Effects of sodium chloride intake on urea-N recycling and renal urea-N kinetics in lactating Holstein cows.

The effects of high (2.5% of DM) versus normal dietary sodium chloride (NaCl) intake on renal urea-N kinetics and urea-N metabolism were investigated in 9 rumen-cannulated and multi-catheterized lactating dairy cows in a crossover design with 21-d periods. It was hypothesized that urinary urea-N excretion would be greater, and blood urea-N concentration lower in response to greater diuresis induced by high NaCl intake. Also, urea-N transport across ruminal and portal drained viscera (PDV) tissues was hypothesized to be affected by dietary sodium intake. A second experiment was conducted using 8 lactating cows in a crossover design with 14-d periods to test high NaCl (2.5% of DM) versus high KCl (3.2% of DM) intake on milk yield and milk urea-N concentrations. Experiment 1 showed that despite greater diuresis there was no effect of high NaCl intake on urinary urea-N excretion or blood urea-N concentration. The high NaCl intake did not affect rumen ammonia concentrations, total rumen VFA concentrations, ruminal venous - arterial concentration differences for ammonia, or ammonia absorption indicating that high NaCl did not adversely affect ruminal fermentation and microbial protein synthesis. High NaCl intake did not affect the total amount of urea-N transport from blood to gut, but ruminal venous - arterial concentration differences for urea-N were lower with high NaCl and ruminal extraction of arterial urea-N was numerically smaller, indicating that the ruminal epithelial urea-N transport was lower with high NaCl. Energy corrected milk yield was greater with high NaCl (3.2 ± 1.5 kg/d); however, milk urea-N concentrations were not affected by treatment. In experiment 2, ECM was greater with NaCl (1.4 ± 0.31 kg/d) compared with KCl (30.2 and 28.8 ± 0.91 kg ECM / d, respectively). Milk urea-N concentration was lower with KCl, suggesting a urea-N lowering effect in milk not evident with high NaCl intake. In conclusion, the present data show that dietary Na intake of 12-13 g/kg DM was followed by greater diuresis but did not impact urea-N excretion or blood urea-N concentration. High NaCl intake did not affect the total amount of urea-N transfer across PDV tissues. Energy corrected milk yield was greater with high NaCl compared with both control and feeding KCl, however, with KCl milk urea-N decreased.

Pharmacokinetics of α-tocopherol stereoisomers in plasma and milk of cows following a single dose injection of all-rac-α-tocopheryl acetate.

Among different stereoisomers of all-rac-α-tocopherol, 2R-stereoisomers have higher biological activities than their 2S-stereoisomers. Two experiments were conducted to study the pharmacokinetics of stereoisomers of all-rac-α-tocopherol and investigated the discrimination and distribution of α-tocopherol stereoisomers in plasma and milk as well as quantitative secretion into milk with lactating Holstein cows after a single dose intramuscular injection of 2.50 g of all-rac-α-tocopheryl acetate. The highest half-life (2.92/h) and lowest elimination rate (0.36/h) were found for RRR-α-tocopherol. After a single dose injection of all-rac-α-tocopherol, highest maximal daily increase (Smax, 8.36 mg/day) and accumulation secretion (AS, 50.8 mg) were observed for milk RRR-α-tocopherol. The majority of the Æ©2S stereoisomers were found in the liver 36 h post injection, while the 2R stereoisomers were more equal distributed between liver and plasma. The present findings showed a clear discrimination between RRR-α-tocopherol, synthetic 2R stereoisomers and Æ©2S stereoisomers in milk and plasma of dairy cows.

Helicobacter pylori infection is not associated with rheumatoid arthritis.

