The objectives of this study were to determine the effects of far-off and close-up diets on prepartum metabolism, postpartum metabolism, and postpartum performance of multiparous Holstein cows. From dry-off to -25 d relative to expected parturition (far-off dry period), cows were fed a control diet to meet National Research Council (NRC) recommendations for net energy for lactation (NE(L)) at ad libitum intake (100NRC; n = 25) or a higher nutrient density diet, which was fed for either ad libitum intake to provide at least 150% of calculated NE(L) requirement (150NRC; n = 25) or at restricted intake to provide 80% of calculated NE(L) requirements (80NRC; n = 24). From -24 d relative to expected parturition until parturition (close-up period), cows were fed a diet that met or exceeded NRC nutrient recommendations at either ad libitum intake (n = 38) or restricted intake (n = 36) to provide 80% of the calculated NE(L) requirement. After parturition, all cows were fed a lactation diet and measurements were made through 56 d in milk (DIM). Prepartum metabolism was consistent with the plane of nutrition. During the first 10 DIM, far-off treatments had significant carryover effects on dry matter intake, energy balance, serum nonesterified fatty acid (NEFA) concentration, and serum beta-hydroxybutyrate concentration. Cows with the lower energy balance during the far-off period (100NRC and 80NRC) had higher dry matter intake and energy balance and lower serum NEFA and beta-hydroxybutyrate during the first 10 DIM. There were no effects of close-up diet and no interactions of far-off and close-up treatments. During the first 56 DIM, there were no residual effects of far-off or close-up diets on dry matter intake, milk yield or composition, body weight, body condition score, serum glucose and insulin concentrations, or muscle lipid concentration. Serum NEFA was higher for 150NRC than 80NRC; 100NRC was intermediate. Thus, the effects of far-off and close-up treatments on postpartum variables diminished as lactation progressed. Overfeeding during the far-off period had a greater negative impact on peripartum metabolism than did differences in close-up period nutrition.

Insulin resistance is a homeorhetic adaptation to parturition in dairy cows transitioning from late pregnancy to early lactation. An increase in prepartum adiposity can predispose periparturient cows to greater lipolysis and insulin resistance, thus increasing the risk for metabolic disease. Mechanisms mediating the development of insulin resistance in overweight peripartal dairy cows may depend on ceramide metabolism. The sphingolipid ceramide accumulates in plasma and tissues of overweight monogastric animals, and facilitates saturated fatty acid-induced insulin resistance. Considering this evidence, we hypothesized that plasma ceramides would be elevated in periparturient dairy cattle and that these sphingolipids would correlate with the magnitude of lipolysis and insulin resistance. To test our central hypothesis, multiparous Holstein cows were allocated into 2 groups according to their body condition score (BCS) at d −30 prepartum: lean (BCS <3.0; n = 10) or overweight (BCS >4.0; n = 11). Blood samples were collected at d −45, −30, −15, and −7, relative to expected parturition, and at d 4 postpartum. Plasma glucose, insulin, nonesterified fatty acids (NEFA), and β-hydroxybutyrate (BHBA) concentrations were measured, and insulin sensitivity was estimated. The concentrations of individual plasma ceramide and glycosylated ceramide were determined using liquid chromatography-based mass spectrometry. Results demonstrated that greater adiposity was associated with a greater loss in body condition during late pregnancy. Overweight cows had greater circulating concentrations of glucose, insulin, and NEFA, and lower insulin sensitivity relative to lean cows. We detected 30 different sphingolipids across 6 lipid classes with acyl chains ranging from 16 to 26 carbons. The most abundant plasma sphingolipids detected were C24:0-ceramide, C24:0-monohexosylceramide, and C16:0-lactosylceramide. Plasma concentrations of total ceramide and monohexosylceramide increased as lactation approached, and saturated ceramide and monohexosylceramide were elevated in cows with greater adiposity relative to those with a lean phenotype. Plasma ceramides (e.g., C24:0-ceramide) were positively correlated with plasma NEFA and inversely correlated with insulin sensitivity. Our data demonstrate a remodeled plasma sphingolipidome in dairy cows transitioning from late pregnancy to lactation characterized by a concomitant increase in plasma ceramides with the development of peripartal insulin resistance.

