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.     

A canonical discriminant analysis to study the association between milk fatty acids of ruminal origin and milk fat depression in dairy cows

Although milk fat depression (MFD) has been observed and described since the beginning of the last century, all the molecular and biochemical mechanisms involved are still not completely understood. Some fatty acids (FA) originating during rumen biohydrogenation have been proposed as causative elements of MFD. However, contradictory results were obtained when studying the effect of single FA on MFD. An alternative could be the simultaneous evaluation of the effect of many FA using a multivariate approach. The aim of this study was to evaluate the relationship between individual milk FA of ruminal origin and MFD using canonical discriminant analysis, a multivariate technique able to distinguish 2 or more groups on the basis of a pool of variables. In a commercial dairy herd, a diet containing 26% starch on a DM basis induced an unintentional MFD syndrome in 14 cows out of 40. Milk yielded by these 14 animals showed a fat content lower than 50% of the ordinary value, whereas milk production and protein content were normal. The remaining 26 cows secreted typical milk fat content and therefore were considered the control group, even though they ate the same diet. The stepwise discriminant analysis selected 14 milk FA of ruminal origin most able to distinguish the 2 groups. This restricted pool of FA was used, as variables, in a run of the canonical discriminant analysis that was able to significantly discriminate between the 2 groups. Out of the 14 FA, 5 conjugated linoleic acid isomers (C18:2 trans-10,trans-12, C18:2 trans-8,trans-10, C18:2 trans-11,cis-13, C18:2 cis-9,cis-11, C18:2 cis-10,cis-12) and C15:0 iso were more related to the control group, whereas C18:2 trans-10,cis-12, C16:1 trans-6–7, C16:1 trans-9, C18:1 trans-6–8, C18:1 trans-9, C18:1 trans-10, C18:1 cis-11, and C18:3n-3 were positively associated with the MFD group, allowing a complete discrimination. On the basis of these results, we can conclude that (1) the shift of ruminal biohydrogenation from C18:1 trans-11 to C18:1 trans-10 seemed to be strongly associated with MFD; (2) at the same time, other C18:1 trans isomers showed a similar association; (3) on the contrary, conjugated linoleic acid isomers other than C18:2 trans-10,cis-12 seemed to be associated with a normal fat secretion. Results confirmed that MFD is the consequence of a combined effect of the outflow of many ruminal FA, which collectively affect mammary fat synthesis. Because the animals of the 2 groups were fed the same diet, these results suggested that factors other than diet are involved in the MFD syndrome. Feeding behavior (i.e., ability to select dietary ingredients in a total mixed ration), rumen environment and the composition of ruminal bacteria are additional factors able to modify the products of rumen biohydrogenation. Results of the present work confirmed that the multivariate approach can be a useful tool to evaluate a metabolic pathway that involves several parameters, providing interesting suggestions about the role of some FA involved in MFD. However, results about the MFD syndrome obtained in the present research require a deep molecular investigation to be confirmed.     

Induction of and recovery from milk fat depression occurs progressively in dairy cows switched between diets that differ in fiber and oil concentration

Milk fat depression (MFD) caused by intermediates of ruminal biohydrogenation commonly occurs in dairy cattle. The time course of recovery from MFD is important to mechanistic investigation and management of the condition. Nine cows were used in a repeated design, allowing analysis of recovery from diet-induced MFD. A high-fiber, low-oil diet was fed during the control and recovery periods, and a low-fiber, high-oil (LFHO) diet was fed during the induction period. Milk yield was not affected by treatment. Milk fat percentage and yield decreased progressively during induction and were lower by d 3 and 5, respectively. Milk fat concentration and yield increased progressively when cows were fed the recovery diet and were not different from control on d 19 and 15, respectively. Yield of de novo synthesized fatty acids (FA) decreased progressively during the induction period and was lower than that of controls by d 5. A biphasic response was seen for milk fat trans isomers, where trans-11 C18:1 and cis-9,trans-11 conjugated linoleic acid (CLA) were elevated initially and trans-10 C18:1 and trans-10,cis-12 CLA increased progressively during the induction period. A similar biphasic response was seen during recovery from MFD, with trans-10 C18:1 and trans-10,cis-12 rapidly decreasing initially and trans-11 C18:1 and cis-9,trans-11 CLA increasing slightly above control levels during the second phase. Recovery from diet-induced MFD occurs gradually with a short lag when dietary fiber and oil concentrations are corrected. This time course provides a framework to identify factors causing MFD and set expectations during recovery from MFD.     

