Effects of storage temperature and time on canine plasma ammonia concentrations

Stability of ammonia in canine plasma was determined, with regard to temperature and time of storage. Heparinized venous blood samples were collected from 8 healthy dogs, immediately placed in an ice water bath, and centrifuged at 5 C. Plasma was harvested from the blood samples, and the initial analysis of each sample for plasma ammonia was performed within 30 minutes after collection. Separate aliquots of the plasma from each dog were stored at 21 C, 4 C, -15 C, or -40 C. Ammonia concentrations of the aliquots stored at the various temperatures were determined at 24, 48, and 96 hours after collection. Statistical analysis of the data from each dog did not indicate a significant relationship between the initial concentration of plasma ammonia and subsequent determinations. Correlation or significance was not found among samples stored at similar temperatures and evaluated at similar times.     

Technical note: Effect of determining baseline plasma urea nitrogen concentrations on subsequent posttreatment plasma urea nitrogen concentrations in 20- to 50-kilogram pigs

Plasma urea N (PUN) has been used as an indicator of AA requirements and efficiency of AA utilization in swine. However, PUN concentrations vary among a population of pigs, even a population with a close range of BW and fed the same diet. Thus, pretreatment or baseline PUN concentrations are used as a covariate to reduce variation of posttreatment PUN. However, this procedure increases experimental costs and stress to the pigs. Data from 14 experiments (26 to 28 d in duration) conducted using PUN as a response variable were compiled into 1 data set. Each experiment had 4 to 6 treatments. The purpose of this technical report was to summarize the effect of determining pretreatment baseline PUN concentrations on subsequent posttreatment PUN concentrations in 20- to 50-kg pigs. In all experiments, pigs were fed corn- and soybean meal-based diets and low-CP diets with various AA additions; pigs were assigned to dietary treatments in a randomized complete block design with a minimum of 4 replicates of 3 to 5 pigs each. Before the start of each experiment, all pigs were fed a common diet for a minimum of 3 d. Blood samples were collected from each pig before allotment to dietary treatments (d 0) and at the end of each experiment. The baseline (d 0) PUN was analyzed as a covariate for posttreatment PUN. Data from each experiment were analyzed without and with baseline PUN in the statistical model. In all experiments combined, there were 768 possible treatment comparisons. The covariate baseline PUN was statistically significant (P < 0.10) in 9 of 14 experiments. However, only 8 treatment differences changed statistical significance as a result of analyzing the data with baseline PUN as a covariate. These 8 treatment differences were in 3 experiments. These results indicate that it is not always necessary to determine baseline PUN concentrations when feeding diets with large differences in AA content.     

Effects of season and sample handling on measurement of plasma alpha-melanocyte-stimulating hormone concentrations in horses and ponies

OBJECTIVE: To investigate effects of sample handling, storage, and collection time and season on plasma alpha-melanocyte-stimulating hormone (alpha-MSH) concentration in healthy equids. ANIMALS: 11 healthy Standardbreds and 13 healthy semiferal ponies. PROCEDURE: Plasma alpha-MSH concentration was measured by use of radioimmunoassay. Effects of delayed processing were accessed by comparing alpha-MSH concentrations in plasma immediately separated with that of plasma obtained from blood samples that were stored at 4 degrees C for 8 or 48 hours before plasma was separated. Effects of suboptimal handling were accessed by comparing alpha-MSH concentrations in plasma immediately stored at -80 degrees C with plasma that was stored at 25 degrees C for 24 hours, 4 degrees C for 48 hours or 7 days, and -20 degrees C for 30 days prior to freezing at -80 degrees C. Plasma alpha-MSH concentrations were compared among blood samples collected at 8:00 AM, 12 noon, and 4:00 PM. Plasma alpha-MSH concentrations were compared among blood samples collected in January, March, April, June, September, and November from horses and in September and May from ponies. RESULTS: Storage of blood samples at 4 degrees C for 48 hours before plasma was separated and storage of plasma samples at 4 degrees C for 7 days prior to freezing at -80 degrees C resulted in significant decreases in plasma alpha-MSH concentrations. A significantly greater plasma alpha-MSH concentration was found in September in ponies (11-fold) and horses (2-fold), compared with plasma alpha-MSH concentrations in spring. CONCLUSIONS AND CLINICAL RELEVANCE: Handling and storage conditions minimally affected plasma alpha-MSH concentrations. Seasonal variation in plasma alpha-MSH concentrations must be considered when evaluating pituitary pars intermedia dysfunction in equids.     

