Feeding 2-hydroxy-4-(methylthio)-butanoic acid to periparturient dairy cows improves milk production but not hepatic metabolism

Forty-eight Holstein cows, entering second or later lactation, were utilized to determine the effects of 2-hydroxy-4-(methylthio)-butanoic acid (HMB) on milk production, hepatic lipid metabolism, and gluconeogenesis during the periparturient period. Cows were fed one of 3 diets as TMR starting 21 d before expected calving. These diets contained 0 (the basal diet), 0.09 (+HMB), or 0.18 (++HMB)% HMB. From parturition to 84 DIM, cows were fed diets that contained 0, 0.13, or 0.20% HMB. Prepartum and postpartum dry matter intakes were similar among cows fed the basal diet, +HMB and ++HMB. There was a quadratic effect on milk yield such that cows fed +HMB had the greatest milk yield; yields of milk by cows fed the basal diet and ++HMB were similar. This led to trends for increased yields of 3.5% fat-corrected milk and total solids when cows were fed +HMB. Percentages of fat, protein, and total solids in milk were not affected by treatment. Despite differences in milk yield, calculated energy balance was not affected by treatment. Plasma concentrations of NEFA, beta-hydroxybutyrate, and glucose were not different among treatments. Liver triglyceride content was similar among treatments on d 1 postpartum and was increased for cows consuming +HMB on d 21 postpartum compared with the other dietary treatments. Capacities for metabolism of [1-14C]palmitate by liver slices in vitro were not affected by treatment; however, conversion of [1-14C]propionate to CO2 and glucose decreased as the amount of HMB consumed by cows increased on d 21 postpartum. Cows consuming +HMB had greater days to first ovulation compared with cows consuming the basal diet and ++HMB as measured by plasma progesterone concentrations. These data suggest that adding HMB to low Met diets to achieve a predicted Met supply of approximately 2.3% of metabolizable protein supply is beneficial for increasing milk production but does not appear to benefit hepatic energy metabolism during early lactation.     

Duration and degree of diet-induced metabolic acidosis prepartum alter tissue responses to insulin in dairy cows

The objective of this experiment was to determine the effects of altering the dietary cation-anion difference (DCAD) fed for the last 21 or 42 d of gestation on glucose metabolism and tissue insulin responsiveness. Ninety parous Holstein cows at 232 d of gestation were assigned randomly to dietary treatments with 2 levels of DCAD (?70 or ?180 mEq/kg) fed for 2 durations (short: the last 21 d of gestation; long: the last 42 d of gestation). For the short treatments, a diet with +110 mEq/kg was fed from 232 to 254 d of gestation. Intravenous glucose tolerance tests (IVGTT) were performed at either 250 or 270 d of gestation by infusing 0.25 g of dextrose/kg of body weight within 1 min. The following day, cows underwent an insulin challenge (IC) and received 0.1 IU of insulin/kg of body weight intravenously. Blood was sampled at min ?15, ?5, and 0 to establish a baseline and from 5 to 180 min relative to infusions; plasma concentrations of glucose, insulin, and fatty acids were determined, and the respective areas under the curves (AUC) were calculated. Liver was sampled after the IVGTT, and adipose tissue was sampled after the IVGTT and IC for quantification of mRNA expression and protein abundance. Reducing the DCAD altered acid–base balance compatible with a compensated metabolic acidosis. At 250 d, reducing the DCAD increased the AUC for glucose and reduced that of insulin following the IVGTT, whereas during the IC, clearance rate decreased and time to half-life of insulin increased with reducing DCAD, resulting in a tendency to a larger AUC for fatty acids. At 270 d, quantitative insulin sensitivity check index and the revised quantitative insulin sensitivity check index were smaller in cows fed the acidogenic diets for the last 42 d of gestation compared with the last 21 d of gestation, thereby suggesting reduced insulin sensitivity. In addition, cows fed for the long duration tended to have greater AUC for glucose but smaller AUC for insulin following an IVGTT than those fed for the short duration, thereby suggesting reduced insulin release and glucose disposal. Treatments did not affect hepatic mRNA expression of G6PC, PCK1, PCK2, and PC or adipose tissue mRNA expression of ATGL, ACC, B2AR, HSL, and PLIN1. On the other hand, for proteins, reducing the DCAD linearly reduced abundance of rabbit anti-mouse protein kinase B (AKT) and tended to reduce rabbit anti-human phosphorylated (Ser-9) glycogen synthase kinase-3 β (pGSK) and the pGSK:rabbit anti-human glycogen synthase kinase-3 β (GSK) ratio in hepatic tissue, whereas a linear increase in rabbit anti-human hormone-sensitive lipase (HSL) and rabbit anti-mouse phosphorylated (Ser-660) hormone-sensitive lipase (pHSL) in adipose tissue was observed after the IVGTT at 250 d. Moreover, reducing the DCAD resulted in a linear reduction of AKT and tended to reduce rabbit anti-human acetyl-CoA carboxylase (ACC) but increased pHSL linearly in adipose tissue after an IC at 250 d. Cows fed acidogenic diets for a short duration tended to have less pHSL in adipose tissue than those fed for a long duration after an IVGTT at 270 d. Associations were observed between blood pH and mRNA and protein abundance in hepatic and adipose tissues. Diet-induced metabolic acidosis altered insulin release and insulin signaling, resulting in a shift in adipose tissue metabolism that would favor lipolysis over lipogenesis.     

