Cis-12 Conjugated Linoleic Acid Research Articles

Conjugated linoleic acid and betaine affect lipolysis in pig adipose tissue explants

Consumers’ demand of leaner meat products is a challenge. Although betaine and conjugated linoleic acid (CLA) have the potential to decrease porcine adipose tissue, their mode of action is poorly understood. The aim of the study was to determine the lipolytic effect of betaine and CLA in the adipose tissue of Iberian pigs. Adipose tissue explants from five pigs (38 kg BW) were prepared from dorsal subcutaneous adipose tissue samples and cultivated for 2 h (acute experiments) or 72 h (chronic experiments). Treatments included 100 µM linoleic acid (control), 100 µM trans-10, cis-12 CLA, 100 µM linoleic acid + 1 mM betaine and 100 µM trans-10, cis-12 CLA + 1 mM betaine (CLABET). To examine the ability of betaine or CLA to inhibit insulin’s suppression of isoproterenol-stimulated lipolysis, test medium was amended with 1 µM isoproterenol ±10 nM insulin. Media glycerol was measured at the end of the incubations. Acute lipolysis (2 h) was increased by CLA and CLABET (85% to 121%; P < 0.05) under basal conditions. When lipolysis was stimulated with isoproterenol (1090%), acute exposure to betaine tended to increase (13%; P = 0.071), while CLA and CLABET increased (14% to 18%; P < 0.05) isoproterenol-stimulated lipolysis compared with control. When insulin was added to isoproterenol-stimulated explants, lipolytic rate was decreased by 50% (P < 0.001). However, supplementation of betaine to the insulin + isoproterenol-containing medium tended to increase (P = 0.07), while CLABET increased (45%; P < 0.05) lipolysis, partly counteracting insulin inhibition. When culture was extended for 72 h, CLA decreased lipolysis under basal conditions (18%; P < 0.05) with no effect of betaine and CLABET (P > 0.10). When lipolysis was stimulated by isoproterenol (125% increase in rate compared with basal), CLA and CLABET decreased glycerol release (27%; P < 0.001) compared with control (isoproterenol alone). When insulin was added to isoproterenol-stimulated explants, isoproterenol stimulation of lipolysis was completely blunted and neither betaine nor CLA altered the inhibitory effect of insulin on lipolysis. Isoproterenol, and especially isoproterenol + insulin, stimulated leptin secretion compared with basal conditions (68% and 464%, respectively; P < 0.001), with no effect of CLA or betaine (P > 0.10). CLA decreased leptin release (25%; P < 0.001) when insulin was present in the media, partially inhibiting insulin stimulation of leptin release. In conclusion, betaine and CLA produced a biphasic response regarding lipolysis so that glycerol release was increased in acute conditions, while CLA decreased glycerol release and betaine had no effect in chronic conditions. Furthermore, CLA and CLABET indirectly increased lipolysis by reducing insulin-mediated inhibition of lipolysis during acute conditions.

Trans10, cis12-conjugated linoleic acid exhibits a stronger antioxidant capacity than cis9, trans11-conjugated linoleic acid in primary cultures of laying hen hepatocytes

The objective of this study consisting of 2 trials was to investigate the antioxidant role of conjugated linoleic acid (CLA) isomers (c9, t11-CLA and t10, c12-CLA) and the underlying mechanism by which they act in modulating redox status in a primary laying hen hepatocyte culture. In trial 1, the cytotoxicity of CLA isomers or linoleic acid (LA) (0, 25, 50, 100, 200, 400, 800 μmol/L) was evaluated by 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay. The concentration of CLA isomers or LA (25, 50, 100 μmol/L) for proper antioxidant activity was evaluated by measuring the antioxidant enzyme activity. In trial 2, there were 5 groups: control group, cells were untreated; H2O2 group, cells were exposed to 4mmol/L H2O2 for 2h; c9, t11 or t10, c12 or LA group, cells were treated with c9, t11-CLA or t10, c12-CLA or LA for 24h and then exposed to 4mmol/L H2O2 for 2h. Trial 1 showed that the non-toxic dose range for CLA isomers was 0 to 200 μmol/L. The optimum concentration of c9, t11-CLA and t10, c12-CLA for trial 2 was 100 μmol/L. In trial 2, pretreatment with t10, c12-CLA but not c9, t11-CLA attenuated the increase in reactive oxygen species (ROS) compared to hydrogen peroxide (H2O2) group (P < 0.05). t10, c12-CLA elevated the superoxide dismutase (SOD) and catalase (CAT) activities compared with the H2O2 group (P < 0.05). In addition, t10, c12-CLA up-regulated the mRNA expression of nuclear factor E2-related factor-2 (Nrf2) as well as its target genes, Cu-Zn superoxide dismutase (SOD1) and CAT (P < 0.05). Pretreatment with t10, c12-CLA but not c9, t11-CLA decreased Nrf2 protein expression in the cytoplasm and increased Nrf2 protein expression in the nucleus compared with the H2O2 group (P < 0.05). The results indicate that t10, c12-CLA exhibits a stronger antioxidant capacity than c9, t11-CLA in primary cultured laying hen hepatocytes. t10, c12-CLA increases the activity and mRNA expression of antioxidant enzymes via facilitating nuclear translocation of Nrf2.

