Conjugated linoleic acid and C18:1 isomers content in milk fat of sheep and their transfer to Pecorino Toscano cheese

The aim of the study was to evaluate the transfer of conjugated linoleic acid (CLA) and of C18:1 isomers from sheep milk to Pecorino Toscano protected denomination of origin cheese and to monitor the stability of these fatty acids during ripening. The total content of C18:1 fatty acids was not affected by milk treatment but there were changes in the isomers profile. In particular, the milk trans11 C18:1 decreased with pasteurization and after starter culture addition. In contrast, trans10 C18:1 and cis9, trans11 C18:2 were enhanced by the thermal treatment of milk. The fatty acid profile of cheese reflected that of milk used for cheese making with the exception of the C18:1 and CLA isomer profile. The cheese ripening affected the percentage of cis9, trans11 C18:2, which increased more than 10% on total content of CLA.     

FATTY ACID COMPOSITION IN TISSUES OF MICE FED DIETS CONTAINING CONJUGATED LINOLENIC ACID AND CONJUGATED LINOLEIC ACID

The influence of 1% alpha-eleostearic acid (α-ESA, cis9,trans11,trans 13-18:3) and 1% punicic acid (PA, cis9,trans11,cis13-18:3) on fatty acid composition in mouse tissues was compared with conjugated linoleic acid (CLA, mixture of primarily cis9,trans11- and trans10,cis12-18:2) in the present study. The content (% total fatty acids) of 18:2n-6 was significantly reduced in the heart and adipose tissues, and total polyunsaturated fatty acids (PUFAs) and n-6 PUFA were significantly reduced in adipose tissue by α-ESA, PA and CLA feeding. The content of 22:6n-3 and total n-3 PUFA were significantly increased in the liver, kidney and heart by PA feeding, but not by α-ESA. In contrast to PA, supplementation with CLA significantly decreased 22:6n-3 in the liver, kidney and heart. The content of 20:4n-6 was significantly decreased in the liver and kidney by CLA feeding, but not by α-ESA and PA. The present results indicate that α-ESA, PA and CLA have differential effects on 22:6n-3 and 20:4n-6 content in mouse tissues.

PRACTICAL APPLICATIONS

Conjugated linolenic acid (CLnA), a group of octadecatrienoic acid isomers with a conjugated triene system, has been reported to exhibit favorable physiological effects, including anticancer properties and regulation of lipid metabolism. Punicic acid and alpha-eleostearic acid, two isomers of CLnA, have been shown to convert into cis 9, trans 11-18:2 in vivo . The effect of CLnA on fatty acid composition in mouse tissues was investigated in comparison with CLA mixtures in the present study. The data obtained here could provide information for the potential application of CLnA-containing seeds as functional food ingredients, a natural source of endogenously formed cis 9, trans 11-18:2 and a dietary feeding strategy to beneficially modify the fatty acid composition of animal tissues.     

Conjugated linoleic acid in ewe milk fat

Ewe milk fat from five different herds was studied to determine the content of conjugated linoleic acid (CLA) isomers. Research was carried out by combining gas chromatography-mass spectrometry (GC-MS) of fatty acid methyl esters (FAME) and 4,4-dimethyloxazolyne derivatives (DMOX) with silver ion-high performance liquid chromatography (Ag+-HPLC). Reconstructed mass spectral profiles of CLA characteristic ions from DMOX were used to identify positional isomers and Ag+-HPLC to quantify them. Total CLA content varied from 0.57 to 0.97 g/100 g of total fatty acids. FAME and DMOX were separated into a complex mixture of minor isomers and major rumenic acid (9-cis 11-trans C18:2) by GC-MS using a 100-m polar capillary column. Rumenic acid would represent more than 75% of total CLA. 11-trans 13-trans, 11-13 cis/trans plus trans/cis and 7-9 cis/trans plus trans/cis were the main CLA isomers after rumenic acid. Minor amounts of 8-10 and 10-12 C18:2 isomers were also found. Although most of the isomers were present in each herd's milk, differences in content were observed for some CLA species.     

