4.25, and 4.89) linearly increased with LSO supplementations. Results from this trial suggest that the increase in milk cis-9, trans-11 CLA was proportional to the amounts of LSO fed. In conclusion, adding LSO to grazing dairy cow diets can improve the nutritional value of milk without compromising milk composition or cow performance.     

Effect of plant oils and camelina expeller on milk fatty acid composition in lactating cows fed diets based on red clover silage

Five multiparous Finnish Ayrshire cows fed red clover silage-based diets were used in a 5 × 5 Latin square with 21-d experimental periods to evaluate the effects of various plant oils or camelina expeller on animal performance and milk fatty acid composition. Treatments consisted of 5 concentrate supplements containing no additional lipid (control), or 29 g/kg of lipid from rapeseed oil (RO), sunflower-seed oil (SFO), camelina-seed oil (CO), or camelina expeller (CE). Cows were offered red clover silage ad libitum and 12 kg/d of experimental concentrates. Treatments had no effect on silage or total dry matter intake, whole-tract digestibility coefficients, milk yield, or milk composition. Plant oils in the diet decreased short- and medium-chain saturated fatty acid (6:0-16:0) concentrations, including odd- and branched-chain fatty acids and enhanced milk fat 18:0 and 18-carbon unsaturated fatty acid content. Increases in the relative proportions of cis 18:1, trans 18:1, nonconjugated 18:2, conjugated linoleic acid (CLA), and polyunsaturated fatty acids in milk fat were dependent on the fatty acid composition of oils in the diet. Rapeseed oil in the diet was associated with the enrichment of trans 18:1 (Δ4, 6, 7, 8, and 9), cis-9 18:1, and trans-7,cis-9 CLA, SFO resulted in the highest concentrations of trans-5, trans-10, and trans-11 18:1, Δ9,11 CLA, Δ10,12 CLA, and 18:2n-6, whereas CO enhanced trans-13-16 18:1, Δ11,15 18:2, Δ12,15 18:2, cis-9,trans-13 18:2, Δ11,13 CLA, Δ12,14 CLA, Δ13,15 CLA, Δ9,11,15 18:3, and 18:3n-3. Relative to CO, CE resulted in lower 18:0 and cis-9 18:1 concentrations and higher proportions of trans-10 18:1, trans-11 18:1, cis-9,trans-11 CLA, cis-9,trans-13 18:2, and trans-11,cis-15 18:2. Comparison of milk fat composition responses to CO and CE suggest that the biohydrogenation of unsaturated 18-carbon fatty acids to 18:0 in the rumen was less complete for camelina lipid supplied as an expeller than as free oil. In conclusion, moderate amounts of plant oils in diets based on red clover silage had no adverse effects on silage dry matter intake, nutrient digestion, or milk production, but altered milk fat composition, with changes characterized as a decrease in saturated fatty acids, an increase in trans fatty acids, and enrichment of specific unsaturated fatty acids depending on the fatty acid composition of lipid supplements.     

Milk fatty acid profile and dairy sheep performance in response to diet supplementation with sunflower oil plus incremental levels of marine algae

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 (SOMA₁), 16 (SOMA₂), or 24 (SOMA₃) 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 SOMA₃. 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 Δ⁹-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 (SOMA₂ and SOMA₃) 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.     

Supplementation with extruded linseed cake affects concentrations of conjugated linoleic acid and vaccenic acid in goat milk

