A model of net amino acid absorption and utilization by the portal-drained viscera of the lactating dairy cow.

A more complete understanding of amino acid (AA) metabolism by the various tissues of the body is required to improve upon current systems for predicting the use of absorbed AA. The objective of this work was to construct and parameterize a model of net removal of AA by the portal-drained viscera (PDV). Six cows were prepared with arterial, portal, and hepatic catheters and infused abomasally with 0, 200, 400, or 600 g of casein daily. Casein infusion increased milk yield quadratically and tended to increase milk protein yield quadratically. Arterial concentrations of a number of essential AA increased linearly with respect to infusion amount. When infused casein was assumed to have a true digestion coefficient of 0.95, the minimum likely true digestion coefficient for noninfused duodenal protein was found to be 0.80. Net PDV use of AA appeared to be linearly related to total supply (arterial plus absorption), and extraction percentages ranged from 0.5 to 7.25% for essential AA. Prediction errors for portal vein AA concentrations ranged from 4 to 9% of the observed mean concentrations. Removal of AA by PDV represented approximately 33% of total postabsorptive catabolic use, including use during absorption but excluding use for milk protein synthesis, and was apparently adequate to support endogenous N losses in feces of 18.4 g/d. As 69% of this use was from arterial blood, increased PDV catabolism of AA in part represents increased absorption of AA in excess of amounts required by other body tissues. Based on the present model, increased anabolic use of AA in the mammary and other tissues would reduce the catabolic use of AA by the PDV.

Rates of production of acetate, propionate, and butyrate in the rumen of lactating dairy cows given normal and low-roughage diets.

Five lactating dairy cows with a permanent cannula in the rumen were given (kg DM/d) a normal diet (7.8 concentrates, 5.1 hay) or a low-roughage (LR) diet (11.5 concentrates, 1.2 hay) in two meals daily in a two-period crossover design. Milk fat (g/kg) was severely reduced on diet LR. To measure rates of production of individual volatile fatty acids (VFA) in the rumen, 0.5 mCi 1-(14)C-acetic acid, 2-(14)C-propionic acid, or 1-(14)C-n-butyric acid were infused into the rumen for 22 h at intervals of 2 to 6 d; rumen samples were taken over the last 12 h. To measure rumen volume, we infused Cr-EDTA into the rumen continuously, and polyethylene glycol was injected 2 h before the morning feed. Results were very variable, so volumes measured by rumen emptying were used instead. Net production of propionic acid more than doubled on LR, but acetate and butyrate production was only numerically lower. Net production rates pooled across both diets were significantly related to concentrations for each VFA. Molar proportions of net production were only slightly higher than molar proportions of concentrations for acetate and propionate but were lower for butyrate. The net energy value (MJ/d) of production of the three VFA increased from 89.5 on normal to 109.1 on LR, equivalent to 55 and 64% of digestible energy, respectively. Fully interchanging, three-pool models of VFA C fluxes are presented. It is concluded that net production rates of VFA can be measured in non-steady states without the need to measure rumen volumes.

Effect of method of application of a fibrolytic enzyme product on digestive processes and milk production in Holstein-Friesian cows.

Four multiparous cows with cannulas in the rumen and proximal duodenum were used in early lactation in a 4 x 4 Latin square experiment to investigate the effect of method of application of a fibrolytic enzyme product on digestive processes and milk production. The cows were given ad libitum a total mixed ration (TMR) composed of 57% (dry matter basis) forage (3:1 corn silage:grass silage) and 43% concentrates. The TMR contained (g/kg dry matter): 274 neutral detergent fiber, 295 starch, 180 crude protein. Treatments were TMR alone or TMR with the enzyme product added (2 kg/1000 kg TMR dry matter) either sprayed on the TMR 1 h before the morning feed (TMR-E), sprayed only on the concentrate the day before feeding (Concs-E), or infused into the rumen for 14 h/d (Rumen-E). There was no significant effect on either feed intake or milk yield but both were highest on TMR-E. Rumen digestibility of dry matter, organic matter, and starch was unaffected by the enzyme. Digestibility of NDF was lowest on TMR-E in the rumen but highest postruminally. Total tract digestibility was highest on TMR-E for dry matter, organic matter, and starch but treatment differences were nonsignificant for neutral detergent fiber. Corn silage stover retention time in the rumen was reduced by all enzyme treatments but postruminal transit time was increased so the decline in total tract retention time with enzymes was not significant. It is suggested that the tendency for enzymes to reduce particle retention time in the rumen may, by reducing the time available for fibrolysis to occur, at least partly explain the variability in the reported responses to enzyme treatment.

