Short communication: Effect of blackberry and pomegranate oils on vaccenic acid formation in a single-flow continuous culture fermentation system

A single-flow continuous culture fermenter system was used to study the effect of blackberry and pomegranate oils on vaccenic acid (trans-11 C18:1; VA) formation. Four continuous culture fermenters were used in a 4 × 4 Latin square design with 4 periods of 10 d each. Diets were (1) control (CON), (2) control plus soybean oil (SBO), (3) control plus blackberry oil (BBO), and (4) control plus pomegranate oil (PMO). Oil supplements were added at 30 g/kg of diet dry matter. Effluents were collected from each fermenter during the last 3 d of each period and analyzed for nutrient and fatty acid composition. The concentration of VA in effluents increased with oil supplements and was greatest with the BBO diet. The concentration of stearic acid (C18:0) increased with the addition of soybean oil but decreased with the addition of pomegranate oil compared with the CON diet. The concentration of cis-9,trans-11 conjugated linoleic acid increased with oil supplements and was greatest with the PMO diet. In conclusion, all 3 oil sources were effective in increasing the production of VA. The effect of PMO and BBO on VA may have resulted in part from inhibition of the final step in the biohydrogenation of VA to stearic acid.     

Short communication: docosahexaenoic acid promotes vaccenic acid accumulation in mixed ruminal cultures when incubated with linoleic acid

Previous studies found that feeding dairy cows a blend of fish and soybean oils enhanced milk vaccenic acid (VA) and conjugated linoleic acid (CLA) concentrations more than when the oils were fed separately. In these studies, the authors concluded that a component in fish oil was stimulating ruminal VA production from other sources of unsaturated fatty acids; however, that component was not identified. The objective of this study was to determine whether docosahexaenoic acid (DHA), an omega-3 fatty acid (FA) in fish oil, is the active component that promotes trans-C18:1 FA, VA in particular, accumulation using cultures of mixed ruminal microorganisms. Treatments consisted of control, control plus 5 mg of DHA (DH), control plus 30 mg of soybean oil (SBO), and control plus 5 mg of DHA and 30 mg of SBO (DHSBO). Treatments were incubated in triplicate in 125-mL flasks, and 5 mL of culture contents was taken at 0 and 24 h for fatty acid analysis by gas-liquid chromatography. After 24 h of incubation, the level of trans-C18:1 FA (14.1 and 11.7 mg/culture) and VA (13.0 and 10.2 mg/culture) increased more with added DHA than with added SBO, respectively. Combining DHA and SBO yielded higher quantities of trans-C18:1 FA (21.3 mg/culture) and VA (19.8 mg/culture) in the cultures than either fat source alone. These data suggest that DHA is the component in fish oil that promotes VA accumulation when incubated with linoleic acid.     

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

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

Effects of chemically or technologically treated linseed products and docosahexaenoic acid addition to linseed oil on biohydrogenation of C18:3n-3 in vitro

Rumen biohydrogenation kinetics of C18:3n-3 from several chemically or technologically treated linseed products and docosahexaenoic acid (DHA; C22:6n-3) addition to linseed oil were evaluated in vitro. Linseed products evaluated were linseed oil, crushed linseed, formaldehyde treated crushed linseed, sodium hydroxide/formaldehyde treated crushed linseed, extruded whole linseed (2 processing variants), extruded crushed linseed (2 processing variants), micronized crushed linseed, commercially available extruded linseed, lipid encapsulated linseed oil, and DHA addition to linseed oil. Each product was incubated with rumen liquid using equal amounts of supplemented C18:3n-3 and fermentable substrate (freeze-dried total mixed ration) for 0, 0.5, 1, 2, 4, 6, 12, and 24 h using a batch culture technique. Disappearance of C18:3n-3 was measured to estimate the fractional biohydrogenation rate and lag time according to an exponential model and to calculate effective biohydrogenation of C18:3n-3, assuming a fractional passage rate of 0.060/h. Treatments showed no differences in rumen fermentation parameters, including gas production rate and volatile fatty acid concentration. Technological pretreatment (crushing) followed by chemical treatment applied as formaldehyde of linseed resulted in effective protection of C18:3n-3 against biohydrogenation. Additional chemical pretreatment (sodium hydroxide) before applying formaldehyde treatment did not further improve the effectiveness of protection. Extrusion of whole linseed compared with extrusion of crushed linseed was effective in reducing C18:3n-3 biohydrogenation, whereas the processing variants were not different in C18:3n-3 biohydrogenation. Crushed linseed, micronized crushed linseed, lipid encapsulated linseed oil, and DHA addition to linseed oil did not reduce C18:3n-3 biohydrogenation. Compared with the other treatments, docosahexaenoic acid addition to linseed oil resulted in a comparable trans11,cis15-C18:2 biohydrogenation but a lesser trans10+11-C18:1 biohydrogenation. This suggests that addition of DHA in combination with linseed oil was effective only in inhibiting the last step of biohydrogenation from trans10+11-C18:1 to C18:0.     