Factors influencing proportion and composition of CLA in beef

Bovine meat is criticised for the bad nutritional image of its lipids and fatty acids. However, with dairy products, beef is the major source of conjugated linoleic acid (CLA) which could have several human health benefits. The present study compared, from data of five nutritional experiments on bovine animals performed by the laboratory, the impact of factors linked to the animals (breed, age, sex, type of muscle) and to feeding conditions (basal diet, lipid supplements) on the CLA proportion and composition in muscles. Among these factors, linseed supplementation was an efficient way to increase CLA proportion in beef (+22% to +36%) but was highly modulated by the nature of the basal diet, and by intrinsic factors (breed, age/sex, type of muscle) since these ones could modulate CLA proportion in beef from 24% to 47%. Moreover, these factors modified also the proportion of cis,trans-CLA, related to cis,cis- and trans,trans-isomers. Specific biological properties of these latter isomers should be determine to understand the consequences of intramuscular CLA isomer variations for the health of consumers.     

Milk yield and composition during abomasal infusion of conjugated linoleic acids in dairy cows

Conjugated linoleic acids refer to a mixture of positional and geometric isomers of linoleic acid with conjugated double bounds. Three supplements of conjugated linoleic acids which differed in isomer enrichment were infused into the abomasum of lactating dairy cows to determine postruminal effect on milk yield and composition. Four Holstein cows were used in a 4 x 4 Latin square design. Treatments were 3-d abomasal infusion of 1) control, 5 kg of skim milk (carrier for conjugated linoleic acid supplements), 2) conjugated linoleic acid supplement 1 (28.8 g/d; 6.9 g of cis/trans 9,11; 6.4 g of cis/trans 8,10), 3) conjugated linoleic acid supplement 2 (48.5 g/d; 7.1 g of cis/trans 9,11; 4.1 g of cis/trans 8,10; 8.3 g of cis/trans 10,12; 5.5 g of cis/trans 11,13), and 4) conjugated linoleic acid supplement 3 (16.3 g/d; 7.1 g of cis/trans 9,11; 7.2 g of cis/trans 10,12). Infusions increased the conjugated linoleic acids content of milk fat from 0.43 g/100 g of fat for the control treatment to 1.02, 1.52, and 0.95 g/100 g of fat for conjugated linoleic acid supplements 1, 2, and 3, respectively. Apparent efficiency of transfer in milk fat was 25.2, 33.5, 21.0, and 28.4% for cis/trans 8,10, cis/trans 9,11, cis/trans 10,12, and cis/trans 11,13, respectively. Infusion of conjugated linoleic acids had no effect on dry matter intake, milk yield, and milk protein content. However, conjugated linoleic acid supplements reduced the content and yield of milk fat by 28 and 25%, respectively. The similarity of response for the different conjugated linoleic acid supplements did not allow us to identify the specific role of different isomers, but the changes in milk fatty acid composition indicated that effects were primarily on de novo fatty acid synthesis and the desaturation process.     

Conjugated linoleic acid conversion by six Lactobacillus plantarum strains cultured in MRS broth supplemented with sunflower oil and soymilk

