Dairy cows are highly susceptible after parturition to developing liver lipidosis and ketosis, which are costly diseases to farmers. A bovine microarray platform consisting of 13,257-annotated oligonucleotides was used to study hepatic gene networks underlying nutrition-induced ketosis. On day 5 postpartum, 14 Holstein cows were randomly assigned to ketosis-induction (n = 7) or control (n = 7) groups. Cows in the ketosis-induction group were fed at 50% of day 4 intake until they developed signs of clinical ketosis, and cows in the control group were fed ad libitum throughout the treatment period. Liver was biopsied at 10-14 (ketosis) or 14 days postpartum (controls). Feed restriction increased blood concentrations of nonesterified fatty acids and beta-hydroxybutyrate, but decreased glucose. Liver triacylglycerol concentration also increased. A total of 2,415 genes were altered by ketosis (false discovery rate = 0.05). Ingenuity Pathway Analysis revealed downregulation of genes associated with oxidative phosphorylation, protein ubiquitination, and ubiquinone biosynthesis with ketosis. Other molecular adaptations included upregulation of genes and nuclear receptors associated with cytokine signaling, fatty acid uptake/transport, and fatty acid oxidation. Genes downregulated during ketosis included several associated with cholesterol metabolism, growth hormone signaling, proton transport, and fatty acid desaturation. Feed restriction and ketosis resulted in previously unrecognized alterations in gene network expression underlying key cellular functions and discrete metabolic events. These responses might help explain well-documented physiological adaptations to reduced feed intake in early postpartum cows and, thus, provide molecular targets that might be useful in prevention and treatment of liver lipidosis and ketosis.

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.

Previous research in our laboratory showed that dietary fat supplementation during the dry period was associated with decreased peripartum hepatic lipid accumulation. However, fat supplementation decreased dry matter (DM) intake and thereby confounded results. Consequently, 47 Holstein cows with body condition scores (BCS) < or = 3.5 at dry-off were used to determine whether source or amount of energy fed to dry cows was responsible for the decreased hepatic lipid content. Moderate grain- or fat-supplemented diets [1.50 Mcal of net energy for lactation (NE(L))/kg] were fed from dry-off (60 d before expected parturition) to calving at either ad libitum (160% of NE(L) requirement) or restricted (80% of NE(L) requirement) intakes. Postpartum, cows were fed a single lactation diet for ad libitum intake and performance was measured for 105 d. Prepartum intakes of DM and NE(L) were significantly lower for feed-restricted cows as designed. During the first 21 d postpartum, previously restricted cows had higher intakes of DM and NE(L). Body weights and BCS were lower prepartum for restricted cows but groups converged to similar nadirs postpartum. Restricted-fed cows had lower concentrations of glucose and insulin and increased concentrations of NEFA in plasma during the dry period. Peripartum NEFA rose markedly for all treatments but were higher postpartum for cows previously fed ad libitum. Plasma concentrations of NEFA and BHBA remained lower in cows restricted-during the dry period. Postpartum concentrations of total lipid and triglyceride in liver were lower in cows previously feed-restricted. Across dietary treatments, activity of carnitine palmitoyltransferase (CPT) in hepatic mitochondria was lowest at - 21 d, highest at 1 d, and decreased at 21 and 65 d relative to parturition. The activity of CPT at d 1 tended to be higher for previously feed-restricted cows; thereafter, CPT activity declined more rapidly than in cows fed ad libitum. Nutrient intake during the dry period had more pronounced effects on peripartal lipid metabolism and DMI than did composition of the prepartum diet.

Long-term molecular adaptations in liver from high-producing dairy cows are virtually unknown. Liver from five Holstein cows was biopsied at -65, -30, -14, +1, +14, +28, and +49 days relative to parturition for transcript profiling using a microarray consisting of 7,872 annotated cattle cDNA inserts. More than 5,000 cDNA elements represented on the microarray were expressed in liver. From this set we identified 62 differentially expressed genes related to physiological state, with a false discovery rate threshold of P = 0.20. The dominant expression pattern consisted of upregulation from day -30 through day +1, followed by downregulation through day +28. There was a threefold decrease from day -65 through day +14 in expression of IGFBP3, GSTM5, and PDPK1. These genes mediate IGF-I transport, oxidative stress, and glucose homeostasis, respectively. IGFBP3, EIF4B, and GSTM5 mRNA levels were positively correlated with blood serum total protein. Correlation analysis showed positive associations between serum nonesterified fatty acids and mRNA expression for SAA1, CPT1A, ACADVL, and TFAP2A. Transcript levels of ACSL1, PPARA, and TFAP2A were positively correlated with serum beta-hydroxybutyrate. Expression patterns for certain genes (e.g., IGFBP3, HNF4A, GPAM) revealed adaptations commencing well ahead of parturition, suggesting they are regulated by factors other than periparturient hormonal environment. Results provide evidence that hepatic inflammatory responses occurring near parturition initiate or augment adipose catabolism. In this context, cytokines, acute-phase proteins, and serum nonesterified fatty acids are key players in periparturient cow metabolism. We propose a model for integrating gene expression, metabolite, and liver composition data to explain physiological events in placenta, adipose, and liver during the periparturient period.

Dairy cows undergo tremendous metabolic and physiological adaptations around parturition to support lactation. The liver 

is central to many of these processes, including gluconeogenesis and metabolism of fatty acids mobilized from adipose tis- 

sue. Fat accumulation may impair normal functions of the liver and increase ketogenesis, which in turn may predispose cows 

to other metabolic abnormalities. Several aspects of dietary management and body condition may alter these adaptations, 

affect dry matter intake, and increase or decrease susceptibility to periparturient health problems. Overfeeding energy dur- 

ing the dry period is a prominent risk factor. Considerable progress has been made in recent years in describing the adap- 

tive changes in the liver and other organs in normal and abnormal states, but this knowledge has not yet identified unequiv- 

ocally the key steps that might be compromised during development of metabolic disorders. The potential role of signaling 

compounds, such as the inflammatory cytokines released in response to environmental stressors, infectious challenge, and 

oxidative stress, in the pathogenesis of periparturient disease is under investigation. New techniques such as functional 

genomics, using cDNA or oligonucleotide microarrays, as well as proteomics and metabolomics, provide additional high- 

throughput tools to determine the effects of nutrition, management, or stressors on tissue function in development of dis- 

ease. Integrative approaches should be fruitful in unraveling the complex interactions of metabol...