V.R. Osborne

Effects of supplementing glycerol and soybean oil in drinking water on feed and water intake, energy balance, and production performance of periparturient dairy cows

The objective of this study was to determine the effects of supplementing glycerol and soybean oil in drinking water on feed and water intake, calculated energy balance, and production performance of periparturient dairy cows. Ninety multiparous Holstein dairy cows were randomly assigned to 1 of 3 treatments: 1) no nutrients supplemented in the drinking water (control); 2) 20 g/L of glycerin supplemented in the drinking water (glycerol); and 3) 10 g/L of soybean oil supplemented in the drinking water (SBO). The trial lasted from 7 d prepartum to 7 d postpartum. Cows were offered a close-up and milking cow TMR for ad libitum intake, pre- and postpartum, respectively. The dry matter intake of cows supplemented with glycerol and SBO was lower than for the control cows throughout the experimental period but not different from each other. Water intake for the control cows was greater than the average for the glycerol and SBO cows prepartum, and greater than for SBO cows but similar to that of glycerol cows postpartum. Glycerol cows consumed more water than SBO cows. There were no differences in energy intake and energy balance of the cows pre- and postpartum. Serum triacylglycerol concentration for glycerol cows was lower than for the control and SBO cows prepartum and was lower than for the SBO cows postpartum. There were no differences in the serum nonesterified fatty acids and glucose concentrations throughout the experiment. There were no differences in the serum beta-hydroxybutyrate (BHBA) concentrations at parturition, but serum BHBA concentration of the glycerol cows was greater than for control and SBO cows during the prepartum period. However, during the postpartum period, serum BHBA concentrations of the control cows were greater than for glycerol and SBO cows. There were no differences in calf birth weights or milk yield and composition. Although the glucogenic property of glycerol supplemented in the drinking water at 20 g/L may not have been sufficient to elicit a milk yield response, it did reduce the concentration of BHBA postpartum.

Genetic and environmental relationships between body condition score and milk production traits in Canadian Holsteins.

The objective of this research was to estimate genetic parameters of first-lactation body condition score (BCS), milk yield, fat percentage (Fat%), protein percentage (Prot%), somatic cell score (SCS), milk urea nitrogen (MUN), lactose percentage (Lact%), and fat to protein ratio (F:P) using multiple-trait random regression animal models. Changes in covariances between BCS and milk production traits on a daily basis have not been investigated before and could be useful for determining which BCS estimated breeding values (EBV) might be practical for selection in the future. Field staff from Valacta milk recording agency (Sainte-Anne-de-Bellevue, QC, Canada) collected BCS from Québec herds several times per cow throughout the lactation. Average daily heritabilities and genetic correlations among the various traits were similar to literature values. On an average daily basis, BCS was genetically unfavorably correlated with milk yield (i.e., increased milk yield was associated with lower body condition). The unfavorable genetic correlation between BCS and milk yield became stronger as lactation progressed, but was equivalent to zero for the first month of lactation. Favorable genetic correlations were found between BCS with Prot%, SCS, and Lact% (i.e., greater BCS was associated with greater Prot%, lower SCS, and greater Lact%). These correlations were strongest in early lactation. On an average daily basis, BCS was not genetically correlated with Fat% or MUN, but was negatively correlated with F:P. Furthermore, BCS at 5 and 50 d in milk (DIM) had the most favorable genetic correlations with milk production traits over the lactation (at 5, 50, 150, and 250 DIM). Thus, early lactation BCS EBV shows potential for selection. Regardless, this study showed that the level of association BCS has with milk production traits is not constant over the lactation. Simultaneous selection for both BCS and milk production traits should be considered, mainly due to the unfavorable genetic correlation between BCS with milk yield.

Selenomethionine increases proliferation and reduces apoptosis in bovine mammary epithelial cells under oxidative stress.

The decline in mammary epithelial cell number as lactation progresses may be due, in part, to oxidative stress. Selenium is an integral component of several antioxidant enzymes. The present study was conducted to examine the effect of oxidative stress and selenomethionine (SeMet) on morphology, viability, apoptosis, and proliferation of bovine mammary epithelial cells (BMEC) in primary culture. Cells were isolated from mammary glands of lactating dairy cows and grown for 3 d in a low-serum gel system containing lactogenic hormones and 0 or 100 μM H2O2 with 0, 10, 20, or 50 nM SeMet. Hydrogen peroxide stress increased intracellular H2O2 to 3 times control concentrations and induced a loss of cuboidal morphology, cell-cell contact, and viability of BMEC by 25%. Apoptotic cell number more than doubled during oxidative stress, but proliferating cell number was not affected. Supplementation with SeMet increased glutathione peroxidase activity 2-fold and restored intracellular H2O2 to control levels with a concomitant return of morphology and viability to normal. Apoptotic BMEC number was decreased 76% below control levels by SeMet and proliferating cell number was increased 4.2-fold. These findings suggest that SeMet modulated apoptosis and proliferation independently of a selenoprotein-mediated reduction of H2O2. In conclusion, SeMet supplementation protects BMEC from H2O2-induced apoptosis and increased proliferation and cell viability under conditions of oxidative stress.