Effects of postrumen starch infusion on milk production and energy metabolism in dairy cows.

Two experiments were conducted to determine effects of postrumen starch infusion on milk production and energy and nitrogen utilization in lactating dairy cows. In experiment 1, four cows in early lactation fed grass silage and concentrates were continuously infused into the duodenum with water or 700, 1400, or 2100 g of purified maize starch daily for 10 to 12 d in a 4 x 4 Latin square design with 2-wk periods. Starch infusion increased milk yield linearly and decreased milk fat concentration in a quadratic manner such that increases in fat-corrected milk and calculated milk energy yield were minimal except at the highest rate of infusion. Changes in milk energy output suggest that even at the highest infusion rate metabolizable energy supplied by infused starch was used for tissue energy or oxidized. In experiment 2 energy and nitrogen balance were measured in four cows in late lactation fed a mixture of dehydrated lucerne, grass silage, and concentrates during the last 6 d of 2-wk abomasal infusions of 1200 g of purified wheat starch daily or water in a balanced switchback design with 5-wk periods. Measurements of fecal starch concentration indicated nearly all the starch infused was digested, but decreased fecal pH and apparent nitrogen digestion suggested an increase in hindgut starch fermentation. Starch infusion decreased urine nitrogen output in part because of increased tissue nitrogen retention but had no effect on milk nitrogen output. In absolute terms, numerical decreases in feed energy intake and energy digestion reduced the recovery of starch energy infused as digestible and metabolizable energy, but in terms of changes in total energy supply with starch infusion, 79% was recovered as metabolizable energy. Starch infusion had no effects on heat or milk energy but increased net energy for lactation due to a numerical increase in tissue energy, implying that in late-lactation cows, starch digested postruminally was used with high efficiency for tissue energy retention as protein and fat.

The effect of energy supplementation on nitrogen utilization in lactating dairy cows fed grass silage diets.

An experiment was conducted to examine the effect that various isoenergetic diets, containing different quantities of soluble carbohydrate and fiber and different types of starch, have on nitrogen (N) balances. Six lactating dairy cows in early to midlactation consuming grass silage diets with not less than 600 g/kg total DMI as forage were used in the experiment. Four concentrates were prepared that had higher amounts of either fiber, soluble sugars, corn (low degradable starch source), or barley (high degradable starch source). Overall N utilization by the cows was poor, rarely exceeding 0.30 g milk N/g of dietary N intake. Fecal N outputs accounted for more than half of total N excreted in all treatments except for diets supplemented with high degradable starch, in which urinary N excretion was significantly higher compared with the other treatments. Milk yield was unaffected by concentrate type, averaging 19.9 kg/d, but milk protein content decreased from 32.9 for starch-based diets to 30.9 and 30.0 g/kg for the soluble sugar- and fiber-based diets, respectively. The efficiency of N utilization improved in the low degradable starch treatment, which had lower N excretion (65%) and higher protein concentration in milk. Furthermore, feeding cows corn-based concentrates reduced urinary N excretion by almost 30% compared with barley-based concentrates; therefore, feeding corn-based diets is recommended for the reduction of nitrogen pollution in lactating dairy cows.

The effect of protein supplementation on nitrogen utilization in lactating dairy cows fed grass silage diets.

The study set out to examine the effects of supplementing grass silage with various levels of protein concentration and degradability on dietary nitrogen (N) excretion in lactating dairy cows consuming at least 60% forage. Six Holstein/Friesian cows in early to midlactation were offered six diets comprising two levels of crude protein (210 and 290 g/kg DM) and three levels of protein degradability in the concentrate achieved using different amounts of untreated or formaldehyde-treated soybean meal. Despite a difference of almost 100 g/d in N intake, apparent fecal and milk N outputs were not significantly affected. Protein degradability also had no effect on N outputs in feces and milk. However, there was a major effect of both level and degradability of CP on urinary N output. Moreover, an interaction between level and degradability of CP was detected, such that the rate at which urinary N increases with increasing CP degradability was higher on the high-CP than on the low-CP diet. A low level of protein (150 g/kg DM in the diet) and medium to low rumen-degradable protein supplements provided a significant reduction in N excretion without compromising lactational performance (mean 24.8 kg/d), in terms of both milk yield and composition. This study also demonstrated that a high efficiency of N utilization could be achieved on low-CP diets (supplying less than 400 g N/d), with feces being the main route of N excretion, whereas an exponential excretion of urinary N was observed as N intake exceeded 400 g N/d.

