Diet‐dependent occurrence of CLA isomers in rumen and duodenal digesta of slaughtered bulls

This study examined the effects of linolenic acid‐rich vs. linoleic acid‐rich feeding system on the occurrence of individual CLA isomers in the rumen and duodenum digesta of German Holstein and German Simmental bulls using Ag⁺‐HPLC/DAD. The diet affected the biosynthesis of individual CLA isomers in the rumen of the bulls of both breeds. The isomer t‐11,c‐13 CLA was detected as the most abundant isomer in the rumen of linolenic acid‐rich diet‐fed bulls, up to six times higher compared to linoleic acid‐rich diet‐fed bulls. However, the main isomer in muscle lipid, c‐9,t‐11 CLA, was produced to a low extent in the rumen of linolenic acid‐rich diet‐fed bulls compared to higher concentrations of this isomer in the rumen of linoleic acid‐rich diet‐fed bulls. The isomers t‐7,c‐9 CLA and t‐8,c‐10 CLA were not present in the rumen samples of bulls fed both diets; however, abundant t‐7,c‐9 CLA was identified in the duodenum. The CLA isomers t‐12,t‐14 CLA and t‐11,t‐13 CLA were identified as the main t,t CLA isomers in the rumen, and were significantly enhanced in the rumen of linolenic acid‐rich diet‐fed compared to linoleic acid‐rich diet‐fed bulls. In contrast to c‐9,t‐11 CLA, the t,t CLA isomers seem to be biosynthesized predominantly in the rumen, further transported via the duodenum and finally deposited in the tissue lipids mainly in linolenic acid‐rich diet‐fed bulls. This was shown earlier for muscle and subcutaneous fat samples from the same animal experiment.     

<Emphasis Type="Italic">Trans</Emphasis>-7,<Emphasis Type="Italic">cis</Emphasis>-9 CLA is synthesized endogenously by Δ<Superscript>9</Superscript>-desaturase in dairy cowsin dairy cows

Cis-9,trans-11 and trans-7,cis-9 CLA are the most prevalent CLA isomers in milkfat. The majority of cis-9,trans-11 CLA is synthesized endogenously by Δ⁹-desaturase. We tested the hypothesis that trans-7,cis-9 CLA originates from endogenous synthesis by inhibiting Δ⁹-desaturase with a source of cyclopropene FA (sterculic oil: SO) or with a trans-10,cis-12 CLA supplement. Experiment 1 (four cows; Latin square) involved four treatments: control, SO, partially hydrogenated vegetable oil (PHVO), and PHVO+SO. Milk, plasma, and rumen fluid were collected. Experiment 2 treatments (four cows) were 0 or 14.0 g/d of 10,12 CLA supplement; milk and plasma were collected. Samples were analyzed by GC and Ag⁺-HPLC to determine FA. In Experiment 1, SO decreased milkfat content of trans-7,cis-9 CLA by 68 to 71% and cis-9,trans-11 CLA by 61 to 65%. In Experiment 2, the 10,12 CLA supplement decreased milkfat content of trans-7,cis-9 CLA and cis-9,trans-11 by 44 and 25%, respectively. Correcting for the extent of treatment-induced inhibition of Δ⁹-desaturase based on changes in myristic and myristoleic acids, endogenous synthesis of trans-7,cis-9 CLA represented 85 and 102% in Experiments 1 and 2, respectively. Similar corrected values were 77 and 58% for endogenous synthesis of cis-9,trans-11 CLA. Thus, milkfat cis-9,trans-11 CLA was primarily from endogenous synthesis with a minor portion from rumen escape. In contrast, trans-7,cis-9 CLA was not present in rumen fluid in significant amounts. Results indicate this isomer in milkfat is derived almost exclusively from endogenous synthesis via Δ⁹-desaturase.     