The effect of rumen digesta inoculation on the time course of recovery from classical diet-induced milk fat depression in dairy cows

Ten ruminally cannulated cows were used in a crossover design that investigated the effect of rumen digesta inoculation from non-milk fat-depressed cows on recovery from classical diet-induced milk fat depression (MFD) characterized by reduced fat yield, reduced de novo milk fat synthesis, and increased alternate trans isomers. Two additional cows fed a high-fiber and low-polyunsaturated fatty acid (FA) diet (31.8% neutral detergent fiber, 4.2% FA, and 1.2% C18:2) were used as rumen digesta donors. Milk fat depression was induced during the first 10 d of each period by feeding a low-fiber and high-polyunsaturated FA diet (induction; 26.1% neutral detergent fiber, 5.8% FA, and 1.9% C18:2), resulting in a 30% decrease in milk fat yield. A recovery phase followed where all cows were switched to the high-forage, low-polyunsaturated FA diet and were allocated to (1) control (no inoculation) or (2) ruminal inoculation with donor cow digesta (8 kg/d for 6 d). Milk yield and composition were measured every 3 d. Milk yield progressively decreased during recovery. Milk fat concentration increased progressively during the recovery phase and no effect of treatment existed at any time point. Also, no treatment effect of milk fat yield was detected. The concentration of milk de novo FA increased progressively during recovery for both treatments and was higher for inoculated compared with control cows on d 6. In agreement, milk fat concentration of trans-10,cis-12 conjugated linoleic acid decreased progressively in both treatments and was lower in inoculated cows on d 3 and 6. Ruminal inoculation from non-milk fat-depressed cows did not change milk fat yield, but slightly accelerated the rate of recovery of de novo FA synthesis and normal ruminal FA biohydrogenation, demonstrating a possible opportunity for other interventions that improve the ruminal environment to accelerate recovery from this condition.     

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.     

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.     

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.     

Effects of intravenous infusion of conjugated diene 18:3 isomers on milk fat synthesis in lactating dairy cows

It has been previously established that trans-10, cis-12 conjugated linoleic acid plays an important role in milk fat depression (MFD). However, in many situations of dietary induced MFD, the reduction in milk fat synthesis is much greater than what would be predicted based on the milk fat concentration of trans-10, cis-12 18:2. These observations suggest that other biohydrogenation intermediates could be implicated in MFD. The objective of this study was to evaluate the effects on milk fat synthesis of an intravenous administration of 2 conjugated diene 18:3 isomers (cis-9, trans-11, cis-15 and cis-9, trans-13, cis-15 18:3), which are intermediates in ruminal biohydrogenation of α-linolenic acid. Three multiparous Holstein dairy cows (days in milk = 189 ± 37 d; body weight = 640 ± 69 kg; mean ± standard deviation), fitted with indwelling jugular catheters, were randomly assigned to a 3 × 3 Latin square design. For the first 5 d of each period, cows were infused intravenously with a 15% lipid emulsion providing 1) cis-9, trans-11, cis-15 18:3 + cis-9, trans-13, cis-15 18:3 + trans-10, cis-12 18:2 (CD18:3 + CLA); 2) cis-9, cis-12, cis-15 18:3 + cis-9, cis-12 18:2 as a control (ALA + LA); or 3) cis-9, cis-12, cis-15 18:3 + trans-10, cis-12 18:2, as a positive control (ALA + CLA). Milk production was recorded, and milk was sampled daily at each milking for analyses of fat, protein, lactose, milk urea nitrogen, and somatic cell count. Dry matter intake, milk yield, and milk protein were not affected by treatment. Over the experimental period, milk fat content was decreased by 7% for cows that received either ALA + CLA or CD18:3 + CLA compared with ALA + LA. The temporal pattern of milk fat content showed a linear decrease during the infusion period for ALA + CLA and CD18:3 + CLA treatment groups. The transfer efficiencies of conjugated diene 18:3 isomers into milk fat averaged 39 and 32% for cis-9, trans-11, cis-15 18:3 and cis-9, trans-13, cis-15 18:3, respectively. The CD18:3 + CLA treatment had no effect on milk fat concentration beyond that attributable to its trans-10, cis-12 18:2 content. In conclusion, results from the current study offered no support for a role of either cis-9, trans-11, cis-15 18:3 or cis-9, trans-13, cis-15 in MFD.     