OBJECTIVE: Rheumatoid arthritis (RA) is an autoinflammatory disease caused by genetic susceptibility and environmental triggers, which include infectious agents. Helicobacter pylori, a bacterium that frequently colonizes the stomach, is associated with the development of certain autoinflammatory disorders. This study examined a possible association between H. pylori infection and RA. METHOD: This cohort study was performed in the Central Denmark Region. Patients were enrolled from primary healthcare centres after a urea breath test (UBT) for H. pylori and followed for a median of 8 years. Nationwide administrative registries provided information about the patients' diagnoses, country of birth, and gender. Comorbidity was determined using the Charlson Comorbidity Index. We compared the prevalence of RA via odds ratios (ORs) and incidences using Cox regression to calculate the hazard ratios (HRs) by comparing H. pylori-positive and H. pylori-negative individuals and adjusting for confounding variables. RESULTS: A total of 56 000 people diagnosed as H. pylori positive or negative had similar rates of comorbidity. No link was found between H. pylori and RA. There was no difference in RA prevalence until time of UBT [OR = 0.91, 95% confidence interval (CI) 0.70-1.19)] or incidence of new RA cases after UBT (HR = 0.80, 95% CI 0.56-1.13) between H. pylori-positive and -negative subjects. Validation via four other RA definitions provided similar results. CONCLUSION: This study found no association between H. pylori infection and RA. This result does not support the involvement of H. pylori in a gut-joint axis of importance for RA development.

Preoperative antithrombotic therapy and risk of blood transfusion and mortality following hip fracture surgery: a Danish nationwide cohort study.

Hip fracture surgery is associated with high risk of bleeding and mortality. The patients often have cardiovascular comorbidity, which requires antithrombotic treatment. This study found that preoperative use of oral anticoagulants was not associated with transfusion or mortality following hip fracture surgery, whereas increased risk may exist for antiplatelet drugs. INTRODUCTION: Hip fracture surgery is associated with high bleeding risk and mortality; however, data on operative outcomes of hip fracture patients admitted while on antithrombotic therapy is sparse. We examined if preoperative antithrombotic treatment was associated with increased use of blood transfusion and 30-day mortality following hip fracture surgery. METHODS: Using data from the Danish Multidisciplinary Hip Fracture Registry, we identified 74,791 hip fracture surgery patients aged ≥ 65 years during 2005-2016. Exposure was treatment with non-vitamin K antagonist oral anticoagulant (NOAC), vitamin K antagonists (VKA), or antiplatelet drugs at admission for hip fracture. Outcome was blood transfusion within 7 days postsurgery and death within 30 days. RESULTS: A 45.3% of patients received blood transfusion and 10.6% died. Current NOAC use was associated with slightly increased risk of transfusion (adjusted relative risk (aRR) 1.07, 95% confidence interval (CI) 1.01-1.14), but similar mortality risk (adjusted hazard ratio (aHR) 0.88, 95% CI 0.75-1.03) compared with non-users. The pattern remained when restricting to patients with short surgical delay (< 24 h). VKA users did not have increased risk of transfusion or mortality. The risks of transfusion (aRR 1.15 95% CI 1.12-1.18) and 30-day mortality (aHR 1.18 95% CI 1.14-1.23) were increased among antiplatelet users compared with non-users. CONCLUSIONS: In an observational setting, neither preoperative NOAC nor VKA treatments were associated with increased risk of 30-day postoperative mortality among hip fracture patients. NOAC was associated with slightly increased risk of transfusion. Preoperative use of antiplatelet drugs was associated with increased risk of transfusion and mortality.

Factors Affecting the Absorption of Subcutaneously Administered Insulin: Effect on Variability.

Variability in the effect of subcutaneously administered insulin represents a major challenge in insulin therapy where precise dosing is required in order to achieve targeted glucose levels. Since this variability is largely influenced by the absorption of insulin, a deeper understanding of the factors affecting the absorption of insulin from the subcutaneous tissue is necessary in order to improve glycaemic control and the long-term prognosis in people with diabetes. These factors can be related to either the insulin preparation, the injection site/patient, or the injection technique. This review highlights the factors affecting insulin absorption with special attention on the physiological factors at the injection site. In addition, it also provides a detailed description of the insulin absorption process and the various modifications to this process that have been utilized by the different insulin preparations available.

Effect of experimentally increased protein supply to postpartum dairy cows on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis.

