Effect of conjugated linoleic acid and acetate on milk fat synthesis and adipose lipogenesis in lactating dairy cows

During biohydrogenation-induced milk fat depression (MFD), nutrients are spared from milk fat synthesis and are available for other metabolic uses. Acetate is the major carbon source spared and it may increase lipid synthesis in adipose tissue during MFD. The objective of this study was to compare the effect of trans-10,cis-12 conjugated linoleic acid (CLA) and the amount of acetate spared during CLA-induced MFD on adipose tissue lipogenesis. Nine multiparous, lactating, ruminally cannulated Holstein cows (244 ± 107 d in milk; 25 ± 8.4 kg of milk/d; mean ± standard deviation) were randomly assigned to treatments in a 3 × 3 Latin square design. Experimental periods were 4 d followed by a 10-d washout. Treatments were control (CON), ruminal infusion of acetate (AC; continuous infusion of 7 mol/d adjusted to pH 6.1 with sodium hydroxide), or abomasal infusion of CLA (10 g/d of both trans-10,cis-12 CLA and cis-9,trans-11 CLA). Dry matter intake, milk yield, and milk protein yield and percentage were not affected by treatments. Compared with CON, milk fat yield decreased 23% and fat percent decreased 28% in CLA, and milk fat yield increased 20% in AC. Concentration and yield of milk de novo synthesized fatty acids (<C16) were reduced and concentration of preformed fatty acids (>C16) was increased by CLA, compared with CON. Yield of de novo synthesized fatty acids and palmitic acid was increased by AC, compared with CON. Lipogenesis capacity of adipose tissue explants was decreased 72% by CLA, but was not affected by AC. Acetate oxidation by adipose explants was not affected by treatments. Treatments had no effect on expression of key lipogenic factors, lipogenic enzymes, and leptin; however, expression of fatty acid binding protein 4 was reduced in CLA compared with CON. Additionally, hormone-sensitive lipase and perilipin 1 were decreased by CLA and acetate. Plasma glucose and glucagon concentrations were not affected by treatments; however, CLA increased nonesterified fatty acids 17.7%, β-hydroxybutyrate 16.1%, and insulin 27.8% compared with CON, and AC increased plasma β-hydroxybutyrate 18%. In conclusion, during CLA-induced MFD in low-producing cow adipose tissue was sensitive to the anti-lipogenic effects of CLA, while spared acetate did not stimulate adipose lipogenesis. However, acetate may play an important role in stimulating lipogenesis and improving energy status in the mammary gland under normal conditions.     

Combined effects of trans-10,cis-12 conjugated linoleic acid, propionate, and acetate on milk fat yield and composition in dairy cows

Diets inducing milk fat depression (MFD) are known to alter ruminal lipid metabolism, leading to the formation of specific isomers [such as trans-10,cis-12 conjugated linoleic acid (CLA)] that inhibit milk fat synthesis in lactating dairy cows. However, ruminal outflow of these isomers does not fully account for the decreases in milk fat synthesis observed during diet-induced MFD. The high-concentrate diets inducing MFD also induce a greater production of propionate, suggesting a possible inhibition of milk fat by propionate associated with trans-10,cis-12-CLA during MFD. The present experiment aimed to study the combined effects of propionate and trans-10,cis-12-CLA (both inhibitors of milk fat synthesis) on milk fat secretion and the effects of the combination of 2 nutrients with opposite effects (acetate and propionate). Six Holstein cows were used in a 6 × 6 Latin square design with 21-d periods (14 d of nutrient infusion). The treatments were control; ruminal infusion of 1,500 g/d of acetate (A); ruminal infusion of 800 g/d of propionate (P); duodenal infusion of 1.60 g/d of trans-10,cis-12-CLA (CLA); ruminal infusion of 750 g/d of acetate + 400 g/d of propionate (A+P); and duodenal infusion of 1.60 g/d of trans-10,cis-12-CLA + ruminal infusion of 800 g/d of propionate (CLA+P). The amounts of nutrients infused were chosen to induce a similar variation in milk fat content. Treatments A and P decreased dry matter intake. Compared with the control, P and CLA treatments decreased milk fat content and yield by 9% and 15% on average. Treatment A increased milk fat content by 6.5% but did not modify milk fat yield (because of a decrease in milk yield). The effects of A and P, and CLA and P on milk fat and fatty acid percentages and yield were additive (A+P and CLA+P treatments). With a same dose of trans-10,cis-12-CLA, the additional supply of propionate induced a decrease in milk fat 40% higher than that induced by trans-10,cis-12-CLA alone. The milk fatty acid profile obtained with CLA+P was similar to those observed with high-concentrate diets inducing MFD. In conclusion, under our experimental conditions, the effects of the 3 nutrients were additive on mammary lipogenesis, regardless of their separate effects. We also show that propionate could contribute to the milk fat reductions unaccounted for by trans-10,cis-12-CLA during MFD induced by high-concentrate diets.     