The objectives of this study were to determine the effects of dietary L-carnitine supplementation on liver lipid accumulation, hepatic nutrient metabolism, and lactation in multiparous cows during the periparturient period. Cows were assigned to treatments at d -25 relative to expected calving date and remained on the experiment until 56 d in milk. Treatments were 4 amounts of supplemental dietary carnitine: control (0 g/d of L-carnitine; n = 14); low carnitine (LC, 6 g/d; n = 11); medium carnitine (MC, 50 g/d; n = 12); and high carnitine (HC, 100 g/d; n = 12). Carnitine was supplied by mixing a feed-grade carnitine supplement with 113.5 g of ground corn and 113.5 g of dried molasses, which was then fed twice daily as a topdress to achieve desired daily carnitine intakes. Carnitine supplementation began on d -14 relative to expected calving and continued until 21 d in milk. Liver and muscle carnitine concentrations were markedly increased by MC and HC treatments. Milk carnitine concentrations were elevated by all amounts of carnitine supplementation, but were greater for MC and HC than for LC during wk 2 of lactation. Dry matter intake and milk yield were decreased by the HC treatment. The MC and HC treatments increased milk fat concentration, although milk fat yield was unaffected. All carnitine treatments decreased liver total lipid and triacylglycerol accumulation on d 10 after calving. In addition, carnitine-supplemented cows had higher liver glycogen during early lactation. In general, carnitine supplementation increased in vitro palmitate beta-oxidation by liver slices, with MC and HC treatments affecting in vitro palmitate metabolism more potently than did LC. In vitro conversion of Ala to glucose by liver slices was increased by carnitine supplementation independent of dose. The concentration of nonesterified fatty acids in serum was not affected by carnitine. As a result of greater hepatic fatty acid beta-oxidation, plasma beta-hydroxybutyric acid was higher for the MC and HC treatments. Serum insulin was greater for all carnitine treatments, although plasma glucose was unaffected. Plasma urea N was lower and plasma total protein was higher for the MC and HC treatments. By decreasing liver lipid accumulation and stimulating hepatic glucose output, carnitine supplementation might improve glucose status and diminish the risk of developing metabolic disorders during early lactation.

It has previously been established that supplementation of trans-10, cis-12 18:2 reduces milk fat content and fat deposition in several species. The objectives of the study were 1) to examine whether potential mechanisms by which trans-10, cis-12 18:2 is reported to affect lipid metabolism in adipose tissue of different species could be partly responsible for the inhibition in milk fat synthesis in lactating dairy cows; and 2) to investigate the effects of trans-10, cis-12 18:2 on the expression of a newly identified isoform of stearoyl-coenzyme A desaturase (SCD) in bovine mammary tissue. Four primiparous Holstein cows in established lactation, fitted with indwelling jugular catheters, were used in a balanced 2 x 2 crossover design. For the first 5 d of each period, cows were infused intravenously with a 15% lipid emulsion providing 10 g/d of either cis-9, cis-12 18:2 (control) or trans-10, cis-12 18:2 (conjugated linoleic acid; CLA). On d 5 of infusion, mammary gland biopsies were performed and tissues were analyzed for mRNA expression of acetyl-coenzyme A carboxylase, fatty acid synthetase, lipoprotein lipase, SCD1, SCD5, sterol regulatory element-binding protein-1, IL6, IL8, and tumor necrosis factor-alpha by real-time PCR. Compared with the control treatment, CLA reduced milk fat concentration and yield by 46 and 38%, respectively, and increased the trans-10, cis-12 18:2 content in milk fat from 0.05 to 3.54 mg/g. Milk yield, milk protein, and dry matter intake were unaffected by treatment. Infusion of the CLA treatment reduced the mRNA expression of acetyl-coenzyme A carboxylase and fatty acid synthetase by 46 and 57%, respectively, and tended to reduce the expression of SCD1 and lipoprotein lipase. Abundance of mRNA for sterol regulatory element-binding protein-1 was reduced by 59% in the CLA treatment group. However, infusing trans-10, cis-12 18:2 did not affect the expression of transcripts for SCD5, tumor necrosis factor-alpha, IL6, and IL8. Results from the current study corroborate the idea that effects of trans-10, cis-12 18:2 reported on adipose tissue in animal models and humans are not part of the response in the inhibition of milk fat synthesis in lactating dairy cows. They also support the hypothesis that SCD1 and SCD5 present important differences in their regulation and physiological roles.