Effect of trans-10, cis-12 conjugated linoleic acid (CLA) on expression of lipogenic genes is related to amount of trans-10, cis-12 CLA per unit of metabolic weight in lactating dairy ewes

This study evaluated the effect of trans-10, cis-12 CLA on the expression of genes involved in decreasing milk fat synthesis in two groups of lactating ewes with different metabolic weights. Twelve Lacaune ewes were alloted, according to metabolic weight (MW), into a Light Metabolic Weight (LMW) and Heavy Metabolic Weigth (HMW) groups, both of which were orally fed 30 g/d of rumen-unprotected CLA supplement containing 29.9% trans-10, cis-12. Milk samples were collected on days 0, 3 and 10 and mammary and adipose tissue biopsies were taken on days 3 and 10. RNA extraction, cDNA synthesis and RT-qPCR were performed and data were analyzed using the MIXED procedure of SAS as repeated measure in a completely randomized design. The trans-10, cis-12 CLA reduced milk fat synthesis between days 3 and 10 (P = 0.001), and the HMW group had a greater expression (P < 0.05) of the genes acetyl-CoA carboxylase alpha - promoter III (ACACA-α PIII), fatty acid synthase (FASN), stearoil CoA desaturase 1 (SCD1) and thyroid hormone responsive spot14 (THRSP) in the mammary gland and of acetyl-CoA carboxylase alpha - promoter I (ACACA-α PI) and leptin (P < 0.05) in adipose tissue. Our findings indicate that the response of animals to trans-10, cis-12 CLA appears to be metabolic weight dependent.

Conjugated linoleic acid production by lactic acid bacteria: a bio-transformation study in media with oil hydrolysates

The unique ability of lactic acid bacteria (LAB) to convert unsaturated fatty acids to saturated fatty acids via the production of unsaturated intermediates is known as bio-hydrogenation, which is responsible for the conversion or the bio-transformation of unsaturated fatty acids like linoleic acid to saturated fatty acids like stearic acid via the intermediate compounds such as conjugated linoleic acid (CLA). Realizing the health benefits of CLA, lactic acid bacteria were isolated from curd in order to study their ability of CLA production. Initially, four bacterial strains isolated from curd and designated as 1s, 13s, 15s, and 17s were screened and found to be gram-positive, catalase negative and acid producers. Out of these four strains, 1s and 13s showed rapid adaptation to the change in the substrate from glucose to linseed oil hydrolysate. The fermentation study conducted in shake flask mode revealed that the maximum concentration of trans-10, cis-12 CLA produced in medium enriched with 0.4% (w/v) castor oil hydrolysate (mCOH) by isolate 1s was 620 mg·L-1 at 4h while that produced by isolate 13s in mCOH was 430mg·L-1 at 8h as assessed by gas chromatography (GC) performed using a flame ionization detector (FID). Thus, the study proved the ability of LAB to synthesize CLA. Furthermore, in order to test the feasibility of CLA production on a large scale, an additional study was conducted using Lactobacillus delbreukii NCIM 2025 in a continuously stirred tank reactor in both batch and continuous modes using linseed oil hydrolysate as the fatty acid substrate. The maximum concentration of CLA produced by Lactobacillus delbreukii NCIM 2025 in medium enriched with 0.4% linseed oil hydrolysate (mLOH) was 380µg·L-1 at 6 h in batch mode and 590 µg·L-1 at 1h in continuous mode as analyzed using highperformance liquid chromatography (HPLC) performed using an ultraviolet (UV) detector at 233nm. Thus, the study also reveals that there is a vast scope for improvement in the scale-up of CLA production from the laboratory scale to a commercial scale.