Decline in mammary translational capacity during intravenous glucose infusion into lactating dairy cows

The objective of this study was to determine effects of glucose on milk protein yield and mammary mammalian target of rapamycin (mTOR) activity in dairy cattle in early lactation. Eight multiparous cows at 73 ± 8 d in milk were randomly assigned to 2 treatments in a crossover design for two 6-d periods. Treatments were jugular infusion of either saline (Sal) or 896 g/d glucose (Glc). All cows were fed a total mixed ration with 42% neutral detergent fiber, had free access to water, and were milked twice a day. Within each period, blood samples were taken (d 5) and mammary tissue was collected by biopsy (d 6) from each hindquarter for Western blot analysis. In addition to Sal and Glc treatments, on d 6, rapamycin dissolved in 50% dimethyl sulfoxide was administered via the teat canals into the left quarters, with a control solution administered into the right quarters. Rapamycin had no effect on milk protein yields or phosphorylation state of mTOR signaling proteins. Infusions of Glc significantly increased milk yield but only tended to increase milk protein yields. Milk fat tended to be decreased in cows infused with Glc, whereas lactose yields were significantly increased. Glucose infusion did not increase plasma glucose levels, but insulin and nonessential AA concentrations increased by 21 and 16%, respectively, branched-chain AA concentrations decreased 24%, and essential AA concentrations tended to decrease by 14%. Infusion of Glc significantly decreased abundances of both phosphorylated and total ribosomal S6 kinase 1 (S6K1) in mammary tissue by 27 and 11%, respectively. Abundance of phosphorylated eukaryotic initiation factor 4E-binding protein 1 (4EBP1) decreased significantly by 25%, whereas total 4EBP1 exhibited a tendency to decrease by 16%. We conclude that the mTOR signaling pathway is not the only regulator of milk protein synthesis. Decreases in essential AA concentrations in plasma suggest that protein synthesis was stimulated in nonmammary tissues of the body, presumably skeletal muscle.

Effect of enhanced whole-milk feeding in calves on subsequent first-lactation performance.

The objective of this study was to determine the effect of enhanced whole-milk (WM) feeding systems in calves from birth to 8wk of age on subsequent first-lactation performance. The experiment was conducted as a completely randomized design consisting of 2 treatment groups. At birth, 152 Holstein heifer calves were randomly assigned to 1 of 2 treatments: (i) 4L of WM/d or (ii) 8L of WM/d. The calves were bucket fed 2 or 4L of WM twice daily at 0700 and 1600h. Each calf was housed individually in temperature-controlled nurseries and had ad libitum access to water and textured calf starter daily. Calves consumed greater volumes of textured calf starter when fed 4 versus 8L of WM/d. Water intakes mirrored starter intakes, leading to greater water consumption at weaning. Calves reared on 8L of WM/d were heavier at d 56 than calves reared on 4L of WM/d. The average daily gain of the calves offered 8L of WM/d from d 0 to 56 was greater than that of calves offered 4L of WM/d. Structural measurements were significantly greater for calves that consumed 8L of WM/d. The differences observed in withers height and live BW due to WM feeding level were not apparent by 3 and 12mo of age, respectively. Rumen pH was higher in calves that consumed 8L of WM/d than in calves that consumed 4L of WM/d. Whole-milk feeding level did not affect age at first calving or milk-production parameters. These results suggest that enhanced WM feeding improved growth performance until 3mo of age. However, first-lactation results indicated no lactation-performance benefits of increased nutrition and growth performance during the milk-fed period in dairy calves.