Differences in CLA isomer distribution of cow's milk lipids

The uniqueness of ruminant milk lipids is based on their high concentration of CLA. Maximal CLA concentrations in milk lipids require optimal conditions of ruminal fermentation and substrate availability, conditions like those present in pasture-fed cows. Our previous work showed that farm management (indoor feeding vs. pasture feeding) markedly influenced the CLA concentration. In this study, the objective was to evaluate the influence of the farm management system as dependent on different locations. Milk samples from different locations (Thuringia and the Alps, representing diverse altitudes) were collected during the summer months and analyzed for FA profile and CLA isomer distribution. The proportion of PUFA and total CLA in milk fat was significantly lower in milk from indoor cows compared with the pasture cows in the Alps. The trans-11 18∶1 in milk fat of Alpine cows was elevated, in contrast to lower values for trans-10 18∶1. Milk from cows grazing pasture in the Alps was higher in EPA and lower in arachidonic acid than milk from indoor-fed cows. The proportion of cis,trans/trans,cis isomers of CLA was 10 times higher from the indoor cows than from the Alpine cows. In addition to the major isomer cis-9,trans-11, this difference also occurred for the trans-11,cis-13 isomer, which represented more than a fourth of the total CLA present in milk fat. This is the first report showing a special isomer distribution in the milk fat of cows living under very natural conditions. We hypothesize that the CLA isomer trans-11,cis-13 is formed in large quantity as a result of grazing mountain pasture, which is rich in α-linolenic acid.     

Evaluating the In Vitro Metabolism of Docosahexaenoic Acid in Sheep Rumen Fluid

Rumen metabolism (e.g., biohydrogenation) of dietary unsaturated fatty acids (FA) is one of the main reasons why ruminant fats tend to be highly saturated and contain many isomerized FA intermediates. The process by which long-chain (20- to 24-carbon FA) polyunsaturated FA (LC-PUFA) are metabolized by rumen bacteria is not as well understood as that of linoleic or linolenic acids. In order to better understand the fate of LC-PUFA in the rumen several concentrations of docosahexaenoic acid (DHA) were evaluated in in vitro batch incubations ranging from 100 to 1,500 μg per 6 mL of incubation volume using rumen fluid from sheep and incubated for 0, 1, 2, 3, and 6 h. From the results, it was shown that DHA was extensively metabolized at low (100 to 300 μg/6 mL incubation volume), but not at high level of inclusion (800 μg). At 300 μg of DHA most of the depleted DHA was recovered as LC-DHA metabolites within the first 6 h of incubation, and at the lowest levels (100 μg of incubation volume) further metabolism is apparent at 6 h. Using SP-2560 GC columns several LC-DHA metabolites were shown to elute after 24:0 and just past DHA, a region generally free of interfering FA. The present in vitro study would appear to be a useful method to evaluate the production of DHA metabolites in combination with its depletion.     

Polyunsaturated fatty acids regulate cytokine and prostaglandin E2 production by respiratory cells in response to mast cell mediators

A protective association between breastfeeding and the development of bronchial asthma has been demonstrated. However, a mechanism remains unclear. FA present in human milk but rare in infant formula have been associated with marked immunological modulation as well as some indications of protection from asthma development. We examined the effect of in vitro manipulation of membrane phospholipid on the production of cytokines and prostaglandin (PG)E₂ by respiratory epithelial cells (A549) in response to stimulation by mast cell mediators of allergic disease [histamine, tumor necrosis factor (TNF)-α, interleukin (IL)-4 and IL-5]. DHA and CLA significantly decreased the production of IL-8 in response to stimulation by TNF-α [2907±970 (DHA) and 6471±1203 (CLA) vs. 12,287±2309 (control) pg/ml; P≤0.05, mean±SEM], whereas both EPA and DHA reduced histamine-stimulated RANTES (regulation on activation, T cell-expressed and-secreted) production [2314±861 (EPA) and 877±326 (DHA) vs. 8526±1118 (control) pg/ml; P≤0.03]. PGE₂ released in response to histamine was decreased by n−3 [1305±399 (α-linolenic acid), 406±73 (EPA), and 265±32 (DHA) vs. 9324±3672 (control) pg/mL; P≤0.05] and increased by n−6 [18,843±4439 (arachidonic acid) vs. 9324±3672 (control) pg/mL; P=0.02], with CLA producing a decrease of the same magnitude as DHA [553±126 (CLA) vs. 9324±3672 (control) pg/mL; P=0.03]. This study demonstrates the potential for immunological manipulation of the respiratory epithelium by FA in situ during allergic responses and suggests that further investigation into FA intervention in infants via human milk or supplemented infant formula, to prevent the development of allergic disease, may be worthwhile.     