Six strains of Lactobacillus plantarum, isolated from traditional dairy products of minority nationalities, were evaluated for their ability to produce conjugated linoleic acid (CLA) from free linoleic acid in vitro. All the 6 strains were found to be capable of converting linoleic acid to CLA when using sunflower oil as substrate or during soymilk fermentation. The inhibitory effect of linoleic acid on the growth of the L. plantarum was also discussed. The production of CLA was increased with adding high concentration of substrate in sunflower oil and IMAU60042 produced the highest CLA both in sunflower oil and soymilk. The CLA was composted by 2 isomers: cis9, trans11-CLA and tran10, cis12-CLA, and cis9, tran s11-CLA covered the most part of the total CLA formed except for L. plantarum P8. The production of CLA was decreased during the storage of fermented soymilk. The CLA contents decreased significantly in the first week, also more quickly in 2 wk. Especially, tran10, cis12-CLA decreased more rapidly than cis9, tran11-CLA. No dramatic change was observed among other 8 fatty acids in soymilk. The proportion of unsaturated fatty acids varied after fermentation with different L. plantrum strains, but all decreased the during storage. The research on the ability of converting CLA of L. plantrum strains could be basis for the future research and development of fermented soymilk products. PRACTICAL APPLICATION: Desirable probiotic traits, such as acid and bile tolerance, aggregation activity, and antibacterial activity, have been proved for the 6 Lactobacillus plantarum strains. The 6 L. plantarum strains might be used in the fermentation of soymilk to produce multifunctional probiotic soymilk products, especially the rich CLA contents.     

Hot topic: Fatty acid and conjugated linoleic acid (CLA) isomer composition of commercial CLA-fortified dairy products: evaluation after processing and storage

Conjugated linoleic acid (CLA) exerts a strong positive influence on human health but intake of these fatty acids is typically too low, and increased consumption of CLA is recommended. A good way to raise the CLA content in the diet without a radical change in eating habits seems to be the enrichment of commonly consumed food products with CLA supplements. This study analyzed the total fatty acid content and the CLA isomer composition of 6 commercially available CLA-fortified dairy products during processing and 10 wk of refrigerated storage. Research was carried out by combining gas chromatography and silver-ion HPLC. The tested samples were a CLA oil supplement, and several skim milk dairy products fortified with the supplement (milk, milk powder, fermented milk, yogurt, fresh cheese, and milk-juice blend). The CLA oil supplement was added such that the consumer received 2.4 g/d of CLA by consuming 2 servings. The predominant isomers present, C18:2 cis-9, trans-11 CLA and C18:2 cis-10, trans-12 CLA, were in at a similar ratio, which ranged from 0.97 to 1.05. These major isomers were not significantly affected by processing but a decrease in total CLA in fresh cheese samples was detected after 10 wk of refrigerated storage. Refrigerated storage and thermal treatment resulted in significant decreases or disappearance of some of the minor CLA isomers and a significant increase of trans, trans isomers from both cis, trans, trans, cis, and cis, cis isomers especially in CLA-fortified milk powder but also in fermented milk, yogurt, and milk-juice blend.     

Role of the conjugated linoleic acid in the prevention of cancer

There are multiple lines of evidence that a variety of natural fatty acids are effective in health promotion. Among these fatty acids, conjugated linoleic acid (CLA)--a collective term referring to a mixture of positional and geometric isomers of linoleic acid (LA, cis-9, cis-12-octadecadienoic acid)--is currently under intensive investigation due to its health-promotion potential. The antitumor activity of CLA is of special interest, since it shows inhibitory effects against multistage carcinogenesis at relatively low dietary levels. Many studies using in vivo and in vitro models have shown that CLA suppresses the development of multistage carcinogenesis at different sites. The research to date on CLA has provided a vast amount of information about the mechanism on how CLA functions in the prevention of cancer. This article discusses characteristics of CLA in the prevention of cancer in both in vivo and in vitro studies and the possible underlying chemoprevention mechanisms.     

Conjugated linoleic and linolenic acid production kinetics by bifidobacteria differ among strains

