Effects of storage time on chemistry results from canine whole blood,heparinized whole blood,serum and heparinized plasma

The stability and storage characteristics of 24 blood constituents from dogs including nine enzymes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, lipase), 15 metabolites and minerals (albumin, bile acids, bilirubin, calcium, cholesterol, creatinine, fructosamine, glucose, magnesium, phosphate, potassium, protein, sodium, triglycerides, urea) were studied. Conditions studied included storing of nonanticoagulated and heparinized whole blood for 3 days (Part A), and storing of serum and heparinized plasma for 3 days (Part B). The storage temperature for both studies was +4 degrees C from day 0 to day 1, and +20 degrees C, from day 1 to day 2 and 3. Eight of 24 analytes showed no significant differences (p > 0.05) for three days in whole blood. However, the stability of all 24 analytes greatly improved by storing serum or heparinized plasma compared to nonanticoagulated or heparinized whole blood. In stored serum or heparinized plasma, 20 of 24 analytes showed no significant differences (p < 0.05) for 3 days. Nine of 24 analytes showed significant differences (p < 0.05) between serum and heparinized plasma, where CK, LDH, GGT, and potassium showed differences of possible clinical importance. This study strongly supports the practice of separating serum/plasma from clot/cells as promptly as possible to achieve improved stability for most analytes under test.     

针灸对脾虚犬体重及部分血清指标的影响

通过注射利血平复制犬脾虚证动物模型,观察犬临床体征、体重及部分血清指标变化情况,并研究针灸对脾虚犬体重及血清指标的影响。结果显示,实验犬出现脾虚证后,其体重显著低于空白对照组（P〈0.05）,血清谷丙转氨酶（ALT）、谷草转氨酶（AST）活性与丙二醛（MDA）含量均显著增高（P〈0.05）;血清超氧化物歧化酶（SOD）活性显著降低（P〈0.05）。经针灸治疗后,实验组犬各项指标均得到明显改善,与阳性对照组比较呈显著性差异（P〈0.05）。试验结果表明,皮下注射利血平复制犬脾虚证动物模型后,选用一定穴位进行针灸,对治疗犬脾虚证有明显疗效。     

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.     

Comparison of amylase and lipase activities in serum and plasma of dogs

BACKGROUND: Amylase and lipase activities are most often determined in serum, although heparinized plasma is more convenient to obtain and is used for many routine biochemical analyses. OBJECTIVE: The purpose of this study was to compare amylase and lipase activities in serum and plasma of dogs and to determine whether either specimen type is acceptable for analysis. METHODS: Serum and heparinized plasma were obtained from 101 randomly selected dogs and analyzed in parallel for alpha-amylase and lipase. Results were compared using Passing-Bablock regression, Bland-Altman difference plots, and correlation analysis. RESULTS: There was a high correlation between the results obtained from serum and those from plasma. Regressions (with 95% confidence intervals in parentheses) were as follows: lipase(plasma) = 0.984 (0.976/0.995) Chi lipase(serum) - 0.9 (2.9/0.7) (r =.999); a-amylase(plasma) = 1.003 (0.977/1.032) Chi alpha-amylase(serum) - 1.9 ( 20.7/23.3) (r =.991). Mean differences (serum - plasma) were 8 U/L and 4 U/L for lipase and alpha-amylase, respectively. Classification of results as normal or abnormal did not differ according to specimen type. CONCLUSION: In dogs, lipase and alpha-amylase activities can be determined with the same level of accuracy in serum and in heparinized plasma.     

Effect of storage conditions on prothrombin time, activated partial thromboplastin time and fibrinogen concentration on canine plasma samples

The present study was to assess the effect of storage conditions on prothrombin time (PT), activated partial thromboplastin time (aPTT) and fibrinogen concentration in blood samples of healthy dogs. Thirty-five dogs of various breeds were included in the study. Citrated blood samples were obtained and plasma was divided into four aliquots to assess selected clotting parameters by means of a coagulometer. The first aliquot was analysed within 1 h after collection, while the remaining 3 were stored at 8℃ for 4, 8 and 24 h, respectively. One-way repeated measures analysis of variance documented a significant decreasing effect on PT at 24 h compared to 8 h and on fibrinogen concentration after 8 and 24 h compared to sampling time and at 4 and 24 h compared to 8 h post sampling. In conclusion, the results of this study indicate that only fibrinogen appears prone to significant decrease. In fact, aPTT is not substantially affected by refrigeration for at least 24 h post sampling and PT showed a statistical difference that does not necessary indicate biological significance as the results obtained were within reference intervals for the dog.     