Physical characteristics, blood hormone concentrations, and plasma lipid concentrations in obese horses with insulin resistance

OBJECTIVE: To compare obese horses with insulin resistance (IR) with nonobese horses and determine whether blood resting glucose, insulin, leptin, and lipid concentrations differed between groups and were correlated with combined glucose-insulin test (CGIT) results. ANIMALS: 7 obese adult horses with IR (OB-IR group) and 5 nonobese mares. PROCEDURES: Physical measurements were taken, and blood samples were collected after horses had acclimated to the hospital for 3 days. Response to insulin was assessed by use of the CGIT, and maintenance of plasma glucose concentrations greater than the preinjection value for > or = 45 minutes was used to define IR. Area under the curve values for glucose (AUC(g)) and insulin (AUC(i)) concentrations were calculated. RESULTS: Morgan, Paso Fino, Quarter Horse, and Tennessee Walking Horse breeds were represented in the OB-IR group. Mean neck circumference and BCS differed significantly between groups and were positively correlated with AUC values. Resting insulin and leptin concentrations were 6 and 14 times as high, respectively, in the OB-IR group, compared with the nonobese group, and were significantly correlated with AUC(g) and AUC(i). Plasma nonesterified fatty acid, very low-density lipoprotein, and high-density lipoprotein-cholesterol (HDL-C) concentrations were significantly higher (86%, 104%, and 29%, respectively) in OB-IR horses, and HDL-C concentrations were positively correlated with AUC values. CONCLUSIONS AND CLINICAL RELEVANCE: Measurements of neck circumference and resting insulin and leptin concentrations can be used to screen obese horses for IR. Dyslipidemia is associated with IR in obese horses.     

Using blood urea nitrogen to predict nitrogen excretion and efficiency of nitrogen utilization in cattle, sheep, goats, horses, pigs, and rats

The objectives of this study were to evaluate the potential for using blood urea N concentration to predict urinary N excretion rate, and to develop a mathematical model to estimate important variables of N utilization for several different species of farm animals and for rats. Treatment means (n = 251) from 41 research publications were used to develop mathematical relationships. There was a strong linear relationship between blood urea N concentration (mg/100 mL) and rate of N excretion (g x d(-1) x kg BW(-1)) for all animal species investigated. The N clearance rate of the kidney (L of blood cleared of urea x d(-1) x kg BW(-1)) was greater for pigs and rats than for herbivores (cattle, sheep, goats, horses). A model was developed to estimate parameters of N utilization. Driving variables for the model included blood urea N concentration (mg/100 mL), BW (kg), milk production rate (kg/d), and ADG (kg/d), and response variables included urinary N excretion rate (g/d), fecal N excretion rate (g/d), rate of N intake (g/d), and N utilization efficiency (N in milk and gain per unit of N intake). Prediction errors varied widely depending on the variable and species of animal, with most of the variation attributed to study differences. Blood urea N concentration (mg/100 mL) can be used to predict relative differences in urinary N excretion rate (g/d) for animals of a similar type and stage of production within a study, but is less reliable across animal types or studies. Blood urea N concentration (mg/100 mL) can be further integrated with estimates of N digestibility (g/g) and N retention (g/d) to predict fecal N (g/d), N intake (g/d), and N utilization efficiency (grams of N in milk and meat per gram of N intake). Target values of blood urea N concentration (mg/100 mL) can be backcalculated from required dietary N (g/d) and expected protein digestibility. Blood urea N can be used in various animal species to quantify N utilization and excretion rates.     

Effects of exercise intensity and duration on plasma beta-endorphin concentrations in horses

OBJECTIVE: To determine the relationship between plasma beta-endorphin (EN) concentrations and exercise intensity and duration in horses. ANIMALS: 8 mares with a mean age of 6 years (range, 3 to 13 years) and mean body weight of 450 kg. PROCEDURE: Horses were exercised for 20 minutes at 60% of maximal oxygen consumption (VO2max) and to fatigue at 95% V02max. Plasma EN concentrations were determined before exercise, after a 10-minute warmup period, after 5, 10, 15, and 20 minutes at 60% VO2max or at the point of fatigue (95% VO2max), and at regular intervals after exercise. Glucose concentrations were determined at the same times EN concentrations were measured. Plasma lactate concentration was measured 5 minutes after exercise. RESULTS: Maximum EN values were recorded 0 to 45 minutes after horses completed each test. Significant time and intensity effects on EN concentrations were detected. Concentrations were significantly higher following exercise at 95% VO2max, compared with those after 20 minutes of exercise at 60% VO2max (605.2 +/- 140.6 vs 312.3 +/- 53.1 pg/ml). Plasma EN concentration was not related to lactate concentration and was significantly but weakly correlated with glucose concentration for exercise at both intensities (r = 0.21 and 0.30 for 60 and 95% VO2max, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: A critical exercise threshold exists for EN concentration in horses, which is 60% VO2max or less and is related to exercise intensity and duration. Even under conditions of controlled exercise there may be considerable differences in EN concentrations between horses. This makes the value of comparing horses on the basis of their EN concentration questionable.     

