The effect of breed,parity, and stage of lactation on conjugated linoleic acid (CLA) in milk fat from dairy cows

Dairy products are the main source of conjugated linoleic acid (CLA), a functional food component with health benefits. The major source of cis-9, trans-11 CLA in milk fat is endogenous synthesis via delta9-desaturase from trans-11 18:1, with the remainder from incomplete rumen biohydrogenation of linoleic acid. Diet has a major influence on milk fat CLA; however, effects of physiological factors have received little attention. Our objectives were to examine milk fat content of CLA and the CLA-desaturase index with regard to: 1) effect of breed, parity, and stage of lactation, and 2) variation among individuals and the relationship to milk and milk fat. Holstein (n = 113) and Brown Swiss (n = 106) cows were fed a single diet and milk sampled on the same day to avoid confounding effects of diet and season. Frequency distributions demonstrated that milk fat content of CLA and CLA-desaturase index varied over threefold among individuals, and this needs to be considered in the design of experiments. Holsteins had a higher milk fat content of CLA and CLA-desaturase index, but breed differences were minor. Parity and days in milk also had little or no relationship to the individual variation for these two CLA variables. Breed, parity, and days in milk accounted for < 0.1, < 0.3, and < 2.0% of total variation in CLA concentration in milk fat, respectively. Milk fat content of CLA and CLA-desaturase index were essentially independent of milk yield, milk fat percent, and milk fat yield. We speculate that the basis for the genetic variation among individuals is related to rumen output of trans-11 18:1 and to a lesser extent cis-9, trans-11 CLA, and to the tissue amount and activity of delta9-desaturase.     

Endogenous synthesis of cis-9, trans-11 conjugated linoleic acid in dairy cows fed fresh pasture

New Zealand Holstein-Friesian cows (n = 4) were used to quantify the importance of endogenous synthesis of cis-9, trans-11 conjugated linoleic acid (CLA) via Delta(9)-desaturase in cows fed a fresh pasture diet. The experiment was a 4 x 4 Latin square design with treatments arranged in a 2 x 2 factorial. Treatments lasted 4 d and were pasture only, pasture plus sterculic oil, pasture plus sunflower oil, and pasture plus sunflower oil plus sterculic oil. Abomasal infusion of sterculic oil inhibited Delta(9)-desaturase and decreased the concentration of cis-9, trans-11 CLA in milk fat by 70%. Using the changes in cis-9 10:1, cis-9 12:1 and cis-9 14:1 to correct for incomplete inhibition of Delta(9)-desaturase, a minimum estimate of 91% of cis-9, trans-11 CLA in milk fat was produced endogenously in cows fed fresh pasture. Dietary supplementation of a pasture diet with sunflower oil increased the proportion of long chain fatty acids in milk fat; however, the increase in vaccenic acid concentration was small (18%) and there was no increase in cis-9, trans-11 CLA concentration. Overall, results show that endogenous synthesis is responsible for more than 91% of the cis-9, trans-11 CLA secreted in milk fat of cows fed fresh pasture. However, the failure of plant oil supplements to increase the concentration of cis-9, trans-11 CLA in milk fat from pasture-fed cows requires further investigation.     

A comparison between feeding systems (pasture and TMR) and the effect of vitamin E supplementation on plasma and milk fatty acid profiles in dairy cows

Unidentified constituents in fresh pasture increase milk fat cis-9, trans-11 conjugated linoleic acid (CLA) concentration, and prevent milk fat depression, even though ruminal conditions conducive to reducing milk fat synthesis exist. One possible explanation is vitamin E (kappa-tocopherol), a constituent high in fresh pasture, but naturally low in conserved/dried forages and cereal grains. Twenty late-lactating dairy cows previously consuming a total mixed ration (TMR) were randomly allocated to one of two dietary treatments for 21 d: TMR (control; n=10); and TMR plus an additional 10,000 i.u. alpha-tocopherol/d (VIT E; n = 10). These cows were simultaneously compared with 13 late-lactation dairy cows previously grazing fresh pasture (PAS) balanced for age, parity and genetic merit. Average daily alpha-tocopherol intakes were approximately 468, 10,520 and 1,590 i.u./cow for the control, VIT E and PAS treatments, respectively. Dietary alpha-tocopherol supplementation (VIT E v. control) slightly increased milk fat content by 0.23 percentage units, but did not significantly alter milk fatty acid composition. Plasma trans-11 18:1 (VA) content tended to increase and trans-10 18:1 levels numerically declined following alpha-tocopherol supplementation suggesting possible changes in rumen biohydrogenation products. In addition, increased alpha-tocopherol intake in TMR-fed cows decreased serum urea levels and tended to alter milk fat 15:0 suggesting changes in rumen microbial populations. However, when compared with cows grazing pasture, TMR-fed cows supplemented with alpha-tocopherol, still produced milk with lower cis-9, trans-11 CLA and VA, and higher trans-10 18:1 concentrations suggesting alpha-tocopherol is not a primary reason for milk fatty acid profile differences between pasture and TMR-fed cows. Therefore, additional unknown pasture constituents favour production of fatty acids originating from the cis-9, trans-11 instead of the trans-10, cis-12 CLA biohydrogenation pathways.     