A conjugated linoleic acid supplement containing trans-10, cis-12 reduces milk fat synthesis in lactating sheep

The efficacy of conjugated linoleic acid (CLA) supplements containing trans-10, cis-12 for reducing milk fat synthesis has been well documented in dairy cows, but studies with other ruminant species are less convincing, and there have been no investigations of this in sheep. Therefore, the current study was designed to determine whether trans-10, cis-12 CLA would inhibit milk fat synthesis in sheep. Twenty multiparous ewes in early lactation were paired and randomly allocated to 2 treatments: grass hay plus concentrate either unsupplemented (control) or supplemented with lipid-encapsulated CLA to provide 2.4 g/d of trans-10, cis-12 CLA. The CLA dose was based on published responses of dairy cows extrapolated to ewes on a metabolic body weight basis. The experimental design was a 2-period crossover with 10-d treatment periods separated by a 10-d interval. Compared with the control, CLA supplementation reduced milk fat content from 6.4 to 4.9% and reduced fat yield from 95 to 80 g/d. The CLA treatment also increased milk yield from 1,471 to 1,611 g/d and increased protein yield from 68 to 73 g/d. Milk protein content and DMI were unaffected by treatment. The reduction in milk fat yield was due to decreases in both de novo fatty acid synthesis and uptake of preformed fatty acids. Milk fat content of trans-10, cis-12 CLA was < 0.01 and 0.12 g/100 g of fatty acids for the control and CLA treatments, respectively. The transfer efficiency of trans-10, cis-12 CLA from the dietary supplement into milk fat was 3.8%. Results of the present study demonstrate that a CLA supplement containing trans-10, cis-12 CLA reduces milk fat synthesis in lactating sheep in a manner similar to dairy cows when fed at an equivalent dose (metabolic body weight basis). Furthermore, the nutrients spared by the reduction in milk fat coincided with an increase in milk and milk protein yield.     

Effect of abomasal infusions of a mixture of octadecenoic acids on milk fat synthesis in lactating cows

Diets causing milk fat depression (MFD) are known to alter ruminal lipid metabolism leading to the formation of specific biohydrogenation intermediates that exert antilipogenic effects. Several isomers of conjugated linoleic acid (CLA), namely trans-10, cis-12 CLA, cis-10, trans-12 CLA, and trans-9, cis-11 CLA, inhibit mammary lipogenesis in the lactating cow, but ruminal outflow of these biohydrogenation intermediates does not account entirely for the reductions in milk fat synthesis during diet-induced MFD. Milk fat trans-10 18:1 concentrations are consistently increased on diets that cause MFD, suggesting a possible role in the regulation of milk fat secretion. Three rumen-fistulated cows in mid lactation were used in a 3 × 3 Latin square to evaluate the effects of a mixture of 18:1 fatty acid methyl esters (FAME) on milk fat synthesis. Experimental treatments consisted of abomasal infusions of ethanol (control), 6 g/d of trans-10, cis-12 CLA (positive control; CLA), or 247 g/d of a mixture of 18:1 FAME containing (% fatty acids) cis-9 (9.45), cis-12 (3.35), trans-10 (37.3), trans-11 (37.4), and trans-12 (2.66) as major isomers (T181 treatment). Administration of the T181 treatment supplied 92.1 g/d of trans-10 18:1. Infusions were conducted over a 5-d period with a 9-d interval between treatments. Treatments had no effect on dry matter intake, milk yield, or milk protein. Relative to the control, abomasal infusion of T181 and trans-10, cis-12 CLA treatments reduced milk fat secretion by 19.5 and 41.5%, respectively. Even though a direct cause and effect on mammary lipogenesis could not be established, comparisons with published data and considerations of the relative abundance of constituent FAME in treatment T181 implicated trans-10 18:1 as the isomer responsible. In conclusion, current data suggest that trans-10 18:1 potentially exerts antilipogenic effects and may contribute to the reduction in milk fat synthesis during diet-induced MFD in the lactating cow.     