Effects of oral cimetidine on plasma concentrations of phenylbutazone in horses

Phenylbutazone was administered to six Thoroughbred horses in a cross-over study in which the horses received cimetidine pretreatment or no cimetidine pretreatment. Blood samples were collected at various times for 48 h after phenylbutazone administration and the plasma was analysed for phenylbutazone. Cimetidine pretreatment elevated phenylbutazone plasma concentrations during the first 8 h after phenylbutazone administration. The absorption rate, maximum phenylbutazone plasma concentrations and AUC were significantly greater with cimetidine pretreatment. The half-life of phenylbutazone did not change with cimetidine pretreatment; however, lower plasma concentrations of the metabolite gamma-hydroxyphenylbutazone were observed with cimetidine pretreatments. Plasma concentrations of the metabolite oxyphenbutazone were unchanged with cimetidine pretreatment compared to control values. Twenty-four-hour plasma concentrations of phenylbutazone were not different from control values with cimetidine pretreatment. This study suggests that concurrent treatment with cimetidine and phenylbutazone 24 h before race time does not result in elevations of plasma phenylbutazone concentrations above control values.     

Relationship of serum and plasma copper and ceruloplasmin concentrations of cattle and the effects of whole blood sample storage

The present study was conducted to determine whether plasma and serum copper and ceruloplasmin concentrations in cattle are different and whether transport of samples with storage on ice before centrifugation affects the measurements. Mean copper and ceruloplasmin values were higher in plasma than in serum. Linear regressions were plasma copper (microgram/ml) = 1.200 serum copper -0.032 (r2 = 0.99), serum ceruloplasmin (mg/dl) = 14.0 serum copper + 2.34 (r2 = 0.48), and plasma ceruloplasmin (mg/dl) = 18.2 plasma copper + 2.1 (r2 = 0.43). The percentage of copper associated with ceruloplasmin was less in serum (55%) than in plasma (66%). Storage of blood samples on ice for 3 days decreased serum copper value by 3.5%. Linear regressions to correct for storage effects were corrected serum copper = 1.11 stored serum copper -0.04 (r2 = 0.94) and corrected plasma copper = 1.22 stored plasma copper -0.17 (r2 = 0.86). A cuproprotein may be involved in the blood clotting process, and some ceruloplasmin and its copper are apparently trapped in the fibrin clot, causing less copper in serum, compared with that in plasma. The difference between plasma and serum copper concentrations of calves was slightly increased by dietary copper supplementation.     

Effects of dexamethasone,glucose infusion,adrenocorticotropin, and propylthiouracil on plasma leptin concentrations in horses

In experiment 1, nine light horse geldings (three 3 x 3 Latin squares) received dexamethasone (DEX; 125 microg/kg BW, i.m.), glucose (0.2 g/kg BW, i.v.), or nothing (control) once per day for 4 days. DEX increased (P < 0.001) glucose, insulin, and leptin concentrations and resulted in a delayed increase (P < 0.001) in IGF-I concentrations. In experiment 2, mares were similarly treated with DEX (n = 6) or vehicle (n = 6). DEX again increased (P < 0.01) glucose, insulin, and leptin concentrations; the delayed elevation in IGF-I concentrations occurred on day 10, 12, and 19, relative to the first day of treatment. In experiment 3, six light horse geldings received either 200 IU of adrenocorticotropin (ACTH) i.m. or vehicle twice daily for 4 days. ACTH increased (P < 0.001) cortisol concentrations. Further, ACTH resulted in increases (P < 0.01) glucose, insulin, and leptin concentrations. In experiment 4, plasma samples from four light horse stallions that were fed 6-n-propyl-2-thiouracil (PTU) at 6 mg/kg BW for 60 days to induce hypothyroidism were compared to samples from control stallions. On day 52, stallions receiving PTU had lower concentrations of thyroxine (P < 0.05) and triiodothyronine (P < 0.01) and higher (P < 0.01) concentrations of TSH. Leptin concentrations were higher (P < 0.01) in PTU-fed stallions from day 10 through 52. In conclusion, circulating concentrations of leptin in horses was increased by administering DEX. Treatment with ACTH increased cortisol and resulted in lesser increases in leptin, glucose, and insulin. In addition, PTU feeding results in lesser increases in leptin concentrations.     