The effect of experimentally increasing the postpartum protein supply on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis was studied using 8 periparturient Holstein cows in a complete randomized design. At calving, cows were assigned to abomasal infusion of water (CTRL) or casein (CAS) in addition to a lactation diet. Casein infusion was gradually decreased from 696 ± 1 g/d at +2 d relative to calving (DRTC) to 212 ± 10 g/d at +29 DRTC to avoid excessive supply. Synthesis rate of plasma proteins was measured at -14, +4, +15, and +29 DRTC by measuring [C]Phe isotopic enrichment in arterial plasma free Phe, total plasma proteins, and albumin after 3, 5, and 7 h of jugular ring[C]Phe infusion. Plasma volume was determined at +4 and +29 DRTC by dilution of a [I]BSA dose. Synthesis rate of tissue protein in biopsied rumen papillae was determined by measuring [C]Phe isotopic enrichment, and mRNA expression of selected genes was measured by real-time qPCR. Total and differential leukocyte counts were performed and immune responsiveness of monocytes was evaluated by tumor necrosis factor ɑ (TNFɑ) concentration on ex vivo whole blood stimulation with Escherichia coli lipopolysaccharide (LPS) and responsiveness of T-lymphocytes by interferon γ (IFNγ) concentration on stimulation with Staphylococcus aureus enterotoxin ß (SEB). Further, ELISA plasma concentrations of IgM, IgA, and IgG were determined. The DRTC affected the majority of investigated parameters as expected. The CAS treatment increased milk protein yield (P = 0.04), and tended to lower TNFɑ (P = 0.06), and lowered IFNγ (P = 0.03) responsiveness per monocyte and lymphocyte, respectively, compared with CTRL. Further, fractional synthesis rate of albumin was greater at +4 DRTC for CAS compared with CTRL but did not differ by +29 DRTC (interaction: P = 0.01). In rumen papillae, synthesis rate of tissue protein was greater for CAS compared with CTRL (P < 0.01) and mRNA expression of genes for cell proliferation tended to be or were greater for CAS compared with CTRL (P ≤ 0.07). In conclusion, increased postpartum protein supply seem to enhance vital body functions as interpreted from increased liver synthesis of albumin, increased rumen papillae proliferation, and stabilized the ex vivo inflammatory responsiveness of leukocytes. Further studies are needed to enlighten the importance of increased postpartum protein supply in periparturient cows.

PharmML in Action: an Interoperable Language for Modeling and Simulation.

PharmML is an XML-based exchange format created with a focus on nonlinear mixed-effect (NLME) models used in pharmacometrics, but providing a very general framework that also allows describing mathematical and statistical models such as single-subject or nonlinear and multivariate regression models. This tutorial provides an overview of the structure of this language, brief suggestions on how to work with it, and use cases demonstrating its power and flexibility.

Body mass index, risk of allogeneic red blood cell transfusion, and mortality in elderly patients undergoing hip fracture surgery.

UNLABELLED: Despite improvements in preoperative and postoperative treatment, hip fracture surgery may lead to blood transfusion. Little is known about the impact of body mass index on transfusion risk and subsequent mortality. Opposite overweight and obese patients, underweight patients had increased risk of transfusion and death within 1 year of surgery. INTRODUCTION: Despite improvements in preoperative and postoperative treatment of hip fracture patients, hip fracture surgery may lead to blood loss. We examined the risk of red blood cell transfusion (as an indirect measure of blood loss) and subsequent mortality by body mass index level in patients aged 65 and over undergoing hip fracture surgery. METHODS: This is a population-based cohort study using medical databases. We included all patients who underwent surgery for hip fracture during 2005-2013. We calculated the cumulative risk of red blood cell transfusion within 7 days of surgery treating death as a competing risk and, among transfused patients, short- (8-30 days postsurgery) and long-term mortality (31-365 days postsurgery). RESULTS: Among 56,420 patients, 47.7 % received at least one red blood cell transfusion within 7 days of surgery. In patients with normal weight, the risk was 48.8 % compared with 57.0 % in underweight patients (adjusted RR = 1.11; CI 1.08-1.15), 42.1 % in overweight patients (adjusted RR = 0.89; CI 0.86-0.91), and 42.2 % in obese patients (adjusted RR = 0.87; CI 0.84-0.91). Among transfused patients, adjusted HRs for short-term mortality were 1.52 (CI 1.34-1.71), 0.70 (CI 0.61-0.80), and 0.58 (CI 0.43-0.77) for underweight, overweight, and obese patients, respectively, compared with normal-weight patients. The corresponding adjusted HRs for long-term mortality were 1.45 (CI 1.33-1.57), 0.80 (CI 0.74-0.86), and 0.58 (CI 0.50-0.69). Similar association between BMI and mortality was observed also among non-transfused patients. CONCLUSIONS: Underweight patients had a higher risk of red blood cell transfusion and death in the first year of surgery than normal-weight patients, even when controlling for age and comorbidity. Opposite findings were seen for overweight and obese patients.