Oxidative stability of conjugated linoleic acids relative to other polyunsaturated fatty acids

Contrary to current opinion, conjugated linoleic acids (CLA) as a mixture of several isomers have been previously shown to function as prooxidants in the form of free fatty acids and methyl esters in heated canola oil. Furthermore, CLA oxidizes considerably faster than linoleic acid. However, stability of CLA relative to other polyunsaturated fatty acids remains undetermined. The present study was therefore undertaken to examine the relative oxidation rate of CLA compared with that of linolenic acid (LNA), arachidonic acid (AA), and docosahexaenoic acid (DHA) in air at 90°C. CLA, both in the form of free fatty acids and triacylglycerols, were extremely unstable to the same extent as DHA, but they oxidized considerably faster than LNA and AA. The mechanism by which CLA were readily decomposed was probably due to formation of the unstable free-radical intermediate.     

Lipids and human milk

To support the growth and development of the breast‐fed infant, human milk provides the dietary essential fatty acids, linoleic acid (LA; 18:2n‐6), α‐linolenic acid (ALA, 18:3n‐3), as well as longer‐chain polyunsaturated fatty acids including arachidonic acid (20:4n‐6) and docosahexanoic (DHA 22:6n‐3). The linoleic acid, alpha‐linolenic acid, DHA and arachidonic acid concentration of pasteurized and unpasteurized human milk remains stable during the first month of storage at –20°C and –80°C. However after the first month, a slow decrease in concentration progresses until the end of 6 months of storage at both temperatures. The levels of n‐6 and n‐3 fatty acids, particularly linoleic acid, alpha‐linolenic acid and DHA, in human milk vary widely within and among different populations, and are readily changed by maternal dietary intake of the respective fatty acid. The present paper reviews recent understanding from key researchers of maternal diet and human milk fat composition and form our work the effect of milk fat composition on storage conditions. It is important to understand that maternal diet can affect human milk fat composition and subsequently infant development and growth.     

Supplemental Conjugated Linoleic Acid Consumption Does Not Influence Milk Macronutrient Contents in all Healthy Lactating Women

The term “conjugated linoleic acid” (CLA) refers to a group of positional and geometric isomers that are derived from linoleic acid and are found primarily in meat and milk products from ruminant animals. Due to the array of putative benefits associated with various forms of CLA, there has been recent interest in supplementing human diets with these fatty acids especially when weight loss is desired. However, in many animal models, CLA has been shown to decrease milk fat production. There is some concern, therefore, that maternal CLA supplementation during lactation might inadvertently decrease nutrient supply to the nursing infant. However, there is only limited research on the effect of CLA consumption on milk fat content in women. Based on previously published work from our laboratory, we hypothesized that CLA supplementation would reduce the milk fat percentage in lactating women in a dose-dependent manner. Breastfeeding women (n = 12) were assigned randomly to treatments of 4 g/day safflower oil (SFO), 2 g/day CLA plus 2 g/day SFO, or 4 g/day CLA in a double blind, 3 × 3 Latin square design. Conjugated linoleic acid supplements contained approximately equal amounts of cis9,trans11–18:2 and trans10,cis12–18:2; the two most common isoforms of CLA. Milk was collected by complete breast expression on the last day (day 5) of each intervention period and analyzed for macronutrient and fatty acid composition. On day 4 of each intervention period, infant milk consumption was estimated by 24 h weighing of the infant. Washout periods were 9 days in length. We observed a dose-dependent increase in the concentrations of cis9,trans11–18:2 and trans10,cis12–18:2 in the milk fat. However, we detected neither a change in overall macronutrient composition nor infant milk consumption. These data do not support those obtained from animal models or our previous human work suggesting that consumption of CLA mixtures necessarily reduces milk fat. It is possible that either (1) the interpretation of our previously published data should be reevaluated, and/or (2) there are important intra- and inter-species differences in this regard.     