The aim of this research was to determine the effect of adding extruded linseed cake to the dry diet of goats on the concentrations of conjugated linoleic acid (CLA) and vaccenic acid (VA) in milk fat. Thirty crossbreed dairy goats were divided into 3 groups. Their diet was supplemented with 0% (control group), 5% (low group), or 10% (high group) of extruded linseed cake (ELC), which supplied 0, 16, and 32 g/d of linseed fat, respectively. The milk fat percentage (overall mean 3.5%) and yield did not differ with the different diets, but fatty acid composition was affected by the ELC supplements. The inclusion of ELC in the diets did not influence the concentration of fatty acids from C6:0 to C12:0. The concentrations of C14:0 and C16:0 decreased as the quantity of ELC supplements increased. The concentrations (mg/100 mg of total fatty acid methyl esters) of VA (0.70, 1.23, and 1.39 in control, low, and high groups respectively) and cis-9,trans-11 CLA (0.63, 0.96, and 1.05 in control, low, and high groups, respectively) were increased by ELC supplements. The milk fat content of VA and cis- 9,trans-11 CLA were closely correlated (R² = 0.82). Desaturation of VA in the mammary gland to produce cis-9,trans-11 CLA was higher in the control group than in the groups with ELC diets. Extruded linseed cake supplementation to lactating goats may enhance the nutritional profile of milk lipids.     

Diet supplementation with fish oil and sunflower oil to increase conjugated linoleic acid levels in milk fat of partially grazing dairy cows

The objective of this study was to determine the long-term effect on milk conjugated linoleic acid (cis-9, trans-11 CLA) of adding fish oil (FO) and sunflower oil (SFO) to the diets of partially grazing dairy cows. Fourteen Holstein cows were divided into 2 groups (7 cows/treatment) and fed either a control or oil-supplemented diet for 8 wk while partially grazing pasture. Cows in group 1 were fed a grain mix diet (8.0 kg/d, DM basis) containing 400 g of saturated animal fat (control). Cows in the second group were fed the same grain mix diet except the saturated animal fat was replaced with 100 g of FO and 300 g of SFO. Cows were milked twice a day and milk samples were collected weekly throughout the trial. Both groups grazed together on alfalfa-based pasture ad libitum and were fed their treatment diets after the morning and afternoon milking. Milk production (30.0 and 31.2 kg/d), milk fat percentages (3.64 and 3.50), milk fat yield (1.08 and 1.09 kg/d), milk protein percentages (2.97 and 2.88), and milk protein yield (0.99 and 0.91 kg/d) for diets 1 and 2, respectively, were not affected by the treatment diets. The concentrations of cis-9, trans-11 CLA (1.64 vs. 0.84 g/100 g of fatty acids) and vaccenic acid (5.11 vs. 2.20 g/100 g of fatty acids) in milk fat were higher for cows fed the oil-supplemented diet over the 8 wk of oil supplementation. The concentration of cis-9, trans-11 CLA in milk fat reached a maximum (1.0 and 1.64 g/100 g of fatty acids for diets 1 and 2, respectively) in wk 1 for both diets and remained relatively constant thereafter. The concentration of vaccenic acid in milk fat followed the same temporal pattern as cis-9, trans-11 CLA. In conclusion, supplementing the diet of partially grazing cows with FO and SFO increased the milk cis-9, trans-11 CLA content, and that increase remained relatively constant after 1 wk of oil supplementation.     

Examination of the persistency of milk fatty acid composition responses to fish oil and sunflower oil in the diet of dairy cows

