Comparison of biochemical values in serum and plasma, fresh and frozen plasma, and hemolyzed samples from orange-winged Amazon parrots (Amazona amazonica)

BACKGROUND: To the authors' knowledge, on the basis of sample type, storage condition, or hemolysis, differences in serum and plasma biochemical values have not been evaluated in orange-winged Amazon parrots (Amazona amazonica). OBJECTIVES: The purpose of this study was to compare values for biochemical analytes in serum vs plasma, fresh vs frozen plasma, and nonhemolyzed vs hemolyzed samples in orange-winged Amazon parrots. We also compared differences in serum and plasma yield from whole-blood aliquots. METHODS: Fifteen biochemical analytes were evaluated in paired serum and plasma, fresh and frozen plasma, nonhemolyzed and hemolyzed serum and plasma samples from orange-winged Amazon parrots (n = 10) using a wet reagent analyzer. Hemolysis was assessed qualitatively (visually) and quantitatively (hemoglobin [Hgb] measured spectrophotometrically). Serum and plasma yields from 500-microl whole-blood aliquots were determined from centrifuged samples. RESULTS: Analyte values significantly differed among sample groups, but were still within published reference intervals, with the exception of increases in potassium concentration in markedly hemolyzed serum and plasma samples. Clinically important changes in hemolyzed serum and plasma samples included increases in potassium, phosphorus, and albumin concentrations and lactate dehydrogenase activity. The degree of hemolysis assigned qualitatively did not correlate with quantitative Hgb concentration. A significantly greater yield of plasma (288 +/- 13 microL) than serum (241 +/- 44 microL) was obtained. CONCLUSIONS: Significant differences may occur in different sample types, however, only changes in potassium, phosphorus, albumin, and lactate dehydrogenase values in hemolyzed samples were considered clinically relevant. Lack of agreement between qualitative and quantitative Hgb concentration indicates the unreliability of visual estimation. Based on higher sample yield, and lack of clinically relevant differences from serum, plasma is a better sample choice for clinical chemistry analysis in birds.     

Measurement of plasma antithrombin III activity in healthy horses

A fluorometric assay was used to determine plasma antithrombin III (AT III) activities in 15 healthy adult horses. Nearly all plasma samples had an initial value of greater than 100% thrombin inhibited, so a 1:1 dilution of the prepared samples was performed. Following dilution, the mean value of the animals was 59.17 +/- 7.4% thrombin inhibited. Mares had significantly greater AT III activity than did geldings (P less than 0.01). The results of this study indicate the horse has more AT III activity than did other domestic species in which AT III activity has been reported.     

Evaluation of a chromogenic assay to measure the factor Xa inhibitory activity of unfractionated heparin in canine plasma

BACKGROUND: Unfractionated heparin (UFH) has a complex pharmacologic profile that necessitates patient monitoring to prevent inadequate anticoagulation or overdosage and hemorrhage. Factor Xa inhibitory assays (to measure anti-Xa activity) are used to adjust UFH dosage and define safe and effective regimens for specific thrombotic disorders in humans. OBJECTIVE: In this study, the accuracy, linearity, and clinical utility of a chromogenic assay were assessed for monitoring UFH anti-Xa activity in canine plasma samples. METHODS: A commercial assay (Rotachrom Heparin, Diagnostica Stago, Parsippany, NJ, USA) was used to measure anti-Xa activity in canine plasma samples spiked with different concentrations of UFH. Background absorbance and assay linearity were compared for canine and human plasmas. Percentage recovery of UFH anti-Xa activity and intra- and interassay imprecisions were investigated by multiple measurements of canine plasma to which known amounts of UFH were added. The spiked plasma samples also were used to determine the heparin sensitivity of an activated partial thromboplastin time (aPTT) test. RESULTS: Canine plasma samples were assayed at a higher dilution than were human plasma samples (3:8 versus 4:8) to eliminate higher background anti-Xa activity in canine plasma. Using this modification, the recovery of anti-Xa activity in canine plasma was linear (R2 > .9) at concentrations of 0 - 0.75 U/mL UFH. Intra- and interassay imprecisions for plasma samples containing 0.5 U/mL UFH were <10%, whereas samples containing 0.25 U/mL UFH had imprecisions of 13% and 24%, respectively. The anti-Xa activity range of 0.5 - 0.75 U/mL caused prolongation of aPTTs to 1.5 - 2.5 times the assay mean. CONCLUSION: Plasma anti-Xa activity of dogs treated with UFH can be accurately monitored using this commercially available chromogenic assay.     