Sulfur-substituted and α-methylated fatty acids as peroxisome proliferator-activated receptor activators

FA with varying chain lengths and an α-methyl group and/or a sulfur in the β-position were tested as peroxisome proliferator-activated receptor (PPAR)α,-δ(β), and-γ ligands by transient transfection in COS-1 cells using chimeric receptor expression plasmids, containing cDNAs encoding the ligand-binding domain of PPARα,-δ, and-γ. For PPARα, an increasing activation was found with increasing chain length of the sulfur-substituted FA up to C₁₄-S acetic acid (tetradecylthioacetic acid=TTA). The derivatives were poor, and nonsignificant, activators of PPARδ. For PPARγ, activation increased with increasing chain length up to C₁₆-S acetic acid. A methyl group was introduced in the α-position of palmitic acid, TTA, EPA, DHA, cis9,trans11CLA, and trans10,cis12 CLA. An increased activation of PPARα was obtained for the α-methyl derivatives compared with the unmethylated FA. This increase also resulted in increased expression of the two PPARα target genes acyl-CoA oxidase and liver FA-binding protein for α-methyl TTA, α-methyl EPA, and α-methyl DHA. Decreased or altered metabolism of these derivatives in the cells cannot be excluded. In conclusion, saturated FA with sulfur in the β-position and increasing carbon chain length from C₉−S acetic acid to C₁₄−S acetic acid have increasing effects as activators of PPARα and-γ in transfection assays. Furthermore, α-methyl FA derivatives of a saturated natural FA (palmitic acid), a sulfur-substituted FA (TTA), and PUFA (EPA, DHA, c9,t11 CLA, and t10,c12 CLA) are stronger PPARα activators than the unmethylated compounds.     

Effect of Linseed Fed as Rolled Seeds,Extruded Seeds or Oil on Fatty Acid Rumen Metabolism and Intestinal Digestibility in Cows

Linseed, a source of linolenic acid, is used in ruminant diets to increase polyunsaturated fatty acids (FA) in animal products. Seed processing is known to have an impact on FA rumen metabolism, but few data are available for linseed. We studied the effect of linseed lipid on ruminal metabolism and intestinal digestibility in cows. Three modes of linseed processing: rolled linseed (RL), extruded linseed (EL) and linseed oil plus linseed meal (LO), supplemented at 7.5% of DM intake, were compared to a control diet (C). Duodenal flows, intestinal digestibility and plasma composition were determined. The duodenal flow of linolenic acid was similar among diets. The sum of t10 and t11-18:1, which were coeluted, was increased with lipid-supplemented diets and represented more than 60% of trans 18:1 for EL and LO diets. The main 18:2 isomers were c9, c12 and t11, c15 among the non-conjugated isomers, and t11, t13 among CLA. Linseed supplementation increased the duodenal flow of unsaturated intermediates of biohydrogenation, and this effect was more pronounced for extruded seeds and oil than for rolled seeds. For most 18-carbon FA, intestinal digestibility was slightly higher for C and LO diets than for RL and EL. Plasma concentrations of non-conjugated 18:2 and linolenic acid were similar among the lipid-supplemented diets. Within diet, profiles of 18:1 isomers (except c9) remained very similar between duodenal and plasma FA.     