The effects of increasing doses of 2 preparations of long-acting insulin on short-term plasma profiles of glucose and insulin in lactating dairy cows

Two experiments were conducted to investigate effects of administering increasing doses of 2 different preparations of long-acting insulin on the 24-h profiles of plasma glucose and insulin concentrations in mid lactation dairy cows. The 2 separately analyzed experiments investigated the effects administering either Humulin N (H), a neutral protamine Hagedorn insulin, or insulin glargine (Lantus, L), an insulin analog, at doses of 0 (control), 0.1, 0.2, and 0.4 IU/kg of body weight in a randomized complete block design. Sixteen cows (237 ± 11 d in milk for H; 213 ± 10 d in milk for L; mean ± SD) were used for each insulin preparation, resulting in n = 4 for each dose within insulin preparation. Cows were fitted with a single jugular catheter on the day before the study. On the day of the study, cows were given treatments by subcutaneous injection of either sterile water or the designated insulin type and dose. Blood samples were taken hourly from the jugular catheter. Subcutaneous injection of both H and L resulted in linear decreases in plasma glucose concentrations, increased area under the curve, and decreased nadir for plasma glucose following administration of the insulin preparations. Plasma insulin concentration linearly increased with increasing dose of H. Though elevated concentrations of insulin were measurable in cows treated with H, they were not measurable in cows treated with L. Attempts to measure overall insulin concentrations and metabolites of L by a commercially available ELISA and a commercially available RIA kit were not successful and did not retrieve values that we felt truly represented the amount of insulin activity exhibited during this treatment. Both long-acting insulin preparations elicited insulin-like activity in lactating dairy cows, as evidenced by reduced plasma glucose concentrations. Given these results, the potential exists to use both H and L to study the effects of insulin in mid lactation dairy cows without the confounding effect of severe hypoglycemia (< 20 mg/dL) or concurrent provision of glucose during treatment.     

Liver metabolism and production of cows fed increasing amounts of rumen-protected choline during the periparturient period

Forty-eight multiparous Holstein cows were fed treatments consisting of either 0, 45, 60, or 75 g/d of a rumen-protected choline (RPC) source in a completely randomized design from 21 d before expected calving to 63 d postpartum to determine whether choline supplementation to the diet would affect hepatic fatty acid and glucose metabolism, key metabolites in plasma, and cow performance. Dry matter intake (DMI), milk yield, body condition score, and body weights (BW) were similar for cows receiving the four treatments. Feeding RPC tended to increase yields of milk fat, 3.5% fat-corrected milk, and total solids. Plasma concentrations of nonesterified fatty acids and beta-hydroxybutyrate were not different among cows fed the four treatments. Concentrations of triglycerides in liver were similar, but concentrations of glycogen in liver increased as cows consumed increasing amounts of RPC. Hepatic capacity for storage of [1-(14)C]palmitate as esterified products within liver slices tended to decrease as the amount of RPC consumed by cows increased; however, effects of treatment on hepatic capacity for oxidation of [1-(14)C]palmitate to CO2 were not significant. These data imply that choline may increase the rate of very low density lipoprotein synthesis and secretion of esterified lipid products from liver. Hepatic capacities for conversion of [1-(14)C] propionate to CO2 and to glucose in liver were similar among cows fed the four treatments. Collectively, these results suggest that hepatic fatty acid metabolism and cow performance are responsive to increasing the supply of choline during the periparturient period.     

Short communication: insulin alters hepatic progesterone catabolic enzymes cytochrome P450 2C and 3A in dairy cows

High proportions of embryonic and early fetal losses in dairy cattle are associated with low peripheral concentrations of progesterone, which could result from increased catabolism, decreased production, or both. Progesterone catabolism occurs primarily in the liver via the cytochrome P450 2C (CYP2C) and cytochrome P450 3A (CYP3A) subfamilies (EC 1.14.14.1; unspecific monooxygenases). Recent observations from our laboratory have shown that the fractional rate constant of progesterone decay can be dramatically reduced by insulin because of a decrease in hepatic CYP2C and CYP3A activity. Little information exists on the regulation of progesterone catabolic enzymes in dairy cows. We hypothesized that elevated insulin concentrations would down-regulate hepatic CYP2C and CYP3A mRNA; therefore, our objectives were to determine the relative abundance of hepatic CYP2C and CYP3A mRNA in dairy cows in response to elevated concentrations of insulin. In the first experiment, 17 mature Holstein cows were drenched daily with 500 mL of water (n = 10) or propylene glycol (a gluconeogenic substrate; n = 7) from 10 d before their expected calving date until d 25 postpartum. Cows drenched with propylene glycol had a 30% increase in peripheral concentrations of insulin. Liver biopsies were collected on d 25 postpartum to determine the relative abundance of CYP2C and CYP3A mRNA. In the second experiment, 19 mature, lactating Holstein cows were randomly assigned to a hyperinsulinemic-euglycemic clamp (0.3 or 1.0 μg of insulin/kg of BW per h; n = 6 each) or remained as controls (saline infused; n = 7) for 96 h beginning on d 10 postpartum. Insulin infusion resulted in a 2.6- or 8- fold increase in peripheral concentrations of insulin, respectively. On d 14 postpartum, a liver biopsy was collected to determine CYP2C and CYP3A mRNA abundance. In experiment 1, the relative abundance of CYP2C mRNA in cows treated with propylene glycol did not differ from controls; however, the relative abundance of CYP3A mRNA in the propylene glycol group was 63% that of controls. For experiment 2, there was a dose-dependent decrease in the relative abundance of both CYP2C and CYP3A mRNA with increasing dosage of insulin. In conclusion, this study demonstrated that, in the cow, either providing a gluconeogenic feed-stuff or treatment with insulin decreased the abundance of mRNA for enzymes responsible for hepatic progesterone catabolism.     