The effects of a probiotic on blood urea nitrogen and creatinine concentrations in large felids

Chronic kidney disease is a common finding in older captive exotic felids. The purpose of this study was to evaluate the effectiveness of a probiotic to reduce blood urea nitrogen and creatinine in large felids. Fifteen adult, large felids (6 tigers [Panthera tigris], 5 lions [Panthera leo], 3 cougars [Puma concolor], and 1 leopard [Panthera pardus]) were administered a probiotic twice daily after a baseline complete blood cell count and plasma chemistry panel was obtained. Plasma chemistry values were rechecked at 2 mo (n = 14) and 6 mo (n = 9). There was no significant change in blood urea nitrogen over time; however, there was a significant change in creatinine over time (P = 0.04). Creatinine concentration decreased significantly between 2 and 6 mo (P = 0.02), and a decrease was seen between 0 and 6 mo, but this change was not significant (P = 0.05). There was no significant difference noted for creatinine concentration between 0 and 2 mo (P = 0.35). This probiotic may be helpful in large felids with elevated creatinine concentrations because of chronic kidney disease; however, further studies are warranted.     

Effect of specimen collection and storage on blood glucose and lactate concentrations in healthy,hyperthyroid and diabetic cats

Abstract: The objective of this study was to compare and investigate differences in glucose and lactate concentrations in sodium fluoride/potassium oxalate (NaF/Ox) plasma and serum in healthy cats and cats with metabolic disease. Glucose and lactate concentrations were determined in routinely processed serum and NaF/Ox plasma obtained from healthy (n = 30), hyperthyroid (n = 27) and diabetic (n = 30) cats, and in samples from 6 healthy cats stored at 25°C or 4°C for 0,1, 2, 4, or 8 hours. The packed cell volume (PCV) of blood collected in NaF/Ox was compared with that of blood collected in EDTA. Mean glucose concentration was significantly (P < .05) lower in NaF/Ox plasma than in serum in all groups of cats, by 0.7–2.5 mmol/L (11–45 mg/dL); the difference was greater in cats with hyperglycemia. Mean lactate concentration was significantly higher in serum than in NaF/Ox plasma in all groups of cats, by 0.4–1.2 mmol/L (3.6–10.8 mg/dL); the difference was greater in hyperthyroid and diabetic cats. In vitro, only serum stored on the clot for ≥ 1hour at 25°C had significantly lower glucose and higher lactate concentrations. The PCV of NaF/Ox-anticoagulated blood was lower than that of EDTA-anticoagulated blood, by 7.0%± 1.4% (P<.01). In conclusion, collection of feline blood in NaF/Ox was necessary to prevent in vitro increases in lactate concentration; however, NaF/Ox artifactually decreased plasma glucose concentration because of RBC shrinkage. The PCV should not be determined on blood collected in NaF/Ox.     

Effects of phase-feeding of crude protein on performance, carcass characteristics, serum urea nitrogen concentrations, and manure nitrogen of finishing beef steers