There is great interest in conjugated linoleic acid (CLA) and conjugated linolenic acid (CLNA) isomers because of their supposed health-promoting properties. Therefore, the differences in production kinetics of CLA and CLNA isomers from linoleic acid (LA) and α-linolenic acid (α-LNA), respectively, by bifidobacteria were investigated. Laboratory fermentations, supplemented with LA or α-LNA in the fermentation medium, were performed with Bifidobacterium bifidum LMG 10645, Bifidobacterium breve LMG 11040, B. breve LMG 11084, B. breve LMG 11613, B. breve LMG 13194, and Bifidobacterium pseudolongum subsp. pseudolongum LMG 11595. Conversion of LA and α-LNA to CLA and CLNA isomers, respectively, started immediately upon addition of the substrate fatty acids. During the active growth phase, the c9, t11-CLA isomer and the putative c9, t11, c15-CLNA isomer were formed. Further fermentation resulted in a reduction in the concentration of c9, t11-CLA and c9, t11, c15-CLNA and the subsequent production of the t9, t11-CLA isomer and the putative t9, t11, c15-CLNA isomer, respectively. Modelling of the growth and metabolite data indicated differences in production kinetics among the strains. Some strains displayed a high specific conversion of LA and α-LNA despite poor growth, whereas other strains grew well but displayed lower conversion. Production of specific CLA and CLNA isomers by bifidobacteria holds potential for the production of functional foods and could contribute to their probiotic properties.     

Influence of slaughter season and muscle type on fatty acid composition, conjugated linoleic acid isomeric distribution and nutritional quality of intramuscular fat in Arouquesa-PDO veal

The effects of the slaughter season and muscle type on lipid and conjugated linoleic acid (CLA) contents, fatty acid composition and isomeric profile of CLA in Arouquesa veal, from calves reared according to the specifications of the protected designation of origin (PDO), were assessed. Arouquesa purebred calves (n = 31) were raised in a traditional production system, slaughtered in early autumn (October) or late spring (June), and the longissimus dorsi and semitendinosus muscles were sampled for analysis. Arouquesa-PDO veal only showed seasonal differences in the levels of some minor fatty acids (16:1c9, 17:1c9, 18:1t, 18:3n − 3, 20:0 and 22:4n − 6) and CLA isomers (t12,t14, t9,t11 and c11,t13). Furthermore, significant interactions between the slaughter season and muscle type were obtained for several fatty acids and CLA isomers, total lipids and CLA, and the PUFA/SFA ratio. In both seasons, veal-PDO depicts values of pasture-fed cattle. From a human nutritional perspective, veal-PDO in both slaughter seasons has relatively high CLA contents and percentages of the c9,t11 CLA isomer, which is favourable, while the n − 6/n − 3 ratios are within the recommended values for the human diet. In conclusion, the results suggest that intramuscular fat in Arouquesa-PDO veal has high nutritional value throughout the year.     

Effect of dietary conjugated linoleic acid (CLA) supplementation on CLA isomers content and fatty acid composition of dry-cured Parma ham

To evaluate the effects of dietary conjugated linoleic acid (CLA) supplementation on cis-9, trans-11 and trans-10, cis-12 isomer content and on the fatty acid composition of subcutaneous adipose tissue and intramuscular lipids of biceps femoris, the lipids of 24 dry cured Parma hams, obtained from Large White pigs fed isoenergetic diets supplemented with either 0 (CON) or 0.25% (T1) CLA from 97 to 172kg live weight, were analysed by Ag(+)-HPLC and capillary GLC. The dietary treatment had no influence on lipid content of BF and subcutaneous adipose tissue but increased (P<0.01) the content of both isomers, more markedly in the latter tissue. CLA supplementation increased (P<0.01) myristic, palmitic, palmitoleic and total saturated fatty acids, and decreased oleic and eicosenoic acids in both tissues. In adipose tissue, an increase (P<0.05) in stearic acid content was observed. Supplementing the finishing diet of heavy pigs with CLA has thus shown to be an effective tool for increasing the CLA content in cured ham and, also, the saturated:unsaturated ratio of the lipids, a positive effect from a technological point of view. From a human nutritional standpoint, the changes occurring in fatty acid composition negatively affect the atherogenic and thrombogenic indexes, but probably these changes are of no particular concern, as they would only partly offset the progressive increase in the degree of unsaturation of heavy pig lipids that has occurred in recent years, as a consequence of reduced adipose depots in carcasses.     

