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.     

Effects of wildfire smoke exposure on innate immunity,metabolism, and milk production in lactating dairy cows

Wildfires are particularly prevalent in the Western United States, home to more than 2 million dairy cows that produce more than 25% of the nation's milk. Wildfires emit fine particulate matter (PM_2.5) in smoke, which is a known air toxin and is thought to contribute to morbidity in humans by inducing inflammation. The physiological responses of dairy cows to wildfire PM_2.5 are unknown. Herein we assessed the immune, metabolic, and production responses of lactating Holstein cows to wildfire PM_2.5 inhalation. Cows (primiparous, n = 7; multiparous, n = 6) were monitored across the wildfire season from July to September 2020. Cows were housed in freestall pens and thus were exposed to ambient air quality. Air temperature, relative humidity, and PM_2.5 were obtained from a monitoring station 5.7 km from the farm. Animals were considered to be exposed to wildfire PM_2.5 if daily average PM_2.5 exceeded 35 µg/m³ and wildfire and wind trajectory mapping showed that the PM_2.5 derived from active wildfires. Based on these conditions, cows were exposed to wildfire PM_2.5 for 7 consecutive days in mid-September. Milk yield was recorded daily and milk components analysis conducted before, during, and after exposure. Blood was taken from the jugular vein before, during, and after exposure and assayed for hematology, blood chemistry, and blood metabolites. Statistical analysis was conducted using mixed models including PM_2.5, temperature-humidity index (THI), parity (primiparous or multiparous), and their interactions as fixed effects and cow as a random effect. Separate models included lags up to 7 d to identify delayed and persistent effects from wildfire PM_2.5 exposure. Exposure to elevated PM_2.5 from wildfire smoke resulted in lower milk yield during exposure and for 7 d after last exposure and higher blood CO₂ concentration, which persisted for 1 d following exposure. We observed a positive PM_2.5 by THI interaction for eosinophil and basophil count and a negative PM_2.5 by THI interaction for red blood cell count and hemoglobin concentration after a 3-d lag. Neutrophil count was also lower with a combination of higher THI and PM_2.5. We found no discernable effect of PM_2.5 on haptoglobin concentration. Effects of PM_2.5 and THI on metabolism were contingent on day of exposure. On lag d 0, blood urea nitrogen (BUN) was reduced with higher combined THI and PM_2.5, but on subsequent lag days, THI and PM_2.5 had a positive interaction on BUN. Conversely, THI and PM_2.5 had a positive interacting effect on nonesterified fatty acids (NEFA) on lag d 0 but subsequently caused a reduction in circulating NEFA concentration. Our results suggest that exposure to high wildfire-derived PM_2.5, alone or in concert with elevated THI, alters systemic metabolism, milk production, and the innate immune system.     

Dietary lipid metabolism in lactating dairy cows

Effects of feeding an oil seed supplement treated with formalin upon lipid patterns of blood and synthesis of milk fat were evaluated. Percentages and yields of fatty acids of milk fat with chain lengths between 6 and 16 carbons were decreased while percentages and yields of stearate and linoleate were increased when the lipid supplement was fed. Calculations in cows fed control and supplement, 60% and 80%, respectively, of fatty acids of milk were derived from lipids of blood were supported by arterial-venous differences. Comparisons of the fatty acid compositions of triacylglycerol of plasma and milk fat suggested that triacylglycerol may not be the sole source of linoleate transferred from blood to milk fat. A preliminary evaluation of supplement effects upon lipoprotein patterns of serum indicated two peaks in the low density lipoprotein class and that the increase in total cholesterol of blood caused by feeding lipid supplement is due to increases in cholesterol content of the low density and high density lipoprotein classes.     

Effect of exogenous bovine somatotropin on mammary lipid metabolism and milk yield in lactating dairy cows