Effect of in vitro docosahexaenoic acid supplementation to marine algae-adapted and unadapted rumen inoculum on the biohydrogenation of unsaturated fatty acids in freeze-dried grass

The objective of this study was to examine the ruminal biohydrogenation of linoleic (18:2n-6) and linolenic (18:3n-3) acid during in vitro incubations with rumen inoculum from dairy cattle adapted or not to marine algae and with or without additional in vitro docosahexaenoic acid (DHA, 22:6n-3) supplementation. Treatments were incubated in 100-mL flasks containing 400 mg of freeze-dried grass, 5 mL of strained ruminal fluid, and 20 mL of phosphate buffer. Ruminal fluid was collected just before the morning feeding from 3 cows receiving a control diet (49% ryegrass silage, 39% corn silage, 1% straw, and 11% concentrate, fresh-weight basis) supplemented with marine algae for 21 d (adapted rumen fluid, aRF) or from the same cows receiving the control diet only for 14 d after marine algae supplementation was stopped (unadapted rumen fluid, uRF). In half of the incubation flasks, pure DHA (5 mg) was added as an oil-ethanol solution (100 mL). Incubations were carried out during 0, 0.5, 1, 2, 4, 6, and 24 h. After 24 h, in vitro addition of DHA resulted in greater amounts (mg/incubation) of 18:3n-3 (0.23, 0.43, 0.26, and 0.34 for aRF, aRF+DHA, uRF, and uRF+DHA), 18:2n-6 (0.14, 0.22, 0.15, and 0.20 for aRF, aRF+DHA, uRF, and uRF+DHA) and trans-11, cis-15-18:2 (0.27, 2.40, 0.06, and 2.21 for aRF, aRF+DHA, uRF, and uRF+DHA), whereas no effect of inoculum source was observed. Trans-11-18:1 accumulated after 24 h when aRF was incubated irrespective of in vitro DHA supplementation, whereas in incubations with uRF, accumulation of trans-11-18:1 only occurred when DHA was added (6.40, 4.35, 1.06, and 3.91 for aRF, aRF+DHA, uRF, and uRF+DHA). The increased amounts of trans-11-18:1 were due to the strong inhibition of the reduction to 18:0 because no 18:0 was formed when trans-11-18:1 accumulated after 24 h. The results of the current experiment shows hydrogenation of trans-11, cis-15-18:2 occurred in the absence of in vitro DHA only, whereas substantial hydrogenation of trans-11-18:1 to 18:0 only took place in incubations without DHA and with unadapted rumen inoculum, confirming the higher sensitivity of the latter process to DHA.     