Abomasal amino acid infusion in postpartum dairy cows: Effect on whole-body, splanchnic, and mammary amino acid metabolism.

Nine Holstein cows with rumen cannulas and indwelling catheters in splanchnic blood vessels were used in a generalized randomized incomplete block design with repeated measures to study the effect of increased early postpartum AA supply on splanchnic and mammary AA metabolism. At calving, cows were blocked according to parity (second and third or greater) and allocated to 2 treatments: abomasal infusion of water (CTRL; n=4) or free AA with casein profile (AA-CN; n=5) in addition to a basal diet. The AA-CN infusion started with half of the maximal dose at the calving day (1 d in milk; DIM) and then steadily decreased from 791 to 226 g/d until 29 DIM. On 5, 15, and 29 DIM, 6 sample sets of arterial, portal, hepatic, and mammary blood were taken at 45-min intervals. Over the whole period, increasing AA supply increased milk (+7.8 ± 1.3 kg/d) and milk protein yields (+220 ± 65 g/d) substantially. The increased milk yield was not supported by greater dry matter intake (DMI) as, overall, DMI decreased with AA-CN (-1.6 ± 0.6 kg/d). Arterial concentrations of essential AA were greater for AA-CN compared with CTRL. The net portal-drained viscera (PDV) release of His, Met, and Phe was greater for AA-CN compared with CTRL, and the net PDV recovery of these infused AA ranged from 72 to 102% once changes in DMI were accounted for. The hepatic removal of these AA was increased equivalently to the increased net PDV release, resulting in an unaltered net splanchnic release. The net PDV release of Ile, Leu, Val, and Lys tended to be greater for AA-CN, and the net PDV recovery of these infused AA ranged from 69 to 73%, indicating increased PDV metabolism with AA-CN. The fractional hepatic removal of these AA did not differ from zero and was unaffected by the increased supply. Consequently, the splanchnic release of these AA was approximately equivalent to their net PDV release for both CTRL and AA-CN. Overall, greater early postpartum AA supply increased milk and milk protein yields substantially based on increased mammary AA uptake. The PDV metabolism of branched-chain AA and Lys were increased, whereas it seemed to be unaffected for other essential AA when the intestinal AA supply was increased. On a net basis, the liver removed more group 1 AA (His, Met, Phe, and Trp) for anabolism and catabolism when the early postpartum AA supply was increased. Thus, increasing the postpartum AA supply increased splanchnic and mammary consumption of AA; hence, the protein deficiency persisted.

The Effects of a GLP-1 Analog on Glucose Homeostasis in Type 2 Diabetes Mellitus Quantified by an Integrated Glucose Insulin Model.