trans-10, cis-12 conjugated linoleic acid decreases lipogenic rates and expression of genes involved in milk lipid synthesis in dairy cows

Feeding conjugated linoleic acid (CLA) reduces milk fat synthesis in lactating dairy cows, and the effect has been shown to be specific for the trans-10, cis-12 CLA isomer. Our objectives were to examine potential mechanisms by which trans-10, cis-12 CLA inhibits milk fat synthesis. Multiparous Holstein cows (n = 4) in late lactation were used in a balanced 2 x 2 crossover design. Treatments consisted of a 5 d abomasal infusion of either skim milk (control) or purified trans-10, cis-12 CLA (13.6 g/d) emulsified in skim milk. On d 5 of infusion, mammary gland biopsies were performed and a portion of the tissue analyzed for mRNA expression of acetyl CoA carboxylase, fatty acid synthetase, delta 9-desaturase, lipoprotein lipase, fatty acid binding protein, glycerol phosphate acyltransferase and acylglycerol phosphate acyltransferase. Lipogenic capacity was evaluated with another portion of the tissue. Infusion of trans-10, cis-12 CLA decreased milk fat content and yield 42 and 48%, respectively and increased the trans-10, cis-12 CLA content in milk fat from < 0.1 to 4.9 mg/g. Reductions in milk fat content of C4 to C16 fatty acids contributed 63% to the total decrease in milk fat yield (molar basis). Analysis of the ratios of specific fatty acid pairs indicated trans-10, cis-12 CLA also shifted fatty acid composition in a manner consistent with a reduction in delta 9-desaturase. Mammary explant incubations with radiolabeled acetate established that lipogenic capacity was decreased 82% and acetate oxidation to CO2 was reduced 61% when cows received trans-10, cis-12 CLA. Infusing trans-10, cis-12 CLA also decreased the mRNA expression of all measured enzymes by 39 to 54%. Overall, data demonstrated the mechanism by which trans-10, cis-12 CLA inhibits milk fat synthesis includes decreasing expression of genes that encode for enzyme involved in circulating fatty acid uptake and transport, de novo fatty acid synthesis, desaturation of fatty acids and triglyceride synthesis.     

Use of principal component analysis to investigate the origin of heptadecenoic and conjugated linoleic acids in milk

The aim of this paper was the application of principal component analysis (PCA) 1) to elucidate mutual metabolic relationships between milk fatty acids (FA) and 2) to illustrate the origin of milk FA, in particular C17:1 and cis-9,trans-11 conjugated linoleic acid. Data were combined from 3 experiments with lactating Holstein-Friesian cows offered diets based on grass or legume silage and concentrates. Loading plots of PCA based on milk FA concentrations showed 4 groups of milk FA, having similar precursors or metabolic pathways in the rumen and/or mammary gland: medium-chain saturated FA, de novo synthesized from acetate and beta-hydroxybutyrate; monoenoic milk FA, products of delta9-desaturase activity in the mammary gland; odd chain FA of rumen microbial origin and C18:0, n-6 C18:2, and n-3 C18:3 of dietary origin or the result of rumen biohydrogenation. Loading plots of PCA based on both milk and duodenal FA concentrations as well as on milk FA yields and duodenal FA flows further illustrated the importance of postabsorptive synthesis of the milk medium chain saturated and monoenoic FA and the direct absorption from the blood stream of odd chain FA, C18:0, n-6 C18:2, and n-3 C18:3. In all loading plots, milk oleic acid (C18:1) appeared intermediate between clusters of 18-carbon FA and monoenoic FA, illustrating its dual (dietary and endogenous production) origin. Milk C17:1 was suggested to be a desaturation product of C17:0, in common with other milk monoenoic FA. Finally, the PCA technique, based on milk FA patterns of one experiment, was applied to investigate factors determining cis-9,trans-11 conjugated linoleic acid concentrations in milk. Within the range of diets and cows studied here, we showed changes in cis-9,trans-11 conjugated linoleic acid to be mainly dependent on vaccenic acid supply and to a lesser extent on variation in desaturase activity.     