Variable liver fat concentration as a proxy for body fat mobilization postpartum has minor effects on insulin-induced changes in hepatic gene expression related to energy metabolism in dairy cows

The liver plays a central role in adaptation for energy requirements around calving, and changes in the effects of insulin on hepatic energy metabolism contribute to metabolic adaptation in dairy cows. Hepatic insulin effects may depend on body fat mobilization. The objective of this study was to investigate the effects of insulin on the hepatic gene expression of enzymes involved in energy metabolism and factors related to nutrition partitioning in cows with high and low total liver fat concentration (LFC) after calving. Holstein cows were retrospectively grouped according to their LFC after calving as a proxy for body fat mobilization. Cows were classified as low (LLFC; LFC <24% fat/dry matter; n = 9) and high (HLFC; LFC >24.4% fat/dry matter; n = 10) fat-mobilizing after calving. Euglycemic-hyperinsulinemic clamps [6 mU/(kg × min) of insulin for 6 h] were performed in wk 5 antepartum (ap) and wk 3 postpartum (pp). Before and at the end of the euglycemic-hyperinsulinemic clamps, liver biopsies were taken to measure the mRNA abundance of enzymes involved in carbohydrate and lipid metabolism, expression related to the somatotropic axis, and adrenergic and glucocorticoid receptors. The mRNA abundance of pyruvate carboxylase, cytosolic phosphoenolpyruvate carboxykinase (PEPCK; PCK1), acyl-CoA-dehydrogenase very long chain (ACADVL), and hydroxyl-methyl-glutaryl-CoA-synthase 1 increased, but the mRNA abundance of solute carrier family 2 (SLC2A2 and SLC2A4), growth hormone receptor 1A (GHR1A), insulin-like growth factor 1 (IGF1), sterol regulatory element binding factor 1, adrenoceptor α 1A, and glucocorticoid receptor decreased from ap to pp. Insulin treatment was associated with decreased PCK1, mitochondrial PEPCK, glucose-6-phosphatase, propionyl-CoA-carboxylase α, carnitine-palmitoyl-transferase 1A, ACADVL, and insulin receptor mRNA, but increased IGF1 and SLC2A4 mRNA ap and pp and GHR1A mRNA pp. The mRNA abundance of SLC2A4 was greater, and the mRNA abundance of GHR1A and IGF1 tended to be lower in LLFC than in HLFC. Administration of insulin, albeit at a supraphysiological dose, was associated with inhibition of gene expression related to glucose production and β-oxidation, but we observed variable effects in the degree of insulin depression of individual genes. Insulin status is important for regulation of nutrient partitioning, but different LFC pp had very little influence on changes in hepatic gene expression following administration of insulin.     

Interrelations between glucose-induced insulin response, metabolic indicators, and time of first ovulation in high-yielding dairy cows

High-yielding dairy cows are more susceptible to metabolic and reproductive disorders than low-yielding cows. Insulin plays a pivotal role in the development of both problems. In the present study, we aimed to assess the glucose-induced insulin responses of dairy cows at different time points relative to calving and to relate this to the metabolic status and the time of first ovulation. Twenty-three healthy, multiparous Holstein-Friesian cows with a high genetic merit for milk yield were studied from 14 d prepartum to 42 d postpartum. Intravenous glucose tolerance tests were performed on −14, 14, and 42 d relative to calving to evaluate the plasma insulin and glucose responses to a glucose load, as estimated by the peak concentration, the area under the curve (AUC), and the clearance rates of insulin and glucose. Blood samples were obtained at 3-d intervals and analyzed for glucose, insulin, and nonesterified fatty acids (NEFA). The time of first ovulation was defined by transrectal ultrasonography and plasma progesterone analysis. Glucose-induced insulin AUC and peak concentration decreased and glucose clearance increased during lactation compared with the dry period. Plasma NEFA concentrations were negatively related to insulin AUC and peak concentrations. Fourteen cows ovulated within 42 d postpartum, and the remaining 9 cows suffered from delayed resumption of ovarian function. Survival analysis demonstrated that cows with lower NEFA concentrations during the dry period tended to have earlier resumption of ovarian activity. In conclusion, our data suggest a decreased plasma insulin response to glucose postpartum in high-yielding dairy cows, possibly contributing to metabolic stress during the early postpartum period. It is hypothesized that NEFA impair glucose-induced insulin secretion in dairy cows. Additionally, our results suggest the importance of lipolysis during the transition period as a risk factor for delayed ovulation.     

Suitability of low-dosage oxytocin treatment to induce milk ejection in dairy cows