Prediction of milk protein concentration from elements of the metabolizable protein system.

Elements of the metabolizable protein system in the United Kingdom were examined for their suitability as potential predictors of milk protein concentration. Models were based on data from 163 cows offered five forage mixtures for ad libitum intake plus concentrates at 3, 6, or 9 kg/d of dry matter. The models were then tested on a separate data set of 100 cows offered seven forage mixtures for ad libitum intake plus concentrates at 6 kg/d of dry matter. To minimize problems with collinearity, variables were arranged hierarchically; successive elements were components of variables at higher element levels. Variables from different element levels were not used in the same models. Models were constructed using ridge regression to minimize problems with collinearity. The fit and precision of prediction were generally poor because these models did not take into account animal variables. Models using undergradable dietary protein performed slightly better than did those using digestible undegraded protein. The use of slowly degradable protein and quickly degradable protein rather than rumen-degradable protein generally resulted in improvements in prediction. Models using neutral detergent fiber and quickly fermented carbohydrate were better than those using total carbohydrate. We concluded that there was little to be gained from using the elements of the metabolizable protein system considered here for the prediction of milk protein concentration.

Hepatic response to increased exogenous supply of plasma amino acids by infusion into the mesenteric vein of Holstein-Friesian cows in late gestation.

The hepatic responses of late gestation, dry dairy cows to acute (6 h) infusions of an amino acid (AA) mixture (Synthamin; 0.0, 1.1, 2.2, 4.4, 8.8 and 17.6 mumol/min) into the mesenteric vein were determined. Neither blood flow nor O2 consumption across the portal-drained viscera (PDV) and liver was significantly altered by infusion. Similarly, there were no effects on net absorption, or hepatic removal, of acetate, propionate, butyrate or NH3. Glucose PDV appearance was unchanged but hepatic glucose production increased (P = 0.032) by 0.2 mumol/min per mumol/min of AA infused. Additional extraction of alanine, glycine (both infused) and glutamine (not infused) by the liver was sufficient to account for most of the extra C required for glucose synthesis. The N that would be liberated from these glucogenic AA would also account for a large proportion of the increase in urea-N produced in response to the AA infusion. This supports the concept of a correlation between gluconeogenesis and ureagenesis. Furthermore, the amide-N liberated from the extracted glutamine would contribute up to 0.17 of hepatic NH3 flux and assist in balancing N inputs into the carbamoyl phosphate and arginosuccinate entry points of the ornithine cycle. Rates of fractional extraction of the various AA by the liver were best fitted by linear equations, indicating that even at the highest rates of administration (approximately twice maximal physiological absorption) the transport systems were not saturated. Hepatic fractional extractions of infused essential AA were highest for methionine (0.83) and phenylalanine (0.87) with the lowest proportion removed observed for valine (0.25), leucine (0.30), lysine (0.31) and isoleucine (0.49). For the non-essential AA, the highest apparent fractional extractions were for glycine (0.73), arginine (0.79) and tyrosine (0.63) followed by alanine (0.54), proline (0.47) and serine (0.37). Hepatic removal of AA-N exceeded the increase in urea-N formation such that, at the highest rate of infusion, approximately 10 mmol/min of the extracted AA was apparently available for hepatic anabolism, more than is required to account for assumed increases in liver mass and export protein synthesis. Similarly, the amount of AA available for peripheral tissue protein gain, when assessed against phenylalanine supply as the limitation, would be the equivalent of a maximum of 0.5 g protein retained/min (6 mmol AA-N/min). This would provide sufficient AA for replenishment of peripheral (muscle) protein stores plus support of the placenta and fetus.

Effect of intravenous amino acid infusion on leucine oxidation across the mammary gland of the lactating goat.