Development of Commercially Viable Method of Conjugated Linoleic Acid Synthesis Using Linoleic Acid Fraction Obtained from Pork By-products.

The purpose of this study was to develop a commercially viable method for synthesis of conjugated linoleic acid (CLA) using the linoleic acid fraction obtained from six pork by-products (liver, lung, heart, stomach, small intestine, and large intestine). The workflow of CLA synthesis from each by-product was as follows: washing→crude fat extraction→fractionation into saturated and unsaturated fatty acids→repeat unsaturated fatty acid fractionation→CLA synthesis. Cis-9, trans-11, and trans-10, cis-12 CLA was synthesized from pork by-products. The yield of CLA synthesis of pork by-products ranged from 1.55 to 11.18 g per 100 g of by-products. The amount of synthesized CLA was the highest in the small intestine and large intestine by-products. Fractionation of pork by-products nearly doubled the yield of CLA. We suggest that commercial fractionation methods could increase the yield of CLA at low cost, reduce waste, and improve the efficiency of by-product utilization.

Conjugated Linoleic Acid Regulates Body Composition and Locomotor Activity in a Sex‐Dependent Manner in Drosophila melanogaster

Conjugated linoleic acid (CLA) has been reported to be a bioactive food component. However, there is limited knowledge on the sex-dependent effects of CLA on energy metabolism. In the present study, Drosophila melanogaster was used to investigate the sex-dependent effects of CLA with respect to body fat, muscle, locomotion, and a key metabolic regulator, AMP-activated protein kinase α (AMPKα). Adult flies were fed a cornmeal-based fly food with 0.5% of CLA oil (50:50 of cis-9,trans-11 and trans-10,cis-12 CLA isomers in triacylglycerol (TAG) form), 0.5% safflower oil (high in linoleic acid [LNA] as control), or 0.5% water (as blank) for 5 days. Accumulation of CLA in tissue was verified using gas chromatography-mass spectrometry. CLA-fed flies had reduced TAG and increased locomotor activity when compared to LNA-fed control flies. In addition, CLA increased the muscle content when compared to the blank. Moreover, following CLA supplementation, increased AMPKα activity was observed in females, but not in males. These sex-dependent metabolic effects of CLA may be due to physiological differences in lipid metabolism and nutrient requirements. In conclusion, CLA promoted the body composition and locomotion behavior in D. melanogaster and regulated the sex-specific metabolism in part via AMPKα. As key physiological processes are conserved between fly and human, information obtained from this research could provide valuable insights into sex-dependent responses to CLA in humans.

Kinetics of trans-10, cis-12-conjugated linoleic acid transfer to plasma and milk following an abomasal bolus in lactating dairy cows.

Trans-10, cis-12-conjugated linoleic acid (CLA) is a potent bioactive fatty acids (FA) that causes milk fat depression in lactating animals. FA are transferred to milk directly through chylomicrons and indirectly by recycling through other tissues. The objective of this study was to characterise the kinetics of trans-10, cis-12 CLA transfer to plasma and milk after a single bolus infusion. Five multiparous mid-lactation cows received a single abomasal bolus infusion of an enriched CLA mixture providing 15 g of trans-10, cis-12 CLA and 15 g of cis-9, trans-11 CLA over a 30-min period. Plasma concentration of trans-10, cis-12 and cis-9, trans-11 CLA peaked 2 h post-bolus, reaching 0·29 and 0·38 % of total plasma FA, respectively, and returned to pre-bolus values at 72 h post-infusion. Milk trans-10, cis-12 CLA yield and concentration peaked 14 h post-bolus (0·25 g/h) and was not detectable in milk after 86 h. Total apparent transfer of trans-10, cis-12 CLA to milk was 41 %, with 73 % transferred to milk through the direct pool (chylomicrons) and the remaining 27 % transferred through the indirect pool (tissue recycling). Compartmental modelling revealed the existence of a transient unavailable pool of trans-10, cis-12 CLA in extravascular tissues represented primarily by the mammary gland, which slowly exchanges with an available pool for secretion in milk fat and transfer to milk. In conclusion, trans-10, cis-12 CLA is predominantly transferred to milk through the direct pathway; however, how this CLA isomer is processed within the mammary gland requires further investigation.