Short communication: Estimates of genetic parameters of body condition score in the first 3 lactations using a random regression animal model

The objective of this research was to estimate the genetic parameters of body condition score (BCS) in the first 3 lactations in Canadian Holstein dairy cattle using a multiple-lactation random regression animal model. Field staff from Valacta milk recording agency (Sainte-Anne-de-Bellevue, QC, Canada) collected BCS from Québec herds several times throughout each lactation. Approximately 32,000, 20,000, and 11,000 first-, second-, and third-parity BCS were analyzed, respectively, from a total of 75 herds. Body condition score was a moderately heritable trait over the lactation for parity 1, 2, and 3, with average daily heritabilities of 0.22, 0.26, and 0.30, respectively. Daily heritability ranged between 0.14 and 0.26, 0.19 and 0.28, and 0.24 and 0.33 for parity 1, 2, and 3, respectively. Genetic variance of BCS increased with days in milk within lactations. The low genetic variance in early lactation suggests that the evolution of the ability to mobilize tissue reserves in early lactation provided cattle with a major advantage, and is, therefore, somewhat conserved. The increasing genetic variance suggests that more genetic differences were related to how well cows recovered from the negative energy balance state. More specifically, increasing genetic variation as lactation progressed could be a reflection of genetic differences in the ability of cows to efficiently control the rate of mobilization of tissue reserves, which would not be crucial in early lactation. The shape of BCS curves was similar across parities. From first to third parity, differences included the progressively deeper nadir and faster rate of recovery of condition. Daily genetic correlations between parities were calculated from 5 to 305 DIM, and were summed and divided by 301 to obtain average daily genetic correlations. The average daily genetic correlations were 0.84 between parity 1 and 2, 0.83 between parity 1 and 3, and 0.86 between parity 2 and 3. Although not 1, these genetic correlations are still strong, so much of the variation observed in BCS was controlled by the same genes for each of the first 3 lactations. If a genetic evaluation for BCS is developed, regular collection of first-lactation BCS records should be sufficient for genetic evaluation.

Selenomethionine stimulates expression of glutathione peroxidase 1 and 3 and growth of bovine mammary epithelial cells in primary culture

This study examined the localization of cellular glutathione peroxidase (GPx1) and extracellular glutathione peroxidase (GPx3) in lactating mammary tissue and in primary cultures of bovine mammary epithelial cells (BMEC). The effect of selenium as selenomethionine (SeMet) on the growth and viability of BMEC and GPx protein expression and activity were also studied. Single mammary epithelial cells were recovered by serial collagenase/hyaluronidase digestion from lactating bovine mammary tissue and cultured in a low-serum collagen gel system enriched with lactogenic hormones and 0, 10, 20, or 50 nM SeMet. Positive immunostaining with anti-cytokeratin and bovine anti-casein confirmed the epithelial nature and differentiated state of BMEC. Addition of SeMet to media facilitated rapid confluence of BMEC and formation of dome structures. Immunohistochemical and immunocytochemical staining revealed that both GPx1 and GPx3 are synthesized by BMEC and localized in the cytoplasm and nucleus. Up to 50 nM SeMet linearly increased BMEC number and viability over 5 d of culture. Bovine mammary epithelial cells cultured in SeMet-supplemented medium also exhibited markedly elevated GPx activity and linear increases in abundance of GPx1 and GPx3 proteins. It is apparent that SeMet degradation to release Se for synthesis of selenoproteins is carried out by BMEC. Results indicate that bovine mammary epithelial cells express GPx1 and GPx3 in vivo and in vitro; SeMet enhances expression of these selenoproteins in vitro and the growth and viability of BMEC.

Mammary blood flow and metabolic activity are linked by a feedback mechanism involving nitric oxide synthesis

To test which, if any, of the major milk precursors can elicit a rapid change in the rate of mammary blood flow (MBF) and to define the time course and magnitude of such changes, 4 lactating cows were infused with glucose, amino acids, or triacylglycerol into the external iliac artery feeding one udder half while iliac plasma flow (IPF) was monitored continuously by dye dilution. Adenosine and saline were infused as positive and negative controls, respectively, and insulin was infused to characterize the response to a centrally produced anabolic hormone. To test the roles of cyclooxygenase, NO synthase and ATP-sensitive K (KATP) channels in nutrient-mediated changes in blood flow, their respective inhibitors-indomethacin, Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), and glibenclamide-were infused simultaneously with glucose. Each day, 1 infusate was given twice to each cow, over a 20-min period each time, separated by a 20-min washout period. In addition, each treatment protocol was administered on 2 separate days. A 73% increase in IPF during adenosine infusion showed that the mammary vasodilatory response was quadratic in time, with most changes occurring in the first 5min. Glucose infusion decreased IPF by 9% in a quadratic manner, most rapidly in the first 5min, indicating that a feedback mechanism of local blood flow control, likely through adenosine release, was operative in the mammary vasculature. Amino acid infusion increased IPF 9% in a linear manner, suggesting that mammary ATP utilization was stimulated more than ATP production. This could reflect a stimulation of protein synthesis. Triacylglycerol only tended to decrease IPF and insulin did not affect IPF. A lack of IPF response to glibenclamide indicates that KATP channels are not involved in MBF regulation. Indomethacin and L-NAME both depressed IPF. In the presence of indomethacin, glucose infusion caused a quadratic 9% increase in IPF. Indomethacin is an inhibitor of mitochondrial function, so the glucose-induced increase in IPF was interpreted as feedback on mammary adenosine release from an anabolic response to glucose. Because NO synthase was not inhibited during indomethacin infusion, the feedback system is postulated to act through endothelial NO synthase. In the presence of L-NAME, glucose infusion had no effect on IPF, indicating that endothelial cyclooxygenase is not involved in glucose-induced changes in MBF.