Changes in the kinetics of leucine in the mammary gland were examined in four lactating goats (25, 38, 45, and 135 DIM) that were given an i.v. infusion of a mixture of 18 AA, not including leucine, to alter the availability of leucine to the gland relative to other AA. Arteriovenous monitoring of [1-13C]leucine kinetics across one-half of the mammary gland was conducted on the last day (d 6 or 7) of the saline (control) and the AA infusion periods. Although blood flow to the mammary gland and the arterial concentration of most AA other than leucine were increased by the AA infusion, milk and protein yields did not change. For goats in early lactation (n = 3), arterial leucine concentrations fell considerably during AA infusion; however, the arteriovenous difference of leucine was maintained, resulting in uncommonly low leucine concentrations in venous plasma (8 microM). Whole body leucine flux (protein synthesis plus oxidation) was unaffected by AA infusion, but, because whole body leucine oxidation was reduced, whole body utilization of leucine for protein synthesis increased. The AA infusion reduced mammary oxidation of leucine to approximately one-third of control values. These results suggest that leucine oxidation can be reduced considerably without affecting milk protein output; thus, leucine oxidation may not be an irrevocable consequence of mammary metabolism. If catabolism of other AA either by the gland or in the whole body can be reduced, then the efficiency of milk yield can be improved.

Responses in milk constituents to intravascular administration of two mixtures of amino acids to dairy cows.

Four Holstein-Friesian cows were used to investigate the effects of intravascular infusions of AA mixtures on milk constituents. Cows were in wk 11 to 28 of lactation and were fed a basal concentrate (142 g of CP/kg of DM) and grass silage (149 g of CP/kg of DM) in a 60:40 ratio (percentage of DM). Cows were fed hourly, and feed intake was fixed at 95% of ad libitum intake for each experimental period. Each cow received a 4-d jugular saline infusion, followed by a 5-d jugular infusion of a mixture of AA. Two mixtures of AA were used in a crossover design. The first mixture contained both the essential AA and non-essential AA found in milk protein (total AA); this mixture was infused at 400 g of AA/d. The other mixture represented the essential AA fraction only and was infused at 208 g/d. Infusion of total AA increased milk protein concentration from 32.4 to 35.0 g/kg, and essential AA increased milk protein concentration from 32.5 to 36.9 g/kg; milk protein yield increased by 87 g/d (total AA) and 143 g/d (essential AA). Intravascular administration of AA specifically stimulated milk protein concentration, and the efficiency with which the AA were used was higher than had been previously reported when AA supply was increased either by dietary supplementation or by abomasal infusion.

The effect of dietary crude protein as protected soybean meal on mammary metabolism in the lactating dairy cow.

Metabolism in the mammary gland was related to changes in milk output in response to changes in dietary protein intake. Three diets of grass silage and concentrate were fed to four lactating dairy cows equipped with intravascular catheters across the mammary gland. Concentrates differed in the inclusion of protected soybean meal and provided 11.3, 15.4, and 20.1% CP, respectively. Blood samples were taken to assess the effect of protein percentage on the nutrient fluxes across the gland and their relationship to milk production. Milk production, milk protein yield, and milk protein concentration were all increased as CP intake increased, although these responses were not linear. Concentrations of urea in milk reflected those in plasma and increased as dietary protein intake increased. Uptake of glucose and BHBA by the mammary gland tended to increase as milk production increased. Arterial supply of essential AA increased as the dietary protein increased. Supply and uptake of nonessential AA were unchanged by dietary treatment, and uptake was insufficient to account for output of nonessential AA residues in milk protein. The supply of essential AA was not limiting for milk protein synthesis, and some alternative mechanism must have existed for the control of milk protein yield.

Kinetics of blood free and milk casein-amino acid labelling in the dairy goat at two stages of lactation.