Time-dependent effect of trans-10,cis-12 conjugated linoleic acid on gene expression of lipogenic enzymes and regulators in mammary tissue of dairy cows

Trans-10,cis-12 conjugated linoleic acid (CLA) has been identified as an intermediate of rumen fatty acid biohydrogenation that caused milk fat depression (MFD) in the dairy cow. Previous studies in cows experiencing CLA- and diet-induced MFD have identified reduced mammary expression of the master lipogenic regulator sterol response element transcription factor 1 (SREBF1) and many of its dependent genes. To distinguish between primary mechanisms regulating milk fat synthesis and secondary adaptations to the reduction in milk fat, we conducted a time-course experiment. Eleven dairy cows received by abomasal infusion an initial priming dose of 6.25 g of CLA followed by 12.5 g/d delivered in multiple pulses per day for 5 d. Cows were milked 3×/d and mammary biopsies were obtained under basal condition (prebolus control) and 12, 30, and 120 h relative to initiation of CLA infusion. Milk fat concentration and yield decreased progressively reaching a nadir at 69 h (1.82% and 38.2 g/h) and averaged 2.03 ± 0.19% and 42.1 ± 4.10 g/h on the last day of treatment (±standard deviation). Expression of fatty acid synthase (FASN) and lipoprotein lipase (LPL) were decreased at 30 and 120 h compared with control. Expression of SREBF1 and THRSP were also decreased at 30 and 120 h compared with control. Additionally, we failed to observe changes in other factors, including peroxisome proliferator-activated receptor γ and liver × receptor β and milk fat globular membrane proteins, during CLA treatment. However, expression of milk fat globular membrane proteins were decreased after 14 d of diet-induced MFD in samples from a previous experiment, indicating a possible long-term response. The rapid decrease in lipogenic enzymes, SREBF1, and THRSP provide strong support for their transcriptional regulation as a primary mechanism of milk fat depression.

Prepartum supplementation of conjugated linoleic acids (CLA) increased milk energy output and decreased serum fatty acids and β-hydroxybutyrate in early lactation dairy cows.

Prepartum supplementation with conjugated linoleic acid (CLA) may influence lipolysis and hyperketonemia in dairy cows. The objective of this study was to examine the effect of prepartum CLA supplementation on lactation performance and serum fatty acids (FA) and β-hydroxybutyrate (BHB) in early lactation dairy cows, and secondarily on reproductive performance. Multiparous cows were enrolled in the study at 18 days prior to expected calving date, and randomly assigned 100 g/day of Lutrell Pure (BASF, Ludwigshafen, Germany; 75% FA), providing 10 g/day of each CLA isomer (trans-10 cis-12 and cis-9 trans-11 CLA) or equivalent amount of rumen inert fatty acids as control (78 g/day of Energy Booster 100; Milk Specialties Global, Eden Prairie, MN). Treatments were top dressed daily to individual cows from enrollment to calving and all cows were offered the same ration. Blood samples were collected on the first day of supplementation, 10 days prepartum, and 1, 7, 14, and 30 days postpartum. Hyperketonemia was defined as serum BHB ≥ 1.2 mM. Milk yield was recorded daily until 60 days postpartum and averaged weekly. Milk samples were obtained weekly for component analysis. Prepartum CLA supplementation tended to increase serum concentration of cis-9, trans-11 CLA and increased trans-10, cis-12 CLA prepartum. Cows supplemented with CLA had increased milk protein yield and tended to have increased milk fat yield and milk yield, which together resulted in greater energy content of milk. Cows supplemented with CLA had lower serum FA on day 1 and 7 postpartum and overall lower serum BHB postpartum, which resulted in decreased prevalence of hyperketonemia on day 14 postpartum. There were no differences in body condition score change, other health disorders, or reproductive outcomes by treatment. Together, these findings indicate that prepartum CLA supplementation may be a plausible strategy to positively influence postpartum performance.

