Dale E. Bauman

Regulation and nutritional manipulation of milk fat: low-fat milk syndrome

Abstract Diet-induced low-fat milk syndrome, milk fat depression (MFD), was first described over a century ago. It continues to be an active research area and we reviewed theories that have been proposed to explain diet-induced MFD. Many theories were based on the concept that reduced milk fat was a consequence of a limited supply of lipid precursors, e.g. the insulin-glucogenic theory; experimental data provide little support for this concept as the basis for diet-induced MFD. Other theories attributed MFD to a direct inhibition of lipid synthesis in the mammary gland. Davis and Brown (In: Phillipson, A.T. (Ed.), Physiology of Digestion and Metabolism in the Ruminant. Oriel Press, Newcastle upon Tyne, UK, 1970, pp. 545–565) noted increased trans -C18:1 fatty acids in milk fat during MFD, and proposed these fatty acids inhibited fat synthesis. We recently established the increase was specific for trans -10 C18:1 and its rumen precursor, trans -10, cis -12 conjugated linoleic acid (CLA). Across a range of diets there was a curvilinear relationship between the reduction in milk fat yield and the increase in milk fat content of trans -10, cis -12 CLA. Furthermore, postruminal infusion of trans -10, cis -12 CLA resulted in a marked inhibition of milk fat synthesis and a shift in fatty acid pattern similar to dietary-induced MFD. Therefore, diets that cause MFD alter rumen biohydrogenation resulting in the production of trans -10, cis -12 CLA, and perhaps other unique fatty acids, that are potent inhibitors of milk fat synthesis. We refer to this as the biohydrogenation theory of MFD and discuss the possibility that it may represent a unifying concept to explain diet-induced MFD.

Modifying Milk Fat Composition of Dairy Cows to Enhance Fatty Acids Beneficial to Human Health

There is increased consumer awareness that foods contain microcomponents that may have beneficial effects on health maintenance and disease prevention. In milk fat these functional food components include EPA, DHA, and CLA. The opportunity to enhance the content of these FA in milk has improved as a result of recent advances that have better defined the interrelationships between rumen fermentation, lipid metabolism, and milk fat synthesis. Dietary lipids undergo extensive hydrolysis and biohydrogenation in the rumen. Milk fat is predominantly TG, and de novo FA synthesis and the uptake of circulating FA contribute nearly equal amounts (molar basis) to the FA in milk fat. Transfer of dietary EPA and DHA to milk fat is very low (<4%); this is, to a large extent, related to their extensive biohydrogenation in the rumen, and also partly due to the fact that they are not transported in the plasma lipid fractions that serve as major mammary sources of FA uptake (TG and nonesterified FA). Milk contains over 20 isomers of CLA but the predominant one is cis-9,trans-11 (75–90% of total CLA). Biomedical studies with animal models have shown that this isomer has anticarcinogenic and anti-atherogenic activities. cis-9,trans-11-CLA is produced as an intermediate in the rumen biohydrogenation of linoleic acid but not of linolenic acid. However, it is only a transient intermediate, and the major source of milk fat CLA is from endogenous synthesis. Vaccenic acid, produced as a rumen biohydrogenation intermediate from both linoleic acid and linolenic acid, is the substrate, and Δ9-desaturase in the mammary gland and other tissues catalyzes the reaction. Diet can markedly affect milk fat CLA content, and there are also substantial differences among individual cows. Thus, strategies to enhance milk fat CLA involve increasing rumen outflow of vaccenic acid and increasing Δ9-desaturase activity, and through these, several-fold increases in the content of CLA in milk fat can be routinely achieved. Overall, concentrations of CLA, and to a lesser extent EPA and DHA, can be significantly enhanced through the use of diet formulation and nutritional management of dairy cows.

Adaptations of Glucose Metabolism During Pregnancy and Lactation

Increased glucose requirements of the gravid uterus during late pregnancy and even greater requirements of the lactating mammary glands necessitate major adjustments in glucose production and utilization in maternal liver, adipose tissue, skeletal muscle, and other tissues. In ruminants, which at all times rely principally on hepatic gluconeogenesis for their glucose supply, hepatic glucose synthesis during late pregnancy and early lactation is increased to accommodate uterine or mammary demands even when the supply of dietary substrate is inadequate. At the same time, glucose utilization by adipose tissue and muscle is reduced. In pregnant animals, these responses are exaggerated by moderate undernutrition and are mediated by reduced tissue sensitivity and responsiveness to insulin, associated with decreased tissue expression of the insulin-responsive facilitative glucose transporter, GLUT4. Peripheral tissue responses to insulin remain severely attenuated during early lactation but recover as the animal progresses through mid lactation. Specific homeorhetic effectors of decreased insulin-mediated glucose metabolism during late pregnancy have yet to be conclusively identified. In contrast, somatotropin is almost certainly a predominant homeorhetic influence during lactation because its exogenous administration causes specific changes in glucose metabolism (and many other functions) of various nonmammary tissues which faithfully mimic normal adaptations to early lactation.