The kinetics of blood free amino acids (AA) transfer into milk casein were compared in goats (n 4) at 61 (SE 5) d (Expt 1; post-peak, 4.51 (SE 0.26) kg milk/d) and at 180 (SE 6) d (Expt 2; late, 2.36 (SE 0.16) kg milk/d) of lactation during non-primed, continuous (Expt 1, 12 h; Expt 2, 16 h) intravenous infusions of mixtures of L-[1-13C]leucine and L-[1-13C]phenylalanine with either L-[1-13C]valine (Expt 1) or L-[5-13C]methionine (Expt 2). The 13C enrichments of blood free and casein-bound AA were fitted to a single exponential model to estimate isotopic plateaux and the fractional rate constant for milk casein labelling. Milk protein output and its contribution to whole-body flux was higher in Expt 1 (post-peak) than in Expt 2 (late lactation), but the kinetics of 13C labelling of the casein-bound AA were similar for all AA tracers in both experiments. At both stages of lactation the delay (6-8 h) between the attainment of isotopic plateau for the blood free AA and the corresponding attainment of plateau for the casein-bound AA indicated that the blood free pool was not the immediate precursor pool for milk casein biosynthesis. Plateau enrichments of casein-bound AA were generally higher than those for the corresponding blood free AA in both experiments. These results indicate that the relative contributions of different AA sources to the immediate precursor pool for milk casein biosynthesis are similar at different stages of lactation despite major changes in the partitioning of whole-body flux towards milk protein output. Non-milk protein fluxes were also similar in post-peak and late lactation.

The effect of supplementary protein on in vivo metabolism of the mammary gland in lactating dairy cows.

Four lactating cows equipped with rumen and duodenal cannulas were fed a diet of grass silage and concentrates containing either 12.4 or 17.2% CP (DM basis) in a change-over design. Additional protein was supplied as white fish meal. Fish meal did not affect molar proportions of VFA in the rumen, but duodenal NAN supply was increased .69 g/g of N in supplementary feed. In Experiment 2, three lactating dairy cows that had been prepared with catheters across the mammary gland were fed the same diets using a switchback design. Blood samples were taken to determine changes in metabolite flux to the mammary gland. In both experiments, milk production and protein yield were nonsignificantly increased by addition of fish meal. Milk urea output was increased from 3.18 to 4.74 g/d by fish meal supplementation, reflecting increased arterial concentrations of urea. Concentrations of glucose, VFA, and BHBA in blood showed no substantial changes because of dietary supplementation of fish meal. Supply of essential AA increased 26% with fish meal supplementation, mammary uptake increased 34%, but milk protein output only increased 5%. The low efficiency of conversion of supplementary protein to milk protein appears to be related to the inability of the gland to utilize the additional AA.

Impact of a clinical pharmacist on cost saving and cost avoidance in drug therapy in an intensive care unit.

The authors performed a study to document the impact of a clinical pharmacist on cost saving and cost avoidance in an intensive care unit, and to evaluate the cost saving and avoidance to justify additional clinical pharmacist positions. Over 13 consecutive 5-day weeks, a clinical pharmacist with 50% teaching responsibility documented time spent and all interventions that impacted the cost of drug therapy. Both cost avoidance and cost saving were documented on change in route, change in dosage, change to another drug, discontinuation of therapy, discontinuation of therapeutic duplication, discontinuation of inappropriate therapy, notification of pharmacy of discrepancy, and improper drug-level monitoring avoidance. Cost analysis was calculated based on acquisition costs. The final cost saving was derived from adding cost avoided and cost saved minus pharmacist salary for the time spent in conducting the study. A total of 310 interventions were documented during the 13 weeks (65 days) of the study. The final cost saving was $79,723, which would extrapolate to an annual savings of $318,891. Although 31.3% of interventions involved change of dosage, interventions involving change to another drug (13.9%) had the largest economic impact ($62,527). The majority (85.4%) of the savings involved costs of medications saved (actual dollars saved rather than avoided). The authors concluded that the clinical pharmacist had a significant impact on the cost of drug therapy in the intensive care unit and that the cost of additional clinical pharmacist positions should be justified.

Use of granulocyte colony-stimulating factor in the treatment of pancytopenia secondary to colchicine overdose.

OBJECTIVE: To report a case of pancytopenia following colchicine overdose and to discuss the use of granulocyte colony-stimulating factor (G-CSF) for treating this severe complication. CASE SUMMARY: A 19-year-old man developed pancytopenia four days after ingestion of approximately 50-60 0.6-mg colchicine tablets in a suicide attempt. His pancytopenia resolved after subcutaneous administration of one 300-micrograms dose of G-CSF. The patient recovered from his other multiorgan disturbances during his hospitalization and was discharged from the hospital with elevated liver enzyme concentrations. CONCLUSIONS: Colchicine overdose is rare, but can be fatal. The use of G-CSF appears to be beneficial in alleviating bone marrow depression in colchicine overdose situations.