Alterations in ruminal bacterial populations at induction and recovery from diet-induced milk fat depression in dairy cows

Ten ruminally cannulated Holstein cows were used in a crossover design that investigated changes in ruminal bacterial populations in response to induction and recovery from diet-induced milk fat depression (MFD). Further, the effect on the ruminal microbiota of the cows with diet-induced milk fat depression inoculated with rumen contents from non-milk fat-depressed donor cows was evaluated. Milk fat depression was induced during the first 10 d of each period by feeding a low-fiber, high-starch, and high-polyunsaturated fatty acid diet (26.1% neutral detergent fiber, 28.1% starch, 5.8% total fatty acids, and 1.9% C18:2), resulting in a 30% decrease in milk fat yield. Induction was followed by a recovery phase, where all cows were switched to a high-fiber, low-starch, and low-polyunsaturated fatty acid diet (31.8% neutral detergent fiber, 23% starch, 4.2% total fatty acids, and 1.2% C18:2) and were allocated to (1) control (no inoculation) or (2) ruminal inoculation with donor cow digesta (8 kg/d for 6 d). Ruminal samples were collected at the end of induction (d 10) and during recovery (d 13, 16, and 28), separated to solid and liquid fractions, extracted for DNA, PCR- amplified for the V1-V2 region of the 16S rRNA gene, and analyzed for bacterial diversity. Results indicated that bacterial communities were different between fractions. In each fraction, differences were significant between the induction (d 10) and recovery (d 13, 16, and 28) periods; however, differences were less apparent with time during the recovery period. The MFD (d 10) was typified by a reduction in the relative sequence abundance of Bacteroidetes and an increase in the relative sequence abundance of Firmicutes and Actinobacteria across both fractions. At the genus level, relative sequence abundance of unclassified Lachnospiraceae, Butyrivibrio, Bulleidia, and Coriobacteriaceae were higher on d 10 and were positively correlated with trans-10,cis-12 CLA and the trans-10 isomer, suggesting their potential role in altered biohydrogenation reactions. A switch to the recovery diet resulted in a sharp increase in the Bacteroidetes lineages and a decrease in Firmicutes members on d 13; however, this shift appears to stabilize by d 28, indicating the restoration process for ruminal bacteria from an altered state is gradual and complex. Inoculation of 10% of rumen contents from non-MFD donor cows to MFD cows revealed this procedure had transient effects on only a few bacterial populations, and such effects disappeared after d 16 following cessation of inoculation. It can be concluded that alterations in milk FA profiles at induction are preceded by microbial alterations in the rumen driven by dietary changes.     

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.     

Changes in fermentation and animal performance during recovery from classical diet-induced milk fat depression using corn with differing rates of starch degradability