Trans-10, trans-12 conjugated linoleic acid does not affect milk fat yield but reduces delta9-desaturase index in dairy cows

Trans-10, cis-12 conjugated linoleic acid (CLA) is a potent inhibitor of milk fat synthesis, and the magnitude of milk fat depression is often correlated with the fat content of this isomer. However, the trans-10, cis-12 CLA content does not always correspond to the extent of milk fat depression, and in some instances, an increase in the milk fat content of trans-10, trans-12 CLA has been observed. We synthesized trans-10, trans-12 CLA (>90% purity) and investigated its effect on milk fat synthesis and incorporation into plasma lipids. Three rumen-fistulated Holstein cows were randomly assigned in a 3 × 3 Latin square experiment. Treatments were a 4-d abomasal infusion of 1) ethanol (control), 2) a trans-10, cis-12 CLA supplement (positive control), and 3) a trans-10, trans-12 CLA supplement; 5 g/d of the CLA isomer of interest was provided. Milk yield, dry matter intake, and milk protein were unaffected by treatment. Treatment with trans-10, trans-12 CLA had no effect on milk fat yield, whereas treatment with trans-10, cis-12 CLA reduced milk fat yield by 28%. Incorporation of CLA was greatest for the plasma triglyceride fraction, and the milk fat content was subsequently elevated within the respective treatment groups. The milk fatty acid composition indicated that Δ⁹-desaturase was reduced significantly for both CLA treatments, but the reduction was greater for the treatment with trans-10, trans-12 CLA. Overall, abomasal infusion of trans-10, trans-12 CLA and trans-10, cis-12 CLA altered the desaturase ratios, but only trans-10, cis-12 CLA reduced milk fat synthesis.     

A conjugated linoleic acid supplement containing trans-10, cis-12 conjugated linoleic acid reduces milk fat synthesis in lactating goats

The effect of conjugated linoleic acid (CLA) supplements containing trans-10, cis-12 for reducing milk fat synthesis has been well described in dairy cows and sheep. Studies on lactating goats, however, remain inconclusive. Therefore, the current study investigated the efficacy of a lipid-encapsulated trans-10, cis-12 CLA supplement (LE-CLA) on milk production and milk fatty acid profile in dairy goats. Thirty multiparous Alpine lactating goats in late lactation were used in a 3 × 3 Latin square design (14-d treatment periods separated by 14-d intervals). Does were fed a total mixed ration of Bermuda grass hay, dehydrated alfalfa pellets, and concentrate. Does were randomly allocated to 3 treatments: A) unsupplemented (control), B) supplemented with 30 g/d of LE-CLA (low dose; CLA-1), and C) supplemented with 60 g/d of LE-CLA (high dose; CLA-2). Milk yield, dry matter intake, and milk protein content and yield were unaffected by treatment. Compared with the control, milk fat yield was reduced 8% by the CLA-1 treatment and 21% by the CLA-2 treatment, with milk fat content reduced 5 and 18% by the CLA-1 and CLA-2 treatments, respectively. The reduction in milk fat yield was due to decreases in both de novo fatty acid synthesis and uptake of preformed fatty acids. Milk fat content of trans-10, cis-12 CLA was 0.03, 0.09, and 0.19 g/100 g of fatty acids for the control, CLA-1, and CLA-2 treatments, respectively. The transfer efficiency of trans-10, cis-12 CLA from the 2 levels of CLA supplement into milk fat was not different between treatments and averaged 1.85%. In conclusion, trans-10, cis-12 CLA reduced milk fat synthesis in lactating dairy goats in a manner similar to that observed for lactating dairy cows and dairy sheep. Dose-response comparisons, however, suggest that the degree of reduction in milk fat synthesis is less in dairy goats compared with dairy cows and dairy sheep.     

Trans-9, cis-11 conjugated linoleic acid reduces milk fat synthesis in lactating dairy cows