Based on the potential benefits of cis-9, trans-11 conjugated linoleic acid (CLA) for human health, there is a need to develop effective strategies for enhancing milk fat CLA concentrations. Levels of cis-9, trans-11 CLA in milk can be increased by supplements of fish oil (FO) and sunflower oil (SO), but there is considerable variation in the response. Part of this variance may reflect time-dependent ruminal adaptations to high levels of lipid in the diet, which lead to alterations in the formation of specific biohydrogenation intermediates. To test this hypothesis, 16 late lactation Holstein-British Friesian cows were used in a repeated measures randomized block design to examine milk fatty acid composition responses to FO and SO in the diet over a 28-d period. Cows were allocated at random to corn silage-based rations (8 per treatment) containing 0 (control) or 45 g of oil supplement/kg of dry matter consisting (1:2; wt/wt) of FO and SO (FSO), and milk composition was determined on alternate days from d 1. Compared with the control, the FSO diet decreased mean dry matter intake (21.1 vs. 17.9 kg/d), milk fat (47.7 vs. 32.6 g/kg), and protein content (36.1 vs. 33.3 g/kg), but had no effect on milk yield (27.1 vs. 26.4 kg/d). Reductions in milk fat content relative to the FSO diet were associated with increases in milk trans-10 18:1, trans-10, cis-12 CLA, and trans-9, cis-11 CLA concentrations (r² = 0.74, 0.57, and 0.80, respectively). Compared with the control, the FSO diet reduced milk 4:0 to 18:0 and cis 18:1 content and increased trans 18:1, trans 18:2, cis-9, trans-11 CLA, 20:5 n-3, and 22:6 n-3 concentrations. The FSO diet caused a rapid elevation in milk cis-9, trans-11 CLA content, reaching a maximum of 5.37 g/100 g of fatty acids on d 5, but these increases were transient, declining to 2.35 g/100 g of fatty acids by d 15. They remained relatively constant thereafter. Even though concentrations of trans-11 18:1 followed the same pattern of temporal changes as cis-9, trans-11 CLA, the total trans 18:1 content of FSO milk was unchanged because of the concomitant increases in the concentration of other isomers (Δ^4-10 and Δ^12-15), predominantely trans-10 18:1. In conclusion, supplementing diets with FSO enhances milk fat cis-9, trans-11 CLA content, but the high level of enrichment declines because of changes in ruminal biohydrogenation that result in trans-10 replacing trans-11 as the major 18:1 biohydrogenation intermediate formed in the rumen.     

Cheese manufacture with milk with elevated conjugated linoleic acid levels caused by dietary manipulation

The objective of this study was to assess the effect of dietary supplementation of cows on pasture with sunflower oil for conjugated linoleic acid (cis-9, trans-11 CLA) enrichment of milk, for the production of CLA-enriched cheese. A group of 40 autumn-calving dairy cows were assigned to either a control group (indoor feeding on grass silage ad libitum and 6 kg/d of a typical indoor concentrate) or an experimental group (on pasture, being fed 6 kg of a supplement containing 100 g/kg of sunflower oil per d). These diets were fed for 16 d, during which time milk was collected for pilot-scale hard cheese manufacture. The pasture-based diet with sunflower oil resulted in a significant effect on the milk fatty acid CLA content. The concentration of cis-9, trans-11 CLA in the milk produced from cows on this diet increased to 2.22 g/100 g of fatty acid methyl esters (FAME) after 14 d, compared with 0.46 g/100 g of FAME in milk produced on the control indoor diet. The content of cis-9, trans-11 CLA in the cheese manufactured from the indoor control milk was 0.78 g/100 g of FAME and that from the pasture-based sunflower oil milk was 1.93 g/100 g of FAME. The cheese was assessed during the ripening period and CLA concentrations were stable throughout the 6 mo of ripening. Other cheese variables (microbiology, composition, flavor, free AA) were monitored during the ripening period, and the cheese with the elevated CLA concentrations compared favorably with the control cheese. Thus, a pasture-based diet supplemented with an oil source rich in linoleic acid resulted in an enhanced CLA content of bovine milk fat, compared with an indoor grass silage-based diet.     

Milk and cheese from cows fed calcium salts of palm and fish oil alone or in combination with soybean products

Twenty cows were used in a randomized block design experiment for 6 wk to determine the influence of feeding partial ruminally inert Ca salts of palm and fish oil (Ca-PFO), alone or in combination with extruded full-fat soybeans or soybean oil, on milk fatty acid (FA) methyl esters composition and consumer acceptability of milk and Cheddar cheese. Cows were fed either a diet containing 44% forage and 56% concentrate (control) or a diet supplemented with 2.7% Ca-PFO (FO), 5% extruded full-fat soybeans + 2.7% Ca-PFO (FOESM), or 0.75% soybean oil + 2.7% Ca-PFO (FOSO). Total dietary FA content in the control, FO, FOESM, and FOSO diets were 4.61, 6.28, 6.77, and 6.62 g/100 g, respectively. There was no difference in nutrient intake, milk yield, or milk composition among treatments. Conjugated linoleic acid (CLA) C_18:2cis-9, trans-11 isomer, C_18:1trans-11 (VA), and total n-3 FA in milk from cows on the control, FO, FOESM, and FOSO treatments were 0.56, 1.20, 1.36, and 1.74; 3.29, 4.66, 6.34, and 7.81; 0.62, 0.69, 0.69, and 0.67 g/100 g of FA, respectively. Concentrations of CLA, VA, and total n-3 FA in cheese were similar to milk. A trained sensory panel detected no difference in flavors of milk and cheese, except for acid flavor below a slightly perceptible level in cheese from all treatments. Results suggest that feeding Ca-PFO alone or in combination with extruded full-fat soybeans or soybean oil enhanced the CLA, VA, total unsaturated and n-3 FA in milk and cheese without negatively affecting cow performance and consumer acceptability characteristics of milk and cheese.     