Biosynthesis of conjugated linoleic acid in ruminants and humans.

Publisher Summary This chapter discusses the biosynthesis of conjugated linoleic acid (CLA) in ruminants and humans. Conjugated linoleic acid is a mixture of positional and geometric isomers of linoleic acid with a conjugated double-bond system. Because of its potential to improve human health, there is great interest to increase the amount of CLA in the human food supply. This has caused great effort to be expended toward increasing the concentration of CLA, and more specifically rumenic acid (RA), in the milk and tissues of ruminant foods because these are the predominant source of CLA in human diets. RA is the predominant CLA isomer present in ruminant products, and the major source of its occurrence is endogenous synthesis via desaturation of vaccenic acid (VA) by ∆-9-desaturase. The chapter focuses on improving the understanding of biohydrogenation in the rumen and examining milk and tissue CLA responses to a range of diets. The diversity of various BH intermediates in digesta, milk, and tissues indicates the complexity of the BH processes as a whole and the population dynamics of the ruminal bacteria involved. Predicting the outcome of changes in the diet is complicated by the interactions of the ruminal environment, substrate supply and forms of dietary lipids, all of which influence the BH process simultaneously.

Recent advances in the regulation of milk fat synthesis

In addition to its economic value, milk fat is responsible for many of milks characteristics and can be markedly affected by diet. Diet-induced milk fat depression (MFD) was first described over a century ago and remains a common problem observed under both intensive and extensive management. The biohydrogenation theory established that MFD is caused by an inhibition of mammary synthesis of milk fat by specific fatty acids (FA) produced as intermediates in ruminal biohydrogenation. During MFD, lipogenic capacity and transcription of key lipid synthesis genes in the mammary gland are down-regulated in a coordinated manner. Our investigations have established that expressions of sterol response element-binding protein 1 (SREBP1) and SREBP-activation proteins are down-regulated during MFD. Importantly, key lipogenic enzymes are transcriptionally regulated via SREBP1. Collectively, these results provide strong evidence for SREBP1 as a central signaling pathway in the regulation of mammary FA synthesis. Spot 14 is also down-regulated during MFD, consistent with a lipogenic role for this novel nuclear protein. In addition, SREBP1c and Spot 14 knock-out mice exhibit reduced milk fat similar to the magnitude and pattern of MFD in the cow. Application of molecular biology approaches has provided the latest chapter in the regulation of milk fat synthesis and is reviewed along with a brief background in nutritional regulation of milk fat synthesis in ruminants.

Nutrigenomics, Rumen-Derived Bioactive Fatty Acids, and the Regulation of Milk Fat Synthesis

Mammary synthesis of milk fat continues to be an active research area, with significant advances in the regulation of lipid synthesis by bioactive fatty acids (FAs). The biohydrogenation theory established that diet-induced milk fat depression (MFD) in the dairy cow is caused by an inhibition of mammary synthesis of milk fat by specific FAs produced during ruminal biohydrogenation. The first such FA shown to affect milk fat synthesis was trans-10, cis-12 conjugated linoleic acid, and its effects have been well characterized, including dose-response relationships. During MFD, lipogenic capacity and transcription of key mammary lipogenic genes are coordinately down-regulated. Results provide strong evidence for sterol response element-binding protein-1 (SREBP1) and Spot 14 as biohydrogenation intermediate responsive lipogenic signaling pathway for ruminants and rodents. The study of MFD and its regulation by specific rumen-derived bioactive FAs represents a successful example of nutrigenomics in present-day animal nutrition research and offers several potential applications in animal agriculture.

Effect of CLA on milk fat synthesis in dairy cows: comparison of inhibition by methyl esters and free fatty acids, and relationships among studies.

CLA is a potent inhibitor of milk fat synthesis, as shown by investigations using mixtures of CLA isomers in FFA form. However, methyl esters of CLA can be initially formed in commercial synthesis, and their use in a supplement has certain manufacturing and cost advantages. Our objective was to compare abomasal infusion of methyl esters of CLA (ME-CLA) and FFA of CLA (FFA-CLA) on milk fat synthesis. Data were also combined with previous investigations to examine broader relationships between trans-10,cis-12 CLA and the reduction in milk fat. Three mid-lactation, rumen-fistulated Holstein cows were used in a 3×3 Latin square design. Treatments were (i) control, (ii) ME-CLA, and (iii) FFA-CLA. The ME-CLA and FFA-CLA treatments (4.2 g/d trans-10,cis-12 CLA) resulted in a comparable reduction in milk fat yield (38 and 39%, respectively) and pattern of reduction in individual FA. In contrast, milk yield, milk protein, and feed intake were unaltered by CLA treatment. Combining data across studies revealed strong correlations relating the reduction in milk fat yield to abomasal dose of trans-10,cis-12 CLA (R2=0.86), milk fat content of trans-10,cis-12 CLA (R2=0.93), and milk fat secretion of trans-10,cis-12 CLA (R2=0.82). Across studies, transfer efficiency of abomasally infused trans-10,cis-12 CLA into milk fat was relatively constant (22%; R2=0.94). Overall, ME-CLA and FFA-CLA were equally potent in reducing milk fat, and either form could be used to formulate a dietary supplement that would induce milk fat depression.