Apparent pseudohyperkalemia in a Chinese Shar Pei dog

Whole blood in a serum clot tube and EDTA-anticoagulated samples from an 8-year-old spayed female Chinese Shar Pei dog were submitted by an external clinic to the diagnostic laboratory at Atlantic Veterinary College for routine biochemical and hematologic analysis prior to entropion surgery. Laboratory abnormalities included mild hyperkalemia (6.3 mmol/L, reference interval 3.6-6.0 mmol/L), mild normocytic, hypochromic, nonregenerative anemia (HCT 0.31 L/L, reference interval 0.37-0.55 L/L; MCHC 290 g/L, reference interval 320-360 g/L), and increased red cell distribution width (RDW; 26.2%, reference interval 11-14%). A small subpopulation of macrocytic, slightly hypochromic erythrocytes was noted on Wright's-Giemsa-stained blood smears. Biochemical and hematologic data obtained from this patient over the previous 7.5 years indicated that serum (and in 1 case, heparinized plasma) potassium concentration was increased (range, 6.3-10.9 mmol/L) in 5 of 8 samples (HCT ranged from 0.31-0.43 L/L, Hgb 91-124 g/L, MCHC 280-312 g/L, and RDW 18.2-26.9%). Clinical signs suggestive of hyperkalemia were not observed at any time, suggesting pseudohyperkalemia as the cause of the increased potassium concentrations. An erythrocyte lysate prepared from a heparinized blood sample had a high potassium concentration (16.8 mmol/L) compared with that of a clinically healthy, non-Shiba control dog (6.7 mmol/L). An osmotic fragility test of the patient's erythrocytes showed 50% hemolysis at 0.57% NaCl, compared with 0.48% NaCl for the control dog, indicating increased fragility. On scanning electron microscopy, a small subpopulation of erythrocytes were large, flattened, and had a tendency to fold. These findings supported the provisional diagnosis of pseudohyperkalemia due to increased intracellular RBC potassium concentration. High-potassium erythrocytes have been reported in Akitas, Shibas, Jindos, other East Asian dog breeds, and occasionally, in mixed-breed dogs. Pseudohyperkalemia should also be considered when an otherwise unexplained elevation in serum or plasma potassium concentration is observed in Chinese Shar Pei dogs, and may be accompanied by increased RDW, low MCHC, and increased osmotic fragility with or without mild anemia.     

One-stage prothrombin time, activated partial thromboplastin time, thrombin time, fibrin degradation product concentration, and antithrombin activity in healthy adult alpacas

Background: Alpacas are increasingly presented to veterinarians for evaluation and care. Reports of alpaca reference intervals for one‐stage prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), concentration of fibrin degradation products (FDP), and antithrombin (AT) activities are scarce or nonexistent. Objective: The aim of this study was to determine values for blood coagulation times (PT, aPTT, and TT), FDP concentrations, and AT activities in healthy adult alpacas. Methods: Of blood samples collected from 35 clinically healthy adult alpacas via jugular venipuncture and placed into sodium citrate and FDP tubes, 29 samples were assayable for coagulation testing. PT, aPTT, and TT were determined by physical (mechanical) clot detection; AT activity was determined using a thrombin‐specific chromogenic substrate end‐point assay; and FDP concentrations were determined by the slide agglutination method. Results: Median values and ranges (minimum–maximum) were determined for PT (8.7 seconds, 6.6–11.2 seconds), aPTT (17.3 seconds, 11.9–22.5 seconds), TT (10.2 seconds, 5.4–16.0 seconds), and AT activity (123.3%, 104.8–144.2%). The mean concentration of FDP was <8 μg/mL. Conclusion: These values for coagulation times, FDP concentration, and AT activity will provide a useful starting point in the diagnostic evaluation of ill adult alpacas.     

