Effects of substrate, passage rate, and pH in continuous culture on flows of conjugated linoleic acid and trans C18:1

A dual-flow continuous culture system consisting of 4 fermenters was used in a 4 x4 Latin square design. The objective of the research was to evaluate the effects of solid dilution rate (SDR), pH, and concentration of linoleic acid (LA) in the feed mixture on the production of conjugated linoleic acid (CLA) and trans-C18:1. The 4 treatments were 1) control = pH 6.5, 1% LA, 4%/h SDR; 2) high solid dilution rate (HSDR) = pH 6.5, 1% LA, 8%/h SDR; 3) high linoleic acid (HLA) = pH 6.5, 3% LA, 4%/h SDR; and 4) low pH (LPH) = pH 5.8, 1% LA, 4%/h SDR. Inoculum was collected 6 h after feeding from a cow fed 40% alfalfa hay and 60% grain. Liquid dilution rate was held at 0.12/h. All treatments except HLA contained 2% tallow. The LA was dissolved in buffer and continuously infused into the fermenters. The CLA flows were 16.5, 20.4, 23.2, and 25.2 mg/d for control, HSDR, HLA, and LPH, respectively. Compared with control, LPH increased flows of CLA, cis-C18:1, and C18:2, and decreased flow of C18:0. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibilities were not affected by pH. The HSDR tended to increase CLA flow compared to control, possibly because a shorter solid retention time led to incomplete biohydrogenation (BH). The NDF and ADF digestibilities and bacterial numbers were reduced by HSDR. With more LA available as a substrate for CLA, HLA resulted in a higher flow of CLA than control. The HLA resulted in the highest acid detergent fiber and fatty acid digestibilities, bacterial numbers, and BH. Increasing solids passage rate, reducing pH, and increasing dietary LA appears to increase in vitro CLA production.     

Evaluating the conjugated linoleic acid and trans 18:1 isomers in milk fat of dairy cows fed increasing amounts of sunflower oil and a constant level of fish oil

The objective was to evaluate different levels of sun-flower oil (SFO) in dairy rations to increase vaccenic (trans-11-18:1) and rumenic acids (cis-9,trans-11-18:2) in milk fat, and assess the content and composition of other trans-octadecenoic (trans-18:1) and conjugated linoleic acids (CLA) isomers. Eighty lactating Holstein cows were fed control diets for 4 wk and then placed on 4 diets for 38 d; milk fat was analyzed after 10 and 38 d. The treatments were: control, 1.5% SFO plus 0.5% fish oil (FO), 3% SFO plus 0.5% FO, and 4.5% SFO plus 0.5% FO. The forage-to-concentrate ratio was 50:50 and consisted of barley/alfalfa/hay silage and corn/barley grain concentrate. There were no differences in milk production. Supplementation of SFO/FO reduced milk fat compared with respective pretreatment periods, but milk protein and lactose levels were not affected. There was a linear decrease in all short- and medium-chain saturated fatty acids (SFA) in milk fat after 10 d (25.5, 24.1, 20.2, and 16.7%) and a corresponding linear increase in total trans-18:1 (5.2, 9.1, 14.1, and 21.3%) and total CLA (0.7, 1.9, 2.4, and 3.9%). The other FA in milk fat were not affected. Separation of trans-18:1 isomers was achieved by combination of gas chromatography (GC; 100-m highly polar capillary column) and prior separation of trans FA by silver ion-thin layer chromatography followed by GC. The CLA isomers were resolved by a combination of GC and silver ion-HPLC. The trans-11- and trans-10-18:1 isomers accounted for ∼50% of the total trans-18:1 increase when SFO/FO diets were fed. On continued feeding to 38 d, trans-11-18:1 increased with 1.5% SFO/FO, stayed the same with 3%, and declined with 4.5% SFO/FO. Rumenic acid showed a similar pattern on continued feeding as trans-11-18:2; levels increased to 0.43, 1.5, 1.9, and 3.4% at 10 d and to 0.42, 2.15, 2.09, and 2.78% at 38 d. Rumenic acid was the major CLA isomer in all 4 diets: 66, 77, 78 and 85%. The CLA isomers trans-7,cis-9-, trans-9,cis-11-, trans-10,cis-12-, trans-11,trans-13-, and trans-9,trans-11-/trans-10,trans-12-18:2 also increased from 0.18 (control) to 0.52% (4.5% SFO/FO). Milk fat produced from 3% SFO/FO appeared most promising: trans-11-18:1 and cis-9,trans-11-18:2 increased 4.5-fold, total SFA reduced 18%, and moderate levels of trans-10-18:1 (3.2%), other trans-18:1 (6.6%) and CLA isomers (0.5%) were observed, and that composition remained unchanged to 38 d. The 4.5% SFO/FO diet produced higher levels of trans-11-18:1 and cis-9,trans-11-18:2, a 28% reduction in SFA, and similar levels of other trans-18:1 (9.2%) and CLA isomers (0.52%), but the higher levels of trans-11-18:1 and cis-9,trans-11-18:2 were not sustained. A stable milk fat quality was achieved by feeding moderate amounts of SFO (3% of DM) in the presence of 0.5% FO that had 4% vaccenic and 2% rumenic acids.     