Chronic use of high oxytocin (OT) dosages can cause a reduced response to endogenous OT. In this study the OT dosages used in the milking practice of 82 dairy cow farms were recorded. The OT dosages per cow used were high, especially when injected i.m. (23 ± 2 IU) compared with i.v. (7 ± 1 IU). In addition, the minimum OT dosages needed to obtain normal milk removal in cows with disturbed milk ejection were investigated. Seventeen cows routinely treated with OT during milking (group T) and 17 cows without previous OT treatment were used (group C). After cessation of spontaneous milk flow, both T and C groups were injected i.v. with a low dosage of OT (0.2 or 0.5 IU/cow). The time from injection until cessation of the OT-induced milk flow was recorded (response phase). The response phase and the amounts of removed milk by effect of the OT injection increased with increasing OT dosage. Values for 0.2 and 0.5 IU/cow of OT injected i.v. were (response phase and amount of milk removed) 198 ± 27 and 302 ± 18 s and 3.4 ± 0.7 kg and 6.5 ± 1.3 kg, respectively, for the C group, and 157 ± 15 and 221 ± 16 s and 3.2 ± 0.5 and 5.5 ± 1.0 kg, respectively, for the T group. Within 20 min of the OT injection, plasma concentrations returned to basal levels. The threshold OT concentration at cessation of milk flow after injection of 0.2 or 0.5 IU/cow of OT was calculated based on the OT plasma half-life. The threshold increased with increasing dosages of OT and was higher in group T (8 ± 1 and 14 ± 1 pg/mL for 0.2 and 0.5 IU/cow, respectively) than in group C (7 ± 1 and 11 ± 1 pg/mL for 0.2 and 0.5 IU/cow, respectively). In conclusion, desensitization of the udder toward OT occurs when the udder is exposed to elevated OT plasma concentrations, both short-term during the actual milking and long-term due to chronic high-dosage OT treatment. However, low-dosage OT treatments to induce normal milk removal can minimize the observed side effects.     

Immune response of postpartum dairy cows fed flaxseed

Thirty Holstein cows were allotted at calving to 10 groups of three cows blocked for similar calving dates to determine the effects of dietary fatty acids on functional properties of immunocompetent cells in early lactation and at breeding. Cows were assigned at calving to one of three isonitrogenous, isoenergetic, and isolipidic supplements based on either calcium salts of palm oil, Megalac, micronized soybeans, or whole flaxseed. On the day of AI and 20 d later, cows were injected with ovalbumin to measure the antibody response. Blood samples were taken at different times after calving (d 5, 21, 42, and 105) and after AI (d 0, 10, 20, and 40) for quantification of serum progesterone, fatty acids, and prostaglandin E2 concentrations. Isolated peripheral blood mononuclear cells were cultured to evaluate the proliferative response to concanavalin A and in vitro productions of interferon-gamma and prostaglandin E2. In general, feeding flaxseed increased serum omega-3 fatty acids concentration compared with feeding Megalac or soybeans, which decreased the omega-6 to omega-3 fatty acids ratio. There was a significant diet x day interaction for the proliferative response of mononuclear cells after calving and AI, indicating that cell responses from cows fed flaxseed were transiently reduced compared with those fed Megalac and soybeans. Moreover, during the breeding period, serum progesterone concentration was significantly greater in cows fed flaxseed compared with those fed Megalac, whereas serum concentration of prostaglandin E2 was significantly lower in cows fed flaxseed than in those fed Megalac or soybeans. Dietary treatments had no effect on the antibody response to ovalbumin and on in vitro productions of interferon-gamma and prostaglandin E2. However, interferon-gamma and prostaglandin E2 were impaired in the first 3 wk after parturition regardless of dietary treatment. These results suggest that changes in fatty acids, progesterone, and prostaglandins E2 concentrations in serum due to dietary treatment and physiological status influenced systemic immunity as shown by reduced proliferative response. However, other mechanisms must be considered and are discussed to explain dietary effect on lymphocyte proliferative response to mitogenic stimulation and other immune functions.     

Effects of rumen-protected Capsicum oleoresin on productivity and responses to a glucose tolerance test in lactating dairy cows

The objective of this experiment was to investigate the effects of rumen-protected Capsicum oleoresin (RPC) supplementation on feed intake, milk yield and composition, nutrient utilization, fecal microbial ecology, and responses to a glucose tolerance test in lactating dairy cows. Nine multiparous Holstein cows were used in a replicated 3 × 3 Latin square design balanced for residual effects with three 28-d periods. Each period consisted of 14 d for adaptation and 14 d for data collection and sampling. Treatments were 0 (control), 100, and 200 mg of RPC/cow per day. They were mixed with a small portion of the total mixed ration and top-dressed. Glucose tolerance test was conducted once during each experimental period by intravenous administration of glucose at a rate of 0.3 g/kg of body weight. Dry matter intake was not affected by RPC. Milk yield tended to increase for RPC treatments compared to the control. Feed efficiency was linearly increased by RPC supplementation. Concentrations of fat, true protein, and lactose in milk were not affected by RPC. Apparent total-tract digestibility of dry matter, organic matter, and crude protein was linearly increased, and fecal nitrogen excretion was linearly decreased by RPC supplementation. Rumen-protected Capsicum oleoresin did not affect the composition of fecal bacteria. Glucose concentration in serum was not affected by RPC supplementation post glucose challenge. However, compared to the control, RPC decreased serum insulin concentration at 5, 10, and 40 min post glucose challenge. The area under the insulin concentration curve was also decreased 25% by RPC. Concentration of nonesterified fatty acids and β-hydroxybutyrate in serum were not affected by RPC following glucose administration. In this study, RPC tended to increase milk production and increased feed efficiency in dairy cows. In addition, RPC decreased serum insulin concentration during the glucose tolerance test, but glucose concentration was not affected by treatment.     