In recent years, several glucagon-like peptide-1 (GLP-1)-based therapies for the treatment of type 2 diabetes mellitus (T2DM) have been developed. The aim of this work was to extend the semimechanistic integrated glucose-insulin model to include the effects of a GLP-1 analog on glucose homeostasis in T2DM patients. Data from two trials comparing the effect of steady-state liraglutide vs. placebo on the responses of postprandial glucose and insulin in T2DM patients were used for model development. The effect of liraglutide was incorporated in the model by including a stimulatory effect on insulin secretion. Furthermore, for one of the trials an inhibitory effect on glucose absorption was included to account for a delay in gastric emptying. As other GLP-1 receptor agonists have similar modes of action, it is believed that the model can also be used to describe the effect of other receptor agonists on glucose homeostasis.

Pharmacometrics Markup Language (PharmML): Opening New Perspectives for Model Exchange in Drug Development.

The lack of a common exchange format for mathematical models in pharmacometrics has been a long-standing problem. Such a format has the potential to increase productivity and analysis quality, simplify the handling of complex workflows, ensure reproducibility of research, and facilitate the reuse of existing model resources. Pharmacometrics Markup Language (PharmML), currently under development by the Drug Disease Model Resources (DDMoRe) consortium, is intended to become an exchange standard in pharmacometrics by providing means to encode models, trial designs, and modeling steps.

Mechanistic model to predict colostrum intake based on deuterium oxide dilution technique data and impact of gestation and prefarrowing diets on piglet intake and sow yield of colostrum.

The aims of the present study were to quantify colostrum intake (CI) of piglets using the D2O dilution technique, to develop a mechanistic model to predict CI, to compare these data with CI predicted by a previous empirical predictive model developed for bottle-fed piglets, and to study how composition of diets fed to gestating sows affected piglet CI, sow colostrum yield (CY), and colostrum composition. In total, 240 piglets from 40 litters were enriched with D2O. The CI measured by D2O from birth until 24 h after the birth of first-born piglet was on average 443 g (SD 151). Based on measured CI, a mechanistic model to predict CI was developed using piglet characteristics (24-h weight gain [WG; g], BW at birth [BWB; kg], and duration of CI [D; min]: CI, g=-106+2.26 WG+200 BWB+0.111 D-1,414 WG/D+0.0182 WG/BWB (R2=0.944). This model was used to predict the CI for all colostrum suckling piglets within the 40 litters (n=500, mean=437 g, SD=153 g) and was compared with the CI predicted by a previous empirical predictive model (mean=305 g, SD=140 g). The previous empirical model underestimated the CI by 30% compared with that obtained by the new mechanistic model. The sows were fed 1 of 4 gestation diets (n=10 per diet) based on different fiber sources (low fiber [17%] or potato pulp, pectin residue, or sugarbeet pulp [32 to 40%]) from mating until d 108 of gestation. From d 108 of gestation until parturition, sows were fed 1 of 5 prefarrowing diets (n=8 per diet) varying in supplemented fat (3% animal fat, 8% coconut oil, 8% sunflower oil, 8% fish oil, or 4% fish oil+4% octanoic acid). Sows fed diets with pectin residue or sugarbeet pulp during gestation produced colostrum with lower protein, fat, DM, and energy concentrations and higher lactose concentrations, and their piglets had greater CI as compared with sows fed potato pulp or the low-fiber diet (P<0.05), and sows fed pectin residue had a greater CY than potato pulp-fed sows (P<0.05). Prefarrowing diets affected neither CI nor CY, but the prefarrowing diet with coconut oil decreased lactose and increased DM concentrations of colostrum compared with other prefarrowing diets (P<0.05). In conclusion, the new mechanistic predictive model for CI suggests that the previous empirical predictive model underestimates CI of sow-reared piglets by 30%. It was also concluded that nutrition of sows during gestation affected CY and colostrum composition.

Abomasal protein infusion in postpartum transition dairy cows: effect on performance and mammary metabolism.