Transcript profiling of the ruminant liver indicates a unique program of transcriptional regulation of ketogenic enzymes during food restriction.

Ruminants absorb little glucose and rely on hepatic gluconeogenesis and ketogenesis in the fed state to convert short-chain fatty acids produced during digestion into glucose and ketone bodies, respectively. In contrast to the non-ruminant response, fluxes through gluconeogenic and ketogenic pathways decrease during food restriction. Transcriptional regulation responsible for these unique food restriction responses has not been established. To determine the hepatic transcriptional response of ruminants to an acute drop in dietary nutrient supply, 102 yearling heifers were assigned to either ad libitum feeding or 24 h of food withdrawal in a randomized block design. Liver biopsies were obtained for microarray and quantitative real-time PCR analyses of gene expression. Plasma concentrations of non-esterified fatty acids were higher in food restricted heifers, while levels of β-hydroxybutyrate, triacylglycerol, and glucose were decreased. Despite a decline in substrate supply and a lower hepatic production of glucose, expression of the key gluconeogenic enzymes pyruvate carboxylase, phosphoenolpyruvate carboxykinase and fructose-1,6-bisphosphatase was upregulated as in non-ruminants. Downregulation of cholesterolgenic genes and upregulation of fatty acid oxidative genes were consistent with SREBP-2 and PPARα control, respectively. Ketogenesis from short-chain fatty acids was downregulated, contrary to the non-ruminant response to food restriction. Short-chain fatty acids may exert transcriptional control in the ruminant liver similar to that demonstrated in the large intestine of non-ruminants.

Addition of glycerol to lactating cow diets stimulates dry matter intake and milk protein yield to a greater extent than addition of corn grain

The objective of this study was to determine if the addition of glycerol to the diet of dairy cows would stimulate milk protein yield in the same manner as the addition of corn grain. Twelve multiparous lactating dairy cows at 81 ± 5 d in milk were subjected to 3 dietary treatments in a replicated 3 × 3 Latin square design for 28-d periods. The diets were a 70% forage diet considered the basal diet, the basal diet with 19% ground and high-moisture corn replacing forages, and the basal diet with 15% refined glycerol and 4% added protein supplements to be isocaloric and isonitrogenous with the corn diet. Cows were milked twice a day and samples were collected on the last 7 d of each period for compositional analysis. Within each period, blood samples were collected on d 26 and 27, and mammary tissue was collected by biopsy on d 28 for Western blot analysis. Dry matter intake increased from 23.7 kg/d on the basal diet to 25.8 kg/d on the corn diet and 27.2 kg/d on the glycerol diet. Dry matter intake tended to be higher with glycerol than corn. Milk production increased from 39.2 kg/d on the basal diet to 43.8 kg/d on the corn diet and 44.2 kg/d on the glycerol diet. However, milk yield did not differ between corn and glycerol diets. Milk lactose yields were higher on the corn and glycerol diets than the basal diet. Milk fat yield significantly decreased on the glycerol diet compared with the basal diet and tended to decrease in comparison with the corn diet. Mean milk fat globule size was reduced by glycerol feeding. Milk protein yield increased 197 g/d with addition of corn to the basal diet and 263 g/d with addition of glycerol, and the glycerol effect was larger than the corn effect. The dietary treatments had no effects on plasma glucose concentration, but plasma acetate levels decreased 27% on the glycerol diet. Amino acid concentrations were not affected by dietary treatments, except for branched-chain amino acids, which decreased 22% on the glycerol diet compared with the corn diet. The decreases in plasma acetate and branched-chain amino acid concentrations with glycerol and the larger effects of glycerol than corn on milk protein and fat yields suggest that glycerol is more glucogenic for cows than corn grain.