Fatty acid composition of milk from multiparous Holstein cows treated with bovine somatotropin and fed n-3 fatty acids in early lactation

Multiparous cows (n = 59) were blocked by expected calving date and previous milk yield and assigned randomly to treatments to determine effects of bovine somatotropin (bST; Posilac, Monsanto Animal Agricultural Group, St. Louis, MO) and source of dietary fat on milk fatty acid composition during the first 140 d in milk. Diets were provided from calving and included whole, high-oil sunflower seeds (SS; 10% of dietary dry matter; n-6/n-3 ratio of 4.6) as a source of linoleic acid or a mixture of Alifet-High Energy and Alifet-Repro (AF; Alifet USA, Cincinnati, OH; 3.5 and 1.5% of dietary dry matter, respectively; n-6/n-3 ratio of 2.6) as a source of protected n-3 fatty acids (15.7% 18:3, 1.3% 20:5, and 1.3% 22:6). Treatments were derived from a 2 × 2 combination of supplemental fat source (SS, AF) and with 0 (SSN, AFN) or 500 (SSY, AFY) mg of bST administered every 10 d from 12 to 70 d in milk and at 14-d intervals thereafter. Milk fatty acid composition was determined in samples collected from 32 cows (8 complete blocks) during wk 2, 8, and 20 of lactation. Data were analyzed as repeated measures using mixed model procedures to determine the effects of diet, bST, week of lactation, and their interactions. Proportions of 18:3 (4.02 vs. 3.59 ± 0.16%), 20:5 (0.52 vs. 0.41 ± 0.02%), and 22:6 (0.11 vs. 0.02 ± 0.02%) were greater and the n-6/n-3 fatty acid ratio (7.40 vs. 8.80 ± 0.30) was reduced in milk from cows fed AF compared with SS. Proportions of de novo-synthesized fatty acids increased and preformed fatty acids decreased as lactation progressed, but bST administration delayed this shift in origin of milk fatty acids. Transfer efficiency of 18:3, 20:5, and 22:6 from AF to milk fat averaged 36.2, 4.9, and 5.2%, respectively. These efficiencies increased as lactation progressed, but were delayed by bST. Apparent mammary Δ⁹-desaturase activity and milk conjugated linoleic acid (cis-9, trans-11 conjugated linoleic acid) content increased through the first 8 wk of lactation. Based on the product-to-substrate ratio of 14:1/14:0 fatty acids in milk, there was an interaction of diet and bST because bST decreased apparent Δ⁹-desaturase activity in SSY cows but increased it in AFY cows (0.10, 0.09, 0.08, and 0.09 ± 0.01 for SSN, SSY, AFN, and AFY, respectively). Feeding Alifet-Repro increased n-3 fatty acids in milk and bST prolonged the partitioning of dietary fatty acids into milk fat.     

Long-term effects of feeding monensin on milk fatty acid composition in lactating dairy cows

The objective of this study was to determine the long-term effects of feeding monensin on milk fatty acid (FA) profile in lactating dairy cows. Twenty-four lactating Holstein dairy cows (1.46 ± 0.17 parity; 620 ± 5.9 kg of live weight; 92.5 ± 2.62 d in milk) housed in a tie-stall facility were used in the study. The study was conducted as paired comparisons in a completely randomized block design with repeated measurements in a color-coded, double blind experiment. The cows were paired by parity and days in milk and allocated to 1 of 2 treatments: 1) the regular milking cow total mixed ration (TMR) with a forage-to-concentrate ratio of 60:40 (control TMR; placebo premix) vs. a medicated TMR [monensin TMR; regular TMR + 24 mg of Rumensin Premix per kg of dry matter (DM)] fed ad libitum. The animals were fed and milked twice daily (feeding at 0830 and 1300 h; milking at 0500 and 1500 h). Milk samples were collected before the introduction of treatments and monthly thereafter for 6 mo and analyzed for FA composition. Monensin reduced the percentage of the short-and medium-chain saturated FA 7:0, 9:0, 15:0, and 16:0 in milk fat by 26, 35, 19, and 6%, respectively, compared with the control group. Monensin increased the percentage of the long-chain saturated FA in milk fat by 9%, total monounsaturated FA by 5%, total n-6 polyunsaturated FA (PUFA) by 19%, total n-3 PUFA by 16%, total cis-18:1 by 7%, and total conjugated linoleic acid (CLA) by 43% compared with the control group. Monensin increased the percentage of docosahexaenoic acid (22:6n-3), docosapentaenoic acid (22:5n-3), and cis-9, trans-11 CLA in milk fat by 19, 13, and 43%, respectively, compared with the control. These results suggest that monensin was at least partly effective in inhibiting the biohydrogenation of unsaturated FA in the rumen and consequently increased the percentage of n-6 and n-3 PUFA and CLA in milk, thus enhancing the nutritional properties of milk with regard to human health.     