Under certain dietary situations, rumen biohydrogenation results in the production of unique fatty acids that inhibit milk fat synthesis. The first of these to be identified was trans-10, cis-12 conjugated linoleic acid (CLA), but others are postulated to contribute to diet-induced milk fat depression (MFD). Our objective was to examine the potential role of trans-9, cis-11 CLA in the regulation of milk fat. In a preliminary study, we used gas-liquid and high-performance liquid chromatography techniques to examine milk fat samples from a diet-induced MFD study and found that an increase in trans-9, cis-11 CLA corresponded to the decrease in milk fat yield. We investigated this further using a CLA enrichment of 9, 11 isomers to examine the biological effect of trans-9, cis-11 CLA on milk fat synthesis. Four rumen-fistulated Holstein cows were randomly assigned in a 4 × 4 Latin square experiment involving 5-d treatment periods and abomasal infusion of 1) ethanol (control), 2) a 9, 11 CLA mix (containing 32% trans-9, cis-11, 29% cis-9, trans-11, and 17% trans-9, trans-11), 3) a trans-9, trans-11 CLA supplement, and 4) a trans-10, cis-12 CLA supplement (positive control). The trans-9, trans-11 CLA and trans-10, cis-12 CLA supplements were of high purity (>90%), and all supplements were infused at a rate to provide 5 g/d of the CLA isomer of interest. Milk yield and dry matter intake did not differ among treatments. Compared with the control treatment, milk fat yield was reduced by 15% for the 9, 11 CLA mixture and by 27% for the trans-10, cis-12 CLA treatment. We also found that trans-9, trans-11 CLA had no effect on milk fat yield, and previous research has shown that milk fat yield is unaltered when cows are infused with cis-9, trans-11 CLA. When all treatments were considered, results suggested that trans-9, cis-11 was the CLA isomer in the 9, 11 CLA mix responsible for the reduction in milk fat synthesis, although the magnitude was less than that observed for trans-10, cis-12 CLA. Interestingly, trans-9, trans-11 CLA altered the milk fat desaturase index, further demonstrating that alterations in desaturase can occur independently of effects on milk fat synthesis. Overall, our investigations identified that an increase in milk fat content of trans-9, cis-11 CLA was associated with diet-induced MFD and provided evidence of a role for this isomer in MFD based on the 15% reduction in milk fat yield with abomasal infusion of a CLA enrichment that supplied 5 g/d of trans-9, cis-11 CLA.     

Principal component and multivariate analysis of milk long-chain fatty acid composition during diet-induced milk fat depression

The objective of this study was to assess the relationship between individual milk fatty acids (FA) and diet-induced milk fat depression (MFD) using principal component analysis (PCA) and multivariate analysis (MA). Cow treatment observations (n = 63) from 3 published feeding experiments with lactating dairy cows were used in the analyses. In the PCA, principal component loading plots 1 (PC1) and 2 (PC2) described 55.9% of the total variation in milk FA and fat concentrations. Saturated FA (14:0, 16:0, and 17:0) and milk fat percentage showed negative loading for PC1. Trans-18:1 isomers (trans-6+7+8 to trans-15), trans-7, cis-9 conjugated linoleic acid (CLA), and trans-10, cis-12 CLA showed positive (opposite) loading, suggesting a negative relationship between these isomers and milk fat percentage. Cis-11, trans-13 CLA and cis-9, trans-11 CLA were associated with the PC2 axes (neutral), indicating that they were not associated with MFD. Multivariate analysis with milk fat percentage as the dependent variable and individual PC1 positive loading variables showed a breakpoint relationship for trans-6+7+8-, trans-9-, trans-10-, and trans-13+14-18:1 and a linear relationship for trans-11-, trans-12-, trans-15-18:1, trans-10, cis-12 CLA, and trans-7, cis-9 CLA. Subsequent MA was conducted on 41 treatment means from 12 independent experiments from the literature, in which concentrations of trans-6+7+8-, trans-9-, trans-10-, and trans-11-18:1, and cis-9 trans;-11, and trans-10, cis-12 CLA were reported. Significant negative effects of trans-9-18:1, trans-10-18:1, and trans-10, cis-12 CLA on milk fat percentage were observed. In this study, the PCA and MA showed that among trans-18:1 isomers, trans-10-18:1 was the most negatively correlated to milk fat percentage. However, the threshold concentration related to maximum MFD indicated that the relative potency was greatest for trans-6+7+8- and lowest for trans-10-18:1. These results suggested that trans-6+7+8-18:1 might be more important than trans-10-18:1 in MFD. Principal component analysis also showed that trans-10, cis-12 and trans-7, cis-9 CLA were the isomers most negatively correlated to milk fat percentage, implying a possible role of trans-7, cis-9 CLA in MFD. Additional experiments are needed to establish whether trans-7-18:1 is involved in MFD or that its effects are mediated via the endogenously synthesized trans-7, cis-9 CLA.     