Conjugated linoleic acid increases in milk when cows fed fish meal and extruded soybeans for an extended period of time

The objective of this study was to determine the effect of feeding a conjugated linoleic acid (CLA) stimulating diet for an extended period of time on milk cis-9, trans-11 CLA and vaccenic acid (VA) concentrations. Twenty cows (16 Holstein and 4 Brown Swiss) were divided into 2 groups (n = 10 per treatment) for a 10-wk study. Cows in group 1 were fed a traditional corn-soybean-basal diet (control), while those in group 2 were fed a blend of 0.5% fish oil from fish meal and 2% soybean oil from extruded soybeans (FMESB) to achieve higher milk fat cis-9, trans-11 CLA and VA. Diets were formulated to contain 18% CP and were composed (dry matter basis) of 50% concentrate mix, 25% corn silage, and 25% alfalfa hay. Dry matter intake was not affected by diet. Milk production increased in cows fed the FMESB diet. Milk fat and milk protein percentages decreased with the FMESB diet; however, milk fat and protein yields were not affected by treatments. Milk fat cis-9, trans-11 CLA and VA concentration (g/100 of fatty acids) and yield (g/d) were 2.5-fold greater for cows fed the FMESB diet over the 10 wk of fat supplementation. For cows fed the FMESB diet, contents of milk fat cis-9, trans-11 CLA and VA gradually increased from the first week of fat supplementation, reached the highest concentrations in wk 3, then gradually decreased during wk 4 and 5 and then remained relatively constant until wk 10. The concentration of cis-9, trans-11 CLA and VA from the control diet was relatively constant over the 10 wk of fat supplementation. Concentrations of cis-9, trans-11 CLA and VA in milk fat can be increased within a week by feeding a blend of fish meal and extruded soybeans, and that increase remains relatively constant after wk 5 of fat supplementation.     

Effect of dose of calcium salts of conjugated linoleic acid (CLA) on percentage and fatty acid content of milk fat in midlactation holstein cows

Increasing conjugated linoleic acid (CLA) content of milk fat from lactating dairy cattle has become a research interest due to the possible health benefits afforded humans consuming CLA. Dietary supplementation of CLA to lactating dairy cows is one potential method by which CLA content of milk and dairy products may be enhanced. Feeding CLA in calcium salt form could potentially deliver CLA to the lower digestive tract through prevention of biohydrogenation by rumen microbes. Milk fat depression (MFD) occurs when cows receive CLA-60, a commercially available CLA source containing numerous CLA isomers, abomasally. Our objectives were to determine the quantity of CLA as calcium salts required to elicit maximal MFD and to evaluate the effects of CLA supplementation on fatty acid composition of milk fat. Five Holstein cows at approximately 93 DIM were utilized in a 5 x 5 balanced Latin square crossover design. Periods were 14-d in length with a 5-d treatment phase and 9-d rest phase. Treatments were 5-d supplementation of 0, 12.5, 25, 50, and 100 g of CLA-60 in calcium salt form. Milk samples were collected on d 5 of CLA supplementation and analyzed for composition and fatty acid profile. Regression analysis of milk fat data suggested that MFD was not maximized over the dose levels investigated, despite delivery of 34.5 g of trans-10, cis-12 CLA in the 100-g dose of CLA. Supplementation with 50 and 100 g of CLA per day resulted in a reduction of milk fat percent of 29 and 34%, respectively. Trend analysis indicated a linear decrease in the milk fat content of caprylic, capric, and lauric acids as the dose of CLA increased. Milk fat content of cis-9, trans-11, and trans-10, cis-12 CLA increased at an increasing rate as dose increased.     

Comparison of calcium salts and formaldehyde-protected conjugated linoleic Acid in inducing milk fat depression

Abomasal infusion studies have shown that trans-10, cis-12 conjugated linoleic acid (CLA) decreases milk fat synthesis. However, supplements of CLA must avoid rumen biohydrogenation for this technology to be applied to ruminants. Rumen protection methods would reduce CLA metabolism in the rumen and increase its supply to the small intestine. Our objective was to compare the efficacy of 2 forms of rumen-protected CLA at inducing milk fat depression. Three mid to late lactation Holstein cows each fitted with a rumen fistula were used in a 3 x 3 Latin square design. Treatments were: 1) control, 2) calcium salts of CLA (Ca-CLA), and 3) formaldehyde-protected CLA (FP-CLA). Supplements were designed to provide 10 g/d of trans-10, cis-12 CLA and were administered intraruminally once per day to ensure exact delivery of amount. Both CLA treatments substantially reduced milk fat yield and content compared with control, with the reductions in milk fat yield averaging 34% for the Ca-CLA treatment and 44% for the FP-CLA treatment. In contrast, milk yield, milk protein yield, and dry matter intake were unaltered by CLA treatment. Efficiency of transfer of trans-10, cis-12 CLA from the supplement into milk fat was 3.2 and 7.0% for Ca-CLA and FP-CLA, respectively. These values are much lower than transfer efficiencies reported for abomasally infused CLA, suggesting that much of the trans-10, cis-12 CLA present in the 2 formulations was biohydrogenated in the rumen. Overall, the extent of the reduction in milk fat yield indicates that both protection formulations are acceptable methods for the formulation of CLA supplements to induce milk fat depression in lactating dairy cows.     