Vaccenic Acid and <Emphasis Type="Italic">cis</Emphasis>-9,<Emphasis Type="Italic">trans</Emphasis>-11 CLA in the Rumen and Different Tissues of Pasture- and Concentrate-Fed Beef Cattle

The objective of present study was the comparison of trans-11 18:1 (VA) and cis-9,trans-11 CLA concentrations in the rumen and different tissues in beef cattle, and to examine the diet and breed effects on the compound concentration and deposition. Sixty-four German Holstein and German Simmental bulls were randomly assigned to two dietary treatments, based on concentrate or pasture. The concentration of cis-9,trans-11 CLA and VA in rumen, duodenal digesta and different tissues was determined by gas chromatography. The results showed that pasture relative to concentrate feeding significantly increased the concentration of VA in duodenal digesta, plasma and erythrocyte phospholipids. Pasture-based feeding resulted in a significant enrichment of cis-9,trans-11 CLA in plasma lipids and erythrocyte phospholipids, but not in rumen and duodenal digesta, compared to concentrate-fed diet. Diet did not affect the cis-9,trans-11 CLA concentrations (mg/100 g fresh tissue) in semitendinosus muscle and subcutaneous fat. There was a breed effect on the deposition of cis-9,trans-11 CLA in longissimus muscle with lower concentration in pasture-fed German Simmental bulls compared to concentrate-fed bulls. However, pasture feeding significantly increased both, the VA and cis-9,trans-11 CLA concentrations in liver and heart tissues. Both diet and breed effects on Δ⁹-desaturase index was observed in muscle and subcutaneous fat tissues. There was a linear relationship between the concentration of VA and cis-9,trans-11 CLA and the coefficients of determination (R ²) varied between 0.29 and 0.87 from rumen to the different tissues.     

Partitioning of Rumen-Protected n-3 and n-6 Fatty Acids is Organ-Specific in Growing Angus Heifers

Dietary polyunsaturated fatty acids (PUFA), especially n-3 and n-6 fatty acids (FA), play an important role in the regulation of FA metabolism in all mammals. However, FA metabolism differs between different organs, suggesting a distinct partitioning of highly relevant FA. For the present study in cattle, a novel technology was applied to overcome rumen biohydrogenation of dietary unsaturated FA. Angus heifers were fed a straw-based diet supplemented for 8 weeks with 450 g/day of rumen-protected oil, either from fish (FO) or sunflower (SO). The FA composition in blood and five important organs, namely heart, kidney, liver, lung, and spleen, was examined. In blood, proportions of polyunsaturated FA were increased by supplementing FO compared to SO. The largest increase of eicosapentaenoic acid (EPA) proportion was found with FO instead of SO in the kidney, the lowest in the lung. Docosahexaenoic acid (DHA) was increased more in the liver than in kidney, lung, and spleen. The heart incorporated seven times more EPA than DHA, which is more than all other organs and described here for the first time in ruminants. In addition, the heart had the highest proportions of α-linolenic acid (18:3n-3) and linoleic acid (18:2n-6) of all organs. The proportions of polyunsaturated FA in the lung and spleen were exceptionally low compared to heart, liver, and kidney. In conclusion, it was shown that the response to FO in the distribution of dietary n-3 FA was organ-specific while proportions of n-6 FA were quite inert with respect to the type of oil supplemented.     

Maternal supplementation with CLA decreases milk fat in humans

CLA refers to isomers of octadecadienoic acid with conjugated double bonds. The most abundant form of CLA (rumenic acid (RA): c9,t11-18∶2) is found in milk and beef fat. Further, CLA supplements containing RA and t10,c12−18∶2 are now available. Consumption of commercially produced CLA has been shown to decrease adipose accretion in growing laboratory and production animals and cause milk fat depression in cows. We tested the hypothesis that CLA supplementation would increase milk CLA concentration and decrease milk fat content in humans. Breastfeeding women (n=9) participated in this double-blind, placebo-controlled, crossover study divided into three periods: intervention l (5 d), washout (7 d), and intervention II (5 d). Women were randomized to treatment order. During each intervention period, women consumed 1.5 g of CLA supplement or placebo (olive oil) daily; during the washout period, no supplements were consumed. Milk was collected by complete breast expression on the final day of each period; milk output was estimated by 24-h weighing on the penultimate day of each intervention period. Milk RA and t10,c12−18∶2 concentrations were greater (P<0.05) during the CLA treatment period as compared to the placebo period. Milk fat content was significantly lower during the CLA treatment, as compared to the placebo treatment (P<0.05). Data indicate no effect of treatment on milk output. Therefore, it would be prudent that lactating women not consume commercially available CLA supplements at this time. This paper was published in part in Masters, N., McGuire, M.A., and McGuire, M.K. (1999) Conjugated Linoleic Acid Supplementation and Milk Fat Content in Humans, FASEB J. 13, A697.     