Pigeon peas as a supplement for lactating dairy cows fed corn silage-based diets

Holstein rumen-cannulated cows [n = 7; initial body weight (BW) 640.56 ± 71.43 kg] were fed a corn silage basal diet with 1 of 3 concentrates (C = control; P10 = 10% pigeon peas; P20 = 20% pigeon peas). Cows were randomly assigned to treatments in a replicated 3 × 3 Latin square and individually fed using Calan gates. Each experimental period was 21 d with 7 d for adaption and 14 d for sample collection. Ruminal fluid samples were taken the last day of each experimental period and analyzed for pH, ammonia, long-chain fatty acids, and volatile fatty acids (VFA). Consecutive a.m. and p.m. milk samples were taken during the last 2 wk of the 21-d period and analyzed for fat, protein, long-chain fatty acids, and somatic cell count. Dry matter intake (kg/d) was reduced during the second period and was greater for P10 diets. Milk protein was greater for cows fed P20 compared with P10. Energy-corrected milk was greater for cows fed the control diet compared with P10. Treatment had no effect on milk yield. Ruminal fluid pH decreased over sampling times; however, pH remained at or above 5.5. Diets did not affect ruminal fluid pH; however, pH was different for sampling periods. Ruminal ammonia decreased until 8 h postfeeding at which time it peaked consistent with changes in ammonia concentrations that usually peak 3 to 5 h postfeeding on diets high in plant proteins. Dietary treatments altered ruminal fluid VFA with reduced concentrations of acetate and greater concentrations of propionate for control diet, resulting in reduced acetate:propionate ratio. Isobutyrate exhibited an hour by treatment interaction, in which isobutyrate decreased until 8 h postfeeding and then tended to be greater for P10 than for other treatments. Animals fed the P10 diet had greater concentrations of ruminal isovalerate. Ruminal cis-9,trans-11 and trans-10,cis-12 conjugated linoleic acid (CLA) isomers were not affected by dietary treatments. The P10 diet had greatest ruminal synthesis of cis-9,trans-11, but control cows had greatest ruminal synthesis of trans-10,cis-12. Milk CLA isomers were similar among treatments. Trends were observed for greater cis-9,trans-11 and trans-10,cis-12 for the P10 diet. Pigeon peas may be used as a protein supplement in dairy diets without affecting milk production, dry matter intake, or ruminal environment when they replace corn and soybean meal.     

Evaluation of the mechanism of action of conjugated linoleic acid isomers on reproduction in dairy cows

The objective of this study was to evaluate the mechanism of action through which conjugated linoleic acid (CLA) beneficially affects reproduction. Lactating Holstein cows (n = 45, 20 ± 1 DIM) were assigned to 1 of 3 treatments: 70 g/d of Ca salts of tallow (control); 63 g/d of lipid-encapsulated CLA providing 7.1 g/d of cis-9, trans-11 CLA and 2.4 g/d of trans-10, cis-12 CLA (CLA 75:25); or 76 g/d of lipid-encapsulated CLA providing 7.1 g/d each of cis-9, trans-11 and trans-10, cis-12 CLA (CLA 50:50). Supplements were top-dressed for 37 d, milk production and DMI were recorded daily, and blood samples were taken 3 times per week. At 30 ± 3 DIM, ovulation was synchronized in all cows with a modified Ovsynch protocol, and on d 15 of the cycle cows received an oxytocin injection; blood samples were obtained frequently to measure 13,14 dihydro, 15-keto PGF_2α. On d 16 of the cycle cows received a PGF_2α injection and ovarian follicular aspiration was performed 54 h later. Follicular fluid was analyzed for fatty acids, progesterone, and estradiol. Endometrial biopsies were taken before and again near the end of the supplementation period for fatty acid analysis. The CLA resulted in decreased milk fat content of 14.1 and 6.1% at wk 5 of treatment of CLA 50:50 and CLA 75:25, respectively. There were no differences in energy balance or plasma nonesterified fatty acids; however, plasma IGF-I was greater in cows supplemented with CLA 50:50. The CLA isomers were not detectable in endometrial tissue, but cis-9, trans-11 CLA tended to be greater in follicular fluid of supplemented cows. Response to the oxytocin challenge was not different among treatments. Progesterone during the early luteal phase and the estradiol:progesterone ratio in follicular fluid tended to be greater in cows supplemented with CLA 50:50. Overall, these results indicate that short periods of CLA supplementation do not alter uterine secretion of PGF_2α. The mechanism through which CLA affects reproduction may involve improved ovarian follicular steroidogenesis and increased circulating concentrations of IGF-I.     