Effects of a single injection of methylprednisolone acetate on serum biochemical parameters in 11 cats

BACKGROUND: Therapeutic use of corticosteroids has been implicated in a variety of serum biochemical abnormalities in dogs; however, the effects in cats are less well characterized. OBJECTIVE: The objective of this brief communication is to report serum biochemical changes in response to methylprednisolone acetate (MPA) in adult cats. METHODS: Serum biochemical profiles were performed on 11 cats with dermatologic diseases at baseline, 3-6 days, and 16-24 days after a single intramuscular dose of 5 mg/kg MPA. RESULTS: Median serum albumin and bicarbonate concentrations and amylase activity were increased compared to baseline at both post-treatment time points; aspartate aminotransferase activity and magnesium concentration were increased at 3-6 days post-treatment only; and alkaline phosphatase activity and total calcium concentration were increased at 16-24 days post-treatment only. Median serum creatinine concentration was decreased compared to baseline at both post-treatment time points. Examination of data from individual cats revealed significant variability in serum biochemical changes in response to MPA. No adverse clinical reactions to the drug were reported. CONCLUSIONS: A single dose of MPA results in significant changes in some serum biochemical values in cats, although individual responses will vary.     

Effects of delayed serum separation and long-term storage on the measurement of thyroid hormones in equine blood samples

Studies were conducted to determine the effects of delaying the separation of serum from the clot and of long-term storage of serum samples on the measurement of thyroid hormones in blood from horses using a fluorescence polarization immunoassay. The measured concentrations of T3 and T4 were not affected by leaving serum on the clot for as long as 24 hours at room temperatures. Storage of serum for 19 to 22 months at -20 degrees C resulted in significant increases of measured T4, but not T3. These studies support previous work demonstrating that thyroid hormones are resistant to degradation, immunologically stable, and reasonably insensitive to potential problems of routine specimen handling when measured with an immunoassay.     

Effect of temperature,storage time,and sample type on sorbitol dehydrogenase activity in llama serum and plasma

Abstract: Serum and heparinized plasma samples were collected from 11 adult, clinically healthy llamas. Aliquots were assayed for sorbitol dehydrogenase (SDH) activity after storage at room temperature (20°C), 4°C, or −20°C for defined time intervals up to 1 week postcollection. Sorbitol dehydrogenase activity in all samples was within reference intervals for our laboratory. No difference was found between serum and plasma SDH activity when measured immediately (within 1 hour) after collection. Sorbitol dehydrogenase activity decreased to 79% of initial activity by 24 hours in serum stored at room temperature; plasma had 94% of initial SDH activity under the same conditions. Sorbitol dehydrogenase activity was stable in both plasma and serum stored for up to 1 week at 4°C or −20°C. With the exception of serum stored at 20°C for > 8 hours, in vitro stability of llama SDH was adequate for its use in diagnostic testing.     

A comparison of liquid and lyophilized egg yolk plasma to low density lipoproteins for freezing of canine spermatozoa

The current study aimed to explore the potential usefulness of liquid or lyophilized egg yolk plasma (EYP) as a substitute for low‐density lipoproteins (LDL) for cryopreservation of canine spermatozoa. In the first experiment, a total of 20 ejaculates harvested from six Beagles were frozen in extenders containing 6% LDL (control) or liquid or lyophilized EYP at one of three concentrations (20%, 40% or 60%). Motility parameters were assessed 10 min after thawing using computer‐assisted sperm analysis. For both liquid and lyophilized EYP, the 40% concentration yielded motility similar (p > 0.05) to that observed with the control extender. In the second experiment, 12 ejaculates collected from the same six dogs were frozen in 6% LDL (Control), 40% liquid EYP or 40% lyophilized EYP extenders. Spermatozoal membrane integrity (hypo‐osmotic swelling test [HOSt] and SYBR14/propidium iodide [PI] staining), acrosome integrity (FITC‐Pisum sativum agglutinin staining) and DNA integrity (acridine orange staining) characteristics were evaluated 10 min after thawing. Both liquid and lyophilized 40% EYP‐based extenders successfully preserved all assessed integrity parameters as efficiently as the control. Results of this study suggest that lyophilized EYP is a viable alternative to LDL in freezing extenders for dog semen.     