Eight lactating Holstein cows were used to study the effect of bovine somatotropin on mammary lipid metabolism, milk production, and milk composition. Eight cows were injected with either excipient (n = 4) or 50 IU somatotropin/d (n = 4). A 2-wk adjustment period preceded a 10-d period when treatments were administered. On d 9, serial blood samples were collected from 2 to 5 h post-injection to determine concentrations of somatotropin and metabolites in plasma. Mammary tissue biopsies were obtained 2 to 3 h after a.m. milking on d 10 to determine lipoprotein lipase activity and in vitro rates of acetate incorporation into fatty acids and acetate oxidation by mammary tissue slices. Activity of lipoprotein lipase in milk and mammary tissue and rate of acetate metabolism by mammary tissue were not affected by treatment. The following means for placebo and somatotropin, respectively, were: milk production (21.5 and 24.8 kg/d), milk fat (3.8 and 4.0%), and 4% FCM (20.7 and 25.1 kg/d). Plasma concentrations of somatotropin (3.0 and 43.4 ng/ml) and free fatty acids (105.6 and 219.5 mu eq/L) were significantly elevated with somatotropin. The injection of somatotropin into lactating dairy cows did not affect the mechanisms of fatty acid synthesis or metabolism evaluated in this study.     

High-oil, high-protein diets and milk secretion by cows

Intakes of grass silage and yield and composition of milk were examined in cows given high-protein diets to which two quantities of soybean oil had been added either as intracellular or extracellular inclusions. Adding extra protein to the diet did not prevent reduction of dry matter intakes of forage when soybean oil was added to the diet of cows. There is a fundamental difference in the mechanism by which high-starch diets reduce intake of forage dry matter from that of high-oil diets. In diets containing extracellular unsaturated oils, concentration of oil in the total ration, not the amount of oil per se, is the primary determinant reducing milk fat percentage.     

Dietary fat and nitrogen composition of milk from lactating cows

Effects of dietary fat on milk composition, particularly milk N, were evaluated in lactating dairy cows at two stages of lactation. Complete mixed diets containing 0, 3.5, or 7% of the diet DM as animal fat were fed to 12 cows in a 3 X 3 Latin square. Cows were divided into two status categories based on stage of lactation resulting in two squares of early and two of late lactation cows. Percentages of fat, solids, lactose, and protein were decreased and ash increased in the milk of cows fed the 7% fat diet. Percent of casein N was elevated while nonprotein N was depressed by fat fed in diet. Percentage of solids and protein was higher and lactose lower for cows in late lactation than for those in early lactation. Dietary fat reduced the proportion of short-chain fatty acids in milk fat and increased C18:0 and C18:1 fatty acids. Digestibility of DM, energy, and fiber was not significantly affected by dietary fat, but estimates decreased with increased dietary fat. Cows in late lactation had a higher digestibility of dietary fractions than early lactation cows even though intakes of DM were similar.     

Influence of sodium chloride or potassium chloride on systemic acid-base status, milk yield, and mineral metabolism in lactating dairy cows

Our objective was to evaluate the response of lactating dairy cows to dietary Na, K, and Cl while holding cation-anion balance constant. Fifteen lactating Holstein cows, blocked according to age and previous milk yield, were assigned randomly to replicated 3 x 3 Latin squares with experimental periods of 3 wk. Diets contained sorghum silage and concentrate in a 40:60 ratio (DM basis) and were formulated to provide +32 meq of [(Na + K) - Cl]/100 g diet DM via one of three variations: 1) basal concentrations of dietary Na, K, and Cl, 2) basal diet with addition of 20 meq of Na and 20 meq of Cl/100 g in the form of 1.17% added NaCl, or 3) basal diet with the addition of 20 meq of K and 20 meq of Cl/100 g in the form of 1.56% added KCl. Free proton concentration in blood was increased by addition of NaCl and KCl; however, this increase did not appear to be physiologically significant, and no other measures of acid-base status were significantly affected. Plasma K was higher and plasma Mg was lower for the diets with supplemental NaCl or KCl than for basal diet. Urine mineral excretion reflected dietary mineral concentration, except Ca and Mg excretion rates were reduced by feeding the KCl diet. Milk yield reflected DM intake, which was lowest with supplemental NaCl. Results of this study indicate that, at a dietary cation-anion balance of +32 meq/100 g of diet DM, the balance of Na and K to Cl in the diet is a more important determinant of dietary impact on systemic acid-base status than actual dietary concentrations of Na, K, and Cl.     