Effects of fatty acid oxidation products (green odor) on rumen bacterial populations and lipid metabolism in vitro

This study investigated the effects of green odor fatty acid oxidation products (FAOP) from cut grass on lipid metabolism and microbial ecology using in vitro incubations of rumen microorganisms. These compounds have antimicrobial roles in plant defense, and we hypothesized that they may influence rumen lipid metabolism. Further, they may partially explain the higher levels of conjugated linoleic acid cis-9, trans-11 in milk from cows grazing pasture. The first of 2 batch culture experiments screened 6 FAOP (1 hydroperoxide, 3 aldehydes, 1 ketone, and 1 alcohol) for effects on lipid profile, and in particular C₁₈ polyunsaturated fatty acid biohydrogenation. Experiment 2 used the most potent FAOP to determine effects of varying concentrations and identify relationships with effects on microbial ecology. Batch cultures contained anaerobic buffer, rumen liquor, and FAOP to a final concentration of 100 μM for experiment 1. Triplicates for each compound and controls (water addition) were incubated at 39°C for 6 h. The hydroperoxide (1,2-dimethylethyl hydroperoxide, 1,2-DMEH) and the long chain aldehyde (trans-2 decenal) had the largest effects on lipid metabolism with significant increases in C_18:0 and C_18:1trans and reductions in C_12:0, C_14:0, C_16:0, C_18:1cis, C_18:2n-6, C_18:3n-3, C_20:0 and total branch and odd chain fatty acids compared with the control. This was associated with significantly higher biohydrogenation of C₁₈ polyunsaturated fatty acid. In experiment 2, 1,2-DMEH was incubated at 50, 100, and 200 μM for 2, 6, and 24 h. Increasing 1,2-DMEH concentration resulted in a significant linear increase in C_18:1trans-10, trans-11, conjugated linoleic acid, and C_18:0 and a linear decrease in C_18:2n-6 and C_18:3n-3, although the scale of this response declined with time. Microbial profiling techniques showed that 1,2-DMEH at concentrations of 100 and 200 μM changed the microbial community from as early as 2 h after addition, though microbial biomass remained similar. These preliminary studies have shown that FAOP can alter fatty acid biohydrogenation in the rumen. This change was associated with changes in the microbial population that were detected through DNA and branched- and odd-chain fatty acid profiling approaches.     

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

In an attempt to develop strategies for enhancing the nutritional value of sheep milk fat, dairy ewe diet was supplemented with 3 incremental levels of marine algae (MA), in combination with sunflower oil, to evaluate the effects of these marine lipids on milk fatty acid (FA) profile and animal performance. Fifty Assaf ewes in mid lactation were distributed in 10 lots of 5 animals each and allocated to 5 treatments (2 lots per treatment): no lipid supplementation (control) or supplementation with 25 g of sunflower oil/kg of DM plus 0 (SO), 8 (SOMA₁), 16 (SOMA₂), or 24 (SOMA₃) g of MA (56.7% ether extract)/kg of DM. Milk production and composition, including FA profile, were analyzed on d 0, 3, 7, 14, 21, and 28 of treatment. Neither intake nor milk yield were significantly affected by lipid addition, but all MA supplements decreased milk fat content from d 14 onward, reaching a 30% reduction after 28 d on SOMA₃. This milk fat depression might be related not only to the joint action of some putative fat synthesis inhibitors, such as trans-9,cis-11 C18:2 and probably trans-10 C18:1, but also to the limited ability of the mammary gland to maintain a desirable milk fat fluidity, that would have been caused by the noticeable increase in trans-C18:1 together with the lowered availability of stearic acid for oleic acid synthesis through Δ⁹-desaturase. Furthermore, all lipid supplements, and mainly MA, reduced the secretion of de novo FA (C6:0-C14:0) without increasing the yield of preformed FA (>C16). Supplementation with sunflower oil plus MA resulted in larger increases in cis-9,trans-11 C18:2 than those observed with sunflower oil alone, achieving a mean content as high as 3.22% of total FA and representing a more than 7-fold increase compared with the control. Vaccenic acid (trans-11 C18:1) was also significantly enhanced (on average +794% in SOMA treatments), as was C22:6 n-3 (DHA) content, although the transfer efficiency of the latter, from the diets to the milk, was very low (5%). However, the highest levels of MA inclusion (SOMA₂ and SOMA₃) reduced the milk n-6:n-3 ratio, but MA supplements caused an important increase in trans-10 C18:1, which would rule out the possibility that this milk has a healthier fat profile before determining the specific role of each individual FA and ensuring that this trans-FA is at least innocuous in relation to cardiovascular disease risk.     