An unprotected conjugated linoleic acid supplement decreases milk production and secretion of milk components in grazing dairy ewes

Feeding conjugated linoleic acid (CLA) in a rumen-inert form to dairy ewes has been shown to increase milk production, alter milk composition, and increase the milk fat CLA content. However, few studies have tested ruminally unprotected CLA sources. The objective of this study was to evaluate the effects of an unprotected CLA supplement (29.8% of cis-9,trans-11 and 29.9% of trans-10,cis-12 isomers as methyl esters) on milk yield and composition of dairy ewes. Twenty-four lactating Lacaune ewes were used in a crossover design and received 2 dietary treatments: (1) control: basal diet containing no supplemental lipid and (2) basal diet plus CLA (30 g/d). The CLA supplement was mixed into the concentrate and fed in 2 equal meals after morning and afternoon milkings. Each experimental period consisted of 21 d: 7 d for adaptation and 14 d for data collection. The CLA supplement decreased milk fat content and yield by 31.3 and 38.0%, respectively. Milk yield and secretion of milk lactose and protein were decreased by 8.0, 9.8, and 5.6%, respectively. On the other hand, milk protein content and linear SCC score were 1.8 and 17.7% higher in ewes fed the CLA supplement. The concentration of milk fatty acids originating from de novo synthesis (C16) was increased by 22.6% in ewes fed the CLA supplement. The CLA supplement decreased C14:1/C14:0, C16:1/C16:0, and C18:1/C18:0 desaturase indexes by 25, 18.7, and 0.1%, respectively, but increased the cis-9,trans-11 CLA/trans-11 C18:1 ratio by 8.6%. The concentrations of trans-10,cis-12 CLA and cis-9,trans-11 CLA in milk fat was 309 and 33.4% higher in ewes fed CLA. Pronounced milk fat depression coupled with the deleterious effects on milk yield, milk SCC, and secretion of all milk solids observed in ewes fed an unprotected CLA supplement is likely to be associated with high doses of trans-10,cis-12 CLA reaching the mammary gland, corroborating previous results obtained with dairy cows.     

Conjugated linoleic acid and vaccenic acid in rumen, plasma, and milk of cows fed fish oil and fats differing in saturation of 18 carbon fatty acids

The objective of this study was to examine the effect of feeding fish oil (FO) along with fat sources that varied in saturation of 18 carbon fatty acids (high stearic, high oleic, high linoleic, or high linolenic acids) on rumen, plasma, and milk fatty acid profiles. Four primiparous Holstein cows at 85 d in milk (+/- 40) were assigned to 4 x 4 Latin squares with 4-wk periods. Treatment diets were 1) 1% FO plus 2% commercial fat high in stearic acid (HS); 2) 1% FO plus 2% fat from high oleic acid sunflower seeds (HO); 3) 1% FO plus 2% fat from high linoleic acid sunflower seeds (HLO); and 4) 1% FO plus 2% fat from flax seeds (high linolenic; HLN). Diets were formulated to contain 18% crude protein and were composed of 50% (dry basis) concentrate mix, 25% corn silage, 12.5% alfalfa silage, and 12.5% alfalfa hay. Milk production, milk protein percentages and yields, and dry matter intake were similar across diets. Milk fat concentrations and yields were least for HO and HLO diets. The proportion of milk cis-9, trans-11 conjugated linoleic acid (CLA; 0.71, 0.99, 1.71, and 1.12 g/100 g fatty acids, respectively), and vaccenic acid (TVA; 1.85, 2.60, 4.14, and 2.16 g/100 g fatty acids, respectively) were greatest with the HLO diet. The proportions of ruminal cis-9, trans-11 CLA (0.09, 0.16, 0.18, and 0.16 g/100 g fatty acids, respectively) were similar for the HO, HLO, and HLN diets and all were higher than for the HS diet. The proportions of TVA (2.85, 4.36, 8.69, and 4.64 g/100 g fatty acids, respectively) increased with the HO, HLO, and HLN diets compared with the HS diets, and the increase was greatest with the HLO diet. The effects of fat supplements on ruminal TVA concentrations were also reflected in plasma triglycerides, (2.75, 4.64, 8.77, and 5.42 g/100 g fatty acids, respectively); however, there were no differences in the proportion of cis-9, trans-11 CLA (0.06, 0.07, 0.06, and 0.07 g/100 g fatty acids, respectively). This study further supports the significant role for mammary delta-9 desaturase in milk cis-9, trans-11 CLA production.     