Peroxisome proliferator-activated receptor gamma (PPARγ) agonist fails to overcome trans-10, cis-12 conjugated linoleic acid (CLA) inhibition of milk fat in dairy sheep

The trans-10, cis-12 conjugated linoleic acid (CLA) causes milk fat depression by downregulating expression of genes and transcription factors involved in lipogenesis and it has been proposed that peroxisome proliferator-activated receptor gamma (PPARγ) can be inhibited by trans-10, cis-12 CLA. The PPARγ is a nuclear receptor activated by natural or synthetic ligands and promotes expression of lipogenic genes and its effect on mammary lipogenesis and the interaction with trans-10, cis-12 CLA in lactating ewes was evaluated using thiazolidinedione (TZD), a chemical PPARγ agonist. A total of 24 lactating ewes were randomly assigned to one of the following treatments for 7 days: (1) Control (5 ml/day of saline solution); (2) TZD (4 mg/kg of BW/day in 5 ml of saline solution); (3) CLA (27 g/day with 29.9% of trans-10, cis-12); (4) TZD+CLA. Compared with Control, milk fat content was not changed by TZD, but was decreased 22.3% and 20.5% by CLA and TZD+CLA treatments. In the mammary gland, TZD increased PPARγ gene expression by 174.8% and 207.8% compared with Control and TZD+CLA treatments, respectively. Conjugated linoleic acid reduced sterol regulatory element-binding transcription protein 1 (SREBP1) gene expression 89.2% and 75.3% compared with Control and TZD+CLA, respectively, demonstrating that TZD fails to overcome CLA inhibition of SREBP1 signaling. In adipose tissue, the expression of SREBP1 and stearoyl CoA desaturase 1 (SCD1) genes were increased by the TZD+CLA treatment, compared with the other treatments. Conjugated linoleic acid decreased milk fat concentration and expression of lipogenic genes, while TZD had no effect on milk fat concentration, expression of lipogenic enzymes or regulators in the mammary gland and failed to overcome the inhibition of these by CLA. Therefore, CLA inhibition of milk fat synthesis was independent of the PPARγ pathway in lactating dairy ewes.

Mexican tropical cream cheese yield using low-fat milk induced by trans-10, cis-12 conjugated linoleic acid: effect of palmitic acid

ABSTRACTThe trans-10, cis-12 isomer of conjugated linoleic acid (CLA) reduces milk fat in dairy cows, causing a lower cheese yield. This could be mitigated by adding palmitic acid (PA) to the diet, as it stimulates fat synthesis in mammary gland. The objective of this study was to evaluate the effect of adding PA to the diet of grazing dairy cows complemented with trans-10, cis-12 CLA on the yield and fatty acid (FA) profile of Mexican tropical cream cheese. The lowest cheese yield was the result of the treatment with CLA alone. Supplementing PA to the cows with low fat concentration in their milk, from the effect of the trans-10, cis-12 isomer, mitigates the decrease in cheese yield. The process of cheese making does not modify the profile of FAs, so the supplemented cis-9, trans-11 CLA isomer transfers to the cheese, giving added value to the Mexican tropical cream cheese.

Polyunsaturated fatty acids influence offspring sex ratio in cows

Dietary polyunsaturated fatty acids (PUFAs) can influence fertility in farm animals. Some evidence in mice and sheep have suggested that PUFAs may influence offspring sex ratio, which may have significant value for cattle production. To test this hypothesis, three groups of Holstein cows were supplemented with either 0%, 3% or 5% protected fat (PF) in the form of calcium salt of fatty acids (rich in omega-6) from 14–21 days pre-partum until conception. Proven-fertile frozen semen from the same ejaculate was used for insemination. Calf sex recorded at birth was 8/19 (42.1%) male offspring in the control group, increasing to 14/20 (70%, P > 0.05) and 17/20 (85%, P < 0.05) in 3% and 5% PF, respectively. To test if this effect was caused by a direct influence on the oocyte, we supplemented bovine cumulus oocyte complexes during in vitro maturation with either omega-3 alpha-linolenic acid (ALA), omega-6 linoleic acid (LA) or trans-10, cis-12 conjugated linoleic acid (CLA). Sex ratio of the produced transferable embryos was determined using PCR of SRY gene. Similar to the in vivo results, sex ratio was skewed to the male side in the embryos derived from LA- and CLA-treated oocytes (79% and 71%) compared to control and ALA-treated oocytes (44% and 54%, respectively). These results indicate that both dietary and in vitro supplementation of omega-6 PUFAs can skew the sex ratio towards the male side in cattle. Further experiments are required to confirm this effect on a larger scale and to study the mechanisms of action that might be involved.