Effect of dietary antioxidant and increasing corn oil inclusion on milk fat yield and fatty acid composition in dairy cattle

The objective of this study was to examine the effect of a dietary synthetic antioxidant on feed intake, yields of milk and milk components and milk fatty acids (FA), in combination with increasing concentrations of dietary corn oil to provide increasing rumen unsaturated fatty acid load (RUFAL) challenges. Twenty-six Holstein cows (177 ± 57 d in milk; mean ± standard deviation) were assigned to treatment in a randomized complete block design. Treatments were a control diet (CON; n = 13 cows) or the same diet supplemented with a synthetic antioxidant (AOX; 6.1 g/d; dry blend of ethoxyquin and propyl gallate, Novus International Inc., St. Charles, MO; n = 13 cows). In period 1 (21 d), no supplemental corn oil was fed; in periods 2, 3, and 4 (14 d each), corn oil was supplemented at 0.7, 1.4, and 2.8% of the diet [dry matter (DM) basis] to incrementally increase RUFAL. For all variables measured, no significant interactions were detected between treatment and period, indicating no differences between the CON and AOX treatments at all levels of oil inclusion. Intake of DM was lower for AOX compared with CON but AOX had no effect on milk yield or milk fat concentration and yield. Milk protein yield and feed efficiency (energy-corrected milk/DM intake) tended to be greater for AOX compared with CON. Increasing dietary corn oil concentration (RUFAL) decreased DM intake, milk yield, milk fat concentration and yield, and feed efficiency. The AOX treatment increased the concentration and yield of 16-carbon milk FA, with no effect on de novo (<16 carbon) or preformed (>16 carbon) milk FA. Milk FA concentration of trans-10 C18:1, trans-10,cis-12 C18:2, and trans-9,cis-11 C18:2 were unaffected by AOX but increased with increasing RUFAL. In conclusion, supplementation with AOX did not overcome the dietary-induced milk fat depression caused by increased RUFAL.     

Effectiveness of oils rich in linoleic and linolenic acids to enhance conjugated linoleic acid in milk from dairy cows

Forty Holstein dairy cows were used to determine the effectiveness of linoleic or linolenic-rich oils to enhance C_18:2cis-9, trans-11 conjugated linoleic acid (CLA) and C_18:1trans-11 (vaccenic acid; VA) in milk. The experimental design was a complete randomized design for 9 wk with measurements made during the last 6 wk. Cows were fed a basal diet containing 59% forage (control) or a basal diet supplemented with either 4% soybean oil (SO), 4% flaxseed oil (FO), or 2% soybean oil plus 2% flaxseed oil (SFO) on a dry matter basis. Total fatty acids in the diet were 3.27, 7.47, 7.61, and 7.50 g/100 g in control, SO, FO, and SFO diets, respectively. Feed intake, energy-corrected milk (ECM) yield, and ECM produced/kg of feed intake were similar among treatments. The proportions of VA were increased by 318, 105, and 206% in milk fat from cows in the SO, FO, and SFO groups compared with cows in the control group. Similar increases in C_18:2cis-9, trans-11 CLA were 273, 150, and 183% in SO, FO, and SFO treatments, respectively. Under similar feeding conditions, oils rich in linoleic acid (soybean oil) were more effective in enhancing VA and C_18:2cis-9, trans-11 CLA in milk fat than oils containing linolenic acid (flaxseed oil) in dairy cows fed high-forage diets (59% forage). The effects of mixing linoleic and linolenic acids (50:50) on enhancing VA and C_18:2cis-9, trans-11 CLA were additive, but not greater than when fed separately. Increasing the proportion of healthy fatty acids (VA and CLA) by feeding soybean or flaxseed oil would result in milk with higher nutritive and therapeutic value.     