High-concentrate diets and polyunsaturated oils alter trans and conjugated isomers in bovine rumen, blood, and milk

Three Holstein cows were fed a high-concentrate diet (65:35 concentrate to forage) supplemented with either 5% sunflower oil (SO), 5% linseed oil (LO), or 2.5% fish oil (FO) to examine effects on biohydrogenation and fatty acid profiles in rumen, blood plasma, and milk. Diets were fed in a 3 × 3 Latin square with 4-wk periods with grass hay as the forage. Milk yield, dry matter intake, and percentages of milk fat (2.64) and protein (3.22) did not differ. All diets resulted in incomplete hydrogenation of unsaturated fatty acids as indicated by the profiles of 18:1 isomers, conjugated 18:2 isomers, nonconjugated 18:2 isomers, and 18:0 in ruminal fluid. Percentages of 8:0-14:0 and 16:0 in milk fat were greater with FO. Percentage and yield of trans10,cis12-18:2 were small and greater in cows fed SO (0.14%, 0.57 g/d) than FO (0.03%, 0.15 g/d) or LO (0.04%, 0.12 g/d). Percentage and yield of trans10-18:1, however, increased with FO (6.16%) and SO (6.47%) compared with LO (1.65%). Dietary FO doubled percentage of cis11-18:1 in rumen, plasma, and milk fat. Despite a lack of difference in ruminal percentage of trans11-18:1 (10.5%), cows fed FO had greater plasma trans11-18:1 (116 vs. 61.5 μg/mL) but this response did not result in greater trans11-18:1 percentage in milk fat, which averaged 5.41% across diets. Percentage (2.2%) and yield (14.3 g/d) of cis9,trans11-18:2 in milk fat did not differ due to oils. Unique responses to feeding LO included greater than 2-fold increases in percentages of trans13+14-18:1, trans15-18:1, trans16-18:1, cis15-18:1, cis9,trans12-18:2 and trans11,cis15 -18:2 in umen, plasma, and milk, and cis9,trans13-18:2 in plasma and milk. Ruminal 18:0 percentage had the highest positive correlation with milk fat content (r = 0.82) across all diets. When compared with previous data with cows fed high-concentrate diets without oil supplementation, results suggest that greater production of trans10-18:1, cis11-18:1, and trans11,cis15-18:2 coupled with low production of 18:0 in the rumen may be associated with low milk fat content when feeding high-concentrate diets and fish oil. In contrast, SO or LO could lead to low milk fat content by increasing ruminal trans10-18:1 (SO) or trans11,cis15-18:2 and trans9,trans12-18:2 (LO) along with a reduction in mammary synthesis of 8:0-16:0. Simultaneous increases in ruminal trans11-18:1 with fish oil, at a fraction of sunflower oil supplementation, may represent an effective strategy to maintain cis9,trans11-18:2 synthesis in mammary while reducing milk fat output and sparing energy.     