Maternal dietary alpine butter intake affects human milk: Fatty acids and conjugated linoleic acid isomers

Consumption of CLA by lactating women affects the composition of their milk, but the pattern of the different CLA isomers is still unknown. We determined the effects of short maternal supplementation with CLA-rich Alpine butter on the occurrence of FA and CLA isomers in human milk. In an open randomized controlled study with a two-period cross-over design, milk FA and CLA isomer concentrations were measured on postpartum days >-20 in two parallel groups of lactating women before, during, and after consumption of defined quantities of Alpine butter or margarine with comparable fat content (10 d of butter followed by 10 d of margarine for one group, and vice versa in the other). In the 16 women who completed the study (8/group), Alpine butter supplementation, increased the C₁₆ and C₁₈ FA, the sum of saturated FA, the 18∶1 trans FA, and the trans FA with CLA. The CLA isomer 18∶2 c9, t11 increased by 19.7%. Significant increases were also found for the isomers t9,t111, t7,c9,t11,c13, and t8,c10 18∶2. The remaining nine of the total 14 detectable isomers showed no changes, and concentrations were <5 mg/100g fat. A breastfeeding mother can therefore modulate the FA/CLA supply of her child by consuming Alpine butter. Further studies will show whether human milk containing this FA and CLA isomer pattern acts as a functional food for newborns.     

Effects of cis-9, trans-11 and trans-10, cis-12 CLA isomers on liver and adipose tissue fatty acid profile in hamsters

The aim of the present study was to investigate the effect of cis-9, trans-11 and trans-10, cis-12 CLA on FA composition of TAG in epididymal adipose tissue and liver, and of hepatic phospholipids PL. Twenty-four Syrian Golden hamsters were randomly divided into three groups of eight animals each and fed semipurified atherogenic diets supplemented with either 0.5 g/100g diet of linoleic acid or cis-9, trans-11 or trans-12, cis-9 CLA for 6 wk. Total lipids were extracted, and TAG and PL were separated by TLC. FA profile in lipid species from liver and adipose tissue, as well as in feces, was determined by GC. Trans-10, cis-12 CLA feeding significantly reduced linoleic and linolenic acids in TAG from both tissues, leading to reduced total PUFA content. Moreover, in the epididymal adipose tissue docosenoic and arachidonic acids were significantly increased. In liver PL, although no changes in individual FA were observed, total saturated FA (SFA) were decreased. No changes in TAG and PL FA profiles were induced by the cis-9, trans-11 CLA. TAG and PL incorporated cis-9, trans-11 more readily than trans-11, cis-12 CLA. This difference was not due to differential intestinal absorption, as shown by the analysis of feces. We concluded that only trans-10, cis-12 CLA induces changes in FA composition. Whereas increased PUFA content was observed in either liver or adipose tissue TAG, decreased SFA were found in liver PL. Incorporation of cis-9, trans-11 CLA in TAG is greater than that of trans-10, cis-12 CLA, but this is not due to differences in intestinal absorption.     