Abomasal infusion of butterfat increases milk fat in lactating dairy cows

The objective of this study was to compare the effects of abomasal infusion of butterfat containing all fatty acids (FA) present in milk, including the short- and medium-chain FA, with infusion of only the long-chain FA (LCFA) present in milk, on the FA composition and milk fat yield in lactating dairy cows. Eight rumen-fistulated Holstein cows, in early lactation (49 ± 20 days in milk) were used in a replicated 4 × 4 Latin square design. Treatments were abomasal infusion of the following: 1) no infusion (control), 2) 400 g/d of butterfat (butterfat), 3) 245 g/d of LCFA (blend of 59% cocoa butter, 36% olive oil, and 5% palm oil) providing 50% of the 16:0 and equivalent amounts of C18 FA as found in 400 g of butterfat, and 4) 100 g/d of conjugated linoleic acid (CLA, negative control), providing 10 g of trans-10, cis-12 CLA. Fat supplements were infused in equal portions 3 times daily at 0800, 1400, and 1800 h during the last 2 wk of each 3-wk experimental period. Daily dry matter intake and milk production were unaffected by the infusion treatments. Butterfat infusion increased milk fat percentage by 14% to 4.26% and milk fat yield by 21% to 1,421 g/d compared with controls (3.74% and 1,178 g/d). Milk fat percentage and fat yield were decreased by 43% by CLA. Milk protein percentage was higher (3.70%) in CLA-infused cows than in control (3.30%), butterfat (3.28%), or LCFA (3.27%) treatments. Although LCFA had no effect on fat synthesis, abomasal infusion of butterfat increased milk fat percentage and yield, suggesting that the availability of short- and medium-chain FA may be a limiting factor for milk fat synthesis.     

Cooling ameliorates decreased milk protein metrics in heat-stressed lactating Holstein cows

Cooling can alleviate the negative consequences of heat stress on multiple milk production metrics in dairy cows. However, it is still controversial whether cooling can increase milk protein content compared with heat-stressed cows. The objective of the present study was to evaluate the relief effect of cooling on the decrease in milk protein concentration during heat stress and elucidate the potential metabolic mechanisms. Thirty lactating multiparous Holstein cows (days in milk = 175 ± 25 d, milk yield = 27.5 ± 2.5 kg/d; mean ± SD) were assigned to 1 of 3 treatments: heat stress (HS; n = 10), cooling (CL; n = 10), and cooling with pair-feeding (PFCL; n = 10). The barns for PFCL and CL cows were equipped with sprinklers and fans, whereas the barn for HS cows were not. The average temperature-humidity index during the experiment ranged from 74 to 83. The spraying was activated automatically 2 times per day (1130–1330 h and 1500–1600 h) with 3 min on and 6 min off during the first 2 wk, and 1.5 min on and 3 min off during the last 2 wk, whereas the fans operated 24 h/d. The experiment lasted for 4 wk in total. Milk, urine, feces, total mixed ration, blood, and rumen fluid samples were collected weekly. Compared with HS, feed efficiency (1.24 and 1.49), milk protein yield (0.82 and 0.94 kg/d), and milk fat yield (0.98 and 1.26 kg/d) were increased in PFCL, whereas the differences between CL and HS were not significant. Compared with HS cows, PFCL and CL cows had a lower respiratory rate (70.6, 59.1, and 60.3 breaths per minute, respectively), rectal temperature (38.95, 38.61, and 38.51°C), and shoulder skin temperature (33.95, 33.25, 33.40°C), and had greater milk protein content (3.41, 3.72, and 3.69%) and milk fat percent (4.08, 4.97, 4.65%). Both the blood activity of catalase (increased by 12.8 and 41.0%) and glutathione peroxidase (12.6 and 40.4%) of PFCL and CL cows were greater than the HS cows. Compared with HS, cooling increased the blood content of glucose, methionine, threonine, and cystathionine by 10.7% and 10.3%, 19.0% and 9.5%, 15.8% and 12.0%, and 9.5% and 23.8% in PFCL and CL, respectively. In conclusion, the results indicated that cooling partially rescued milk protein synthesis induced by heat stress, and the potential mechanism may have been due to increased antioxidant ability, blood glucose, and key AA. Consequently, in addition to modifying the environment, nutritional and physiological strategies designed to influence carbohydrate, AA, and oxidative homeostasis may be an opportunity to maintain or correct low milk protein content during the warm summer months.     