Changes in milk fatty acid profile and animal performance in response to fish oil supplementation, alone or in combination with sunflower oil, in dairy 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.     

Effect of fish oil and sunflower oil on rumen fermentation characteristics and fatty acid composition of digesta in ewes fed a high concentrate diet

Studies in ruminants have shown that supplementing the diet with a mixture of fish oil (FO) and sunflower oil (SO) enhances the concentration of cis-9, trans-11 conjugated linoleic acid (CLA), 20:5 n-3, and 22:6 n-3 in milk because of alterations in ruminal biohydrogenation, but the intermediates formed under these conditions are not well characterized. Five ewes fitted with rumen cannula and fed a high concentrate diet were used to examine the effect of a mixture (30 g/kg of DM) of FO and SO (1:2, wt/wt) on temporal changes in rumen fermentation characteristics and the relative abundance of biohydrogenation intermediates in ruminal digesta collected after 0, 3, and 10 d on diet. Appearance and identification of biohydrogenation intermediates was determined based on complementary gas-liquid chromatography and Ag+-HPLC analysis of fatty acid methyl esters and gas chromatography-mass spectrometry analysis of corresponding 4,4-dimethyloxazoline derivatives. Inclusion of FO and SO in the diet had no effect on rumen pH, volatile fatty acid concentrations, or nutrient digestion, but altered the fatty acid composition of ruminal digesta, changes that were characterized by time-dependent decreases in 18:0 and 18:2 n-6 and the accumulation of trans 16:1, trans 18:1, 10-O-18:0, and trans 18:2. Lipid supplements enhanced the proportion of 20:5 n-3 and 22:6 n-3 in digesta and resulted in numerical increases in cis-9, trans-11 conjugated linoleic acid concentrations, but decreased the relative abundance of trans-10, cis-12 conjugated linoleic acid. Furthermore, detailed analysis revealed the appearance of several unique 20:1, 20:2, 22:1, 22:3, and 22:4 products in ruminal digesta that accumulated over time, providing the first indications of 20 and 22 carbon fatty acid intermediates formed during the biohydrogenation of long-chain unsaturated fatty acids in sheep. In conclusion, FO and SO in a high concentrate diet caused a time-dependent inhibition of the complete biohydrogenation of 16 and 18 carbon unsaturated fatty acids, and resulted in the accumulation of trans 16:1, trans 18:1, and trans 18:2, 20, and 22 carbon metabolites in ruminal digesta of sheep, with no evidence of a shift in ruminal biohydrogenation pathways toward trans-10 18:1 formation.     

Formation of trans fatty acids in edible oils during the frying and heating process