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

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

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.     

Production and metabolic responses of periparturient Holstein cows to dietary conjugated linoleic acid and trans-octadecenoic acids

Thirty-eight multiparous Holstein cows were utilized in a completely randomized design to examine the effect of feeding calcium salts of conjugated linoleic acid (CLA) and trans-octadecenoic acids (trans-C18:1) on animal performance and lipid and glucose metabolism during the transition to lactation. Dietary treatments were initiated approximately 28 d prior to expected calving dates and continued through d 49 postpartum. Prepartum treatments consisted of 1) a basal diet (Control), 2) basal diet + 150 g/d of CLA mix (CLA), and 3) basal diet + 150 g/d of trans-C18:1 mix (TRANS). Amounts of calcium salts of CLA and trans-C18:1 mixes were adjusted to 225 g/d during the 49-d postpartum treatment period. All diets were offered as a total mixed ration. Prepartum fat supplementation had no detectable effects on dry matter intake, body weight, or body condition score. After parturition, cows in the TRANS group consumed less dry matter at wk 4, 5, and 6 of lactation than did cows in the control group. Cows fed the trans-C18:1 supplement were in a more severe negative energy balance than those fed the control diet at 1 wk of lactation. Periparturient fat supplementation had no detectable effects on milk yield during wk 1 to 7 of lactation. Milk fat was not affected during wk 1 to 4, but was reduced after wk 4 of lactation by dietary CLA. Feeding calcium salts of CLA decreased short- to medium-chain fatty acid (C4 to C14) concentrations and increased both linoleic and linolenic acid concentrations in milk fat. Concentrations of nonesterified fatty acids and beta-hydroxybutyric acid in blood were greater in cows fed the CLA-supplemented diet than in those fed the control diet at 1 wk of lactation. In spite of small numerical tendencies, hepatic lipid and triacylglycerol concentrations did not vary significantly among dietary treatments. Periparturient fat supplementation had no detectable effects on plasma glucose and insulin concentrations. Steady-state concentrations of hepatic mRNA encoding pyruvate carboxylase and phosphoenolpyruvate carboxykinase were greater for the TRANS treatment group than the control and CLA groups. Results indicate that dietary CLA and trans-C18:1 fatty acids may affect lipid and glucose metabolism in early postpartum Holstein cows through distinct mechanisms.     

Augmentation of vaccenate production and suppression of vaccenate biohydrogenation in cultures of mixed ruminal microbes

To increase ruminal outflow of trans-vaccenic acid (t-VA), a new strain of Butyrivibrio fibrisolvens (MDT-10) was isolated that has a great ability to hydrogenate linoleic acid (LA) to t-VA. When strain MDT-10 was added to the batch cultures of mixed ruminal microbes (1% of the total number of viable ruminal bacteria), LA conversion to t-VA increased greatly; after 3 h, t-VA levels were > 4-fold higher than the control. By 10 h, all of the t-VA was hydrogenated to stearic acid. However, when a new strain of Bifidobacterium adolescentis (HF-11), which has a high capacity for incorporation of t-VA, was added in conjunction with MDT-10 (1% of the total number of ruminal bacteria), t-VA levels after 10 h were 6 times higher than with MDT-10 alone. These results suggest that t-VA produced by MDT-10 was incorporated into HF-11 cells, resulting in protection of t-VA from t-VA-hydrogenating microbes. Similar results were obtained in a continuous culture of mixed ruminal microbes in which addition of HF-11 simultaneously with MDT-10 increased the amount of t-VA in the effluent 2.5-fold. Both MDT-10 and HF-11 appeared to grow readily in the presence of mixed ruminal microbes. Sixty-two percent of t-VA incorporated by HF-11 was present in the free form, whereas 19, 15, and 3%, respectively, were incorporated into monoacylglycerol, glycerophospholipid, and diacylglycerol fractions. Because these lipids can be digested in the small intestine, it is likely that most t-VA in HF-11 cells is absorbed. Thus, introduction of MDT-10 and HF-11 simultaneously to the rumen might increase the amount of t-VA absorbed and might consequently increase the conversion of t-VA to conjugated linoleic acid in tissue.     