Fatty acid profiles of milk and rumen digesta from cows fed fish oil,extruded soybeans or their blend

Four fistulated primiparous cows (two Holstein and two Brown Swiss) averaging 102 DIM were used in a 4 x 4 Latin square with 3-wk periods to determine the effect of feeding fish oil, extruded soybeans, or their combination on fatty acid profiles of milk and rumen digesta. Experimental diets consisted of: 1) control diet; 2) a diet with 2% (DM basis) added fat from menhaden fish oil; 3) a diet with 2% added fat from extruded soybeans; and 4) a diet with 1% added fat from fish oil and 1% fat from extruded soybeans. All diets consisted of 25% corn silage, 25% alfalfa hay, and 50% concentrate. Milk yields (28.6, 29.7, 29.2, and 28.1 kg/d for control, fish oil, extruded soybeans, and combination diets, respectively) were similar for all fat supplements and control. Milk fat and protein percentages (3.49, 3.08; 3.25, 2.96; 3.47, 3.01; 3.48, 2.99 for diets 1, 2, 3, and 4, respectively) were not affected by fat supplements compared with control. Dry matter intake (23.0, 21.6, 22.7, and 21.6 kg/d) was reduced when diets containing fish oil were fed. Concentrations of conjugated linoleic acid [CLA; cis-9, trans-11 CLA, 0.40, 0.88, 0.87, and 0.80 g/100 g fatty acids (FA)] and transvaccenic acid (TVA, 1.02, 2.34, 2.41, and 2.06 g/100 g of FA) were increased in milk fat by all fat supplements, with no differences in milk CLA and TVA observed among fat supplements. As with milk fat, proportions of ruminal CLA (0.09, 0.26, 0.18, and 0.21 g/100 g of FA) and TVA (2.61, 4.56, 4.61, and 4.39 g/100 g of FA) increased with fat supplements. The effects of fat supplements on ruminal TVA and CLA concentrations were also reflected in rumen FA-salts, free fatty acids, and neutral lipids. The higher TVA to CLA ratio in the rumen compared with milk indicated that fat supplements increased milk CLA concentration mainly by increasing ruminal production of TVA, which also implied the significant role that mammary delta-9 desaturase plays in milk CLA concentrations.     

Milk conjugated linoleic acid response to fish oil supplementation of diets differing in fatty acid profiles

The objective of this experiment was to examine the effect of feeding fish oil (FO) along with fat sources that varied in their fatty acid compositions (high stearic, high oleic, high linoleic, or high linolenic acids) to determine which combination would lead to maximum conjugated linoleic acid (cis-9,trans-11 CLA) and transvaccenic acid (TVA) concentrations in milk fat. Twelve Holstein cows (eight multiparous and four primiparous cows) at 73 (+/- 32) DIM were used in a 4 x 4 Latin square with 4-wk periods. Treatment diets were 1) 1% FO plus 2% fat source high in stearic acid (HS), 2) 1% FO plus 2% fat from high oleic acid sunflower seeds (HO), 3) 1% FO plus 2% fat from high linoleic acid sunflower seeds (HLO), and 4) 1% FO plus 2% fat from flax seeds (high linolenic; HLN). Diets formulated to contain 18% crude protein were composed of 50% (dry basis) concentrate mix, 25% corn silage, 12.5% alfalfa haylage, and 12.5% alfalfa hay. Milk production (35.8, 36.3, 34.9, and 35.0 kg/d for diets 1 to 4) was similar for all diets. Milk fat percentages (3.14, 2.81, 2.66, and 3.08) and yields (1.13, 1.02, 0.93, and 1.08 kg/d) for diets 1 to 4 were lowest for HLO. Milk protein percentages (3.04, 3.03, 3.10, and 3.08) and dry matter intake (DMI) (25.8, 26.0, 26.2, and 26.2 kg/d) for diets 1 to 4 were similar for all diets. Milk cis-9,trans-11 CLA concentrations (0.70, 1.04, 1.70, and 1.06 g/100 g fatty acids) for diet 1 to 4 and yields (7.7, 10.7, 15.8, and 11.3 g/d) for diets 1 to 4 were greatest with HLO and were least with HS. Milk cis-9,trans-11 CLA concentrations and yields were similar for cows fed the HO and the HLN diets. Similar to milk cis-9,trans-11 CLA, milk TVA concentration (1.64, 2.49, 3.74, and 2.41 g/100 g fatty acids) for diets 1 to 4 was greatest with the HLO diet and least with the HS diet. Feeding a high linoleic acid fat source with fish oil most effectively increased concentrations and yields of milk cis-9,trans-11 CLA and TVA.     