The effect of increasing the postpartum metabolizable protein (MP) supply on performance and mammary metabolism was studied using 8 Holstein cows in a complete randomized design. At parturition, cows were assigned to abomasal infusion of water (CTRL) or casein (CAS). Arterial and epigastric venous blood samples were taken 14 d before expected parturition and at 4, 15, and 29 d in milk (DIM). To compensate previously estimated deficiency of essential AA and to avoid oversupply, casein protein infusion was graduated with 696±1, 490±9, and 212±10g/d at 4, 15 and 29 DIM, respectively. Dry matter intake was unaffected by CAS. Compared with CTRL, MP supply was greater at 4 DIM with CAS but did not differ by 29 DIM. Milk yield was greater with CAS (+7.2±1.3kg/d from 1 to 29 DIM). Milk protein yield was greater with CAS at 4 DIM and averaged 1,664±39g/d compared with 1,212±86g/d for CTRL, but did not differ at 29 DIM (1,383±48g/d). The ratio of MP total supply to requirement was numerically greater at 4 DIM for CAS compared with CTRL, indicating less postpartum protein deficiency. In contrast, a greater net energy deficiency tended to be induced with CAS, but the greater milk yield allowed a large part of mobilized fat to be secreted in milk. Arterial concentration of total essential AA increased sharply after parturition for CAS compared with slight decreases for CTRL. The patterns of arterial concentrations combined with arterial-mammary venous concentration differences indicated that Lys, Leu, and Tyr were the first-limiting AA at 4 DIM with CTRL. Mammary plasma flow was unaffected by treatment, indicating similar perfusion of mammary tissue. The greater milk yield with CAS was associated with greater mammary uptake of individual essential AA, tendencies to greater uptake of glucose, lactate, and ß-hydroxybutyrate, whereas uptakes of volatile fatty acids were unaffected. Despite similar MP supply by 29 DIM, milk and lactose yields were greater with CAS indicating a persistent response to increased postpartum MP supply. In conclusion, the postpartum MP deficiency can have a substantial negative effect in dairy cows as the major outcome of increasing the postpartum MP supply was increased milk, milk protein, and lactose yield, as well as an enhanced MP balance. Potential positive effects for other body functions than milk synthesis are discussed. Future investigations are needed to delineate how to transfer the effect into practical feeding strategies.

Methods for Predicting Diabetes Phase III Efficacy Outcome From Early Data: Superior Performance Obtained Using Longitudinal Approaches.

The link between glucose and HbA1c at steady state has previously been described using steady-state or longitudinal relationships. We evaluated five published methods for prediction of HbA1c after 26/28 weeks using data from four clinical trials. Methods (1) and (2): steady-state regression of HbA1c on fasting plasma glucose and mean plasma glucose, respectively, (3) an indirect response model of fasting plasma glucose effects on HbA1c, (4) model of glycosylation of red blood cells, and (5) coupled indirect response model for mean plasma glucose and HbA1c. Absolute mean prediction errors were 0.61, 0.38, 0.55, 0.37, and 0.15% points, respectively, for Methods 1 through 5. This indicates that predictions improved by using mean plasma glucose instead of fasting plasma glucose, by inclusion of longitudinal glucose data and further by inclusion of longitudinal HbA1c data until 12 weeks. For prediction of trial outcome, the longitudinal models based on mean plasma glucose (Methods 4 and 5) had substantially better performance compared with the other methods.

Impact of silage additives on aerobic stability and characteristics of high-moisture maize during exposure to air, and on fermented liquid feed.

AIMS: To (i) measure the aerobic stability- and describe the characteristics, during aeration, of high-moisture maize (HMM) treated with various additives, and (ii) describe the microbial characteristics of fermented liquid feed (FLF) added HMM. METHODS AND RESULTS: Four treatments were prepared with each of three HMM samples: (i) The HMM as is (CONTROL); and the control added (ii) acids (ACID); (iii) heterofermentative lactic acid bacteria (HETERO); or (iv) homofermentative lactic acid bacteria (HOMO). After ensiling, aerobic stability was measured (Aim 1) and FLF prepared (Aim 2). The ACID treatment improved the aerobic stability of samples 1 and 3 from 9 to 14 h in the CONTROL to 67-115 h. All additives improved aerobic stability of sample 3 from 32 h in the CONTROL to 104-168 h. No proliferation of Enterobacteriacaea was detected during incubation of FLF. CONCLUSION: The microbial profile during aeration- and impact of additives on the aerobic stability of HMM depended on the characteristics of the samples. No blooming of Enterobacteriaceae was observed in FLF containing c. 20 g HMM 100 g(-1) . SIGNIFICANCE AND IMPACT OF THE STUDY: The impact of silage additives on aerobic stability of HMM should be tested in samples with varying characteristics. Inclusion of HMM could be a way of improving biosafety of FLF.