Duodenal availability of conjugated linoleic acids after supplementation to dairy cow diets

The objective of the present study was to investigate the effects of a lipid‐encapsulated CLA preparation on rumen metabolism and the actual post‐ruminal bioavailability of the applied CLA isomers. In the rumen, the CLA supplementation modified the molar proportions of VFA. In period CLA‐1 the rumen fermentation shifted toward more butyric acid at the expense of acetic acid. The highest CLA supplementation resulted in increased amounts of isobutyric, isovaleric, and valeric acid. The apparent ruminal digestibility of starch increased in period CLA‐2. The ruminal protein degradation was higher after CLA supplementation, while the efficiency of the use of the RDP for microbial protein synthesis declined. The duodenal flow of trans‐10,cis‐12 CLA amounted to 16 and 5% of the intake in periods CLA‐1 and CLA‐2, respectively. The transfer of trans‐10,cis‐12 CLA from duodenum into milk was 36 and 48% in periods CLA‐1 and CLA‐2, respectively. Overall, the observed effects of the supplementation of lipid‐encapsulated CLA on the parameters of rumen metabolism were negligible. The actual low post‐ruminal bioavailability of trans‐10,cis‐12 CLA suggest that most of the applied fat supplement was biohydrogenated.     

Consumption of c9,t11-18:2 or t10,c12-18:2 enriched dietary supplements does not influence milk macronutrients in healthy, lactating women

Substantial research suggests that the t10,c12–18:2, but not the c9,t11–18:2, isomer of conjugated linoleic acid (CLA) reduces milk fat synthesis in lactating bovine and rodent species. Because fat is the major energy-yielding component in human milk, we were interested in whether this is true for women as well. Thus, the effects of c9,t11–18:2 and t10,c12–18:2 on milk fat were examined in breast-feeding women (n = 12) in a double-blind, placebo-controlled, crossover study with latin-square design. The study was divided into six periods: baseline (3 days), three intervention periods (5 days each), and two washout periods (9 days each). During each intervention period, women consumed 750 mg/day of a supplement containing predominantly c9,t11–18:2, t10,c12–18:2, or 18:1 (olive oil placebo). Milk was collected by complete breast expression on the final day of each period. Infant milk consumption was estimated by 24 h weighing on the penultimate day of each intervention and washout period, and maternal adiposity (% body fat) was determined at baseline using dual energy X-ray absorptiometry. Milk c9,t11–18:2 and t10,c12–18:2 concentrations were greater (P < 0.05) during the corresponding CLA treatment periods as compared to the placebo period, providing strong evidence of subject compliance. Both CLA isomers were transferred into milk fat at relatively high efficiency; average transfer efficiency was estimated to be 23.3%. Compared to the placebo treatment, milk fat content was not reduced during either CLA treatment. Data indicate that body fatness did not modify any putative effect of isomeric CLA consumption on milk fat concentration. The evidence from this study suggests that the sensitivity of lactating women’s mammary tissue to an anti-lipogenic effect of the t10,c12–18:2 isoform of CLA may be less than previously hypothesized.     

Effect of abomasal infusions of geometric isomers of 10,12 conjugated linoleic acid on milk fat synthesis in dairy cows

The trans-10, cis-12 isomer of conjugated linoleic acid (CLA) decreases TAG accumulation in 3T3-L1 adipocytes, reduces lipid accretion in growing animals, and inhibits milk fat synthesis in lactating mammals. However, there is evidence to suggest that other FA may also exert antilipogenic effects. In the current experiment, the effects of geometric isomers of 10,12 CLA on milk fat synthesis were examined using four Holstein-British Friesian cows in a 4×4 Latin Square experiment with 14-d periods. Treatments consisted of abomasal infusions of skim milk, or skim milk containing trans-10, cis-12 CLA (T1), trans-10, trans-12 CLA (T2), or a mixture of predominantly 10,12 isomers containing (g/100 g) trans-10, cis-12 (35.0), cis-10, trans-12 (23.2), trans-10, trans-12 (14.9), and cis-10, cis-12 (5.1). CLA supplements were prepared from purified ethyl linoleate and infused as nonesterified FA. Infusions were conducted over a 4-d period with a 10-d interval between treatments and targeted to deliver 4.5 g/d of 10,12 CLA isomers. Compared with the control, trans-10, trans-12 CLA had no effect (P>0.05) on milk fat yield, whereas treatments T1 and T3 depressed (P<0.05) milk fat content (19.8 and 22.9%, respectively) and decreased milk fat output (20.8 and 21.3%, respectively). Comparable reductions in milk fat synthesis to 4.14 and 1.80 g trans-10, cis-12/d supplied by treatments T1 and T3 indicate that other 10,12 geometric isomers of CLA have the potential to exert antilipogenic effects. The relative abundance of cis-10, trans-12 CLA in treatment T3 and the low transfer efficiency of this isomer into milk suggest that cis-10, trans-12 CLA was the active component.     