Feeding fish meal and extruded soybeans enhances the conjugated linoleic acid (CLA) content of milk

Twelve multiparous Holstein cows averaging 65 (33 to 122) DIM were used in a 4 x 4 Latin square for 4-wk periods to determine whether feeding fish oil as fish meal would stimulate increased amounts of milk conjugated linoleic acid (cis-9, trans-11 C18:2; CLA) and transvaccenic acid (trans-11 C18:1; TVA) when the cows were fed extruded soybeans to supply additional linoleic acid. Treatment diets were 1) control; 2) 0.5% fish oil from fish meal; 3) 2.5% soybean oil from extruded soybeans; and 4) 0.5% fish oil from fish meal and 2% soybean oil from extruded soybeans. Diets were formulated to contain 18% crude protein and were composed (dry basis) of 50% concentrate mix, 25% corn silage, and 25% alfalfa hay. Intake of DM was not affected by diet. Milk production was increased by diets 2, 3, and 4 compared with diet 1 (control). Milk fat and milk protein percentages decreased with diets 3 and 4. Milk fat yield was not affected by treatments, but yield of milk protein was increased with supplemental fish meal and extruded soybeans or their blend. When diets 2, 3, or 4 were fed, concentrations of cis-9, trans-11 CLA in milk fat increased by 0.4-, 1.4-, and 3.2-fold, and TVA concentrations in milk fat increased by 0.4-, 1.8-, and 3.5-fold compared with the control milk fat. Increases in TVA and cis-9, trans-11 CLA were 91 to 109% greater when a blend of fish meal and extruded soybeans was fed than the additive effect of fish meal and extruded soybeans. This suggested that fish oil increased the production of CLA and TVA from other dietary sources of linoleic acid such as extruded soybeans.     

Technical note: production of butter with enhanced conjugated linoleic acid for use in biomedical studies with animal models

Cancer models utilize massive doses of carcinogen so that investigations of anticarcinogenic effects require equally large doses. Conjugated linoleic acids (CLA), predominately consumed in dairy products, are thought to be anticarcinogenic. Our objective was to naturally produce a CLA-enhanced butter for use in biomedical studies with animal models. To do this, we fed cows a low forage diet supplemented with sun-flower oil. This resulted in increases in content of CLA of milk fat, but the markedly elevated concentrations were transient and declined over a 3-wk period. By collecting milk fat over the first few days on the diet (d 7 to 10) and selecting cows with the greatest CLA concentrations, we were able to produce a butter in which CLA content was enhanced sevenfold over control butter (41 vs. 5 mg/g of fatty acids) and the cis-9, trans-11 isomer predominated (91%). Thus, butter produced by this method can be used to investigate the preventive role of CLA in natural foods with biomedical models of different types of cancer. Furthermore, the butter allows examination of the other beneficial health effects of CLA reported with animal models.     

Effects of week of lactation and genetic selection for milk yield on milk fatty acid composition in Holstein cows

Control (CL) and select line (SL) dairy cows (n = 22) managed identically but differing in milk yield (>4100 kg/305 d) were used to determine differences in milk fatty acid profile as lactation progressed. Milk yield was recorded daily and milk samples were collected during wk 1, 4, 8, 12, and 16 postpartum for milk composition analysis. Milk samples from wk 1, 8, and 16 were also analyzed for fatty acid composition. Select-line cows produced more milk (44.4 vs. 31.2 kg/d) and milk components than CL cows during the 16-wk period. There was no difference in rate of milk yield increase, but peak milk yield for SL cows was greater and occurred later in lactation. There were no differences in milk SCC or milk fat, protein, or lactose content. Selection for milk yield did not affect the content of most individual milk fatty acids; however, compared with CL, SL cows had a reduced Delta(9)-desaturase system and tended to produce milk with lower monounsaturated fatty acid content. Selection for milk yield did not affect milk fatty acid origin but the percentage of de novo fatty acids increased and preformed fatty acids decreased as lactation progressed. Milk fat trans-11 18:1 and cis-9,trans-11 conjugated linoleic acid increased with progressing lactation (10.7 vs. 14.1 and 3.1 vs. 5.4 mg/g, or 31 and 76%, respectively) and were correlated strongly among wk 1, 8, and 16 of lactation. Temporal changes in the Delta(9)-desaturase system occurred during lactation but these changes were not correlated with milk fat cis-9,trans-11 conjugated linoleic acid content. Results indicate prolonged genetic selection for milk yield had little effect on milk fatty acid composition, but milk fatty acid profiles varied markedly by week of lactation.     