The evaluation of lycopene and cysteamine supplementation effects on sperm and oxidative stress parameters during chilled storage of canine semen

The aim of this study was to evaluate the effects of different concentrations of lycopene and cysteamine on characteristics of sperm, liquid peroxidation and enzymatic activities in seminal plasma of canine semen preserved at 5°C for 72 hr. The semen samples were divided into eight aliquots: control, control sham (dimethyl sulfoxide 5%), lycopene groups (250, 500 and 750 µg/ml) and cysteamine groups (2.5, 5 and 10 mM). Motility, viability, membrane integrity, DNA integrity, total antioxidant capacity (TAC) and malondialdehyde (MDA) levels were evaluated. Progressive motility and total motility were higher with the 500 and 750 µg/ml lycopene concentrations, respectively, compared to the control group and the cysteamine groups following 72 hr of storage in the liquid storage. Motility characteristics, viability and hypo-osmotic swelling test (HOST) percentages were significantly improved in 500 µg/ml lycopene compared to other groups. The 500 and 750 µg/ml lycopene concentrations, respectively, showed significantly reduced percentages of spermatozoa with DNA integrity compared to the control group. The 500 and 750 µg/ml lycopene concentrations, respectively, led to the significant decrease of MDA levels. The 500 µg/ml lycopene enhanced TAC levels after 48 and 72 hr that was not observed in other groups. In conclusion, the findings of this study showed that lycopene supplementation in canine semen extenders improved canine semen parameters and TAC levels and decreased MDA levels in the chilling process.     

Effect of weaning on behavior and serum parameters in dairy goat kids

This study aimed at investigating the effects of weaning kids abruptly at an average of 55 ± 13 days of age on intake, behavioral and serum parameters and, lasted for a total of six weeks; two weeks pre-weaning and four weeks post-weaning. Sixteen single kids with equal gender were used. Kids were only allowed to stay with their mothers for suckling (45 min/period) both in the morning and in the evening period during pre-weaning. Grower concentrate and hay were offered ad libitum . The duration of the study was divided into three periods for the sampling of behavioral and serum parameters; (i) pre-weaning period lasting for two weeks (P-BW) (ii) early post-weaning period lasting for one week (P-AW1) and (iii) late post-weaning period lasting for three weeks (P-AW2). Daily weight gain of kids gradually decreased as the observation period progressed ( P  = 0.001). However concentrate feed intake increased from 0.154 kg/day in P-BW to 0.479 kg/day in P-AW1 and 0.499 kg/day in P-AW2. Water intake, rumination and standing behaviors decreased in P-AW2 ( P  < 0.001), whereas activity towards concentrate feed (CF) ( P  < 0.001) and roughage ( P  = 0.012) increased as compared to P-BW and P-AW1. Abnormal oral activity was not affected by the periods ( P  = 0.906). CF was significantly higher in females ( P  = 0.003), whereas males displayed higher lying behavior ( P  = 0.007). Glucose, total protein, urea, cholesterol, HDL-cholesterol, LDL-cholesterol concentrations ( P  = 0.001) and ALP activity ( P  = 0.003) were significantly affected by the periods. The results of the present study suggest that behavioral and serum parameters across the periods describe changes in the nutritional conditions as a result of the transition from milk to solid feed in association with weaning.     

Effects of storage on concentration of hydrocortisone (cortisol) in canine serum and plasma

A specific radioimmunoassay was used to measure concentrations of hydrocortisone (cortisol) in the serum and plasma of 4 dogs. Differences (P greater than 0.05) in concentrations of cortisol were not found between serum and plasma (from EDTA-treated and heparinized blood samples). Differences (P greater than 0.05) in serum or plasma concentrations of cortisol were not found between samples stored at 4 C for various times (10 minutes, 10 hours, 40 hours) after collection, but before removal of RBC. In a study designed to determine the stability of cortisol in serum samples stored at room temperature, degradation was dependent on the initial serum concentrations of cortisol. Decreases (P greater than 0.05) did not occur in concentrations of cortisol in serum samples stored up to 15 days when initial concentrations of cortisol were less than 15 ng/ml. However, when initial concentrations of cortisol were approximately 55 ng/ml and 80 ng/ml, significant (P greater than 0.05) degradation occurred after 9 and 5 days of storage, respectively. Results of this investigation indicate that either serum or plasma of dogs is suitable for radioimmunoassay of cortisol and that samples (with and without added coagulants) incubated at 4 C may be left uncentrifuged for up to 40 hours without cortisol degradation. However, prolonged storage of serum at room temperature is detrimental, particularly for samples having large concentrations of cortisol.     