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

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

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

Ruminant diet supplementation with sunflower oil (SO) and fish oil (FO) has been reported as a good strategy for enhancing some milk fat compounds such as conjugated linoleic acid (CLA) and n-3 polyunsaturated fatty acids in dairy cows, but no information is available regarding dairy sheep. In this work, ewe diet was supplemented with FO, alone or in combination with SO, with the aim of improving milk nutritional value and evaluating its effect on animal performance. Sixty-four Assaf ewes in mid lactation, fed a high-concentrate diet, were distributed in 8 lots of 8 animals each and assigned to 4 treatments (2 lots/treatment): no lipid supplementation (control) or supplementation with 20 g of SO/kg (SO), 10 g of FO/kg (FO), or 20 g of SO plus 10 g of FO/kg (SOFO). Milk production and composition, including a complete fatty acid profile, were analyzed on d 0, 3, 7, 14, 21, and 28 of treatments. Supplementation with FO tended to reduce dry matter intake compared with the control treatment (−15%), and its use in combination with SO (SOFO) resulted in a significant decrease in milk yield as well (−13%). All lipid supplements reduced milk protein content, and FO also reduced milk fat content by up to 21% alone (FO) and 27% in combination with SO (SOFO). Although the mechanisms involved in FO-induced milk fat depression are not yet well established, the observed increase in some milk trans-FA that are putative inhibitors of milk fat synthesis, such as trans-9,cis-11 CLA, and the 63% decrease in C18:0 (consistent with the theory of reduced milk fat fluidity) may be involved. When compared with the control, lipid supplementation remarkably improved the milk content of rumenic acid (cis-9,trans-11 CLA; up to 4-fold increases with SO and SOFO diets), whereas FO-containing diets also increased milk n-3 polyunsaturated fatty acids, mainly docosahexaenoic acid (with mean contents of 0.29 and 0.38% of total fatty acids for SOFO and FO, respectively), and reduced the n-6:n-3 FA ratio to approximately half the control value. All lipid supplements resulted in high levels of some trans-FA, mainly trans-11 C18:1 (vaccenic acid) but also trans-10 C18:1.     

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

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

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: III. Fatty acid distribution