Ruminal and intermediary metabolism of propylene glycol in lactating Holstein cows

Four lactating Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, and hepatic vein were used in a cross-over design to study the metabolism of propylene glycol (PG). Each cow received 2 treatments: control (no infusion) and infusion of 650 g of PG into the rumen at the time of the morning feeding. Propylene glycol was infused on the day of sampling only. Samples of arterial, portal, and hepatic blood as well as ruminal fluid were obtained at 0.5 h before feeding and at 0.5, 1.5, 2.5, 3.5, 5, 7, 9, and 11 h after feeding. Infusion of PG did not affect ruminal pH or the total concentration of ruminal volatile fatty acids, but did decrease the molar proportion of ruminal acetate. The ruminal concentrations of PG, propanol, and propanal as well as the molar proportion of propionate increased with PG infusion. The plasma concentrations of PG, ethanol, propanol, propanal, glucose, l-lactate, propionate, and insulin increased with PG and the plasma concentrations of acetate and β-hydroxybutyrate decreased. The net portal flux of PG, propanol, and propanal increased with PG. The hepatic uptake of PG was equivalent to 19% of the intraruminal dose. When cows were dosed with PG, the hepatic extraction of PG was between 0 and 10% depending on the plasma concentration of PG, explaining the slow decrease in arterial PG. The increased net hepatic flux of l-lactate with PG could account for the entire hepatic uptake of PG, which suggests that the primary hepatic pathway for PG is oxidation to l-lactate. The hepatic uptake of propanol increased with PG, but no effects of PG on the net hepatic and net splanchnic flux of glucose were observed. Despite no effect of PG on net portal flux and net hepatic flux of propionate, the net splanchnic flux of propionate increased and the data suggest that propionate produced from hepatic metabolism of propanol is partly released to the blood. The data suggest that PG affects metabolism of the cows by 2 modes of action: 1) increased supply of l-lactate and propionate to gluconeogenesis and 2) insulin resistance of peripheral tissues induced by increased concentrations of PG and propanol as well as a decreased ratio of ketogenic to glucogenic metabolites in arterial blood plasma.     

Excretion of deoxynivalenol and its metabolite in milk, urine, and feces of lactating dairy cows

Corn contaminated with deoxynivalenol was added to the diets of three dairy cows for 5 d and milk, urine, and 3 d following feeding of the diets. Dietary concentrations of deoxynivalenol averaged 66 mg/kg. Following exposure to deoxynivalenol, unconjugated deepoxydeoxynivalenol, a metabolite of deoxynivalenol, was present in milk at concentrations up to 26 ng/ml. Deoxynivalenol was not detected in the milk. Approximately 20% of the deoxynivalenol fed was recovered in the urine and feces in the unconjugated forms as deepoxydeoxynivalenol (96%) and deoxynivalenol (4%). After incubating urine with beta-glucuronidase, the concentration of unconjugated deepoxydeoxynivalenol increased by 7 to 15-fold whereas unconjugated deoxynivalenol increased 1.6 to 3-fold. Detectable concentrations of unconjugated deepoxydeoxynivalenol were found in urine and feces up to 72 h after the last oral exposure. Thus, urine and feces are the diagnostic specimens of choice for the determination of deoxynivalenol exposure in cows. Feeding deoxynivalenol-contaminated diets for 5 d did not alter feed intake or milk production nor were the milk concentrations of calcium, phosphorus, sodium, potassium, magnesium, or nitrogen altered.     

Temporal changes in milk fatty acid composition during diet-induced milk fat depression in lactating cows

Diet-induced milk fat depression (MFD) in lactating cows has been attributed to alterations in ruminal lipid metabolism leading to the formation of specific fatty acid (FA) biohydrogenation intermediates that directly inhibit milk fat synthesis. However, the mechanisms responsible for decreased lipid synthesis in the mammary gland over time are not well defined. The aim of this study was to evaluate the effect of diet on milk FA composition and milk fat production over time, especially during MFD, and explore the associations between MFD and FA biohydrogenation intermediates in omasal digesta and milk. Four lactating Finnish Ayrshire cows used in a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments and 35-d experimental periods were fed diets formulated to cause differences in ruminal and mammary lipid metabolism. Treatments consisted of an iso-nitrogenous total mixed ration based on grass silage with a forage to concentrate ratio of 65:35 or 35:65 without added oil, or with sunflower oil at 50 g/kg of diet dry matter. The high-concentrate diet with sunflower oil (HSO) induced a 2-stage drop in milk fat synthesis that was accompanied by specific temporal changes in the milk FA composition. The MFD on HSO was associated especially with trans-10 18:1 and also with trans-9,cis-11 conjugated linoleic acid (CLA) in milk and omasal digesta across all diets and was accompanied by the appearance of trans-10,cis-15 18:2. Trans-10,cis-12 CLA was increased in HSO, but milk fat secretion was not associated with omasal or milk trans-10,cis-12 CLA. The temporal changes in milk fat content and yield and milk FA composition reflect the shift from the predominant ruminal biohydrogenation pathway to an alternative pathway. The ambiguous role of trans-10,cis-12 CLA suggests that trans-10 18:1, trans-9,cis-11 CLA and trans-10,cis-15 18:2 or additional mechanisms contributed to the diet-induced MFD in lactating cows.     