Diet and plasma evaluation of the main isomers of conjugated linoleic acid and trans-fatty acids in a population sample from Mediterranean north-east Spain

The aim of the present study was to describe the dietary pattern of a representative sample of 516 adult participants (203 men and 313 women) from Catalonia, a Spanish Mediterranean region, to assess their current dietary and plasma levels of trans C18:1, the major trans-fatty acid (TFA), and cis-9, trans-11 CLA, and trans-10, cis-12 CLA, the two major conjugated linoleic acid (CLA) isomers, and to evaluate their correlation with several cardiovascular disease risk factors. The population was a random sample derived from the Catalan Nutrition Survey. Plasma levels of the CLA isomers were determined in a subsample of 100 volunteers. The Catalan diet seemed to maintain some traits of the ‘traditional’ Mediterranean diet, although other components were lost. The dietary intakes of saturated fatty acids (SFA), TFA, cis-9, trans-11 CLA, and trans-10, cis-12 CLA were 12.3%, 0.84% (2.0 g/d), 0.030% (71.5 mg/d), and 0.0015% (3.4 mg/d) of the energy intake, respectively. Trans C18:1 accounted for 0.19% of the total plasma fatty acids, while the sum of cis-9, trans-11and trans-10, cis-12 CLA isomers represented about 0.09% of the plasma fatty acids. Trans C18:1 isomers correlated significantly with the intake of French fries and pastries, while cis-9, trans-11 CLA significantly correlated with the intake of dairy products and ruminant meat. None of the cardiovascular disease risk factors were found to be associated with the plasma levels of TFA or CLA. The results of this study suggest that monounsaturated fatty acids (MUFA) are the main dietary fat source in the Catalan population, due to their regular olive oil consumption. Moreover, plasma levels of the main TFA and CLA suggest that the Catalan diet is not at present strongly influenced by the occidental dietary patterns. However, a reduction of the intake of SFA in the Catalan population should be recommended.     

Temporal changes in milk fatty acid composition during diet-induced milk fat depression in lactating cows

Diet-induced milk fat depression (MFD) in lactating cows has been attributed to alterations in ruminal lipid metabolism leading to the formation of specific fatty acid (FA) biohydrogenation intermediates that directly inhibit milk fat synthesis. However, the mechanisms responsible for decreased lipid synthesis in the mammary gland over time are not well defined. The aim of this study was to evaluate the effect of diet on milk FA composition and milk fat production over time, especially during MFD, and explore the associations between MFD and FA biohydrogenation intermediates in omasal digesta and milk. Four lactating Finnish Ayrshire cows used in a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments and 35-d experimental periods were fed diets formulated to cause differences in ruminal and mammary lipid metabolism. Treatments consisted of an iso-nitrogenous total mixed ration based on grass silage with a forage to concentrate ratio of 65:35 or 35:65 without added oil, or with sunflower oil at 50 g/kg of diet dry matter. The high-concentrate diet with sunflower oil (HSO) induced a 2-stage drop in milk fat synthesis that was accompanied by specific temporal changes in the milk FA composition. The MFD on HSO was associated especially with trans-10 18:1 and also with trans-9,cis-11 conjugated linoleic acid (CLA) in milk and omasal digesta across all diets and was accompanied by the appearance of trans-10,cis-15 18:2. Trans-10,cis-12 CLA was increased in HSO, but milk fat secretion was not associated with omasal or milk trans-10,cis-12 CLA. The temporal changes in milk fat content and yield and milk FA composition reflect the shift from the predominant ruminal biohydrogenation pathway to an alternative pathway. The ambiguous role of trans-10,cis-12 CLA suggests that trans-10 18:1, trans-9,cis-11 CLA and trans-10,cis-15 18:2 or additional mechanisms contributed to the diet-induced MFD in lactating cows.     

Rapeseed or linseed in grass-based diets: Effects on conjugated linoleic and conjugated linolenic acid isomers in milk fat from Holstein cows over 2 consecutive lactations