Our objective was to examine effects of exogenous bovine somatotropin (bST), pregnancy, and dietary fatty acids on fatty acid distribution in various tissues of lactating dairy cows. Two diets were fed, starting about 17 d in milk (DIM), in which oil of whole cottonseed (control diet) was compared with a calcium salt of fish oil-enriched lipid (FO; 1.9% of dietary DM). Starting at 44 ± 5 DIM, ovulation was synchronized with a presynchronization plus Ovsynch protocol (d 0 = time of synchronized ovulation). Some cows were inseminated (77 ± 12 DIM) to create a pregnant group. On d 0 and 11, cows received bST (500 mg) or no bST, and were killed on d 17 (94 ± 12 DIM). Number of cows in control group was 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows; and for the FO diet: 4 bST-treated (bST-FO-C) and 5 non-bST-treated cyclic (no bST-FO-C) cows. At slaughter, samples of endometrium, liver, muscle, s.c. adipose, internal adipose, and mammary gland were collected. Milk was collected at 75 ± 5 DIM. Gas chromatography was used to determine fatty acid percentages in tissues and milk fat. Endometrium from the cows fed FO had increased proportions of C20:5 and C22:6, whereas C20:4 was decreased. Injections of bST reduced both C18:2 and the n-6:n-3 ratio, but increased C22:6 in endometrium of cyclic control-fed, but not pregnant cows. In addition, FO decreased the n-6:n-3 ratio in all tissues and milk fat except for s.c. and internal adipose tissue. Cows fed FO also had increased C18:3, C20:5, and C22:6 in the liver and mammary tissue, and C18:3 and C22:6 were increased in the milk fat. The FO diet decreased the Δ⁹-desaturase index [(product of Δ⁹-desaturase]/(product of Δ⁹-desaturase + substrate of Δ⁹-desaturase]; DIX) in muscle and s.c. tissues, accompanied by an increase in saturated fatty acid (SFA) percentage. In addition, FO diet decreased DIX in the endometrium. In mammary and internal adipose tissues, bST increased DIX in cyclic control-fed cows, whereas bST decreased DIX in FO-fed cows, with no difference in the concentration of SFA and UNSFA. Cis-9, trans-11 conjugated linoleic acid was increased in milk fat, but decreased in the muscle and s.c. adipose tissue of FO-fed cows. The FO-enriched lipid, bST treatment, and early pregnancy can alter fatty acid percentages and distributions that may alter tissue functionality and functional nutrients of consumer products.     

Uptake and utilization of trans octadecenoic acids in lactating dairy cows

Trans fatty acids (FA) arise in ruminant-derived foods as a consequence of rumen biohydrogenation and are of interest because of their biological effects and potential role in chronic human diseases. Our objective was to compare 2 trans FA, elaidic acid (EA; trans-9 18:1) and vaccenic acid (VA; trans-11 18:1), with oleic acid (OA; cis-9 18:1) relative to plasma lipid transport and mammary utilization for milk fat synthesis. Three ruminally cannulated, Holstein dairy cows, 259 ± 6 DIM (mean ± SEM), were randomly assigned in a 3 × 3 Latin square design. Treatments were a 4-d abomasal infusion of 1) OA (45.5 g/d), 2) EA (41.7 g/d), and 3) VA (41.4 g/d). Milk samples were collected at each milking and blood samples were collected at the start and end of each treatment period. The proportions of total plasma FA associated with each plasma lipid fraction at baseline (pretreatment) were 62.6 ± 0.6% phospholipids, 26.1 ± 0.6% cholesterol esters, 9.8 ± 0.4% triglycerides, and 1.5 ± 0.1% nonesterified fatty acids; these values were unaffected by treatment. There were striking differences in the FA composition of the individual plasma lipid fractions and in the distribution of specific 18-carbon FA among the lipid fractions. Infusion of treatment isomers caused their specific increase in the various plasma lipid fractions but had no effect on milk production variables, including milk fat yield and content. Transfer efficiency of infused OA, EA, and VA to milk fat averaged 65.5 ± 3.0%, 59.7 ± 1.5%, and 54.3 ± 0.6%, respectively. For the VA infusion, 24.6 ± 1.1% of the transfer was accounted for by the increased yield of cis-9, trans-11 conjugated linoleic acid in milk fat, consistent with its endogenous synthesis from VA via the mammary enzyme Δ⁹-desaturase. Notably, linoleic acid (18:2n-6) and linolenic acid (18:3n-3) accounted for 47.7% of total plasma FA, but only 2.6% of FA in milk. Overall, results demonstrate clear differences in plasma transport and mammary uptake and utilization of 18-carbon FA, and these relate to the location, orientation, and number of double bonds.     