Dopamine antagonist alters serum cortisol and prolactin secretion in lactating Holstein cows

The role of dopamine in regulating glucocorticoid and prolactin secretion was investigated in lactating Holstein cows by characterizing serum cortisol and prolactin responses to fluphenazine, a dopamine receptor antagonist. Twelve anovulatory cows received an intravenous bolus injection of either saline (n = 6) or 0.3 mg of fluphenazine/kg of body weight (n = 6) in wk 2 postpartum. Blood samples were collected every 30 min for 4 h before and 4 h after saline or fluphenazine injection. Serum progesterone concentration was 0.13 ± 0.1 ng/mL and did not differ between groups. No difference in serum cortisol concentrations was detected between groups before treatments. Fluphenazine increased serum cortisol concentrations within 30 min after fluphenazine administration (>30 ng/mL) and concentrations remained elevated throughout the sampling period. Cortisol remained unchanged in saline-treated cows (<10 ng/mL). Prolactin concentrations also increased after fluphenazine administration (103.1 ± 3.1 ng/mL), but were unaffected by saline (18 ± 3.1 ng/mL). Prolactin concentrations remained elevated throughout the sampling period in fluphenazine-treated cows. Our results indicated that a dopamine antagonist increased cortisol, suggesting that endogenous dopamine, at least in part, regulates cortisol and prolactin secretion. These effects are regulated through dopamine receptors in anovulatory lactating dairy cows during the early postpartum period.     

Effects of hot,humid weather on milk temperature,dry matter intake,and milk yield of lactating dairy cows

Lactating cows were exposed to moderate and hot, humid weather to determine the effect of increasing ambient temperature, relative humidity, or temperature-humidity index (THI) on intake, milk yield, and milk temperature. Minimum and maximum temperatures averaged 17.9 and 29.5 degrees C (cool period) and 22.5 and 34.4 degrees C (hot period), and minimum and maximum THI averaged 63.8 and 76.6 (cool period) and 72.1 and 83.6 (hot period). Environmental conditions had minor effects on intake and milk yield during the cool period. During the hot period, the THI 2 d earlier and mean air temperature 2 d earlier had the greatest impact on milk yield and DMI, respectively. Both breeds maintained milk temperature within normal ranges during the cool period, but Holstein and Jersey p.m. milk temperatures averaged 39.6 and 39.2 degrees C during the hot period. Current day mean air temperature during the hot period had the greatest impact on cow p.m. milk temperature, and minimum air temperature had the greatest influence on a.m. milk temperature. Dry matter intake and milk yield declined linearly with respective increases in air temperature or THI during the hot period and milk temperature increased linearly with increasing air temperature. Dry matter intake and milk yield both exhibited a curvilinear relationship with milk temperature. Environmental modifications should target the effects of high temperatures on cow body temperature and should modify the environment at critical times during the day when cows are stressed, including morning hours when ambient temperatures are typically cooler and cows are not assumed to be stressed.     

Relationship between level of milk production and multiple ovulations in lactating dairy cows