Inflammatory and metabolic responses to an intramammary lipopolysaccharide challenge in early lactating cows supplemented with conjugated linoleic acid.

Supplementation of dairy cows with trans-10, cis-12 conjugated linoleic acid (CLA) allows nutrient repartitioning despite an energy deficiency in early lactation, which might be a benefit for the immune system, too. In this study, we investigated potential nutrient sparing effects of CLA in early lactating cows with low plasma glucose concentrations exposed to an intramammary lipopolysaccharide (LPS) challenge. Fifteen multiparous Holstein cows were exposed to an intramammary LPS challenge in week 4 p.p. Eight cows (CLA) were supplemented daily with 70g of lipid-encapsulated CLA (6.8g trans-10, cis-12 and 6.6g of the cis-9, trans-11 CLA isomer; CLA) and seven cows with 56g of control fat (CON). Blood samples were obtained every 30min along with rectal temperature, heart and respiratory rate, and milk samples were taken hourly until 10hr after the LPS application. Plasma was analysed for concentrations of glucose, free fatty acids, beta-hydroxybutyrate (BHB), cortisol, insulin and glucagon. In milk, somatic cell count and activity of lactate dehydrogenase were determined. Initial plasma glucose concentration was lower in CLA than in CON. During the immunostimulation, CLA had higher glucose concentrations than CON, and BHB decreased distinctly in CLA, whereas CON cows maintained BHB concentration at a lower level. Body temperature in CLA increased earlier, the difference between peak and basal temperature was higher, and the decline thereafter occurred earlier. In conclusion, CLA supplementation of early lactating cows exposed to an intramammary LPS challenge affected local and systemic immune responses. We assume that CLA supplementation triggered glycogen storage. Cows supplemented with CLA provided more glucose and preferentially used BHB as an energy source during the immune response. The more intense metabolic and more concentrated endocrine responses support an immunomodulatory effect of CLA supplementation.

Effect of palmitic acid on the mitigation of milk fat depression syndrome caused by trans-10, cis-12-conjugated linoleic acid in grazing dairy cows

ABSTRACTThe objective of the study was to evaluate the effect of adding protected palmitic acid (PA) to the ration of grazing dairy cows supplemented with protected conjugated linoleic acid (CLA) on milk production, chemical composition and fat profile. Six cows were used, 3/4 American Swiss × Zebu, under a rotational grazing system in a mixed sward with Cynodon plectostachyus, Brachiaria decumbens and Brachiaria brizantha. Furthermore, each cow received daily 4 kg concentrates and 8 kg sorghum silage, which made up the basal diet. The cows were distributed into three two-cow groups. Three treatments were randomly assigned to the groups, using a cross design: (1) control (basal diet), (2) basal diet + CLA (50 g/d) and (3) basal diet + CLA (50 g/d) + PA (412 g/d). The following variables were evaluated: forage intake, milk production, protein, fat and lactose concentration in milk, and milk fatty acid (FA) profile. There were no differences in forage intake between treatments; however, there were differences in milk production, protein, fat and lactose yield and fat concentration, which increased significantly in group CLA + PA when compared with group CLA. The concentration of FA synthesised de novo was lower when PA was included in the diet. Adding PA to the diet of grazing cows mitigates the milk fat decline caused by including trans-10, cis-12 CLA in the diet.

Ruminal Biohydrogenation Kinetics of Defatted Flaxseed and Sunflower Is Affected by Heat Treatment.