Induced hyperketonemia affects the mammary immune response during lipopolysaccharide challenge in dairy cows

Metabolic adaptations during negative energy and nutrient balance in dairy cows are thought to cause impaired immune function and hence increased risk of infectious diseases, including mastitis. Characteristic adaptations mostly occurring in early lactation are an elevation of plasma ketone bodies and free fatty acids (nonesterified fatty acids, NEFA) and diminished glucose concentration. The aim of this study was to investigate effects of elevated plasma β-hydroxybutyrate (BHBA) at simultaneously even or positive energy balance and thus normal plasma NEFA and glucose on factors related to the immune system in liver and mammary gland of dairy cows. In addition, we investigated the effect of elevated plasma BHBA and intramammary lipopolysaccharide (LPS) challenge on the mammary immune response. Thirteen dairy cows were infused either with BHBA (HyperB, n = 5) to induce hyperketonemia (1.7 mmol/L) or with a 0.9% saline solution (NaCl, n = 8) for 56 h. Two udder quarters were injected with 200 μg of LPS after 48 h of infusion. Rectal temperature (RT) and somatic cell counts (SCC) were measured before, at 48 h after the start of infusions, and hourly during the LPS challenge. The mRNA abundance of factors related to the immune system was measured in hepatic and mammary tissue biopsies 1 wk before and 48 h after the start of the infusion, and additionally in mammary tissue at 56 h of infusion (8 h after LPS administration). At 48 h of infusion in HyperB, the mRNA abundance of serum amyloid A (SAA) in the mammary gland was increased and that of haptoglobin (Hp) tended to be increased. Rectal temperature, SCC, and mRNA abundance of candidate genes in the liver were not affected by the BHBA infusion until 48 h. During the following LPS challenge, RT and SCC increased in both groups. However, SCC increased less in HyperB than in NaCl. Quarters infused with LPS showed a more pronounced increase of mRNA abundance of IL-8 and IL-10 in HyperB than in NaCl. The results demonstrate that an increase of plasma BHBA upregulates acute phase proteins in the mammary gland. In response to intramammary LPS challenge, elevated BHBA diminishes the influx of leukocytes from blood into milk, perhaps by via modified cytokine synthesis. Results indicate that increased ketone body plasma concentrations may play a crucial role in the higher mastitis susceptibility in early lactation.     

Glucose and insulin responses to an intravenous glucose tolerance test administered to feed-restricted dairy cows receiving folic acid and vitamin B12 supplements

The present experiment was conducted to determine whether, during periods of negative energy balance, the increase in glucose availability, despite similar DMI and greater milk production, induced by a combined supplement of folic acid and vitamin B₁₂ was related to effects of insulin on metabolism. Sixteen multiparous Holstein cows averaging 45 days in milk (standard deviation: 3) were assigned to 8 blocks of 2 animals each according to their milk production (45 kg/d; standard deviation: 6) during the week preceding the beginning of the experiment. Within each block, they received weekly intramuscular injections of either saline (CON) or folic acid and vitamin B₁₂ (VIT) during 5 consecutive weeks. During the last week, the cows were fed 75% of their ad libitum intake during 4 d. Blood samples were taken the morning before starting the feed restriction and on the third day of feed restriction. On the fourth day of feed restriction, the daily meal was not served and an intravenous glucose tolerance test was performed. During the 4 wk preceding the feed restriction, milk production and DMI were not affected by treatments. During the feed restriction, the vitamin supplement tended to decrease milk fat concentration and increase milk concentration of lactose. Plasma concentrations of homocysteine, Ile, Leu, Val, and branched-chain AA increased in VIT cows during the restriction but not in CON cows. During the glucose tolerance test, insulin peak height was lower and insulin incremental positive area under the curve tended to be lower for VIT than for CON [83 (95% confidence interval, CI: 64–108) vs. 123 (95% CI: 84–180) µg·180 min/L, respectively]. Free fatty acid nadir was reached earlier for VIT than for CON [34 (95% CI: 26–43) vs. 46 (95% CI: 31–57) min, respectively]. Glucose area under the curve, clearance rate and peak height, insulin time to reach the peak and clearance rate, and free fatty acid nadir did not differ between VIT and CON. The reduction in insulin release during a glucose tolerance test without changes in glucose clearance rate or area under the curve suggests that the vitamin supplement improved insulin sensitivity in feed-restricted lactating dairy cows.     

Effects of weekly regrouping of prepartum dairy cows on innate immune response and antibody concentration