Pilot-scale production of conjugated linoleic acid-rich soy oil by photoirradiation

ABSTRACT:  Conjugated linoleic acid (CLA) is found naturally in dairy and beef products at levels of 0.2% to 2% of the total fat. A more concentrated source of dietary CLA, low in saturated fat, would be highly desirable to obtain optimum CLA levels of about 3 g/d. We recently reported photoisomerization of soy oil with iodine catalysis to be a simple way of producing CLA in laboratory without high-energy input or expensive enzymes and microorganisms. However, a long irradiation time of 144 h has been a limitation for this technique to be of practical value. The objectives of this study were to build a pilot plant unit to rapidly produce high-CLA soy oil by photoirradiation and optimize the processing parameters to obtain high-CLA soy oil. Degassed oil with dissolved-iodine catalyst was irradiated by UV lamps in an illuminated laminar flow unit (ILFU). The ILFU consists of 2 borosilicate glass plates in a silicone lined stainless steel frame. The static mode of operation yielded 5.7% of total CLA isomers and performed twice as well than the continuous mode with 2.5% of total CLA. Irradiating oil in a static mode with reflective surfaces increased the CLA yields 3-fold to 16.4%. About 22% of total CLA isomers can be rapidly produced from soy oil linoleic acid with 0.35% iodine catalyst in a 0.5-cm-thick oil layer maintained at 48 °C for 12 h. The peroxide value and GC-MS analysis did not identify any volatile compounds characteristic of lipid oxidation. This study is a definitive step toward the commercialization of large-scale production of CLA-rich soy oil.     

Dilution rate and pH effects on the conversion of oleic acid to trans C18:1 positional isomers in continuous culture

In a previous in vitro study, mixed ruminal microorganisms converted oleic acid to a variety of trans monenes when grown in batch cultures under constant environmental conditions. To determine whether a similar conversion occurs under environmental conditions more typical of the rumen, conversion of ¹³C-labeled oleic acid to biohydrogenation intermediates was determined in ruminal microorganisms grown in continuous culture at two pH (5.5 and 6.5) and liquid dilution rates (0.05 and 0.10/h) arranged factorially. After each morning feeding of the dual-flow continuous cultures, 250 mg of oleic acid in 5 mL of ethanol were injected into each culture. On d 10, 250 mg of oleic-1-¹³C replaced the unlabelled oleic acid in ethanol. Trans fatty acids were isolated from culture samples by solid phase extraction, and ¹³C enrichment and identity of double bond position was determined by gas chromatography-mass spectroscopy. At pH 6.5 and 0.10/h dilution rate, ¹³C enrichment was detected in all trans-C_18:1 isomers having double bond positions from C₆ through C₁₆ in the acyl chain. However, when pH or dilution rate in fermentors was lowered, no ¹³C enrichment was detected in any trans isomer with a double bond position beyond C₁₀. Enrichment in stearic acid increased by reducing culture pH from 6.5 to 5.5, but decreased when dilution rate dropped from 0.10 to 0.05/h. The stearic acid carbons that originated from oleic acid biohydrogenation increased from 30 to 72% when pH dropped from 6.5 to 5.5. The ¹³C enrichment of trans-10 was reduced under low pH and dilution rate conditions. The results of this study confirm that ruminal microorganisms are capable of converting oleic acid to a wide variety of trans-C_18:1 positional isomers when ruminal conditions are favorable (such as the pH 6.5 and 0.10/h dilution rate treatment). However, at low pH and dilution rate, the conversion of oleic acid to trans-C_18:1 still occurs, but positional isomers produced are restricted to double bond positions from C₆ to C₁₀. Low pH conditions also increased the conversion of oleic acid to stearic acid.     