Longitudinal Modeling of the Relationship Between Mean Plasma Glucose and HbA1c Following Antidiabetic Treatments.

Late-phase clinical trials within diabetes generally have a duration of 12-24 weeks, where 12 weeks may be too short to reach steady-state glycated hemoglobin (HbA1c). The main determinant for HbA1c is blood glucose, which reaches steady state much sooner. In spite of this, few publications have used individual data to assess the time course of both glucose and HbA1c, for predicting HbA1c. In this paper, we present an approach for predicting HbA1c at end-of-trial (24-28 weeks) using glucose and HbA1c measurements up to 12 weeks. The approach was evaluated using data from 4 trials covering 12 treatment arms (oral antidiabetic drug, glucagon-like peptide-1, and insulin treatment) with measurements at 24-28 weeks to evaluate predictions vs. observations. HbA1c percentage was predicted for each arm at end-of-trial with a mean prediction error of 0.14% [0.01;0.24]. Furthermore, end points in terms of HbA1c reductions relative to comparator were accurately predicted. The proposed model provides a good basis to optimize late-stage clinical development within diabetes.CPT: Pharmacometrics & Systems Pharmacology (2013) 2, e82; doi:10.1038/psp.2013.58; advance online publication 30 October 2013.

Technical note: A method for quantification of saliva secretion and salivary flux of metabolites in dairy cows.

Salivary flow and net jugular flux of metabolites were studied during resting and rumination in 3 lactating dairy cows (BW 548 ± 17.2 kg, days in milk 113 ± 4 d). The method was based on the concentration difference between arterial and jugular blood, and jugular blood flow measured by downstream dilution of p-aminohippuric acid (pAH). Cows were surgically prepared with a permanent arterial catheter in A. intercostales dorsales before the trial. On sampling days, cows were prepared with left and right side jugular, and ear vein catheters for blood sampling and infusion of pAH, respectively. Blood was sampled simultaneously from the 2 jugular veins and artery during periods of rest and rumination. Secretion of saliva was set equal to the net water extraction calculated from the increased hemoglobin concentration in jugular blood compared with arterial blood. Arterial and jugular blood flow summed for both sides of the head doubled (P < 0.001) during rumination (437 ± 19, 424 ± 18 L/h, respectively) compared with resting (210 ± 19, 202 ± 18 L/h, respectively), consequently doubling the saliva secretion (P < 0.001, resting = 7.6 ± 0.8 L/h, rumination = 13.8 ± 0.8 L/h). The extraction of inorganic phosphate (Pi) from arterial blood during resting periods was greater compared with rumination (P = 0.004; resting = 21.7% ± 0.9%; rumination = 15.6% ± 0.9%), resulting in a greater Pi concentration in saliva secreted during resting. The concentrations of Pi in saliva were 4.5 ± 0.3 and 3.7 ± 0.3 times the arterial concentration during resting and rumination (P = 0.09), respectively. The urea concentration in saliva was 0.63 ± 0.04 times the arterial level, showing that urea is less efficiently transferred from blood than water, resulting in a greater numerical urea concentration in jugular compared with arterial blood. The water extraction method presented in the present paper offers an alternative way of estimating saliva secretion without the chewing activity constraints associated with other methods, for example, allowing for determination of saliva flow during rumination.

Ergot alkaloids from endophyte-infected tall fescue decrease reticuloruminal epithelial blood flow and volatile fatty acid absorption from the washed reticulorumen.