As cattle mature, the dietary protein requirement, as a percentage of the diet, decreases. Thus, decreasing the dietary CP concentration during the latter part of the finishing period might decrease feed costs and N losses to the environment. Three hundred eighteen medium-framed crossbred steers (315 +/- 5 kg) fed 90% (DM basis) concentrate, steam-flaked, corn-based diets were used to evaluate the effect of phase-feeding of CP on performance and carcass characteristics, serum urea N concentrations, and manure characteristics. Steers were blocked by BW and assigned randomly to 36 feedlot pens (8 to 10 steers per pen). After a 21-d step-up period, the following dietary treatments (DM basis) were assigned randomly to pens within a weight block: 1) 11.5% CP diet fed throughout; 2) 13% CP diet fed throughout; 3) switched from an 11.5 to a 10% CP diet when approximately 56 d remained in the feeding period; 4) switched from a 13 to an 11.5% CP diet when 56 d remained; 5) switched from a 13 to a 10% CP diet when 56 d remained; and 6) switched from a 13 to an 11.5% CP diet when 28 d remained. Blocks of cattle were slaughtered when approximately 60% of the cattle within the weight block were visually estimated to grade USDA Choice (average days on feed = 182). Nitrogen volatilization losses were estimated by the change in the N:P ratio of the diet and pen surface manure. Cattle switched from 13 to 10% CP diets with 56 d remaining on feed or from 13 to 11.5% CP with only 28 d remaining on feed had lower (P < 0.05) ADG, DMI, and G:F than steers fed a 13% CP diet throughout. Steers on the phase-feeding regimens had lower (P = 0.05) ADG and DMI during the last 56 d on feed than steers fed 13.0% CP diet throughout. Carcass characteristics were not affected by dietary regimen. Performance by cattle fed a constant 11.5% CP diet did not differ from those fed a 13% CP diet. Serum urea N concentrations increased (P < 0.05) with increasing dietary CP concentrations. Phase-feeding decreased estimated N excretion by 1.5 to 3.8 kg/steer and nitrogen volatilization losses by 3 to 5 kg/steer. The results suggest that modest changes in dietary CP concentration in the latter portion of the feeding period may have relatively small effects on overall beef cattle performance, but that decreasing dietary CP to 10% of DM would adversely affect performance of cattle fed high-concentrate, steam-flaked, corn-based diets.     

Reliability of using reagent test strips to estimate blood urea nitrogen concentration in dogs and cats

OBJECTIVE: To evaluate the clinical accuracy of reagent test strips used to estimate BUN concentration in dogs and cats. DESIGN: Prospective study. ANIMALS: 116 dogs and 58 cats. PROCEDURE: Blood samples were collected at the time of admission to the hospital. Estimates of BUN concentration obtained with reagent test strips (category 1 [5 to 15 mg/dL], 2 (15 to 26 mg/dL], 3 [30 to 40 mg/dL], or 4 [50 to 80 mg/dL]) were compared with SUN concentrations measured with an automated analyzer. For dogs, category 1 and 2 test strip results were considered a negative result (nonazotemic) and category 3 and 4 test strip results were considered a positive result (azotemic). For cats, category 1, 2, and 3 test strip results were considered a negative result (nonazotemic) and category 4 test strip results were considered a positive result (azotemic). RESULTS: On the basis of SUN concentration, 40 of the 174 (23%) animals (20 dogs and 20 cats) were classified as azotemic. One dog and 2 cats had false-negative test strip results, and 1 dog had a false-positive result. Sensitivity and specificity were 95% (20/21) and 99% (94/95), respectively, for dogs and 87% (13/15) and 100% (43/43), respectively, for cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that reagent test strips are a reliable method for rapidly estimating BUN concentrations in dogs and cats. Because test strip results are only semiquantitative and there remains a potential for misclassification, especially in cats, urea nitrogen concentration should ultimately be verified by means of standard chemistry techniques.     

Effects of dietary cysteine on blood sulfur amino acid, glutathione, and malondialdehyde concentrations in cats

OBJECTIVE: To determine effects of dietary cysteine on blood sulfur amino acids (SAA), reduced glutathione (GSH), oxidized glutathione (GSSG), and malondialdehyde (MDA) concentrations in cats. ANIMALS: 12 healthy adult cats. PROCEDURE: Cats were fed diets with a nominal (0.50 g/100 g dry matter [DM]), moderate (1.00 g/100 g DM), or high (1.50 g/100 g DM) cysteine content in a 3 X 3 Latin square design with blocks of 8 weeks' duration. Venous blood samples were collected after each diet had been fed for 4 and 8 weeks, and a CBC and serum biochemical analyses were performed; poikilocyte, reticulocyte, and Heinz body counts were determined; and MDA, GSH, GSSG, and SAA concentrations were measured. RESULTS: Blood cysteine and MDA concentrations were not significantly affected by dietary cysteine content. Blood methionine, homocysteine, and GSSG concentrations were significantly increased when cats consumed the high cysteine content diet but not when they consumed the moderate cysteine content diet, compared with concentrations obtained when cats consumed the nominal cysteine content diet. Blood GSH concentrations were significantly increased when cats consumed the moderate or high cysteine content diet. CONCLUSIONS: Increased dietary cysteine content promotes higher blood methionine, homocysteine, GSH, and GSSG concentrations in healthy cats. CLINICAL RELEVANCE: Supplemental dietary cysteine may be indicated to promote glutathione synthesis and ameliorate adverse effects of oxidative damage induced by disease or drugs.