The composition of bovine milk lipids: January 1995 to December 2000

Data from recent publications on bovine milk lipids are presented and discussed. This includes extraction of lipids, triacylglycerols, phospholipids, other complex lipids, sterols, isoflavones, and fatty acids. Improved gas-liquid and high performance liquid chromatography were used. Data on the trans and cis isomers of fatty acid and of conjugated linoleic acids are given, and the analyses are described. Papers about the lipids in milks and dairy products from the United States are few; where with the exception of trans-fatty acid isomers and conjugated linoleic acids, almost no research has been reported.     

新型食品营养强化剂-共轭亚油酸

共轭亚油酸 (Conjgatedlinoleicacid ,CLA)是一类含有共轭双键的亚油酸的异构体 ,由于具有特殊的脂肪链结构使得其具有多种生理功能。c9t11和t10cl2异构体是共轭亚油酸的主要活性异构体。本文对共轭亚油酸的生理功能、代谢、安全性等进行综述。     

Conjugated linoleic acid content and isomer distribution during ripening in three varieties of cheeses protected with designation of origin

Cheeses have been identified as important sources of conjugated linoleic acid (CLA), a mixture of positional and geometric isomers with potential anticarcinogenic activity and other beneficial properties. The objectives of this study were to examine the effects of ripening on the overall CLA content as well as on the isomers profile using GC and Ag⁺-HPLC. Three Spanish cheeses Protected with Designation of Origin (Mahón, Manchego and Cabrales) were manufactured in different cheesemaking plants and monitored at different times during the ripening period. Total CLA content varied from 3 to 9 mg/g of total fatty acids and rumenic acid (9-cis, 11-trans C18:2, RA) represented more than 75% of total CLA. After RA, 7–9 (cis/trans plus trans/cis), 11-trans, 13-trans and 11-trans, 13-cis C18:2 were the main CLA isomers. The results achieved confirm that the effect of ripening on the total CLA concentration and isomer distribution was negligible.     

Systematic evaluation on the effectiveness of conjugated linoleic acid in human health

The term CLA (conjugated linoleic acid) corresponds to a mixture of positional and geometric isomers of linoleic acid. Two of these isomers (9c, 11t and 10t, 12c) have biological activity. The milk and dairy products are the most abundant source of conjugated linoleic acid, which refers to a group of positional and geometric isomers of CLA (CLA 18:2 cis-9, cis-12). The following research aims to approach aspects regarding the CLA, as well as its relationship with diseases. Conjugated linoleic acids have been studied for their beneficial effects in the prevention and treatment of many diseases, including obesity, cancer, diabetes, and cardiovascular diseases. Scientific information put together the physiological properties of CLA, which serves as inputs to claim their potential as functional ingredients to be used in the prevention and control of several chronic metabolic disorders.     

乙醇胁迫提高植物乳杆菌p-8的共轭亚油酸(CLA)转化率和相关酶转录水平

该文研究了在MRS培养基中添加0.05 mg/mL LA（Linoleic acid,LA）和不同浓度的乙醇时植物乳杆菌p-8的CLA（Conjugated linoleic acid,CLA）转化率和CLA合成相关酶转录水平的差异情况。结果显示,发酵液中的三种CLA异构体转化率都是在添加0.50%乙醇时最高,其中转化cis9,trans11-CLA（t9,t11-CLA）异构体最高,为2.49%,比不添加乙醇增加2.37倍。添加不同浓度乙醇的发酵液中trans10,cis12-CLA（t10,c12-CLA）转化率都是最低的。菌体中产生的CLA非常少,但规律与发酵液的基本一致。添加0.50%乙醇菌体中t9,t11-CLA转化率最高,其转化率仅为0.05%,比不添加乙醇增加了5倍。当乙醇浓度高于0.50%时,各种不同CLA异构体的转化率却都减少。结果表明CLA是在胞液内产生后再被运转到胞外的,一定浓度范围内的乙醇胁迫通过提高CLA合成相关的酶基因转录水平,进而促进了CLA的转化,可见CLA合成相关酶基因转录水平是造成CLA转化率差异的主要原因。结果为阐明植物乳杆菌p-8产CLA的分子机制和寻找有效提高CLA生成的调控手段奠定了基础。     