To assess an impact of heated edible oils on intake of trans fat, the formations of trans fatty acids (TFAs) in cooking conditions was estimated by a frying and heating model system. For the frying model, sliced raw potatoes (10% of the frying oil (w/w)) were fried in commercially available canola oil at 160, 180 and 200 °C, and the 10 frying cycles were performed. The TFAs contained both in fried potatoes and in frying oils were measured by gas chromatography (GC). Lipids content of raw potatoes was about 0.1% (w/w) and TFAs in the raw potatoes were negligible. On the other hand, fried potatoes contained lipids at the level of 8.8%–9.2% and their fatty acid composition was mostly in correspondence with that of the frying oil. The TFAs amount of potatoes fried by the tenth frying operation was at the level of 0.99–1.05 g/100 g lipids. When 100 g potatoes fried in this process were consumed, the TFAs intake was estimated at less than 0.1 g. After 10 frying operations, TFAs content, acid values and peroxide values of the frying oils were measured and compared with those of corresponding heated canola oils without food. The amounts of trans 18:1 FAs contained both in the frying oil and in heated oil were less than the quantitative limit (0.047 g/100 g oil). The increases of trans 18:2 FAs and trans 18:3 FAs of the used frying oil were 0.02 g/100 and 0.05 g/100 g, respectively, compared with those of the fresh oil. trans 18:2 FAs accumulation in the heated oil was slightly less than that in the frying oil. To elucidate TFAs accumulation in various edible oils during cooking, six kinds of commercially available edible vegetable oils were heated to 180 °C in glass test tubes. Small changes in TFAs amounts were observed after four hours heating. These results suggested that an ordinary frying process using unhydrogenated edible oils has little impact on TFAs intake from edible oils.     

Effects of dietary sources of vegetable oils on performance of high-yielding lactating cows and conjugated linoleic acids in milk

This study was conducted to examine the effects of dietary supplementation with vegetable oils on performance of high-yielding lactating cows and conjugated linoleic acid (CLA) content in milk fat. Twelve lactating Holstein cows in early lactation (30 to 45 d postpartum) were used in a triple 4 × 4 Latin square design. In each period, the cows in each group were fed the same basal diet and received one of the following treatments: 1) control (without oil), 2) 500 g of cottonseed oil, 3) 500 g of soybean oil, and 4) 500 g of corn oil. Each experimental period lasted for 3 wk, with the first 2 wk used for adaptation to the diet. Supplementation with vegetable oils tended to increase milk yield, with the highest milk yield in the cottonseed oil group (35.0 kg/d), compared with the control (34.4 kg/d). Milk fat percentage was decreased, but there were few effects on percentage and yield of milk protein as well as milk fat yield. The cows fed added soybean oil produced milk with the highest content of trans-11 C_18:1 (23.8 mg/g of fat), which was twice that of the control (12.6 mg/g of fat). Content of cis-9, trans-11 CLA in milk fat increased from 3.5 mg/g in the control to 6.0, 7.1, and 10.3 mg/g for the cows fed oils from cottonseed, corn, and soybean, respectively. A significant linear relationship existed between trans-11 C_18:1 and cis-9, trans-11 CLA. Supplementation with oils doubled the content of total fatty acids in blood plasma, with little difference between different vegetable oil sources. Octadecenoic acid content was significantly higher in blood plasma of animals fed added oils from cottonseed and soybean than those fed with corn oil and control. The plasma trans-11 C_18:1 content was significantly higher in the oil-added animals than in control. Supplementation of vegetable oils tended to improve milk production of lactating cows, and the CLA content in milk fat was significantly increased. Soybean oil seemed to be the optimal source to increase CLA production.     

Milk composition, milk fatty acid profile, digestion, and ruminal fermentation in dairy cows fed whole flaxseed and calcium salts of flaxseed oil