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

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

Biohydrogenation, duodenal flow, and intestinal digestibility of trans fatty acids and conjugated linoleic acids in response to dietary forage:concentrate ratio and linseed oil in dairy cows

Duodenal flows of hydrogenation intermediates in response to changes in dietary forage:concentrate ratio (F:C) and linseed oil were evaluated using 4 lactating Holstein cows fed a low (65:35 forage to concentrate) or high (35:65) concentrate diet without (LC, HC) added oil or with linseed oil (LCO, HCO) at 3% of DM. A 4 x 4 Latin square design was implemented for 5 wk. Lower hydrogenation of 18:2n-6 and 18:3n-3 was observed with HC, but it increased with LCO or HCO. Duodenal flow of total conjugated linoleic acids (CLA) increased by 1.40 (LCO) to 3.01 (HCO) g/d with linseed oil. This response was associated with greater flows of cis9,trans11- (+0.21 to +0.55 g/d), trans11,cis13- (+0.33 to +0.36), trans11,trans13- (+1.01 to +1.15 g/d), and trans,trans-CLA (+0.12 to +0.72 g/d). Trans10,cis12-CLA flow averaged 0.08 g/d and was not affected by F:C or oil. trans11,cis15-18:2 flow increased by 8.5 (LCO) to 62 (HCO) g/d in response to linseed oil. Total trans-18:1 flow was 37 g/d in cows fed LC and increased to 81 g/d with HC. Feeding oil increased total trans-18:1 to the greatest extent with HCO. Flow of trans10-18:1 was lower with LC than with HC (1.46 vs. 20 g/d). Linseed oil increased trans11-18:1 flow by 40 (LCO) to 113 g/d (HCO). Feeding LCO and HCO also increased flows of trans6+7+8-, trans13+14-, trans15-, and trans16-18:1. Apparent intestinal digestibility of trans-18:1 isomers was largely unaffected by concentrate level and ranged between 67 and 95%. Linseed oil increased digestibility of nearly all isomers by 3 to 16 percentage units. Digestibility of cis9,trans11-CLA was greater in cows fed HC (55%) compared with cows fed LC (32%) and was not affected by linseed oil. Data suggest that high concentrate diets enhanced ruminal outflow of trans10-18:1. We provide initial in vivo evidence that supplemental 18:3n-3 is hydrogenated to trans11,cis15-18:2, trans11-18:1, trans13+14-18:1, trans15-18:1, trans6+7+8-18:1, and trans16-18:1 primarily.     

Effects of dry matter intake, addition of buffer, and source of fat on duodenal flow and concentration of conjugated linoleic acid and trans-11 C18:1 in milk

The primary objective of the study was to investigate the effects of DM intake, addition of buffer, and fish vs. soybean oil on duodenal flows and milk concentrations of conjugated linoleic acid (CLA) and trans-11 C18:1. Four ruminally and duodenally cannulated multiparous cows averaging 106 +/- 17 d in milk at the start of the trial were used in a 4 x 4 Latin square design with treatments as follows: 1) control=diet contained 2% fish oil and fed ad libitum, 2) buffer addition (BUFF)=control diet with 0.8% of NaHCO3 added, 3) low DM intake (LDMI)=DMI restricted to 80% of the control but concentration of fish oil was increased to 2.5% to provide for similar fatty acids (FA) intake, and 4) soybean oil (SBO)=same as control except 2% soybean oil was substituted for fish oil. The diets consisted of 36.2% forage and 63.8% concentrate. Each period consisted of 18 d, with the last 7 d devoted to data collection and the first 4 d used to determine the appropriate amount of feed to be offered to the cow on LDMI. Duodenal flows of CLA and trans-C18:1 were lower for SBO than for diets with fish oil. Feeding buffer did not affect ruminal pH or duodenal flows of trans-11 C18:1 and CLA. Restriction of DMI decreased duodenal flow of trans-11 C18:1 but did not decrease duodenal flow of CLA compared with control. In milk, CLA concentration was lower for SBO (24.5, 17.9, 18.5, and 10.1 mg/g of FA for control, BUFF, LDMI, and SBO, respectively). Cows fed fish oil had higher duodenal flow and milk concentration of n-3 polyunsaturated fatty acids than the cows fed SBO. Compared with SBO, fish oil is more effective in increasing duodenal flows of CLA and trans-11 C18:1, and thus, concentration of CLA in milk.     