Diet-induced milk fat depression (MFD) is a multifactorial disorder that can be triggered by a variety of conditions. Feeding high amounts of starch and unsaturated fatty acids has been shown to reduce milk fat yield and composition, as well as alter ruminal biohydrogenation patterns. However, little is known about how starch degradability in the rumen influences recovery from diet-induced MFD and if production of milk fat–inhibiting isomers will persist following an episode of MFD. The objective of this study was to evaluate production performance and ruminal fermentation in cows recovering from MFD when corn with a low or high starch degradability is fed. Six ruminally fistulated Holstein cows were used in a crossover design with 2 periods. During each period, MFD was induced for 10 d by feeding a diet with low fiber, high starch, and high unsaturated fatty acid. The polyunsaturated fatty acid concentration of the diet during the induction phase was modified primarily through inclusion of soybean oil. Following induction, cows were switched to either a high degradable starch recovery diet (HDS) or a low degradable starch recovery diet (LDS) for 18 d. The 7-h starch degradability was 66.5% for LDS and 87.8% for HDS. Milk was collected every 3 d for component and fatty acid analysis. On d 0, 4, 7, 10, 16, 22, and 28 of each period, ruminal pH and rumen fluid were collected every 2 h. Milk fat yield and composition was reduced during MFD induction and progressively increased by day in both HDS and LDS during recovery. Dry matter intake was similar among treatments and increased steadily over time during recovery. Preformed fatty acids were greater for HDS-fed animals, and de novo fatty acid in milk fat was greater for LDS-fed animals. Milk trans-10 C18:1 tended to be greater for HDS, and trans-10,cis-12 conjugated linoleic acid was significantly greater for HDS. cis-9,trans-11 conjugated linoleic acid was not affected by starch degradability during recovery. Total volatile fatty acids, butyrate, and valerate tended to differ or differed with recovery treatment, but ruminal pH and ammonia concentration were unaffected. The HDS diet responded similarly to the LDS diet during recovery with regard to milk fat percentage, but milk and fat yield tended to consistently be lower in HDS. When considering approaches to ameliorate diet-induced MFD, the degradability of the starch within rations should be evaluated. Although animal performance was similar, some trans fatty acid isomers were persistent in the milk through the recovery phase with HDS-fed animals, suggesting that milk fat synthesis might be potentially inhibited and biohydrogenation pathways modified in the rumen following an episode of MFD.     

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.     

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.     

Effect of 2-hydroxy-4-(methylthio)butanoate (HMTBa) on milk fat,rumen environment and biohydrogenation,and rumen protozoa in lactating cows fed diets with increased risk for milk fat depression

Biohydrogenation-induced milk fat depression (MFD) is a reduction in milk fat synthesis caused by bioactive fatty acids (FA) produced during altered ruminal microbial metabolism of unsaturated FA. The methionine analog 2-hydroxy-4-(methylthio)butanoate (HMTBa) has been shown to reduce the shift to the alternate biohydrogenation pathway and maintain higher milk fat yield in high-producing cows fed diets lower in fiber and higher in unsaturated FA. The objective of this experiment was to verify the effect of HMTBa on biohydrogenation-induced MFD and investigate associated changes in rumen environment and fermentation. Twenty-two rumen cannulated high-producing Holstein cows [168 ± 66 d in milk; 42 ± 7 kg of milk/d (mean ± standard deviation)] were used in a randomized design performed in 2 blocks (1 = 14 cows, 2 = 8 cows). Treatments were control (corn carrier) and HMTBa (0.1% of diet dry matter). The experiment included a 7-d covariate period followed by 3 phases that fed diets with increasing risk of MFD. The diet during the covariate and low-risk phase (7 d) was 32% neutral detergent fiber with no additional oil. The diet during the moderate-risk phase (17 d) was 29% neutral detergent fiber with 0.75% soybean oil. Soybean oil was increased to 1.5% for the last 4 d. The statistical model included the random effect of block and time course data were analyzed with repeated measures including the random effect of cow and tested the interaction of treatment and time. There was no effect of block or interaction of block and treatment or time. There was no overall effect of treatment or treatment by time interaction for dry matter intake, milk yield, and milk protein concentration and yield. Overall, HMTBa increased milk fat percent (3.2 vs. 3.6%) and yield (1,342 vs. 1,543 g/d) and there was no interaction of treatment and dietary phase. Additionally, HMTBa decreased the concentration of trans-10 18:1 in milk fat and rumen digesta. Average total ruminal concentration of volatile FA across the day and total-tract dry matter and fiber digestibility were not affected by HMTBa, but HMTBa increased average rumen butyrate and decreased propionate concentration and increased total protozoa abundance. Additionally, HMTBa increased the fractional rate of α-linoleic acid clearance from the rumen following a bolus predominantly driven by a difference in the first 30 min. Plasma insulin was decreased by HMTBa. In conclusion, HMTBa prevented the increase in trans FA in milk fat associated with MFD through a mechanism that is independent of total volatile FA concentration, but involves modification of rumen biohydrogenation. Decreased propionate and increased butyrate and ruminal protozoa may also have functional roles in the mechanism.     