Conjugated linoleic acid isomer concentrations in milk from high‐ and low‐input management dairy systems

BACKGROUND: Different conjugated linoleic acid (CLA) isomers are known to have contrasting physiology or health effects and there is growing evidence that the profile of natural isomers in milk is influenced by the production system. This survey is the first to compare feeding regimes and concentrations of 14 CLA isomers in milk from three production systems in the UK. RESULTS: Total CLA and seven isomers (including C18:2 c9t11 which comprised > 80% of total) were significantly higher in milk from both organically certified and non‐certified low input (LI) systems compared with milk from conventional high input farms. Sampling date also affected concentrations of total CLA and nine isomers; being lowest in March and highest in August. Seasonal differences were greater in milk from LI herds, thought to be due to changes in herbage and/or stage of lactation. Multivariate analysis showed a strong positive relationship between several CLA isomers and increasing levels of fresh forage in the diet. CONCLUSIONS: These results add to the evidence on how management adjustment may improve the profile of CLA isomers in milk fat, although animal or human intervention studies are required to identify the effects of consuming milk with different CLA levels and isomer profiles on human health. Copyright © 2009 Society of Chemical Industry     

Innovations in beef production systems that enhance the nutritional and health value of beef lipids and their relationship with meat quality

Consumers are becoming more aware of the relationships between diet and health and this has increased consumer interest in the nutritional value of foods. This is impacting on the demand for foods which contain functional components that play important roles in health maintenance and disease prevention. For beef, much attention has been given to lipids. This paper reviews strategies for increasing the content of beneficial omega-3 polyunsaturated fatty acids (PUFA) and conjugated linoleic acid (CLA) and reducing saturated fatty acids (SFA) in beef. Particular attention is given to intramuscular fat (IMF) and the relationships between fatty acid composition and key meat quality parameters including colour shelf life and sensory attributes. Despite the high levels of ruminal biohydrogenation of dietary PUFA, nutrition is the major route for increasing the content of beneficial fatty acids in beef. Feeding grass or concentrates containing linseed (rich in α-linolenic acid, 18:3n-3) in the diet increases the content of 18:3n-3 and its longer chain derivative eicosapentaenoic acid (EPA, 20:5n-3) in beef muscle and adipose tissue, resulting in a lower n-6:n-3 ratio. Grass feeding also increases docasahexaenoic acid (DHA, 22:6n-3). Feeding PUFA rich lipids which are protected from ruminal biohydrogenation result in further enhancement of the PUFA in meat with concomitant beneficial improvements in the ratio of polyunsaturated:saturated fatty acids (P:S ratio) and n-6:n-3 ratio. The main CLA isomer in beef is CLA cis-9, trans-11 and it is mainly associated with the triacylglycerol lipid fraction and therefore is positively correlated with level of fatness. The level of CLA cis-9, trans-11 in beef is related to (1) the amount of this isomer produced in the rumen and (2) synthesis in the tissue, by delta-9 desaturase, from ruminally produced trans vaccenic acid (18:1 trans-11; TVA). Feeding PUFA-rich diets increases the content of CLA cis-9, trans-11 in beef. Trans-fatty acids in foods are of rising importance and knowledge of the differential effects of the individual trans isomers is increasing. TVA is the major trans 18:1 isomer in beef and as the precursor for tissue CLA in both animals and man should be considered as a neutral or beneficial trans-isomer. Increasing the content of n-3 PUFA in beef can influence colour shelf life and sensory attributes of the meat. As the content of n-3 PUFA increases then sensory attributes such as "greasy" and "fishy" score higher and colour shelf life may be reduced. Under these situations, high levels of vitamin E are necessary to help stabilise the effects of incorporating high levels of long chain PUFA into meat. However, grass feeding not only increases n-3 PUFA and CLA but, due to its high content of vitamin E, colour shelf life is improved. It is evident that opportunities exist to enhance the content of health promoting fatty acids in beef and beef products offering opportunities to add value and contribute to market differentiation. However, it is imperative that these approaches to deliver "functional" attributes do not compromise on the health value (lipoperoxidation) or the taste of beef products.     