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.     

Milk conjugated linoleic acid response to fish oil and sunflower oil supplementation to dairy cows managed under two feeding systems

Earlier research showed that conjugated linoleic acid (CLA) content in milk fat is highest when cows’ diets are supplemented with a blend of fish oil (FO) and linoleic acid-rich oils. The objective of this study was to compare the effect of FO and sunflower oil (SFO) supplementation on milk cis-9, trans-11 CLA when dairy cows managed on pasture or in confinement. Fourteen Holstein cows were assigned into 2 treatment groups: cows grazed on alfalfa-grass pasture (PAS) or were fed corn silage-alfalfa hay mix ad libitum (LOT). Both groups were supplemented with a 8.2 kg/d grain supplement containing 640 g of FO and SFO (1:3 wt/wt). Grain supplement was fed in 2 equal portions after each milking, for a period of 3 wk. Milk samples were collected during the last 3 d of the experimental period. Milk yield was greater with the LOT diet (23.1 kg/d) compared with the PAS diet (19.4 kg/d). Milk fat percentages (2.51 and 2.95 for the LOT and PAS, respectively) and yields (0.57 and 0.51 kg/d) were similar for the 2 diets. Milk protein percentages were not affected by diets (3.34 and 3.35 for the LOT and PAS diets, respectively), but protein yields were lower for the PAS diet (0.61 kg/d) compared with the LOT diet (0.75 kg/d). Treatment diets had no effect on milk trans C18:1 concentrations [10.64 and 9.82 g/100 g of total fatty acids (FA) for the LOT and PAS, respectively] or yields (60.65 and 64.01 g/d), but did affect isomers distributions. Concentration (g/100 g of total FA) of vaccenic acid was lower with the LOT diet (2.15) compared with the PAS diet (4.52), whereas concentration of trans-10 C18:1 was greater with the LOT diet (4.99) compared with the PAS diet (1.69). Milk cis-9, trans-11 CLA concentration was greater with the PAS diet (1.52) compared with the LOT diet (0.84). In conclusion, the increase in milk cis-9, trans-11 CLA content was greater when pasture-based diets were supplemented with FO and SFO. The lower cis-9, trans-11 CLA concentration in milk from the confinement-fed cows resulted from trans-10 C18:1 replacing vaccenic acid as the predominant trans C18:1 isomer.     

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.     

棉籽油碱异构化合成共轭亚油酸研究

以棉籽油为原料,研究溶剂选择、反应时间、反应温度、碱与油质量比,溶剂与油质量比对制备共轭亚油酸影响,并通过正交实验确定最优反应条件:即反应时间2 h、反应温度170℃、碱与油质量比0.55∶1、溶剂与油质量比1.6∶1;在此条件下,棉籽油共轭亚油酸转化率为98.2%。该法较简单、产物易处理,且有机溶剂丙二醇无毒性,较适用于CLA功能性食品和保健品开发。     

Short communication: effect of production variables on the cis-9, trans-11 conjugated linoleic acid content of cows' milk

Although there have been numerous studies investigating effects of nutrition and individual variation on the concentration of cis-9, trans-11 conjugated linoleic acid (rumenic acid; RA) in milk, there is limited information on relationships among RA content of milk and production variables. The objective of the current analysis was to examine the effects of production variables on RA content and desaturase index of milk fat. A total of 430 samples were collected from cows fed a commercial total mixed ration in winter and grazing in summer. Across a >6-fold range in production variables, RA content of milk ranged from 1 to 32 mg/g of fatty acids and desaturase index ranged from 0.03 to 0.15. Days in milk, milk yield, milk fat content, and milk fat yield had minimal or no effect on RA content of milk fat or desaturase index (R² values all <0.08). Thus, whereas nutrition and individual variation are major factors affecting RA content and desaturase index of milk fat, these values are minimally affected by days in milk, milk yield, milk fat content, and milk fat yield. Differences in these parameters do not need to be considered, therefore, when designing management strategies to increase RA content of milk fat.     