Our objective was to evaluate factors associated with spontaneous multiple ovulations in lactating dairy cows. Ovaries of cows [n = 267; >50 days in milk (DIM)] were evaluated weekly using ultrasound to determine spontaneous (i.e., no hormonal treatment) ovulation rate starting at 50 DIM and continuing until pregnancy diagnosis. Cows were fitted with a transmitter to record standing activity during estrus, and serum progesterone concentration was assessed weekly starting at wk 1 postpartum for all cows. Overall, 76 (28.5%) cows were anovular and 191 (71.5%) were ovular by 71 DIM. Incidence of anovulation was not associated with level of milk production but was associated with lower body condition. For anovular cows (n = 41) that spontaneously recovered, the multiple ovulation rate at first ovulation was 46.3%. For second and subsequent ovulations (n = 463), the level of milk production for 14 d preceding estrus was associated with increased ovulation rate. To illustrate, incidence of multiple ovulations was 1.6% (2/128), 16.9% (32/189), and 47.9% (70/146) for ovulations when cows were producing <35, 35 to <45, and ≥45 kg/d, respectively. Among cows for which estrous behavior was recorded, those with multiple ovulations (n = 48) had shorter duration of estrus (4.3 ± 0.7 vs. 9.9 ± 0.5 h) and higher production (47.2 ± 0.9 vs. 38.1 ± 0.5 kg/d) than cows with single ovulations (n = 237). Circulating concentrations of estradiol were lower (5.5 ± 0.3; n = 15 vs. 7.8 ± 0.4 pg/mL; n = 71) during periods of estrus with multiple ovulations despite a greater preovulatory follicular volume (4136 ± 123 vs. 3085 ± 110 mm³). Similarly, serum progesterone concentration 7 d after estrus was lower for cows with multiple than single ovulations (2.5 ± 0.3 vs. 3.2 ± 0.1 ng/mL) despite a greater luteal volume (8291 ± 516 vs. 6405 ± 158 mm³). In summary, the first spontaneous ovulation in anovular cows and a higher level of milk production for 14 d preceding estrus were associated with increased multiple ovulation rate. Additionally, cows with multiple ovulations had lower estradiol at estrus, a shorter duration of estrus, and lower progesterone at 7 d after estrus than cows with single ovulations.     

Reevaluation of polychlorinated biphenyl concentrations in milk and body fat of lactating cows

Polychlorinated biphenyls occur as mixtures of congeners, each of which may be absorbed, excreted, or metabolized differently. Quantification of polychlorinated biphenyls by packed-column GLC has been difficult. Many quantification techniques, generally based on quantification of a selected group of the congeners present, have been used by researchers. Results of these studies have not provided a consistent basis to determine the relationship between the amounts of polychlorinated biphenyls consumed and residual polychlorinated biphenyls in milk and animal tissues. In the present study, we hypothesized that if a standardized quantification procedure was applied, consistent dose:residue predictions could be made. Weighted percentage of each peak in the polychlorinated biphenyl mixture procedures was used as a standard method to quantify polychlorinated biphenyls in milk fat from published and unpublished studies where lactating cows were fed between 3.5 and 1000 mg/d (Aroclor 1254) for greater than or equal to 15 d. When steady state occurred by 60 d of dosing, the relationship between concentration in milk fat (microgram/g) and daily dose as mg and as mg/kg BW were described by the equations: [polychlorinated biphenyls, microgram/g]milk fat = .28 (daily dose, mg).82, and [polychlorinated biphenyls, microgram/g]milk fat = 50.2 (daily dose, mg/kg BW).81. Similar equations described the relationships between daily dose and concentrations in adipose tissue and blood plasma. Concentrations of polychlorinated biphenyls residues in milk fat of 79 animals that were dosed with known quantities of polychlorinated biphenyls for 15 to 107 d confirmed that the equations could be valuable in predicting exposure over a wide range of exposure durations and concentrations.     

Pharmacokinetics of progesterone in lactating dairy cows: Gaining some insights into the metabolism from kinetic modeling

Progesterone pharmacokinetics were analyzed for plasma hormone concentrations ranging from linear to saturated metabolism in lactating Holstein cows with differing daily milk yields. The adequacy of 2-coupled first-order (bi-exponential equation), hyperbolic (Michaelis-Menten equation), and sigmoidal (Hill equation) kinetic models to describe the experimental progesterone pharmacokinetic profiles was examined on a statistical basis. After nonlinear regression and statistical analysis of the data-fitting capability, a simple one-compartment model based on Hill equation proved to be most adequate. This model indicates an enzyme-catalyzed metabolism of progesterone involving cooperative substrate-binding sites, resulting from allosteric effects that yield a sigmoidal saturation rate curve. Kinetic parameters were estimated for 2 groups of lactating Holstein cows with different daily milk yields. We found, for the first time, a remarkable quantitative agreement of the Hill coefficient value with that reported in pharmacokinetic studies involving cytochrome P450, family 3, subfamily A (CYP3A)-mediated reactions in other mammals, humans included. It seems that positive cooperativity makes enzymes much more sensitive to plasma progesterone concentration, and their activities can undergo significant changes in a narrow range of concentration as characteristic of sigmoidal behavior. Therefore, the values of classical pharmacokinetic parameters, such as the elimination constant, half-life, and clearance rate, were found to be highly dependent on the plasma progesterone concentration.     