Susceptibility to milk fat depression in dairy sheep and goats: Individual variation in ruminal fermentation and biohydrogenation

Small ruminants are susceptible to milk fat depression (MFD) induced by marine lipid supplementation. However, as observed in dairy cows, there is wide individual variation in the response to MFD-inducing diets, which may be due to individual differences in ruminal processes. Therefore, we compared the ruminal responses of goats and sheep with varying degrees of MFD extent to improve our understanding of this complex syndrome. Our specific aims were to attempt to elucidate whether pre-existing variations in ruminal fermentation and biohydrogenation determine a higher tolerance or susceptibility to MFD, and whether the severity of MFD depends exclusively on the response to the diet. The trial was conducted with 25 does and 23 ewes fed a basal diet without lipid supplementation for 3 wk (control period). Then, 2% fish oil (FO) was added to the same diet for 5 additional weeks (MFD period). Based on the extent of the elicited MFD (i.e., the percentage variation between milk fat concentrations recorded at the end of the control and MFD periods), the 5 most responsive (RESPON+) and the 5 least responsive (RESPON?) animals were selected within each species. On the last day of each period, ruminal fluid samples were collected to examine fermentation parameters and fatty acid profiles. In general, the individual degree of MFD in sheep and goats did not seem to be predetermined by traits related to ruminal fermentation and biohydrogenation, including fatty acids that may serve as biomarkers of microorganisms. Regarding differences in the response to FO, the results suggest no link between MFD susceptibility and concentration of biohydrogenation intermediates such as trans-10-containing C18, C20, and C22 metabolites. The explanation for individual responses based on a shortage of ruminal acetate and 18:0 for mammary uptake also seems to be dismissed, based on the lack of variation in these compounds between RESPON+ and RESPON?. However, the concentration of unsaturated fatty acids provided by FO (e.g., cis-9 16:1, cis-11 18:1, and 20:5n-3) was higher in the rumen of RESPON+ than RESPON? ewes and does. Thus, although further research is needed, the extent of biohydrogenation of these fatty acids might be associated with tolerance or susceptibility to MFD.     

Interaction of unsaturated fat or coconut oil with monensin in lactating dairy cows fed 12 times daily. II. Fatty acid flow to the omasum and milk fatty acid profile

Feeding animal-vegetable (AV) fat or medium-chain fatty acids (FA) to dairy cows can decrease ruminal protozoal counts. However, combining moderate to large amounts of AV fat with monensin (tradename: Rumensin, R) could increase the risk for milk fat depression (MFD), whereas it is not known if diets supplemented with coconut oil (CNO; rich in medium-chain FA) with R would cause MFD. In a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of treatments, 6 rumen-cannulated cows were fed diets without or with R (12 g/909 kg) and either control (no fat), 5% AV fat, or 5% CNO. Diets were balanced to have 21.5% forage neutral detergent fiber, 16.8% crude protein, and 42% nonfiber carbohydrates. Omasal flows of FA were characterized by an increased percentage of trans 18:1 for AV fat and CNO diets compared with the control, a higher percentage of 12:0 and 14:0 for CNO, and higher cis 18:1 for AV fat. Milk FA composition reflected the changes observed for omasal FA digesta flow. The de novo FA synthesis in the mammary gland was decreased by the main effects of R compared without R (averaged over fat treatments) and for added fat (AV fat and CNO) versus control (averaged over R). The percentages of 6:0, 8:0, and 10:0 in milk fat were lower for R and for AV fat and CNO compared with the control. The percentage of trans 18:1 FA in milk fat also higher for AV fat and CNO compared with the control. Against our hypotheses, the feeding of CNO did not prevent MFD, and few interactions between R and fat source were detected. The feeding of CNO did compromise ruminal biohydrogenation, with accumulation of trans 18:1 in the rumen and in milk fat.     