Objectives were to evaluate the effects of a stable prepartum grouping strategy on innate immune parameters, antibody concentration, and cortisol and haptoglobin concentrations of Jersey cows. Cows (253 ± 3 d of gestation) were paired by gestation length and assigned randomly to the stable (all-in-all-out; AIAO) or traditional (TRD) treatment. In the AIAO treatment, groups of 44 cows were moved into a pen where they remained for 5 wk, whereas in the TRD treatment, approximately 10 cows were moved into a pen weekly to maintain stocking density (44 cows for 48 headlocks). Pens were identical in size and design and each pen received each treatment a total of 3 times (6 replicates; AIAO, n = 259; TRD, n = 308). A subgroup of cows (n = 34/treatment) was selected on wk 1 of each replicate from which blood was sampled weekly from d −14 to 14 (d 0 = calving) to determine polymorphonuclear leukocyte (PMNL) phagocytosis, oxidative burst, and expression of CD18 and L-selectin, hemogram, cortisol and glucose concentrations, and haptoglobin concentration. Another subgroup of cows (n = 40/treatment) selected on wk 1 of each replicate was treated with chicken egg ovalbumin on d −21, −7, and 7 and had blood sampled weekly from d −21 to 21 for determination of immunoglobulin G anti-ovalbumin. All cows (n = 149) had blood sampled weekly for nonesterified fatty acid (NEFA) and β-hydroxybutyrate (BHBA) concentrations from d −21 to 21. Treatment did not affect percentage of PMNL positive for phagocytosis and oxidative burst (AIAO = 64.3 ± 2.9 vs. TRD = 64.3 ± 2.9%) and intensity of phagocytosis [AIAO = 2,910.82 ± 405.99 vs. TRD = 2,981.52 ± 406.87 geometric mean fluorescence intensity (GMFI)] and oxidative burst (AIAO = 7,667.99 ± 678.29 vs. TRD = 7,742.70 ± 682.91 GMFI). Similarly, treatment did not affect the percentage of PMNL expressing CD18 (AIAO = 96.3 ± 0.7 vs. TRD = 97.8 ± 0.7%) and L-selectin (AIAO = 44.1 ± 2.8 vs. TRD = 45.1 ± 2.8%) or the intensity of expression of CD18 (AIAO = 3,496.2 ± 396.5 vs. TRD = 3,598.5 ± 396.9 GMFI) and L-selectin (AIAO = 949.8 ± 22.0 vs. TRD = 940.4 ± 22.3 GMFI). Concentration of immunoglobulin G anti-ovalbumin was not affected by treatment (AIAO = 0.98 ± 0.05 vs. TRD = 0.98 ± 0.05 OD). The percentage of leukocytes classified as granulocyte (AIAO = 38.9 ± 1.5 vs. TRD 38.2 ± 1.5%) and the granulocyte:lymphocyte ratio (AIAO = 0.75 ± 0.04 vs. TRD = 0.75 ± 0.04) were not affected by treatment. Concentrations of cortisol (AIAO = 14.95 ± 1.73 vs. TRD = 18.07 ± 1.73 ng/mL), glucose (AIAO = 57.6 ± 1.5 vs. TRD = 60.0 ± 1.5 ng/mL), and haptoglobin (AIAO = 3.09 ± 0.48 vs. TRD = 3.51 ± 0.49 OD) were not affected by treatment. According to the current experiment, a stable prepartum grouping strategy does not improve innate immune parameters or antibody concentration compared with weekly prepartum regrouping.     

Long-acting insulins alter milk composition and metabolism of lactating dairy cows

This study investigated the effect of 2 different types of long-acting insulin on milk production, milk composition, and metabolism in lactating dairy cows. Multiparous cows (n = 30) averaging 88 d in milk were assigned to one of 3 treatments in a completely randomized design. Treatments consisted of control (C), Humulin-N (H; Eli Lilly and Company, Indianapolis, IN), and insulin glargine (L). The H and L treatments were administered twice daily at 12-h intervals via subcutaneous injection for 10 d. Cows were milked twice daily, and milk composition was determined every other day. Mammary biopsies were conducted on d 11, and mammary proteins extracted from the biopsies were analyzed by Western blot for components of insulin and mammalian target of rapamycin signaling pathways. Treatment had no effect on dry matter intake or milk yield. Treatment with both forms of long-acting insulin increased milk protein content and tended to increase milk protein yield over the 10-d treatment period. Analysis of milk N fractions from samples collected on d 10 of treatment suggested that cows administered L tended to have higher yields of milk protein fractions than cows administered H. Milk fat content and yield tended to be increased for cows administered long-acting insulins. Lactose content and yields were decreased by treatment with long-acting insulins. Administration of long-acting insulins, particularly L, tended to shift milk fatty acid composition toward increased short- and medium-chain fatty acids and decreased long-chain fatty acids. Plasma concentrations of glucose and urea N were lower for cows administered long-acting insulins; interactions of treatment and sampling time were indicative of more pronounced effects of L than H on these metabolites. Concentrations of nonesterified fatty acids and insulin were increased in cows administered long-acting insulins. Decreased concentrations of urea N in both plasma and milk suggested more efficient use of N in cows administered long-acting insulins. Western blot analysis of mammary tissue collected by biopsy indicated that the ratios of phosphorylated protein kinase b (Akt) to total Akt and phosphorylated ribosomal protein S6 (rpS6) to total rpS6 were not affected by long-acting insulins. Modestly elevating insulin activity in lactating dairy cows using long-acting insulins altered milk composition and metabolism. Future research should explore mechanisms by which either insulin concentrations or insulin signaling pathways in the mammary gland can be altered to enhance milk fat and protein production.     

Effect of a high cornstarch diet on hepatic cytochrome P450 2C and 3A activity and progesterone half-life in dairy cows

In the cow, inadequate concentrations of progesterone during gestation may lead to an abrupt termination of pregnancy. The primary organ involved in progesterone catabolism is the liver, which contains an abundance of cytochrome P450 isozymes (EC 1.14.14.1; mixed-function monooxygenases). The objectives of the current experiment were to determine the effect of feeding 2 isoenergetic and isonitrogenous diets, formulated to cause divergent insulin secretion, on hepatic cytochrome P450 2C (CYP2C) and 3A (CYP3A) activity as well as the resulting biological half-life of progesterone. Twenty-two Holstein cows averaging 80 ± 7 d in milk were randomly assigned to either a high cornstarch diet or a high fiber diet in a crossover experimental design consisting of two 14-d periods. Dry matter intake, milk yield, milk lactose yield, and milk lactose percentage were similar between the 2 diets. Milk fat yield and milk fat percentage were higher in cows fed the high fiber diet, whereas milk protein yield tended to be higher and milk protein percentage was higher in cows fed the high cornstarch diet. Energy balance tended to be improved by 57% in cows consuming the high cornstarch diet. Insulin concentrations at the time of liver biopsy (3.16 ± 0.04 h post-feeding) were increased by 44% in cows consuming the high cornstarch diet compared with cows consuming the high fiber diet. Cytochrome P450 2C activity was decreased by 45%, whereas CYP3A activity tended to be lowered by 34% in cows consuming the high cornstarch diet. Cytochrome P450 2C mRNA expression tended to be decreased by 21% in cows fed the high cornstarch diet, whereas CYP3A mRNA expression was not different between the dietary treatments. The fractional rate constant of progesterone decay was not different between the 2 diets; however, the half-life of progesterone tended to be longer in cows fed the high cornstarch diet compared with cows fed the high fiber diet (85 vs. 64 min, respectively). In summary, cows consuming the high cornstarch diet had increased insulin concentrations and lower hepatic CYP2C and CYP3A activity and tended to have a longer progesterone half-life compared with cows consuming the high fiber diet. Feeding diets that stimulate insulin secretion could alter progesterone clearance during lactation, when dairy cows have increased rates of progesterone inactivation because of high energy demands and increased DMI.     