Four ruminally lactating Holstein cows averaging 602 ± 25 kg of body weight and 64 ± 6 d in milk at the beginning of the experiment were randomly assigned to a 4 × 4 Latin square design to determine the effects of feeding whole flaxseed and calcium salts of flaxseed oil on dry matter intake, digestibility, ruminal fermentation, milk production and composition, and milk fatty acid profile. The treatments were a control with no flaxseed products (CON) or a diet (on a dry matter basis) of 4.2% whole flaxseed (FLA), 1.9% calcium salts of flaxseed oil (SAL), or 2.3% whole flaxseed and 0.8% calcium salts of flaxseed oil (MIX). The 4 isonitrogenous and isoenergetic diets were fed for ad libitum intake. Experimental periods consisted of 21 d of diet adaptation and 7 d of data collection and sampling. Dry matter intake, digestibility, milk production, and milk concentrations of protein, lactose, urea N, and total solids did not differ among treatments. Ruminal pH was reduced for cows fed the CON diet compared with those fed the SAL diet. Propionate proportion was higher in ruminal fluid of cows fed CON than in that of those fed SAL, and cows fed the SAL and CON diets had ruminal propionate concentrations similar to those of cows fed the FLA and MIX diets. Butyrate concentration was numerically higher for cows fed the SAL diet compared with those fed the FLA diet. Milk fat concentration was lower for cows fed SAL than for those fed CON, and there was no difference between cows fed CON and those fed FLA and MIX. Milk yields of protein, fat, lactose, and total solids were similar among treatments. Concentrations of cis-9 18:1 and of intermediates of ruminal biohydrogenation of fatty acids such as trans-9 18:1 were higher in milk fat of cows fed SAL and MIX than for those fed the CON diet. Concentration of rumenic acid (cis-9, trans-11 18:2) in milk fat was increased by 63% when feeding SAL compared with FLA. Concentration of α-linolenic acid was higher in milk fat of cows fed SAL and MIX than in milk of cows fed CON (75 and 61%, respectively), whereas there was no difference between FLA and CON. Flaxseed products (FLA, SAL, and MIX diets) decreased the n-6 to n-3 fatty acid ratio in milk fat. Results confirm that flax products supplying 0.7 to 1.4% supplemental fat in the diet can slightly improve the nutritive value of milk fat for better human health.     

Rumen biohydrogenation-derived fatty acids in milk fat from grazing dairy cows supplemented with rapeseed, sunflower, or linseed oils

The effects of supplementation with rapeseed, sunflower, and linseed oils (0.5 kg/d; good sources of oleic, linoleic, and linolenic acids, respectively) on milk responses and milk fat fatty acid (FA) profile, with special emphasis on rumen-derived biohydrogenation intermediates (BI), were evaluated in a replicated 4 × 4 Latin square study using 16 grazing dairy cows. The dietary treatments were 1) control diet: 20-h access to grazing pasture supplemented with 5 kg/d of corn-based concentrate mixture (96% corn; CC); 2) RO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of rapeseed oil; 3) SO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of sunflower oil; and 4) LO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of linseed oil. Milk fatty acids were converted to methyl esters and analyzed by gas-liquid chromatography and silver-ion HPLC. Dietary treatments had no effect on milk production or on milk protein content and milk protein production. Supplementation with rapeseed and sunflower oils lowered milk fat content and milk fat production, but linseed oil had no effect. Inclusion of dietary vegetable oils promoted lower concentrations of short-chain (including 4:0) and medium-chain FA (including odd- and branched-chain FA) and 18:3n-3, and higher concentrations of C₁₈ FA (including stearic and oleic acids). The BI concentration was higher with the dietary inclusion of vegetable oils, although the magnitude of the concentration and its pattern differed between oils. The RO treatment resulted in moderate increases in BI, including trans 18:1 isomers and 18:2 trans-7,cis-9, but failed to increase 18:1 trans-11 and 18:2 cis-9,trans-11. Sunflower oil supplementation resulted in the highest concentrations of the 18:1 trans-10, 18:1 cis-12, and 18:2 trans-10,trans-12 isomers. Concentrations of 18:1 trans-11 and 18:2 cis-9,trans-11 were higher than with the control and RO treatments but were similar to the LO treatment. Concentration of BI in milk fat was maximal with LO, having the highest concentrations of some 18:1 isomers (i.e., trans-13/14, trans-15, cis-15, cis-16), most of the nonconjugated 18:2 isomers (i.e., trans-11,trans-15, trans-11,cis-15, cis-9,cis-15, and cis-12,cis-15), and conjugated 18:2 isomers (i.e., trans-11,cis-13, cis-12,trans-14, trans-11,trans-13, trans-12,trans-14, and trans-9,trans-11), and all conjugated 18:3 isomers. The LO treatment induced the highest amount and diversity of BI without decreasing milk fat concentration, as the RO and SO treatments had, suggesting that the BI associated with 18:3n-3 intake may not be the major contributors to inhibition of mammary milk fat synthesis.