Determination of uric acid in canine serum and urine by high performance liquid chromatography

A reproducible high performance liquid chromatography (HPLC) method was developed for analysis of uric acid in canine serum and urine. The method consists of precipitating serum proteins with phosphotungstic acid prior to HPLC analysis. Urine is analyzed after dilution with buffer. Chromatography is performed on a reversed-phase C-18 column with UV detection at 292 nm. Sensitivity of the method will allow reproducible measurement of uric acid at concentrations of 0.05 mg/dl in serum and 0.1 mg/dl in urine. The HPLC method has been used to quantify hundreds of canine serum and urine samples. The method is superior to UV absorption or colorimetric methods because its lower limit of detection allows measurement of uric acid at concentrations found in canine serum and urine.     

Hemolysis as a factor in clinical chemistry and hematology of the dog

Serum biochemical, hemostatic, and hematologic analytes were determined by four laboratories on dog serum or plasmas containing increasing amounts of hemolysate. From the results of testing, interferographs were prepared to aid in decision making and to enhance visualization of the effects of hemolysis on the determination of the analytes. Heniktsus consistently interfered with the analysis of creatinine phosphokinase, lactic dehydrogenase, aspartate aminotransferase, lipase, and albumin, all of which appeared to increase with increasing hemolysis. The results for the remainder of the biochemical, hemostatic, and hematologic analytes varied greatly among analyzers or methods, rarely in a predictable manner, indicating that each laboratory should evaluate the effects of hemolysis on each analyzer and for each method used, in order to make informed decisions on the use of hemolyzed but irreplaceable samples.     

Comparison of two dry chemistry analyzers and a wet chemistry analyzer using canine serum

Canine serum was used to compare seven chemistry analytes on two tabletop clinical dry chemistry analyzers, Boehringer's Reflotron and Kodak's Ektachem. Results were compared to those obtained on a wet chemistry reference analyzer, Roche Diagnostic's Cobas Mira. Analytes measured were urea nitrogen (BUN), creatinine, glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholesterol and bilirubin. Nine to 12 canine sera with values in the low, normal, and high range were evaluated. The correlations were acceptable for all comparisons with correlation coefficients greater than 0.98 for all analytes. Regression analysis resulted in significant differences for both tabletop analyzers when compared to the reference analyzer for cholesterol and bilirubin, and for glucose and AST on the Kodak Ektachem. Differences appeared to result from proportional systematic error occurring at high analyte concentrations.     

Effects of high ambient temperature on plasma metabolomic profiles in chicks

Exposure to high ambient temperature is detrimental to the poultry industry. To understand the influence from a metabolic perspective, we investigated the effects of exposure to high ambient temperature on plasma low‐molecular‐weight metabolite levels in chicks using gas chromatography/mass spectrometry‐based non‐targeted metabolomic analysis. Heat exposure for 4 days suppressed growth and food intake. Of the 92 metabolites identified, the levels of 29 decreased, whereas the levels of nine increased. We performed an enrichment analysis on the identified metabolites and found 35 candidates for metabolic processes affected by heat exposure. Among them, the sulfur amino acid metabolic pathway was clearly detected and the levels of the following related metabolites were decreased: cystathionine, cysteine, cystine, homocysteine and hypotaurine. Changes in the kynurenine pathway in tryptophan metabolism, which is linked to the immune system and oxidative stress, were also observed: kynurenine and quinolinic acid levels increased, whereas nicotinamide levels decreased. These results suggest the possible involvement of various metabolic processes in heat‐exposed chicks. Some of these metabolites would be important to understand the mechanism of biological responses to high ambient temperature in chicks.     

Effect of repeated freeze-thaw cycles on routine plasma biochemical constituents in canine plasma

BACKGROUND: The effect of repeated freeze-thaw cycles on plasma constituents has not been assessed in dogs, although such a procedure is not uncommon to use in routine laboratory practice. OBJECTIVE: To assess the effect of freeze-thaw cycles on routine plasma constituents in healthy dogs. METHODS: Six healthy adult dogs were used. Blood was sampled and placed in heparinized tubes. After centrifugation, plasma was separated into 5 aliquots. One aliquot was considered as the reference aliquot and used immediately for the assay of all of the biochemical constituents. All of the other aliquots were stored at 20 degrees C. Three aliquots underwent 1, 2, or 3 freeze-thaw cycles during a 1- to 3-day period. The last aliquot remained at 20 degrees C throughout the study and was thawed on the third day. The following biochemical constituents were assayed: glucose, urea, creatinine, total proteins, sodium, potassium, chloride, calcium, phosphates, aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), and alkaline phosphatase (ALP). RESULTS: No clinically relevant change was observed between the different aliquots for all of the constituents. CONCLUSION: Repeated freeze-thaw cycles do not cause changes in the biochemical constituents studied in canine plasma.