In vitro biohydrogenation of 13C-labeled α-linolenic acid in response to ruminal alterations associated with diet-induced milk fat depression in ewes

The basis for marine lipid-induced milk fat depression (MFD) has not been established yet, but recent reports suggest the putative contribution of shifts in the ruminal metabolism of α-linolenic acid (ALA). To test this hypothesis, an isotopic tracer approach was used in batch cultures of rumen microorganisms with inoculum collected from cannulated ewes fed either a total mixed ration without lipid supplementation (control inoculum) or the same diet supplemented with 2% of fish oil, which is known to cause MFD in lactating sheep (FO-MFD inoculum). The [1-¹³C]ALA was added at a dose of 1% of incubated dry matter and the proportions of ¹³C-labeled fatty acids (FA) were examined after 24 h of incubation, using complementary gas chromatography and gas chromatography-combustion isotope ratio mass spectrometry (GC-C-IRMS) analyses. Expected differences in FA profiles were confirmed between control and FO-MFD inocula (e.g., large decreases in 18:0 and increases in most 18:1 and 18:2 intermediates, particularly trans isomers, to fish oil supply). The biohydrogenation of ¹³ALA was extensive and yielded multiple metabolites, with a total of 48 chromatographic peaks showing ¹³C enrichment, regardless of the inoculum type. However, although ALA was biohydrogenated through common pathways under standard or MFD conditions, large changes in the accumulation of ¹³C-labeled FA suggest important differences in the relative contribution of each specific route. First, increased accumulation of trans-11-containing FA in FO-MFD incubations was accompanied by a general repression of the trans-13/14 pathway (supported by lower trans-13+14 18:1 or trans-11,trans-13 18:2 proportions), together with a lower production of cis FA (e.g., cis-9, -12, and -15 18:1 and some cis,cis 18:2). Results also downplayed the relevance of the trans-11 to trans-10 shift as an effective marker of diet-induced MFD in sheep, and challenged the involvement of some trans-10 intermediates (e.g., trans-10 18:1 and trans-10,cis-15 18:2) in the low-fat milk syndrome in this species. Conversely, increased abundance of most 18:3 intermediates (including some unidentified isomers) might be representative of ruminal alterations related to fish oil supplementation in ewes. Further research is necessary to examine the potential association between these findings and MFD in lactating animals.     

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.     

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.     

The effect of dietary fiber level on milk fat concentration and fatty acid profile of cows fed diets containing low levels of polyunsaturated fatty acids

The objective of this study was to investigate the effect of dietary fiber level on milk fat concentration, yield, and fatty acid (FA) profile of cows fed diets low in polyunsaturated fatty acid (PUFA). Six rumen-fistulated Holstein dairy cows (639 ± 51 kg of body weight) were used in the study. Cows were randomly assigned to 1 of 2 dietary treatments, a high fiber (HF; % of dry matter, 40% corn silage, 27% alfalfa silage, 7% alfalfa hay, 18% protein supplement, 4% ground corn, and 4% wheat bran) or a low fiber (LF; % of dry matter, 31% corn silage, 20% alfalfa silage, 5% alfalfa hay, 15% protein supplement, 19% ground wheat, and 10% ground barley) total mixed ration. The diets contained similar levels of PUFA. The experiment was conducted over a period of 4 wk. Ruminal pH was continuously recorded and milk samples were collected 3 times a week. Milk yield and dry matter intake were recorded daily. The rumen fluid in cows receiving the LF diet was below pH 5.6 for a longer duration than in cows receiving the HF diet (357 vs. 103 min/d). Neither diet nor diet by week interaction had an effect on milk yield (kg/d), milk fat concentration and yield, or milk protein concentration and yield. During wk 4, milk fat concentration and milk fat yield were high and not different between treatments (4.30% and 1.36 kg/d for the HF treatment and 4.31% and 1.33 kg/d for the LF treatment, respectively). Cows receiving the LF diet had greater milk concentrations (g/100 g of FA) of 7:0; 9:0; 10:0; 11:0; 12:0; 12:1; 13:0; 15:0; linoleic acid; FA <C16; and PUFA; and lower concentrations of iso 15:0; 18:0; trans-9 18:1; cis-9, trans-11 conjugated linoleic acid (CLA); trans-9, cis-12 18:2; 20:0; and cis-9 20:1 compared with cows receiving the HF diet. Milk concentrations (g/100 g of FA) of total trans 18:1; trans-10 18:1; trans-11 18:1; trans-10, cis-12 CLA, and trans-9, cis-11 CLA were not different between treatments. The study demonstrated that cows fed a diet low in fiber and low in PUFA may exhibit subacute ruminal acidosis and moderate changes to milk fatty acid profile but without concomitant milk fat depression. The changes in FA profile may be useful for the diagnosis of SARA even in the absence of milk fat depression.