Use of the ADVIA 120 for differentiating extracellular and intracellular hemoglobin

BACKGROUND: The use of cell-free hemoglobin (Hgb) solutions, such as Oxyglobin (Biopure Corp, Cambridge, MA, USA), as a blood substitute for the treatment of acute anemias is increasing in veterinary medicine. These solutions interfere with colorimetric tests, which do not discriminate between cellular Hgb (Hgb-cell) from the patient and extracellular Hgb (Hgb-delta) from the Oxyglobin, and therefore make the monitoring of anemia, based on Hgb concentration, difficult. The ADVIA 120 hematology analyzer (Bayer Diagnostics, Tarrytown, NY, USA) evaluates Hgb by 2 methods, a standard cyanmethemoglobin colorimetric method and flow cytometry, and therefore might provide the means to differentiate extracellular and intracellular Hgb. OBJECTIVE: The objective of this study was to determine the accuracy and precision of the ADVIA 120 in differentiating extracellular from intracellular Hgb. METHODS: Anticoagulated whole blood samples from 10 healthy dogs were analyzed in triplicate on the ADVIA 120. Hgb-delta concentration was determined by adding Oxyglobin (13 g/dL) to the whole blood samples at dilutions of 1:1, 1:2, 1:4, 1:8, 1:16, and 1:32. Hgb-cell and Hgb-total values were calculated and compared with actual values by linear regression. Analyses were done in triplicate and repeated 9 consecutive times to evaluate intra-assay precision of Hgb-total and Hgb-cell determinations. RESULTS: Correlation between Hgb values obtained by colorimetric (Hb-total) and flow cytometric (Hgb-cell) methods on whole blood samples was high (R(2) = .99; n = 10) with a slope of 0.96 and intercept of 0. Correlation between actual and predicted Hgb-cell values also was high (R(2) = .99), with a small positive bias (0.289 +/- 0.185; n = 60). Intra-assay precisions were high, with most coefficients of variation <2%. CONCLUSION: The ADVIA 120 is capable of differentiating Hgb-cell from Hgb-delta. The flow cytometric method is accurate and precise when compared with the cyanmethemoglobin method. A small bias between the results is unlikely to be clinically significant but may affect the ability of the ADVIA to differentiate small quantities (<0.3 g/dL) of Hgb-delta.     

Overestimation of canine albumin concentration with the bromcresol green method in heparinized plasma samples

Albumin concentrations are routinely measured in dogs with bromcresol green (BCG)-binding assays on automated chemistry analyzers. Several variables affect this assay, including the length of reaction time, sample type, and lack of specificity of BCG for albumin. We observed that albumin concentrations measured with BCG appeared higher in heparinized plasma samples in sick dogs. The objective of this study was to determine the effect of anticoagulant and assay procedure on BCG albumin concentrations in clinically ill dogs. We hypothesized that albumin concentrations would be overestimated in heparinized plasma compared with serum because of the combination of heparin and fibrinogen. Furthermore, we hypothesized that the overestimation would be influenced by assay parameters. Blood was collected from 32 clinically ill dogs into tubes containing heparin, citrate, or no anticoagulant. Citrate was chosen to assess the effect of fibrinogen in the absence of heparin. Albumin concentration was measured in all 3 sample types from each dog using 2 different BCG procedures on an automated chemistry analyzer. The BCG procedures (standard and modified) differed in the wavelengths used for absorbance readings (standard, 600/700; modified, 570/505) and the time point at which absorbance was measured (standard, 100 seconds; modified, 40 seconds). In addition, the modified method incorporated a sample blank. Globulin fractions, fibrinogen concentration, and indices of lipemia, hemolysis, and icterus were evaluated for their contribution to the overestimation of albumin concentration in heparinized plasma compared with serum samples. Albumin concentrations were significantly higher (P 相似文献   