Effects of heat stress on energetic metabolism in lactating Holstein cows

Heat stress has an enormous economic impact on the global dairy industry, but the mechanisms by which hyperthermia negatively affect systemic physiology and milk synthesis are not clear. Study objectives were to evaluate production parameters and metabolic variables in lactating dairy cows during short-term heat stress or pair-fed conditions coupled with bST administration. Twenty-two multiparous Holstein cows were subjected to 3 experimental periods: 1) thermoneutral conditions with ad libitum intake for 7 d (P1); 2) heat stress (HS) with ad libitum intake (n = 10) or pair-fed (PF) in thermoneutral conditions (n = 12) for 7 d (P2), and 3) 7 d of HS or PF in conditions as described in P2 with recombinant bovine somatotropin administered on d 1 (P3). All cows received an intravenous glucose tolerance test (GTT) on d 5 of each period. Heat stress conditions were cyclical and temperatures ranged from 29.4 to 38.9°C. Rectal temperatures and respiration rates increased during heat stress (38.6-40.4°C and 44-89 breaths/min, respectively). Heat stress reduced dry matter intake by 30% and by design PF cows had similar intake reductions (28%). During heat stress and pair-feeding, milk yield decreased by 27.6% (9.6 kg) and 13.9% (4.8 kg), respectively, indicating that reduced feed intake accounted for only 50% of the decreased milk production. Milk yield increased with recombinant bovine somatotropin in both HS (9.7%) and PF (16.1%) cows. Cows in both groups were in positive energy balance (3.95 Mcal/d) during P1 but entered negative energy balance during P2 and P3 (−5.65 Mcal/d). Heat stress and pair-feeding treatments decreased (9.3%) basal glucose concentrations. Heat stress conditions had no effect on basal NEFA levels during P2; however, PF cows (despite a similar calculated energy balance) had a 2-fold increase in basal NEFA concentrations. Both groups had increased plasma urea nitrogen levels during P2 and P3 compared with P1. Basal insulin levels increased (37%) during P2 and P3 in HS cows but did not differ between periods in PF cows. During P2 and compared with P1, PF cows had a decreased rate of glucose disposal, whereas HS cows had a similar disposal rate following the GTT. During P2 and compared with P1, PF cows had a reduced insulin response whereas HS cows had a similar insulin response to the GTT. In summary, reduced nutrient intake accounted for only 50% of heat stress-induced decreases in milk yield, and feed intake-independent shifts in postabsorptive glucose and lipid homeostasis may contribute to the additional reduction in milk yield.     

Effect of glutamine supplementation on splanchnic metabolism in lactating dairy cows

The suggestion that glutamine (Gln) might become conditionally essential postpartum in dairy cows has been examined through increased postruminal supply of Gln. Net nutrient flux through the splanchnic tissues and mammary gland was measured in 7 multiparous Holstein cows receiving abomasal infusions of water or 300 g/d of Gln for 21 d in a crossover design. Milk yield increased significantly (by 3%) in response to Gln supplementation, but the 2.4% increase in milk protein yield was not statistically significant. Glutamine treatment had no effect on portal or hepatic venous blood flows. Net portal appearance of Gln and Glu was increased by Gln supplementation, accounting for 83% of the infused dose with, therefore, only limited amounts available to provide additional energy to fuel metabolism of the portal-drained viscera. The extra net portal appearance of Gln was offset, however, by a corresponding increase in hepatic removal such that net Gln splanchnic release was not different between treatments. Nonetheless, the Gln treatment resulted in a 43% increase in plasma Gln concentration. Infusions of Gln did not affect splanchnic flux of other nonessential amino acids or of essential amino acids. Glutamine supplementation increased plasma urea-N concentration and tended to increase net hepatic urea flux, with a numerical increase in liver hepatic O₂ consumption. There were no effects on glucose in terms of plasma concentration, net portal appearance, net liver release, or postliver supply, suggesting that Gln supplementation had no sparing effect on glucose metabolism. Furthermore, mammary uptake of glucose and amino acids, including Gln, was not affected by Gln supplementation. In conclusion, this study did not support the hypothesis that supplemental Gln would reduce glucose utilization across the gut or increase liver gluconeogenesis or mammary glutamine uptake to increase milk protein synthesis.