Effects of dietary supplementation of rumen-protected conjugated linoleic acid in dairy cows during established lactation

Short-term studies (< 5 d) involving abomasal infusion of a mixture of CLA isomers or pure trans-10, cis-12 CLA have demonstrated that supplements of conjugated linoleic acids (CLA) reduce milk fat synthesis during established lactation in dairy cows. Our objective was to assess longer term effects of supplementation during established lactation using a dietary supplement of rumen-protected CLA. Thirty Holstein cows were blocked by parity and received a dietary fat supplement of either Ca-salts of palm oil fatty acids (control) or a mixture of Ca-salts of palm oil fatty acids plus Ca-salts of CLA (CLA treatment). Supplements provided about 90 g/d of fatty acids and were topdressed on the TMR. The CLA supplement provided 30.4 g/d of CLA in which the predominant isomers were: trans-8, cis-10 (9.2%), cis-9, trans-11 (25.1%), trans-10, cis-12 (28.9%), and cis-11, trans-13 (16.1%). All cows were pregnant; treatments were initiated on d 79 of pregnancy (approximately 200 d prepartum) and continued for 140 d until dry off. Twenty-three cows completed the study; those receiving CLA supplement had a lower milk fat test (2.90 versus 3.80%) and a 23% reduction in milk fat yield (927 versus 1201 g/d). Intake of DM, milk yield, and the yield and content of true protein and lactose in milk were unaffected by treatment. Milk fat analysis indicated that the CLA supplement reduced the secretion of fatty acids of all chain lengths. However, effects were proportionally greater on short and medium chain fatty acids, thereby causing a shift in the milk fatty acid composition to a greater content of longer-chain fatty acids. Changes in body weight gain, body condition score, and net energy balance were not significant and imply no differences in cows fed the CLA supplement in replenishment of body reserves in late lactation. Likewise, maintenance of pregnancy, gestation length, and calf birth weight were unaffected by treatment. Overall, feeding a dietary supplement of rumen-protected CLA to pregnant cows over the last 140 d of the lactation cycle resulted in a marked reduction in milk fat content and yield, and a shift in milk fatty acid composition, but other milk components, DMI, maintenance of pregnancy, and cow well-being were unaffected.     

Changes in milk fat in response to dietary supplementation with calcium salts of trans-18:1 or conjugated linoleic fatty acids in lactating dairy cows

Milk fat was investigated in lactating dairy cows fed diets supplemented with Ca salts of trans fatty acids (Ca-tFA) or Ca salts of conjugated linoleic acids (Ca-CLA). Forty-five Holstein cows (115 days in milk) were fed a control diet (51% forage; dry matter basis) supplemented with 400 g of EnerG II (Ca salts of palm oil fatty acids) for 2 wk; subsequently, 5 groups of 9 cows each were assigned for 4 wk to the control diet or diets containing 100 g of Ca-CLA or 100, 200, or 400 g of Ca-tFA in a randomized block design. Treatments had no effect on dry matter intake, milk production, protein, lactose, or somatic cell count. Milk fat percentage was reduced from 3.39% in controls to 3.30, 3.04, and 2.98%, respectively, by the Ca-tFA diets and to 2.54% by the Ca-CLA diet. Milk fat yield (1.24 kg/d in controls) was decreased by 60, 130, and 190 g/d with increasing dose of Ca-tFA and by 290 g/d with the Ca-CLA supplement. Consistent with increased endogenous synthesis of cis-9-containing CLA from precursors provided by the Ca-tFA diets, total CLA were similar in milk of cows fed Ca-CLA or Ca-tFA. Compared with controls, the Ca-CLA diet increased trans-10, cis-12-18:2 yield in milk, without altering levels of trans-18:1 isomers. In contrast, yields of most trans-18:1 isomers were elevated in milk of cows fed Ca-tFA diets, whereas yields of trans-10, cis-12-18:2 remained similar to control values. We conclude that milk fat depression can occur without an increase in trans-10, cis-12-18:2 in milk and that other components, perhaps the trans-10-18:1 isomer, may be involved.     