Changes in the distribution of conjugated linoleic (CLA) and conjugated linolenic (CLnA) acid isomers in milk from Holstein cows in response to 4 different oilseed supplements rich in either cis-9 18:1 or 18:3n-3 were determined over 2 consecutive lactations in 58 and 35 cows during the first and second years, respectively. For the first 5 wk of the first lactation, all cows were fed the same diet. Thereafter, cows received 1 of 5 treatments for 2 consecutive lactations, including the prepartum period. Treatments comprised the basal diet with no additional lipid, or supplements of extruded linseeds (EL), extruded rapeseeds (ER), cold-pressed fat-rich rapeseed meal, or whole unprocessed rapeseeds to provide 2.5 to 3.0% of additional oil in diet dry matter. During indoor periods, cows were housed and received a mixture (3:1, wt/wt) of grass silage and hay, whereas cows were at pasture during outdoor periods. Over the entire study, EL resulted in the enrichment of ?11,13 CLA, ?12,14 CLA, trans-9,trans-11 CLA, trans-13,trans-15 CLA, ?9,11,15 CLnA, and cis-9,trans-11,trans-13 CLnA (identified for the first time in bovine milk fat) in milk fat, whereas ER and cold-pressed fat-rich rapeseed meal in particular, increased milk fat trans-7,cis-9 CLA concentration. With the exception of the first indoor period, whole unprocessed rapeseeds decreased cis-9,trans-11 CLA, trans-9,cis-11 CLA, and trans-10,trans-12 CLA abundance. During the second indoor period, EL increased milk trans-9,cis-11 CLA and trans-10,cis-12 CLA concentrations, but the increases in cis-9,trans-11 CLA, cis-12,trans-14 CLA, trans-11,cis-13 CLA, and cis-9,trans-11,cis-15 CLnA concentrations to EL and ER were lower for the second than first indoor period. In contrast to the indoor periods, EL and ER decreased milk cis-9,trans-11 CLA, trans-9,cis-11 CLA, and trans-10,cis-12 CLA concentrations at pasture. The extent of changes in the relative distribution and abundance of CLA and CLnA isomers in milk fat were related to the nature (rapeseed or linseed) and form of oilseed (extruded, cold-pressed fat-rich meal or whole unprocessed) supplement and their interactions with the composition of the basal diet (conserved grass or pasture and dietary starch content). Furthermore, milk fat CLA and CLnA responses to treatments were repeatable between both outdoor periods. Variations in milk fat content and yield measured during the entire study were significantly and inversely associated with milk trans-10 18:1, trans-10,cis-12 CLA, and in particular, trans-9,cis-11 CLA concentrations.     

Effects of dietary supplementation of rumen-protected conjugated linoleic acid to grazing cows in early lactation

Conjugated linoleic acids (CLA) are potent anticarcinogens in animal and in vitro models as well as inhibitors of fatty acid synthesis in mammary gland, liver, and adipose tissue. Our objective was to evaluate long-term CLA supplementation of lactating dairy cows in tropical pasture on milk production and composition and residual effects posttreatment. Thirty crossbred cows grazing stargrass (Cynodon nlemfuensis Vanderyst var. nlemfüensis) were blocked by parity and received 150 g/d of a dietary fat supplement of either Ca-salts of palm oil fatty acids (control) or a mixture of Ca-salts of CLA (CLA treatment). Supplements of fatty acids were mixed with 4 kg/d of concentrate. Grazing plus supplements were estimated to provide 115% of the estimated metabolizable protein requirements from 28 to 84 d in milk (treatment period). The CLA supplement provided 15 g/d of cis-9,trans-11 and 22 g of cis-10,trans-12. Residual effects were evaluated from 85 to 112 d in milk (residual period) when cows were fed an 18% crude protein concentrate without added fat. The CLA treatment increased milk production but reduced milk fat concentration from 2.90 to 2.14% and fat production from 437 to 348 g/d. Milk protein concentration increased by 11.5% (2.79 to 3.11%) and production by 19% (422 to 504 g/d) in the cows fed CLA. The CLA treatment decreased milk energy concentration and increased milk volume, resulting in unchanged energy output. Milk production and protein concentration and production were also greater during the residual period for the CLA-treated cows. The CLA treatment reduced production of fatty acids (FA) of all chain lengths, but the larger effect was on short-chain FA, causing a shift toward a greater content of longer chain FA. The CLA treatment increased total milk CLA content by 30% and content of the trans-10,cis-12 CLA isomer by 88%. The CLA treatment tended to decrease the number of days open, suggesting a possible effect on reproduction. Under tropical grazing conditions, in a nutritionally challenging environment, CLA-treated cows decreased milk fat content and secreted the same amount of milk energy by increasing milk volume and milk protein production.     

Response to conjugated linoleic acid in dairy cows differing in energy and protein status