The effect of heat treatment on biohydrogenation of linoleic acid (LA) and linolenic acid (LNA) and formation of stearic acid (SA), cis-9, trans-11 conjugated LA (CLA), trans-10, cis-12 CLA and trans-vaccenic acid (VA) was studied in in vitro incubations with diluted rumen fluid as inoculum and partly defatted flaxseed (DF) and partly defatted sunflower (DS) as test feeds. Feeds were heated in a laboratory oven at 110 °C for 0 (unheated), 45, or 90 min. Michaelis-Menten kinetics was applied for quantifying biohydrogenation rate. The DF heated for 90 min showed the lowest biohydrogenation rate of LNA and LA, indicated by the lowest Vmax value (P < 0.04 and P < 0.03, respectively). The DS heated for 45 min had the lowest biohydrogenation rate of LNA, indicated by the lowest Vmax value (P < 0.04). In conclusion, heat treatment decreased biohydrogenation of LA and LNA in DF and LNA in DS.

Effect of dietary conjugated linoleic acid on vitamin A status of lactating rats and their offspring.

The absorption and metabolism of vitamin A is linked with that of lipids. It is known that conjugated linoleic acid (CLA) affects the lipid metabolism in growing and lactating animals. In the present study, the hypothesis was investigated that dietary CLA influences vitamin A status of lactating rats and their pups during the suckling period. For this purpose, Wistar Han rats were fed either a control diet (control group, n=14) or a diet containing 0.87% of cis-9, trans-11 and trans-10, cis-12 (1:1) CLA (CLA group, n=14) during pregnancy and lactation. Vitamin A concentrations in various body tissues were determined 14days after delivery in dams and 1, 7 and 14days after birth in pups, and expression of selected genes involved in metabolism of retinoids was determined in dams. Vitamin A concentrations in liver, plasma and muscle were similar in control and CLA-fed dams. Expression of genes involved in retinoid transport, storage and degradation in liver and adipose tissue in dams was also not different between control and CLA-fed dams. Vitamin A concentrations in milk curd, sampled at d 1, 7 and 14 of lactation were not different between control and CLA-fed dams. Vitamin A concentrations in liver, lung and adipose tissue were also not different in pups from control dams and pups from CLA-fed dams. In conclusion, we show for the first time that dietary CLA has little effect on vitamin A concentrations and vitamin A metabolism in lactating rat dams and, moreover, does not influence tissue vitamin A concentrations in their newborn and suckling pups.

The trans-10,cis-12 conjugated linoleic acid increases triacylglycerol hydrolysis in yeast Saccharomyces cerevisiae.

The trans-10,cis-12 conjugated linoleic acid (CLA) is known for its antilipogenic effect but the mechanism is not fully clear. In this study, the potential of yeast (Saccharomyces cerevisiae) metabolism to offer evidence for the mechanism was investigated. The inhibitory effect of CLA on lipid accumulation was studied by analysing the transcript abundance of selected genes involved in triacylglycerol synthesis (LRO1, DGA1, ARE1 and ARE2) in the presence of the two bioactive CLA isomers: trans-10,cis-12 and the cis-9,trans-11 CLA. None of the enzymes was reduced in transcription but the expression of ARE2 was induced by trans-10,cis-12 CLA. However, the ARE2 overexpression did not contribute to lipid accumulation. The expression of the Δ9 desaturase gene, OLE1, was reduced by the cis-9,trans-11 but not by the trans-10,cis-12 isomer. In the TGL3/TGL4-knockout strain the triacylglycerol content also remained high in the CLA fed cells. Triacylglycerol hydrolysis rather than synthesis was the most probable reason for the reduced lipid content in yeast induced by CLA. This study revealed new aspects of the functionality of CLA in eukaryotic lipid metabolism. Yeast was proven to be an applicable model to study further the mechanism of trans-10,cis-12 CLA functionality on lipid metabolism.

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

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

Effect of pre- and postpartum supplementation with lipid-encapsulated conjugated linoleic acid on reproductive performance and the growth hormone–insulin-like growth factor-I axis in multiparous high-producing dairy cows