Effect of a high-palmitic acid fat supplement on milk production and apparent total-tract digestibility in high- and low-milk yield dairy cows

The effect of a high-palmitic acid fat supplement was tested in 12 high-producing (mean = 42.1 kg/d) and 12 low-producing (mean = 28.9 kg/d) cows arranged in a replicated 3 × 3 Latin square design. Experimental periods were 21 d, with 18 d of diet adaptation and 3 d of sample collection. Treatments were (1) control (no supplemental fat), (2) high-palmitic acid (PA) supplement (84% C16:0), and (3) Ca salts of palm fatty acid (FA) supplement (Ca-FA). The PA supplement had no effect on milk production, but decreased dry matter intake by 7 and 9% relative to the control in high- and low-producing cows, respectively, and increased feed efficiency by 8.5% in high-producing cows compared with the control. Milk fat concentration and yield were not affected by PA relative to the control in high- or low-producing cows, although PA increased the yield of milk 16-C FA by more than 85 g/d relative to the control. The Ca-FA decreased milk fat concentration compared with PA in high-, but not in low-producing cows. In agreement, Ca-FA dramatically increased milk fat concentration of trans-10 C18:1 and trans-10, cis-12 conjugated linoleic acid (>300%) compared with PA in high-producing cows, but not in low-producing cows. No effect of treatment on milk protein concentration or yield was detected. The PA supplement also increased 16-C FA apparent digestibility by over 10% and increased total FA digestibility compared with the control in high- and low-producing cows. During short-term feeding, palmitic acid supplementation did not increase milk or milk fat yield; however, it was efficiently absorbed, increased feed efficiency, and increased milk 16-C FA yield, while minimizing alterations in ruminal biohydrogenation commonly observed for other unsaturated fat supplements. Longer-term experiments will be necessary to determine the effects on energy balance and changes in body reserves.     

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

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

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

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

Effects of extruded linseed supplementation on n-3 fatty acids and conjugated linoleic acid in milk and cheese from ewes

The objective of this study was to assess the effects of dietary supplementation of extruded linseed on animal performance and fatty acid (FA) profile of ewe milk for the production of n-3 FA- and conjugated linoleic acid-enriched cheeses. A Manchega ewe flock (300 animals) receiving a 60:40 forage:concentrate diet was divided into 3 groups supplemented with 0, 6, and 12 g of extruded linseed/100 g of dry matter for the control, low, and high extruded linseed diets, respectively. Bulk and individual milk samples from 5 dairy ewes per group were monitored at 7, 14, 28, 45, and 60 d following supplementation. Manchego cheeses were made with bulk milk from the 3 treatment groups. Milk yield increased in dairy ewes receiving extruded linseed. Milk protein, fat, and total solids contents were not affected by linseed supplementation. Milk contents of α-linolenic acid increased from 0.36 with the control diet to 1.91% total FA with the high extruded linseed diet. Similarly, cis-9 trans-11 C18:2 rose from 0.73 to 2.33% and its precursor in the mammary gland, trans-11 C18:1, increased from 1.55 to 5.76% of total FA. This pattern occurred with no significant modification of the levels of trans-10 C18:1 and trans-10 cis-12 C18:2 FA. Furthermore, the high extruded linseed diet reduced C12:0 (−30%), C14:0 (−15%) and C16:0 (−28%), thus significantly diminishing the atherogenicity index of milk. The response to linseed supplementation was persistently maintained during the entire study. Acceptability attributes of n-3-enriched versus control cheeses ripened for 3 mo were not affected. Therefore, extruded linseed supplementation seems a plausible strategy to improve animal performance and nutritional quality of dairy lipids in milk and cheese from ewes.     

Effect of Dietary Starch or Micro Algae Supplementation on Rumen Fermentation and Milk Fatty Acid Composition of Dairy Cows