Pharmacokinetics of single-dose intravenous levetiracetam administration in normal dogs

Objective: To determine plasma pharmacokinetics of levetiracetam after a single intravenous dose (60 mg/kg) in normal dogs using a high‐performance liquid chromatography assay validated for canine plasma. Design: Pharmacokinetic study. Setting: A university‐based canine research facility. Animals: Six healthy adult dogs. Interventions: Intravenous drug administration, multiple blood sample procurement. Measurements and main results: There were no obvious adverse effects associated with the intravenous (IV) bolus administration of levetiracetam in any of the dogs. Plasma levetiracetam concentrations remained above or within the reported therapeutic range for humans (5–45 μg/mL) for all dogs, for all time periods evaluated. Mean and median (in parentheses) values for pharmacokinetic parameters included the following: maximum plasma concentration, 254 μg/mL (254 μg/mL); half‐life, 4.0 hours (4.0 hours); volume of distribution at steady state, 0.48 L/kg (0.48 L/kg); clearance, 1.4 mL/kg/min (1.5 mL/kg/min); and median residence time, 6.0 hours (6.0 hours). Conclusions: In normal dogs, a 60 mg/kg IV bolus dose of levetiracetam is well tolerated and achieves plasma drug concentrations within or above the therapeutic range reported for humans for at least 8 hours after administration. Based on the favorable pharmacokinetics and tolerability demonstrated for IV levetiracetam in this study, in addition to previously demonstrated efficacy of oral levetiracetam, IV levetiracetam may be a useful treatment option for emergency management of canine seizure activity.     

Enzymeimmunoassay of oestrone sulphate concentrations in faeces for non-invasive pregnancy determination in mares

AIM: To determine the suitability of measuring faecal oestrone sulphate (OS) by enzymeimmunoassay as a means of determining pregnancy status in mares bred under New Zealand conditions. METHODS: An antibody-coated microtitre plate-based enzymeimmunoassay was used to determine the concentration of OS in faecal and plasma samples obtained from pregnant and non-pregnant mares. RESULTS: In non-pregnant mares, the mean faecal OS concentration was 34 ng/g, and the value three standard deviations above this was 80 ng/g. None of 427 faecal samples collected from 116 non-pregnant mares over a l-year period had an OS concentration >80 ng/g. Only five samples from three mares had an OS concentration >65 ng/g, the value two standard deviations above the mean non-pregnant value. Analysis of faecal OS concentrations in 532 faecal samples collected from 39 pregnant mares showed that as pregnancy progressed, an increasing proportion of faecal samples had OS concentrations >80 ng/g. None of the mares 150 days or more pregnant had faecal OS concentrations <50 ng/g, and 204/220 samples obtained from these mares had faecal OS concentrations >80 ng/g. Following foaling or foetal death, elevated faecal OS concentrations returned quickly to non-pregnant levels. The mean +/- s.e.m. plasma level of OS in five mares bled daily throughout one oestrous cycle was 1.7 +/- 0.2 ng/ml. Sixty-eight blood samples from pregnant mares bled up to five times between 92 days after mating and foaling all had plasma OS concentrations >30 ng/ml, with 64/68 being >50 ng/ml. CONCLUSIONS: This study shows that measuring faecal OS concentrations by enzymeimmunoassay offers a convenient, accurate, non-invasive means of determining pregnancy status in mares from 150 days after mating onwards. Mares with faecal OS concentrations <50 ng/g can be considered not pregnant, while mares with faecal OS concentrations >80 ng/g can be considered pregnant. Those few mares returning a faecal OS concentration between 50 and 80 ng/g should be retested to obtain a conclusive result. Measuring plasma OS concentrations allows pregnancy status to be determined earlier (from 100 days after mating). Moreover, the discrimination between non-pregnant and pregnant levels is greater for OS in plasma than in faeces. CLINICAL RELEVANCE: Measurement of OS concentrations in faeces provides an alternative and non-invasive means of determining pregnancy status in mares from 150 days after mating.     