The trans-10, cis-12 conjugated linoleic acid (CLA) isomer inhibits milk fat synthesis, whereas milk yield and synthesis of other milk components generally remain unchanged in established lactation. However, in some CLA studies increases in milk yield, milk protein yield, or both have been observed in cows limited in energy, either in early lactation or when grazing pasture. Our objective was to evaluate the performance and monitor peripheral tissue responses to homeostatic signals regulating lipolysis and glucose uptake with CLA supplementation when cows were limited in metabolizable energy in combination with moderate or excess metabolizable protein supply. Holstein cows (n = 48; 112 ± 5 d in milk; mean ± SE) were provided ad libitum access to a diet that met energy and protein requirements for a 16-d standardization interval. Based on performance during this interval, the Cornell Net Carbohydrate and Protein System was used to design energy-limiting rations that provided 80% of metabolizable energy requirements, and these were fed throughout the treatment periods. Cows were randomly allocated to 4 treatments, in a 2-period crossover design. Treatments were 1) moderate metabolizable protein (MP) supply, 2) moderate MP supply + CLA, 3) excess MP supply, and 4) excess MP supply + CLA. Moderate and excess MP supply were at 88 and 117%, respectively, of the MP requirement established during the standardization period, as estimated by the Cornell Net Carbohydrate and Protein System. Each experimental period comprised 16 d, with crossover of CLA within each protein level. The lipid-encapsulated CLA supplement provided 12 g/d of trans-10, cis-12 CLA. Conjugated linoleic acid treatment reduced milk fat yield by 21% but increased milk yield and milk protein yield by 2.6 and 2.8%, respectively. Milk yield and content and yield of both milk protein and fat were unaltered by either protein treatment alone or in combination with CLA. Basal concentrations of glucose, insulin, and nonesterified fatty acids were unaffected by CLA supplementation. The fractional rate of glucose clearance in response to an insulin challenge and the nonesterified fatty acid response to an epinephrine challenge were also not altered by either CLA treatment or MP supply. Overall, the results demonstrate that CLA supplementation when cows are energy-limited allows for repartitioning of nutrients, resulting in increased yields of milk and milk protein, and this can occur without changes in whole-body glucose homeostasis and adipose tissue response to lipolytic stimuli.     

Milk production, conjugated linoleic acid content, and in vitro ruminal fermentation in response to high levels of soybean oil in dairy ewe diet

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.     

c9t11‐Conjugated linoleic acid‐rich oil fails to attenuate wasting in colon‐26 tumor‐induced late‐stage cancer cachexia in male CD2F1 mice

Scope: Cancer cachexia is characterized by muscle and adipose tissue wasting caused partly by chronic, systemic inflammation. Conjugated linoleic acids (CLAs) are a group of fatty acids with various properties including anti‐inflammatory cis9, trans11 (c9t11)‐CLA and lipid‐mobilizing trans10, cis12 (t10c12)‐CLA. The purpose of this study was to test whether dietary supplementation of a c9t11‐CLA‐rich oil (6:1 c9t11:t10c12) could attenuate wasting of muscle and adipose tissue in colon‐26 adenocarcinoma‐induced cachexia in mice. Methods and results: Loss of body weight, muscle and adipose tissue mass caused by tumors were not rescued by supplementation with the c9t11‐CLA‐rich oil. In quadriceps muscle, c9t11‐CLA‐rich oil exacerbated tumor‐induced gene expression of inflammatory markers tumor necrosis factor‐α, IL‐6 receptor and the E3 ligase MuRF‐1 involved in muscle proteolysis. In epididymal adipose tissue, tumor‐driven delipidation and atrophy was aggravated by the c9,t11‐CLA‐rich oil, demonstrated by further reduced adipocyte size and lower adiponectin expression. However, expression of inflammatory cytokines and macrophage markers were not altered by tumors, or CLA supplementation. Conclusion: These data suggest that addition of c9t11‐CLA‐rich oil (0.6% c9t11, 0.1% t10c12) in diet did not ameliorate wasting in mice with cancer cachexia. Instead, it increased expression of inflammatory markers in the muscle and increased adipose delipidation.     

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.     

Effects of different forage:concentrate ratios in dairy ewe diets supplemented with sunflower oil on animal performance and milk fatty acid profile

The aim of this study was to evaluate the effect of different forage:concentrate (FC) ratios in dairy ewe diets supplemented with sunflower oil (SO) on animal performance and milk fatty acid (FA) profile, particularly focusing on trans C18:1 FA and conjugated linoleic acid (CLA). Sixty lactating Assaf ewes were randomly assigned to 6 treatments in a 3 × 2 factorial arrangement: 3 FC ratios (30:70, 50:50, and 70:30) and 2 levels of SO addition (0 and 20 g/kg of dry matter). Both the diet FC ratio and SO supplementation affected milk yield, but differences between treatments were small. Although the proportion of concentrate induced limited changes in milk FA profile, dietary SO significantly decreased saturated FA and enhanced total CLA. Furthermore, the incorporation of SO in ewe diets decreased the atherogenicity index value by about 25% and doubled the contents of potentially healthy FA such as trans-11 C18:1 and cis-9,trans-11 CLA. However, the inclusion of SO in a high-concentrate diet (30:70) could switch linoleic acid biohydrogenation pathways, resulting in a significant increase in trans-10 C18:1, trans-9,cis-11 C18:2, and trans-10,cis-12 C18:2 milk fat percentages.