Variations in blood flow to and from the bovine mammary gland measured using transit time ultrasound and dye dilution.

Blood flow across the lactating bovine mammary gland was measured by two techniques. The use of transit time flow probes appeared to give flows which correlated well with dye dilution in only one of five cows, although the relative changes in flow were similar between the techniques in four of the cows. Further studies were made on the effect of posture on mammary blood flow using both techniques. The crossover of venous blood from one side of the mammary gland was also studied using the dye dilution technique, and revealed large differences between animals and also with posture. These observations suggest that particular care should be taken when sampling blood from the milk vein of cows, if a representative sample is required. Changes in blood flow with posture may be indicative of a repartitioning of flow within the body, and the physiology of such a mechanism would be of interest in itself. The control of this mechanism may be useful in modifying blood flow to the mammary gland and thus milk yield, since blood flow is related to the level of milk production.

Rumen fill and digesta kinetics in lactating Friesian cows given two levels of concentrates with two types of grass silage ad lib.

Four lactating Friesian cows with permanent cannulas in the rumen and proximal duodenum were given early (EC)- or late (LC)-cut grass silage ad lib., each with either 3 or 9 kg concentrate dry matter (DM)/d in a 4 x 4 Latin square design starting about 10 weeks after calving. Feed was offered twice daily at 08.30 hours and 15.30 hours. Periods lasted 5 weeks and measurements were made in the last 15 d. The higher amount of concentrates increased total DM intake but reduced silage DM intake and the fractional rate of degradation of silage-fibre DM. Later cutting date of silage had no effect on DM intake or the fractional rate of degradation of silage-fibre DM but reduced potential degradability of silage fibre. Dilution rate of CoEDTA in rumen fluid was greater during the day (eating period) than during the night (resting period). Dilution rates measured at the duodenum were lower than those measured in the rumen, but neither was affected by diet. Silage-particle passage rates were measured by use of ytterbium-labelled silage fibre (Yb-fibre) and chromium-mordanted faecal particles (Cr-faeces) and samples were taken at the duodenum and in the faeces. Values for slower rate constant (k1) and transit time were higher and for faster rate constant (k2) were lower for Yb-fibre than for Cr-faeces, but differences in total mean retention time were inconsistent. Values for k1 for both markers and k2 for Yb-fibre only were similar at both sampling sites, but values for k2 for Cr-faeces were lower in the faeces. No diet effects were established with Yb-fibre but, with Cr-faeces, k1 was reduced by more concentrates and EC-silage. Daily mean weights of wet digesta, liquid, neutral-detergent fibre (NDF) and indigestible NDF in the rumen were greater with LC-silage but were unaffected by the amount of concentrates whereas weight of rumen DM was increased by the amount of concentrates only. Maximum rumen fill occurred at 18.00 hours with all diets. Particle-size distribution of rumen contents did not vary markedly during the day. Mean particle size was generally greater with LC-silage than EC-silage. Very small particles, passing through the 0.3 mm screen, constituted about half the rumen DM. It is concluded that rumen fill could have limited intake of LC-silage but not EC-silage.(ABSTRACT TRUNCATED AT 400 WORDS)

The influence of insulin and amino acid supply on amino acid uptake by the lactating bovine mammary gland.

Four lactating dairy cows received arterial infusions of insulin (1.41 U/h), an AA mixture (threonine, methionine, leucine, phenylalanine, and lysine at 5.87, 1.90, 3.55, 2.17, and 4.21 mmol/h, respectively), and a combination of the two in a 4 x 4 Latin square. The infusions were performed over a 3-d period directly into the extra pudic artery on both sides of the mammary gland, and samples were taken simultaneously of the downstream extra pudic arterial blood and also of subcutaneous abdominal venous blood. Blood flow was measured by dye dilution using p-amino-hippuric acid and was increased by 37% by infusion of insulin plus AA (P less than .05). Infusions of AA tended to increase the arteriovenous difference and uptake of the infused AA (P less than .05 for phenylalanine) and had varying effects on the uninfused AA. Inclusion of insulin in the AA infusion tended to increase uptake of infused AA, whereas infusions of insulin alone tended to decrease uptake. There were no significant effects of infusion on milk yield or composition.