An experiment was conducted to determine if ergot alkaloids affect blood flow to the absorptive surface of the rumen. Steers (n=8) were pair-fed alfalfa cubes and received ground endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum; E+) seed (0.015 mg ergovaline·kg BW(-1)·d(-1)) or endophyte-free tall fescue (E-) seed via the rumen cannula 2x daily for 7 d at thermoneutral (TN; 22°C) and heat stress (HS; 32°C) conditions. On d 8, the rumen was emptied and rinsed. A buffer containing VFA was incubated in the following sequence: control (CON), 15 µg ergovaline·kg BW(-1) (1×EXT) from a tall fescue seed extract, and 45 µg ergovaline·kg BW(-1) (3×EXT). For each buffer treatment there were two 30-min incubations: a 30-min incubation of a treatment buffer with no sampling followed by an incubation of an identical sampling buffer with the addition of Cr-EDTA and deuterium oxide (D2O). Epithelial blood flow was calculated as ruminal clearance of D2O corrected for influx of physiological water and liquid outflow. Feed intake decreased with dosing E+ seed at HS but not at thermoneutral conditions (TN; P<0.02). Dosing E+ seed decreased serum prolactin (P<0.005) at TN. At HS, prolactin decreased in both groups over the 8-d experiment (P<0.0001), but there was no difference in E+ and E- steers (P=0.33). There was a seed treatment×buffer treatment interaction at TN (P=0.038), indicating that E+ seed treatment decreased reticuloruminal epithelial blood flow at TN during the CON incubation, but the two groups of steers were not different during 1×EXT and 3×EXT (P>0.05). Inclusion of the extract in the buffer caused at least a 50% reduction in epithelial blood flow at TN (P=0.004), but there was no difference between 1×EXT and 3×EXT. There was a seed × buffer treatment interaction at HS (P=0.005), indicating that the reduction of blood flow induced by incubating the extract was larger for steers receiving E- seed than E+ seed. Volatile fatty acid flux was reduced during the 1×EXT and 3×EXT treatments (P<0.01). An additional experiment was conducted to determine the effect of time on blood flow and VFA flux because buffer sequence could not be randomized. Time either increased (P=0.05) or did not affect blood flow (P=0.18) or VFA flux (P>0.80), indicating that observed differences are due to the presence of ergot alkaloids in the rumen. A decrease in VFA absorption could contribute to the signs of fescue toxicosis including depressed growth and performance.

Precursors for liver gluconeogenesis in periparturient dairy cows.

The review is based on a compiled data set from studies quantifying liver release of glucose concomitant with uptake of amino acids (AA) and other glucogenic precursors in periparturient dairy cows. It has become dogma that AAs are significant contributors to liver gluconeogenesis in early lactation, presumably accounting for the observed lack of glucogenic precursors to balance estimated glucose need. Until recently, there has been paucity in quantitative data on liver nutrient metabolism in the periparturient period. Propionate is the quantitatively most important glucogenic precursor throughout the periparturient period. However, the immediate post partum increment in liver release of glucose is not followed by an equivalent increment in propionate uptake, because of the lower rate of increment in feed intake compared with the rate of increment in requirements for milk synthesis. The quantitative data on liver metabolism of AA do not support the hypothesis that the rapid post partum increase in net liver release of glucose is supported by increased utilisation of AA for gluconeogenesis. Only alanine is likely to contribute to liver release of glucose through its role in the inter-organ transfer of nitrogen from catabolised AA. AAs seem to be prioritised for anabolic purposes, indicating the relevance of investigating effects of supplying additional protein to post partum dairy cows. Combining data from quantitative and qualitative experimental techniques on L-lactate metabolism point to the conclusion that the quantitatively most important adaptation of metabolism to support the increased glucose demand in the immediate post partum period is endogenous recycling of glucogenic carbon through lactate. This is mediated by a dual site of adaptation of metabolism in the liver and in the peripheral tissues, where the liver affinity for L-lactate is increased and glucose metabolism in peripheral tissues is shifted towards L-lactate formation over complete oxidation.