The objective of the study was to evaluate the effect of prepartum and postpartum (PP) supplementation with 2 isomers of conjugated linoleic acid (CLA) on reproductive parameters and some related metabolic factors in dairy cows. High-producing, multiparous Holstein Friesian cows (n = 60) were allotted to 3 treatment groups: the CLA1 group (n = 20) was supplemented with 70 g of lipid-encapsulated CLA providing 7 g each of cis-9,trans-11 and trans-10,cis-12 CLA from d 21 (d 21) before expected calving until d 7 after artificial insemination (AI), that is, until 77 to 91 d PP; the CLA2 group (n = 20) was supplemented with the same amount of CLA beginning at calving until d 7 after AI; and the control group (n = 20) received an isocaloric, isonitrogenous, and isolipidic diet. Blood samples were taken weekly to measure glucose, insulin, insulin-like growth factor-I (IGF-I), and leptin. Liver biopsy was performed in 10 cows per group for growth hormone receptor 1A and IGF-I mRNA analyses. At d 49 to 63 PP, ovulation was synchronized with the Pre-Synch protocol followed by fixed-time AI. Milk progesterone was monitored from calving until d 35 post-AI. Cows returning to estrus following AI were inseminated. Supplementation with CLA before calving improved the recovery of plasma leptin levels in the early PP period (from the day of calving until wk 3 PP; treatment effect). Later PP (wk 5), plasma IGF-I, and leptin remained significantly higher in both CLA1 and CLA2 groups compared with control, although hepatocellular IGF-I mRNA was not different among groups. Plasma IGF-I levels remained higher in both CLA-treated groups on the day of AI. Growth hormone receptor 1A mRNA levels in hepatic tissue decreased in all groups, reaching a nadir in the first week PP. Days to first PP ovulation did not differ between groups; however, both supplemented groups conceived earlier compared with control (d 97 ± 19, d 97 ± 23, and d 113 ± 30 for CLA1, CLA2, and control, respectively). Plasma progesterone concentration was higher in both supplemented groups on d 2 to 5 following the synchronized ovulation than in controls. We concluded that CLA supplementation around calving alters PP metabolic signals as reflected by higher plasma leptin and IGF-I levels. Conjugated linoleic acid stimulated early luteal function and reduced the PP interval to conception.

The effect of conjugated linoleic acid, acetate, and their interaction on adipose tissue lipid metabolism in nonlactating cows

During biohydrogenation-induced milk fat depression, adipose tissue lipogenesis may be increased through nutrients spared from milk fat synthesis. However, the direct effect of trans-10,cis-12 conjugated linoleic acid (CLA) and the indirect effect of spared nutrients on adipose tissue lipogenesis during milk fat depression is not clear. The objective of this study was to determine the direct effect of CLA, spared acetate, and their interaction on adipose tissue lipogenesis using nonlactating dairy cows as an experimental model, which allows separation of the effect of CLA and nutrient sparing. Eight ruminally cannulated, multiparous nonlactating and pregnant Holstein cows were randomly assigned to treatments in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Factors were CLA and acetate, and treatments were control (CON), rumen acetate infusions (Ac; continuous infusion of 7 mol/d adjusted to pH 6.1 with sodium hydroxide), abomasal infusion of trans-10,cis-12 CLA (CLA; 10 g/d of both trans-10,cis-12 CLA and cis-9,trans-11 CLA), and Ac + CLA (Ac + CLA). Dry matter intake was not affected by Ac, but tended to decrease by CLA. Plasma trans-10,cis-12 CLA concentration as a percentage of fatty acids was increased by 0.3 percentage points by CLA. No treatment effect was observed on plasma glucose and β-hydroxybutyrate, but an interaction was observed of CLA and Ac on plasma insulin and nonesterified fatty acids. Insulin was increased 24% by CLA, but not by Ac + CLA, and nonesterified fatty acids were increased 55% by Ac + CLA, but not by CLA alone. Lipogenesis and oxidation capacity of adipose tissue explants were not affected by treatments. Adipose expression of key lipogenic factors (peroxisome proliferator-activated receptor γ2 and sterol response element binding protein 1c) were reduced by CLA, by the interaction of Ac and CLA (sterol response element binding protein 1c), and tended to be reduced with Ac (S14 and peroxisome proliferator-activated receptor γ1). Expression of several adipose lipogenic enzymes (fatty acid synthase, acetyl-CoA carboxylase, and stearoyl-CoA desaturase 1) was reduced by CLA and Ac. An interaction was observed of Ac and CLA for fatty acid binding protein 4, which was decreased by Ac, but not Ac + CLA. In conclusion, in the nonlactating cow, adipose tissue is sensitive to the anti-lipogenic effects of trans-10,cis-12 CLA at the transcription level and acetate does not stimulate lipogenesis.