Evaluation of a cow-side immunoassay kit for assessing IgG concentration in colostrum

OBJECTIVE: To determine sensitivity and specificity of a cow-side immunoassay kit for assessing IgG concentration in colostrum. DESIGN: Prospective study. ANIMALS: 76 dairy and 11 beef cows of various parities. PROCEDURE: Colostrum from first, second, and third milkings and milk samples were collected, and IgG concentration was determined by means of radial immunodiffusion. The immunoassay was performed according to the manufacturer's instructions, and sensitivity and specificity were calculated by comparing results of the immunoassay (positive vs negative) with results of immunodiffusion (< 50 g/L vs > or = 50 g/L). RESULTS: 135 colostrum or milk samples were collected. Mean +/- SD colostral IgG concentrations, determined by means of radial immunodiffusion for dairy and beef cows were 65.4 +/- 51.4 g/L and 114.8 +/- 42.7 g/L, respectively. Mean IgG concentrations for first-, second-, and third-milking colostrum samples and for milk samples were 92 +/- 49.0 g/L, 74.6 +/- 45.1 g/L, 47.5 +/- 32 g/L, and 6.8 +/- 3.8 g/L, respectively. Sensitivity of the immunoassay (ie, percentage of samples with IgG concentration < 50 g/L with a positive immunoassay result) was 93%, and specificity (ie, percentage of samples with IgG concentration > or = 50 g/L with a negative immunoassay result) was 76%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the immunoassay kit was an acceptable cow-side test to identify colostrum samples with IgG concentrations < 50 g/L. The immunoassay kit should be useful in screening colostrum for adequate IgG concentration before feeding to calves or storage.     

高效液相色谱法测定牛奶中糠氨酸含量

检测牛奶中的糠氨酸含量可以评价牛奶的热处理强度。本文建立高效液相色谱法,在波长280 nm下测定牛奶中糠氨酸的含量。试验结果表明：采用外标法定量,当浓度为0.06~4.00 mg/L,吸光度与糠氨酸质量浓度线性关系良好,相关系数R²=0.999 7;糠氨酸在5 mg/L、10 mg/L、15 mg/L、30 mg/L添加水平的回收率在91.48%~91.73%之间,RSD值在0.13%~0.37% 之间;方法的检出限为1.5 mg/100 g蛋白质,测得结果与第三方机构检测结果偏差小于1.00%,适用于乳品企业对糠氨酸的日常检测。     

Assessment of three automated assays for C-reactive protein determination in dogs

OBJECTIVE: To determine the characteristics of an automated canine C-reactive protein (CRP) assay and evaluate 2 human CRP assays for use in dogs. Animals-56 client-owned dogs with pyometra and 11 healthy control dogs. PROCEDURES: Samples from 11 dogs with high (> 100 mg/L) or low (< 10 mg/L) CRP concentrations (determined by use of a canine ELISA) were evaluated by use of the automated canine CRP assay. Intra- and interassay imprecision was determined (by use of those 2 plasma pools), and assay inaccuracy was assessed by use of logistic regression analysis of results obtained via ELISA and the automated canine CRP assay. Two automated human CRP assays were used to measure plasma CRP concentration in 10 dogs. RESULTS: By use of the ELISA, mean +/- SD plasma CRP concentration was 96.1 +/- 38.5 mg/L and 10.1 +/- 23.2 mg/L in dogs with pyometra and control dogs, respectively. The automated canine assay had intra-assay coefficients of variation (CVs) of 7.8% and 7.9%, respectively, and interassay CVs of 11.1% and 13.1%, respectively. Results from the automated assay were highly correlated with results obtained via ELISA. The human assay results did not exceed 0.4 mg/L in any dog. CONCLUSIONS AND CLINICAL RELEVANCE: The automated canine CRP assay had less interassay imprecision, compared with the ELISA. The 2 human CRP assays were not suitable for analysis of canine plasma samples. The automated canine CRP assay was more precise than the ELISA for serial evaluations of plasma CRP concentration in dogs.     