Effect of slaughter season and muscle type on the fatty acid composition, including conjugated linoleic acid isomers, and nutritional value of intramuscular fat in organic beef

BACKGROUND: Consumer awareness regarding the intake of beef of organic origin is strongly associated with the beneficial outcomes to human health, the environment and animal welfare. In this paper the effects of slaughter season and muscle type on the fatty acid composition, conjugated linoleic acid (CLA) isomeric profile, total cholesterol, α‐tocopherol and β‐carotene contents and nutritional quality of intramuscular fat in organic beef (n = 30) are reported for the first time. RESULTS: Organic beef showed a very low total lipid content, with seasonal changes in the levels of some fatty acids, CLA isomers, n‐6/n‐3 polyunsaturated fatty acid (PUFA) ratio, total cholesterol and β‐carotene. In addition, differences between longissimus lumborum (relatively red) and semitendinosus (relatively white) muscles were found for many fatty acids, specific CLA contents, many CLA isomers and both PUFA/saturated fatty acid (SFA) and n‐6/n‐3 ratios. However, in spite of the seasonal and carcass variations, all organic meats analysed had values of beef similar to pasture‐fed cattle. CONCLUSION: From a nutritional perspective, organic meat from both slaughter seasons seems to have high CLA contents, PUFA/SFA and n‐6/n‐3 indices within the recommended values for the human diet. The data indicate that intramuscular fat in organic meat has a high nutritional value throughout the year. Copyright © 2012 Society of Chemical Industry     

Effects of dietary supplementation of rumen-protected conjugated linoleic acid in dairy cows during established lactation

Short-term studies (< 5 d) involving abomasal infusion of a mixture of CLA isomers or pure trans-10, cis-12 CLA have demonstrated that supplements of conjugated linoleic acids (CLA) reduce milk fat synthesis during established lactation in dairy cows. Our objective was to assess longer term effects of supplementation during established lactation using a dietary supplement of rumen-protected CLA. Thirty Holstein cows were blocked by parity and received a dietary fat supplement of either Ca-salts of palm oil fatty acids (control) or a mixture of Ca-salts of palm oil fatty acids plus Ca-salts of CLA (CLA treatment). Supplements provided about 90 g/d of fatty acids and were topdressed on the TMR. The CLA supplement provided 30.4 g/d of CLA in which the predominant isomers were: trans-8, cis-10 (9.2%), cis-9, trans-11 (25.1%), trans-10, cis-12 (28.9%), and cis-11, trans-13 (16.1%). All cows were pregnant; treatments were initiated on d 79 of pregnancy (approximately 200 d prepartum) and continued for 140 d until dry off. Twenty-three cows completed the study; those receiving CLA supplement had a lower milk fat test (2.90 versus 3.80%) and a 23% reduction in milk fat yield (927 versus 1201 g/d). Intake of DM, milk yield, and the yield and content of true protein and lactose in milk were unaffected by treatment. Milk fat analysis indicated that the CLA supplement reduced the secretion of fatty acids of all chain lengths. However, effects were proportionally greater on short and medium chain fatty acids, thereby causing a shift in the milk fatty acid composition to a greater content of longer-chain fatty acids. Changes in body weight gain, body condition score, and net energy balance were not significant and imply no differences in cows fed the CLA supplement in replenishment of body reserves in late lactation. Likewise, maintenance of pregnancy, gestation length, and calf birth weight were unaffected by treatment. Overall, feeding a dietary supplement of rumen-protected CLA to pregnant cows over the last 140 d of the lactation cycle resulted in a marked reduction in milk fat content and yield, and a shift in milk fatty acid composition, but other milk components, DMI, maintenance of pregnancy, and cow well-being were unaffected.