Dietary Fat Does Not Overcome trans-10, cis-12 Conjugated Linoleic Acid Inhibition of Milk Fat Synthesis in Lactating mice

Trans-10, cis-12 conjugated linoleic acid (CLA) is a potent inhibitor of milk fat synthesis in the cow and similarly reduces milk fat in rodents. The objective of this study was to determine whether dietary fat can overcome CLA inhibition of milk fat concentration in lactating mice. Wild type C57Bl/6J mice (n = 31) were fed semipurified diets containing either low fat (LF; 4% fat) or high fat (HF; 23.6% fat) starting 4–6 days postpartum. Dietary fat was increased by inclusion of high oleic sunflower oil. After 2 days on the experimental diets, lactating dams were orally dosed with either water (control) or trans-10, cis-12 CLA (20 mg/day) for 5 days. CLA treatment decreased pup growth similarly in both HF and LF diets. Milk fat percent was increased over 16% by the HF diet and decreased over 12% by CLA, but there was no interaction of dietary fat and CLA. Both CLA and the HF diet reduced the proportion of short- and medium-chain fatty acids that originate from de novo synthesis, and there was no interaction of diet and CLA. CLA had no effect on the percent of preformed fatty acids, but the HF diet increased their abundance. Dietary fat and CLA both modified mammary expression of lipogenic enzymes and regulators, but no interactions were observed. In conclusion, CLA reduced milk fat concentration and litter growth, but these effects were not overcome by increased dietary fat from high oleic sunflower oil. CLA inhibition of milk fat in the mammary gland is not substrate dependent, and the mechanism is independent from dietary supply of oleic acid.     

Rosiglitazone,a PPAR-γ Agonist,Fails to Attenuate CLA-Induced Milk Fat Depression and Hepatic Lipid Accumulation in Lactating Mice

Our objective was to investigate the combination of rosiglitazone (ROSI) and conjugated linoleic acid (CLA) on mammary and hepatic lipogenesis in lactating C57Bl/6 J mice. Twenty-four lactating mice were randomly assigned to one of four treatments applied from postpartum day 6 to day 10. Treatments included: (1) control diet, (2) control plus 1.5 % dietary CLA (CLA) substituted for soybean oil, (3) control plus daily intra-peritoneal (IP) rosiglitazone injections (10 mg/kg body weight) (ROSI), and (4) CLA plus ROSI (CLA-ROSI). Dam food intake and milk fat concentration were depressed with CLA. However, no effects were observed with ROSI. The CLA-induced milk fat depression was due to reduced expression for mammary lipogenic genes involved in de-novo fatty acid (FA) synthesis, FA uptake and desaturation, and triacyglycerol synthesis. Liver weight (g/100 g body weight) was increased by CLA due to an increase in lipid accumulation triggering a compensatory reduction in mRNA abundance of hepatic lipogenic enzymes, including acetyl-CoA carboxylase I and stearoyl-CoA desaturase I. On the contrary, no effects were observed with ROSI on hepatic and mammary lipogenic gene and enzyme expression. Overall, feeding CLA to lactating mice induced milk fat depression and increased hepatic lipid accumulation, probably due to the presence of trans-10, cis-12 CLA isomer, while ROSI failed to significantly attenuate both hepatic steatosis and reduction in milk fat content.