Effects of sample handling on adrenocorticotropin concentration measured in canine plasma, using a commercially available radioimmunoassay kit.

A commercially available radioimmunoassay (RIA) kit for measurement of human adrenocorticotropin (hACTH) was validated for use in dogs. Assay sensitivity was 3 pg/ml. Intra-assay coefficient of variation (x 100; CV) for 3 canine plasma pools was 3.0 (mean +/- SD, 33 +/- 0.99 pg/ml), 4.2 (71 +/- 2.4 pg/ml) and 3.7 (145 +/- 3.7 pg/ml) %. Interassay CV for 2 plasma pools measured in 6 assays was 9.8 (37 +/- 3.6 pg/ml) and 4.4 (76 +/- 3.4 pg/ml) %, respectively. Dilutional parallelism was documented by assaying 2 pools of canine plasma at 3 dilutions and correcting the measured result for dilution. Corrected mean concentrations for the first pool were 33 (+/- 0.99), 36 (+/- 4.3), and 33 (+/- 6.8) pg/ml; corrected mean concentrations for the second pool were 145 (+/- 5.4), 141 (+/- 10.8) and 125 (+/- 3.4) pg/ml. Recovery of 1-39hACTH added to canine plasma (6.25, 12.5, 25.0, 50.0, and 100.0 pg/ml) was linear and quantitative (slope = 0.890, R2 = 0.961). To test whether anticoagulant or the protease inhibitor, aprotinin, influences ACTH concentration in canine plasma, ACTH was measured in canine blood collected in 4 tubes containing anticoagulant: heparin (H), heparin + 500 kallikrein inhibitor units (KIU) of aprotinin/ml (HA), EDTA (E), and EDTA + aprotinin (EA). Plasma ACTH concentration was the same when samples containing H and HA, or HA and E were compared, and was significantly (P less than 0.01) lower in samples containing EA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Blood acid-base curve nomogram for immature domestic pigs

The purpose in this study was to characterize the acid-base status of arterial blood from healthy young domestic swine and to construct an acid-base curve nomogram appropriate to such animals. Accordingly, 40 immature, 20- to 31-kg domestic pigs were used to establish acid-base characteristics for arterial blood. Samples were collected from chronically implanted catheters while the animals were maintained under steady-state, near-basal conditions. At a measurement temperature of 38 C, pH averaged 7.496; PCO2, 40.6 mm Hg; [HCO3-], 31.6 mEq/L; PO2, 79.1 mm Hg; hemoglobin, 9.65 g/dl; hematocrit, 0.29; plasma albumin, 25.3 g/L; plasma globulin, 32.3 g/L; and plasma buffer base, 45.4 mEq/L. Hourly measurements over a 6-hour period in 6 of these pigs showed a small, but significant decrease in PO2 with time, but no significant change in acid-base status. The data showed that nomograms or other procedures based on blood characteristics of men were invalid when used to estimate base excess concentration of blood from young pigs. The normal pH of arterial blood was higher in immature pigs than in men; thus, reference values defining zero base excess were not equivalent in men and pigs. Constant PCO2 titrations were performed on arterial samples taken from 10 additional pigs, and the data were used to construct an acid-base curve nomogram in which zero base excess was defined for blood with a pH of 7.50 and a PCO2 of 40 mm Hg.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hematologic, biochemical, and protein electrophoretic values in captive tawny owls (Strix aluco)