Two experiments with rumen-fistulated dairy cows were conducted to evaluate the effects of feeding docosahexaenoic acid (DHA; C22:6 n-3)-enriched diets or diets provoking a decreased rumen pH on milk fatty acid composition. In the first experiment, dietary treatments were tested during 21-d experimental periods in a 4 × 4 Latin square design. Diets included a control diet, a starch-rich diet, a bicarbonate-buffered starch-rich diet, and a diet supplemented with DHA-enriched micro algae [Schizochytrium sp., 43.0 g/kg of dry matter intake (DMI)]. Algae were supplemented directly through the rumen fistula. The total mixed ration consisted of grass silage, corn silage, soybean meal, and a standard or glucogenic concentrate. The glucogenic and buffered glucogenic diet had no effect on rumen fermentation and milk fatty acid composition because, unexpectedly, no reduced rumen pH was detected. The algae diet had no effect on rumen pH but provoked decreased butyrate and increased isovalerate molar proportions in the rumen. In addition, algae supplementation affected rumen biohydrogenation of linoleic and linolenic acid as reflected in the modified milk fatty acid composition toward increased conjugated linoleic acid (CLA) cis-9 trans-11, CLA trans-9 cis-11, C18:1 trans-10, C18:1 trans-11, and C22:6 n-3 concentrations. Concomitantly, on average, a 45% decrease in DMI and milk yield was observed. Based on these drastic and impractical results, a second animal experiment was performed for 20 d in which 9.35 g/kg of total DMI of algae were incorporated in the concentrate and supplemented to 3 rumen-fistulated cows. Algae concentrate feeding increased rumen pH, which was associated with decreased rumen short-chain fatty acid concentrations. Moreover, a different shift in rumen short-chain fatty acid proportions was observed compared with the first experiment because molar proportions of butyrate, isobutyrate, and isovalerate increased, whereas acetate molar proportion decreased. The milk fatty acid profile changed as in experiment 1. However, the decrease in DMI and milk yield was less pronounced (on average 10%) at this algae supplementation level, whereas milk fat percentage decreased from 47.9 to 22.0 g/kg of milk after algae treatment. In conclusion, an algae supplementation level of about 10 g/kg of DMI proved effective to reduce the milk fat content and to modify the milk fatty acid composition toward increased CLA cis-9 trans-11, C18:1 trans, and DHA concentrations.     

Docosahexaenoic acid elevates trans-18:1 isomers but is not directly converted into trans-18:1 isomers in ruminal batch cultures

Pathways of docosahexaenoic (DHA) biohydrogenation are not known; however, DHA is metabolized by ruminal microorganisms. The addition of DHA to the rumen alters the fatty acid profile of the rumen and milk and leads to increased trans-18:1 isomers, particularly trans-11 18:1. This study included 2 in vitro experiments to identify if the increase in trans-11 C18:1 was due to DHA being converted into trans-11 18:1 or if DHA stimulated trans-11 products from biohydrogenation of other fatty acids. In each experiment, ruminal microorganisms collected from a lactating Holstein cow were incubated in 10-mL batch cultures for 0, 6, 24, and 48 h and a uniformly ¹³C-labeled DHA was added to the cultures at 0 h as a metabolic tracer. Experiment 1 tested 0.5% DHA supplementation and experiment 2 examined 1, 2, and 3% DHA supplementation to determine if the level of DHA effected its conversion into trans-11 18:1. In both experiments, any fatty acid that was enriched with the ¹³C label was determined to arise from DHA. Palmitic (C16:0), stearic (C18:0), all trans-18:1, eicosanoic (C20:0), and docosanoic (C22:0) acids were examined for enrichment. In experiment 1, the amount of trans-18:1 isomers increased 0.415 mg from 0 to 48 h; however, no label was found in trans-18:1 at any time. Docosanoic acid was highly enriched at 24 h and 48 h to 20.2 and 16.3%. Low levels of enrichment were found in palmitic and stearic acids. In experiment 2, trans-18:1 isomers increased 185, 256, and 272% from 0 to 48 h when DHA was supplemented at 1, 2, and 3%, respectively; however, as in experiment 1, no enrichment occurred of any trans-18:1 isomer. In experiment 2, low levels of label were found in palmitic and stearic acids. Enrichment of docosanoic acid decreased linearly with increased DHA supplementation. These studies showed that trans-18:1 fatty acids are not produced from DHA, supporting that DHA elevates trans-18:1 by modifying biohydrogenation pathways of other polyunsaturated fatty acids.