Flavor and stability of pasteurized milk with elevated levels of conjugated linoleic acid and vaccenic acid

The objectives of this study were to determine if flavor differences between 2% fat pasteurized milks with and without naturally enhanced vaccenic acid (VA) and cis-9, trans-11 conjugated linoleic acids (CLA) levels could be detected over the commercial shelf life of the product and to determine if milk with elevated VA and cis-9, trans-11 CLA levels was more susceptible to development of light-induced oxidative flavor defects. Cows were fed a control diet or the same ration supplemented with 2% soybean oil and 1% fish oil (CLA diet). The milk, standardized to 2% fat, was pasteurized, homogenized, and stored in plastic containers at 4 degrees C. Oxidation was induced by exposing half of the containers to light. Testing was conducted at 1, 7, and 14 d postpasteurization. Average cis-9, trans-11 CLA content of the milks from the control and CLA diet groups was 0.52 and 4.74 g/100 g of fatty acids, respectively (8-fold increase). Average VA content of the milk from the control and CLA diet groups was 1.43 and 12.06 g/100 g of fatty acids, respectively (7.5-fold increase). Together, VA plus CLA represented almost 17% of the total milk fatty acids. There was no effect of light exposure on fatty acid composition initially or over the 14-d storage period. Although VA, cis-9, trans-11 CLA, and degree of unsaturation were significantly elevated in the milk from the CLA diet group, untrained panelists were unable to detect flavor differences initially or over time in 15 of 16 triangle test evaluations. Similarly, sensory results indicated no difference in susceptibility to the development of oxidized off-flavors between the milk from the control and CLA diet groups, even when oxidation was induced by light exposure.     

Use of conjugated linoleic acid (CLA) enrichments to examine the effects of trans-8, cis-10 CLA, and cis-11, trans-13 CLA on milk-fat synthesis

Conjugated linoleic acid (CLA) supplements have typically been comprised of 4 isomers (trans-8, cis-10; cis-9, trans-11; trans-10, cis-12; and cis-11, trans-13 CLA). Abomasal infusion of pure isomers has shown that trans-10, cis-12 CLA is a potent inhibitor of milk-fat synthesis, whereas cis-9, trans-11 CLA has no effect. However, there appear to be additional fatty acids that inhibit milk-fat synthesis, and the objective of this study was to investigate the effects of additional CLA isomers present in CLA supplements. Four rumen fistulated Holstein cows (141+/-8 DIM, mean+/-SE) were randomly assigned in a 4 x 4 Latin square experiment. Treatments were abomasal infusion of (1) skim milk (negative control), (2) trans-10, cis-12 CLA supplement (positive control), (3) trans-8, cis-10 CLA supplement, and (4) cis-11, trans-13 CLA supplement. Treatments 2 through 4 were targeted to provide 4 g/d of the CLA isomer of interest. The trans-8, cis-10 CLA supplement had no effect on milk-fat yield, whereas the trans-10, cis-12 CLA supplement reduced milk-fat yield by 35%. The cis-11, trans-13 CLA supplement contained some trans-10, cis-12 CLA, and when data were compared to the positive control treatment group, it was obvious that cis-11, trans-13 CLA also had no effect on milk-fat synthesis. Milk-fat content of specific CLA isomers was significantly elevated within respective treatment groups. Milk yield, DMI, and milk protein yield were unaffected by treatment. Overall, trans-10, cis-12 CLA reduced milk-fat synthesis, whereas the other major isomers present in CLA supplements (trans-8, cis-10 CLA and cis-11, trans-13 CLA) had no effect.     

Short communication: Postruminal infusion of conjugated linoleic acids negatively impacts milk synthesis in Holstein cows

In view of the potential of rumen-protected conjugated linoleic acid (CLA) as a means to increase the CLA content of bovine milk, a study was undertaken to evaluate the effect of synthetic CLA on milk production and composition. Four Holstein cows received abomasal infusion of: 1) control, no lipid infusion, 2) 150 g/d of synthetic CLA, 31.7% cis-9, trans-11; 30.4% trans-10, cis-12, 3) 150 g/d of safflower oil, and 4) 150 g/d of tallow. Infusion was carried out for 20 to 22 h/d for 11-d periods in a 4 x 4 Latin square design. The milk fat concentration of cis-9, trans-11 and trans-10, cis-12 isomers of CLA was significantly increased with infusion of CLA. However, CLA infusion had other unexpected effects on milk production and composition. Milk yield dropped significantly during the period of CLA infusion. Furthermore, as well as the typical depression in milk fat reported with trans-10 isomers of CLA, other negative effects specific to CLA infusion were observed including a drop in lactose concentration and yield, a drop in protein yield, and an elevated somatic cell count. The important difference between synthetically produced CLA and CLA produced naturally in the cow is the much higher proportion of trans-10 isomers of CLA in the former. The results of this study suggest that the extent of enrichment possible for trans-10 isomers of CLA, and hence the usefulness of synthetic CLA for this purpose, may be limited because of unacceptable effects on milk yield and composition.