No abstract

Reduced insulin action is a key adaptation that facilitates glucose partitioning to the mammary gland for milk synthesis and enhances adipose tissue lipolysis during early lactation. The progressive recovery of insulin sensitivity as cows advance toward late lactation is accompanied by reductions in circulating nonesterified fatty acids (NEFA) and milk yield. Because palmitic acid can promote insulin resistance in monogastrics through sphingolipid ceramide-dependent mechanisms, palmitic acid (C16:0) feeding may enhance milk production by restoring homeorhetic responses. We hypothesized that feeding C16:0 to mid-lactation cows would enhance ceramide supply and ceramide would be positively associated with milk yield. Twenty multiparous mid-lactation Holstein cows were enrolled in a study consisting of a 5-d covariate, 49-d treatment, and 14-d posttreatment period. All cows were randomly assigned to a sorghum silage-based diet containing no supplemental fat (control; n=10; 138±45 d in milk) or C16:0 at 4% of ration dry matter (PALM; 98% C16:0; n=10; 136±44 d in milk). Blood and milk were collected at routine intervals. Liver and skeletal muscle tissue were biopsied at d 47 of treatment. Intravenous glucose tolerance tests (300mg/kg of body weight) were performed at d -1, 24, and 49 relative to start of treatment. The plasma and tissue concentrations of ceramide and glycosylated ceramide were determined using liquid chromatography coupled with tandem mass spectrometry. Data were analyzed as repeated measures using a mixed model with fixed effects of treatment and time, and milk yield served as a covariate. The PALM treatment increased milk yield, energy-corrected milk, and milk fat yield. The most abundant plasma and tissue sphingolipids detected were C24:0-ceramide, C24:0-monohexosylceramide (GlcCer), and C16:0-lactosylceramide. Plasma concentrations of total ceramide and GlcCer decreased as lactation advanced, and ceramide and GlcCer were elevated in cows fed PALM. Palmitic acid feeding increased hepatic ceramide levels, a response not observed in skeletal muscle tissue. Plasma ceramides (e.g., C24:0-ceramide) were positively correlated with plasma NEFA and milk yield, and positively correlated with NEFA levels following a glucose challenge. Our data demonstrate a remodeled plasma and hepatic sphingolipidome in mid-lactation dairy cows fed PALM. The potential involvement in ceramide in homeorhetic nutrient partitioning to support lactation requires further consideration.

Reduced insulin action develops naturally during the peripartum to ensure maternal nutrient delivery to the fetus and neonate. However, increased insulin resistance can facilitate excessive lipolysis which in turn promotes metabolic disease in overweight dairy cattle. Increased fatty acid availability favors the accumulation of the sphingolipid ceramide and is implicated in the pathogenesis of insulin resistance, however, the relationship between sphingolipid metabolism and insulin resistance during the peripartum remains largely unknown. Our objectives were to characterize temporal responses in plasma and tissue sphingolipids in lean and overweight peripartal cows and to establish the relationships between sphingolipid supply and lipolysis, hepatic lipid deposition, and systemic insulin action. Twenty-one multiparous lean and overweight Holstein cows were enrolled in a longitudinal study spanning the transition from gestation to lactation (d -21 to 21, relative to parturition). Plasma, liver, and skeletal muscle samples were obtained, and sphingolipids were profiled using LC/MS/MS. Insulin sensitivity was assessed utilizing intravenous insulin and glucose challenges. Our results demonstrated the following: first, insulin resistance develops postpartum concurrently with increased lipolysis and hepatic lipid accumulation; second, ceramides and glycosylated ceramides accumulate during the transition from gestation to lactation and are further elevated in overweight cows; third, ceramide accrual is associated with lipolysis and liver lipid accumulation, and C16:0- and C24:0-ceramide are inversely associated with systemic insulin sensitivity postpartum; fourth, plasma sphingomyelin, a potential source of ceramides reaches a nadir at parturition and is closely associated with feed intake; fifth, select sphingomyelins are lower in the plasma of overweight cows during the peripartal period. Our results demonstrate that dynamic changes occur in peripartal sphingolipids that are influenced by adiposity, and are associated with the onset of peripartal insulin resistance. These observations are in agreement with a putative potential role for sphingolipids in facilitating the physiological adaptations of peripartum.

Modification of hypothalamic fatty acid (FA) metabolism can improve energy homeostasis and prevent hyperphagia and excessive weight gain in diet-induced obesity (DIO) from a diet high in saturated fatty acids. We have shown previously that C75, a stimulator of carnitine palmitoyl transferase-1 (CPT-1) and fatty acid oxidation (FAOx), exerts at least some of its hypophagic effects via neuronal mechanisms in the hypothalamus. In the present work, we characterized the effects of C75 and another anorexigenic compound, the glycerol-3-phosphate acyltransferase (GPAT) inhibitor FSG67, on FA metabolism, metabolomics profiles, and metabolic stress responses in cultured hypothalamic neurons and hypothalamic neuronal cell lines during lipid excess with palmitate. Both compounds enhanced palmitate oxidation, increased ATP, and inactivated AMP-activated protein kinase (AMPK) in hypothalamic neurons in vitro. Lipidomics and untargeted metabolomics revealed that enhanced catabolism of FA decreased palmitate availability and prevented the production of fatty acylglycerols, ceramides, and cholesterol esters, lipids that are associated with lipotoxicity-provoked metabolic stress. This improved metabolic signature was accompanied by increased levels of reactive oxygen species (ROS), and yet favorable changes in oxidative stress, overt ER stress, and inflammation. We propose that enhancing FAOx in hypothalamic neurons exposed to excess lipids promotes metabolic remodeling that reduces local inflammatory and cell stress responses. This shift would restore mitochondrial function such that increased FAOx can produce hypothalamic neuronal ATP and lead to decreased food intake and body weight to improve systemic metabolism.