Short communication: Survey of hepatic copper concentrations in Midwest dairy cows

Previous research from our laboratory and others indicates that liver copper concentrations in dairy cattle are commonly well above those recognized as adequate for the nutritional needs of the animal. It has also been speculated that hepatic copper concentrations have been increasing in recent years. Unlike other species, the threshold at which elevated liver copper concentrations becomes deleterious to hepatocytes is not known for cattle. Therefore, the objectives of this study were 3-fold: (1) to delineate differences in the range and mean dry matter hepatic copper concentration for dairy cattle in a retrospective analysis (January 1, 2007, to December 31, 2015), (2) to investigate hepatic copper concentrations in Midwest cull dairy cattle, and (3) to evaluate histologic changes in hepatocellular morphology in the context of copper concentration in cull cows. Furthermore, microscopic changes in hepatocellular morphology or architecture were examined and scored for evidence of inflammation, fibrosis, necrosis, and abundance of rhodanine-stained granules using hematoxylin and eosin and rhodanine staining. The retrospective analysis found copper concentrations within a range of 3 to 1,963 µg/g, with a mean of 473 µg/g. Hepatic copper concentrations in our retrospective study did not increase with time. In our abattoir analysis, copper concentrations ranged from 15 to 978 μg/g, with a mean of 390 μg/g. This study found that the range and mean hepatic copper concentrations were comparatively less in the current abattoir study than copper concentrations in our retrospective analysis. There was no evidence for hepatocellular changes associated with increased copper burdens in this study population.     

Hepatic triacylglycerol associations with production and health in dairy cows

The objectives were to evaluate the associations between hepatic triacylglycerol content and production, blood metabolites, incidence of diseases, reproduction, and survival in Holstein cows. Data were collected from 4 experiments including 329 cows in which hepatic tissue was sampled with a mean (± standard deviation) of 8.3 ± 1.5 d postpartum (6 to 11 d) and analyzed for triacylglycerol concentration. The mean (± standard deviation) concentration was 4.4 ± 2.8% on a wet basis and ranged from 0.4 to 16.1%. Intakes of dry matter (DM), energy balance, body weight (BW), body condition (BCS), productive performance, and incidence of diseases were evaluated for the first 105 d postpartum, whereas blood metabolites were assayed in the first 21 d postpartum. Reproductive performance and survival were monitored in the first 300 d postpartum. Mixed models were fitted to the data to investigate the linear and quadratic associations of hepatic triacylglycerol concentration with responses of interest. Increased concentration of hepatic triacylglycerol was associated with a quadratic increase in yields of milk, energy-corrected milk (ECM), and milk components. A change in hepatic triacylglycerol from 2.5 to 7.5% of the wet tissue was associated with an increase in yield of ECM of 1.8 kg/d, and with 0.2 kg more ECM per kg of DM intake. However, the increased efficiency was accompanied by decreases in DM intake, BCS, more exacerbated losses of BW, and a more negative body energy change. Increased concentration of hepatic triacylglycerol was associated with a quadratic increase in blood fatty acids and a linear increase in blood β-hydroxybutyrate concentrations, concurrent with linear decreases in concentrations of glucose and total Ca in blood. Moreover, a change in hepatic triacylglycerol from 2.5 to 7.5% was associated with linear increases in the relative risk of hyperketonemia by 2.5 times (15.2 vs. 37.5%), hypocalcemia by 1.7 times (30.3 vs. 52.4%), metritis by 2.1 times (12.5 vs. 25.7%), and diagnosis of multiple diseases postpartum by 2.4 times (8.7 vs. 21.1%). Survival in the herd by 300 d postpartum tended to decrease from 91.1 to 86.3% with an increase in hepatic triacylglycerol from 2.5 to 7.5% of the wet tissue, but no association was observed between hepatic triacylglycerol and measures of reproduction in the first 300 d postpartum. Concentrations of hepatic triacylglycerol in early lactation varied substantially, and increments resulted in quadratic association with productive performance, but at the expense of tissue reserves as those cows had increased tissue catabolism and risk of diseases that reduced survival.     