Conjugated linoleic acid: Biosynthesis and nutritional significance

The term conjugated linoleic acid (CLA) refers to a mixture of positional and geometric isomers of linoleic acid with a conjugated double bond system; milk fat can contain over 20 different isomers of CLA. CLA isomers are produced as transient intermediates in the rumen biohydrogenation of unsaturated fatty acids consumed in the diet. However, cis-9, trans-11 CLA, known as rumenic acid (RA), is the predominant isomer (up to 90% of total) because it is produced mainly by endogenous synthesis from vaccenic acid (VA). VA is typically the major biohydrogenation intermediate produced in the rumen and it is converted to RA by Δ;9-desaturase in the mammary gland and other tissues.

Efficacy of conjugated linoleic acid for improving reproduction: a multi-study analysis in early-lactation dairy cows.

The feeding of conjugated linoleic acid (CLA) supplements to early-lactation dairy cows has been shown to decrease milk fat synthesis and possibly improve reproductive performance. However, previously reported studies used too few animals to clearly establish the effect of CLA on reproduction. Our objective was to combine data from these studies to evaluate the association of CLA with time to first ovulation and time to conception using methods of survival analysis and overall success of pregnancy by logistic regression. A database was compiled of individual animal data (n = 212) from 5 controlled studies in which CLA had been supplemented to early-lactation dairy cows. Survival analysis incorporated both semi-parametric models (Cox proportional hazards) and parametric models (log-normal). The probability of cows becoming pregnant increased in a nonlinear manner as trans-10, cis-12 CLA dose increased, with the optimal dose predicted to be 10.1 g/d. At the optimal dose, the probability of pregnancy was increased by 26% compared with those animals receiving no CLA (probability = 91% and 72%, respectively). Similarly, the log-normal model predicted that time to conception was decreased in a nonlinear manner with increasing trans-10, cis-12 CLA dose. The predicted optimal dose was 10.5 g of trans-10, cis-12 CLA/d and at this dose the median time to conception was decreased by 34 d when compared with those cows not receiving CLA (117 vs. 151 d in milk, respectively). The log-normal model was also the best-fit model for time to first ovulation. Overall, this multi-study analysis demonstrated a strong concordance between the nature of the dose response and the predicted optimal dose of trans-10, cis-12 CLA across the 3 reproductive variables evaluated. These results indicate that reproductive performance of dairy cows may be improved by feeding of CLA supplements during early lactation.

Survey of the fatty acid composition of retail milk in the United States including regional and seasonal variations.

Consumers are increasingly aware that food components have the potential to influence human health maintenance and disease prevention, and dietary fatty acids (FA) have been of special interest. It has been 25 years since the last survey of US milk FA composition, and during this interval substantial changes in dairy rations have occurred, including increased use of total mixed rations and byproduct feeds as well as the routine use of lipid and FA supplements. Furthermore, analytical procedures have improved allowing greater detail in the routine analysis of FA, especially trans FA. Our objective was to survey US milk fat and determine its FA composition. We obtained samples of fluid milk from 56 milk processing plants across the US every 3 mo for one year to capture seasonal and geographical variations. Processing plants were selected based on the criteria that they represented 50% or more of the fluid milk produced in that area. An overall summary of the milk fat analysis indicated that saturated fatty acids comprised 63.7% of total milk FA with palmitic and stearic acids representing the majority (44.1 and 18.3% of total saturated fatty acids, respectively). Unsaturated fatty acids were 33.2% of total milk FA with oleic acid predominating (71.0% of total unsaturated fatty acids). These values are comparable to those of the previous survey in 1984, considering differences in analytical techniques. Trans FA represented 3.2% of total FA, with vaccenic acid being the major trans isomer (46.5% of total trans FA). Cis-9, trans-11 18:2 conjugated linoleic acid represented 0.55% total milk FA, and the major n-3 FA (linolenic acid, 18:3) composed 0.38%. Analyses for seasonal and regional effects indicated statistical differences for some FA, but these were minor from an overall human nutrition perspective as the FA profile for all samples were numerically similar. Overall, the present study provides a valuable database for current FA composition of US fluid milk, and results demonstrate that the milk fatty acid profile is remarkably consistent across geographic regions and seasons from the perspective of human dietary intake of milk fat.