嗜酸乳杆菌转化菜籽油生成共轭亚油酸条件的优化

文中对嗜酸乳杆菌静息细胞转化菜籽油生成CLA工艺进行了优化研究,通过单因素试验,确定了最适转化液介质、介质浓度、pH、温度、转化时间和细胞浓度,同时,研究了不同金属离子对亚油酸异构酶活性的影响。在该基础上,利用Box-Benhnken中心组合设计和响应面法对转化时间、pH、温度、细胞浓度进行了工艺优化分析,确定了在菜籽油浓度为7.5 mg/mL、脂肪酶的加入量为2 mg/30 mL时,以0.1 mol/L柠檬酸-柠檬酸钠缓冲液为转化液介质条件下,最优转化工艺条件是时间25 h,pH6.5,温度36℃,细胞浓度40 mg/mL,经优化后CLA的生成量可以达到230.12±7.52μg/mL。     

Monensin by fat interactions on trans fatty acids in cultures of mixed ruminal microorganisms grown in continuous fermentors fed corn or barley

In previous studies, monensin (M) and unsaturated plant oils independently increased trans fatty acid concentrations in cultures of mixed ruminal microorganisms. This study was conducted to determine if combining M with plant oil yielded interactions on trans fatty acid concentrations in cultures of mixed ruminal microorganisms or their effects were additive. Four continuous fermentors were fed 14 g of dry feed per day (divided equally between two feedings), consisting of alfalfa hay pellets (30% of DM) and either a high corn (HC) or a high barley (BB) concentrate (70% of DM) in each of two fermentors. Within each grain type, one fermentor was supplemented with M (25 ppm), and the other fermentor was supplemented with 5% soybean oil (SBO) during d 5 to 8. Monensin and SBO were added together in all fermentors during d 9 to 12. Samples were taken at 2 h after the morning feeding on the last day of each period and analyzed for fatty acids by gas-liquid chromatography. A second run of the fermentors followed the same treatment sequence to give additional replication. Average pH across all treatments was 6.15, which was reduced by M but not affected by SBO. Monensin reduced the ratio of acetate to propionate (A:P), which averaged 2.03 across all treatments; fat decreased A:P in cultures not receiving M but increased it in the presence of M. Monensin and SBO altered the concentration of several trans fatty acids, but the only interaction was a grain x M x SBO interaction for trans-10 C18:1. The increase in trans-10 C18:1 by the M and SBO combination exceeded the sum of increases in trans-10 C18:1 for each individual feed additive, but only for KB. For the HC diet, M increased trans-10 C18:1 more than fat alone and more than the M and SBO combination. The results of this study show that M and SBO effects are additive for all trans FA except for trans-10 C18:1. In the case of trans-10 C18:1, M and SBO interacted to give higher trans-10 C18:1 concentrations in ruminal contents than would be expected simply by adding their individual effects, but only for HB. Because some trans fatty acid isomers have been associated with milk fat depression in dairy cows, these results suggest more severe depressions in milk fat content when cows are fed M along with unsaturated plant oils.     

异构化棉油甲酯中共轭亚油酸的定量分析研究

研究了棉油甲酯在碘的催化下发生异构化反应的规律,运用气质联用仪和紫外光谱对异构化产物的成分进行了分析.在最佳的异构化条件下,异构化棉油甲酯中共轭亚油酸质量分数可达31.2%,共轭转化率56%;根据不同质量浓度样品的紫外吸收曲线找出样品质量浓度与吸光度之间的线性关系,并参照气质联用分析的共轭亚油酸质量分数结果,得出紫外光谱法分析计算异构化产物中共轭亚油酸质量分数的经验公式为:ω(CLA)=O.00801A/C,吸光系数为O.00801,适宜的质量浓度为0.02～0.04g/L,误差小于5%.实验证实紫外光谱法是分析测定棉油甲酯异构化产物中共轭亚油酸的一种简便、快捷的方法.