Clinical responses to intramammary endotoxin infusion in dairy cows subjected to feed restriction

Nonpregnant, midlactation primiparous Holstein cows were fed ad libitum (n = 12) or at 80% of maintenance energy requirements (n = 12) to determine whether feed restriction influences clinical response to endotoxin-induced mastitis. After 2 wk of ad libitum or restricted feeding, one mammary quarter per cow was infused with 100 microg of endotoxin. Within 3 to 6 h of intramammary infusion, endotoxin increased mean rectal temperature, heart rate, and milk somatic cell count and immunoglobulin (IgG) concentration; and decreased blood leukocyte count and rumen motility in both restricted and ad libitum-fed cows. Mean serum and milk tumor necrosis factor-alpha (TNF-alpha) concentrations showed only modest increases following endotoxin infusion. Restricted fed cows had slightly different acute fever responses and significantly increased heart and respiration rates than ad libitum fed cows. However, feed restriction did not influence mean total leukocyte count, rumen motility, serum TNF-a concentrations or milk IgG and TNF-alpha concentrations. Thus, results of this study suggest that energy balance does not significantly alter clinical symptoms following acute endotoxin-induced mastitis, at least in midlactation cows. As such, negative energy balance may not underlie the increases in severe coliform mastitis commonly observed in periparturient dairy cows.     

Short-term response in milk production,dry matter intake,and grazing behavior of dairy cows to changes in postgrazing sward height

Postgrazing sward height (PGSH) can be altered to adjust the allowance of grass in the dairy cow’s diet. This study aimed to investigate the short-term dairy cow response to a change in postgrazing height in early lactation. Ninety Holstein Friesian spring-calving cows were randomly assigned across 3 postgrazing height treatments (n = 30): 2.7 cm (severe), 3.5 cm (low), and 4.2 cm (moderate) from February 14 to April 24, 2011. From April 25, animals were rerandomized within each treatment to graze across 2 postgrazing heights: 3.5 cm (low) or 4.5 cm (high). Animal production measurements were taken from April 4 to 24 (measurement period 1; M1) and from April 25 to May 15 (measurement period 2; M2). The 6 treatments (n = 15) of M2 were severe-low, severe-high, low-low, low-high, moderate-low, and moderate-high. During M1, increasing postgrazing height from severe to low to moderate linearly increased daily milk yield (21.5, 24.6 and 25.8 kg/cow per day) and grass dry matter intake (GDMI; 13.2, 14.9, and 15.8 kg of DM/cow per day). Milk solids yield was reduced in the severe (−1,518 g/cow per day) treatment when compared with the low and moderate cows (1,866 g/cow per day, on average). The milk yield (MY) response to change in PGSH between M1 and M2 (V_M1−M2) was established using V_M1−M2 MY = −1.27 − 1.89 × PGSH_M1 + 1.51 × PGSH_M2 (R² = 0.64). The MY response associated with each treatment between M1 and M2 (3 wk) were −1.03 kg/cow for severe-low, 0.68 kg/cow for severe-high, −2.56 kg/cow for low-low, −1.11 kg/cow for low-high, −4.17 kg/cow for moderate-low, and −2.39 kg/cow for moderate-high. The large increase in energy intake in severe-high between M1 and M2 was achieved through higher GDMI per minute and GDMI per bite, which supported the positive change in MY. Treatments low-high, moderate-low, and moderate-high recorded the highest overall cumulative milk yield (74 kg of milk solids/cow) over the 6-wk period, whereas severe-low and severe-high had the lowest (65 kg of MS/cow). From the animal responses observed in the present study, imposing a postgrazing height of 3.5 cm in early spring provides the opportunity to increase postgrazing height thereafter; the cow increases GDMI accordingly and converts the additional energy intake into milk output. The equations established in this paper provide a decision tool for dairy farmers to anticipate the animal response when postgrazing height is altered or maintained around the tenth week of lactation.     

Effects of dietary energy allowance and decline in dry matter intake during the dry period on responses to glucose and insulin in transition dairy cows

We assessed whether high energy intake during the early dry period [144% of metabolizable energy (ME) requirements/d] followed by a gradual restriction of energy intake in the close-up dry period (119% of ME/d; HEI) impaired whole-body insulin sensitivity compared with a controlled energy intake (100% of ME/d; CEI) throughout the 6-wk dry period. Multiparous Ayrshire dairy cows (n = 16) were blocked by body weight, body condition score, and expected date of parturition and were used in a randomized complete block design until 10 d after parturition. Cows were fed either HEI or CEI diets based on grass silage during the first 3 wk of the dry period and grass silage supplemented with a commercial concentrate (30% of ME intake) during the final 3 wk of gestation. After calving, all cows were fed grass silage ad libitum and an increasing amount of commercial concentrate (maximum 9 kg at d 10 postpartum). Intravenous glucose tolerance tests (IVGTT) and intravenous insulin challenges were performed ?10 ± 5 d (n = 15) and +10 ± 1 d (n = 14) relative to parturition. Following glucose injection, we did not find any treatment effects on glucose and insulin responses. The prepartal nonesterified fatty acid (NEFA) response of the HEI group was blunted, basal NEFA and the decrement of NEFA were smaller, and the area under the response curve (AUC) of NEFA was less negative in HEI cows than in CEI cows. The NEFA response reversed after parturition; the NEFA AUC of the HEI group was more negative than that of the CEI group. We did not find similar responses after insulin injection. Across the treatments, NEFA AUC correlated strongly with the basal NEFA concentration during the IVGTT pre- and postpartum. Calculated and model-based indices characterizing the overall glucose tolerance and β-cell function and the insulin sensitivity were higher after parturition than during the dry period. Consistent with the lower basal insulin, the acute insulin release after the glucose infusion was smaller in postpartal IVGTT than in prepartal IVGTT. The results suggest that whole-body insulin sensitivity of the cows increased after parturition. However, the role of peripheral insulin sensitivity in the regulation of glucose partitioning seems to be minor relative to the major change in insulin secretion and clearance during the periparturient period.