BACKGROUND: The tawny owl (Strix aluco) is a protected species in Italy. Orphaned, injured, and ill owls often are sheltered and treated in rehabilitation centers, where hematologic and biochemical analyses would be helpful to evaluate and monitor the status of their health. OBJECTIVES: The major aim of this work was to assess hematologic and biochemical constituents together with protein electrophoretic fractions in healthy tawny owls. In addition, we compared laboratory methods for determining hemoglobin (Hgb), total protein, and albumin concentrations. METHODS: Heparinized blood samples were collected from 10 clinically healthy adult captive tawny owls between March 2001 and November 2003 for CBC, routine biochemical analysis, and protein electrophoresis. Alternate methods for Hgb (estimation as HCT/3 vs spectrophotometry), total protein (biuret vs refractometry), and albumin (bromcresol green vs electrophoresis) concentrations were compared in 34 samples from 16 unhealthy adult owls and 8 nestlings. RESULTS: Results were reported as mean, median, and range (minimum-maximum). Significant differences and poor concordance were observed between methods for Hgb, total protein, and albumin. CONCLUSIONS: Hematologic and plasma biochemical values in captive tawny owls may be useful in evaluating and monitoring the health of this species in captivity.     

Effect of extraction time and acid concentration on the separation of proglycogen and macroglycogen in horse muscle samples

The objective of this study was to determine whether the concentrations of proglycogen (PG) and macroglycogen (MG) in biopsy samples of horse muscle are influenced by extraction time or perchloric acid (PCA) concentration. In study 1, individual muscle-biopsy samples from 10 horses were divided into 4 parts each and then randomly subjected to 4 periods of extraction (10, 20, 60, or 120 min) with 1.5 M PCA. In study 2, individual muscle-biopsy samples from 6 horses were divided into 24 pieces each and then randomly subjected to 12 combinations of extraction time (10, 20, 30, or 40 min) and PCA concentration (0.5, 1.5, or 3.0 M). The results from study 1 indicated that PG and MG concentrations are affected only after extraction for 120 min; the PG concentration decreased significantly (P < 0.05), and the MG concentration increased (not significantly). In study 2, extraction in 3.0 M PCA yielded significantly lower PG and higher MG concentrations (P < 0.05) than extraction in 0.5 or 1.5 M PCA with each of the extraction times. The results of this study further support the existence of 2 glycogen pools and demonstrate that they are not an extraction artifact. The study also suggests that the 2 pools are stable during extraction over a range of extraction times and acid concentrations. However, if the exposure to acid is very long and, or, the acid concentration is high, some of the insoluble PG appears to be hydrolyzed and to enter the MG pool.     

Plasma/muscle ratios of sulfadimethoxine residues in channel catfish (Ictalurus punctatus)

Channel catfish ( n = 84) maintained at a water temperature of 27°C were used in a feeding study to determine the plasma to muscle concentration ratios of sulfadimethoxine (SDM) and 4-N-acetylsulfadimethoxine residues. Sulfadimethoxine medicated feed was provided free choice at 42 mg SDM/kg body weight once daily for 5 days and the plasma and muscle concentrations of SDM were determined at selected withdrawal times (6, 12, 24, 48, 72, and 96 hours) following the last dose. Considerable variation in total SDM tissue concentration among fish within a sampling period was observed. For fish ( n = 12) at six hours post-dose, total SDM concentrations ranged from 1.4–24.8 μg/mL and 0.6–12.6 μg/g, with mean total SDM concentrations of 9.1 μg/mL and 5.3 μg/g for plasma and muscle, respectively. However, a mean plasma:muscle concentration ratio of 1.8:1 ± 0.3:1 was obtained over all concentrations and sampling periods. The plasma:muscle 95% t distribution interval for individual fish was 1.2:1 to 2.4:1. A correlation coefficient of 0.967 was obtained for the relationship between plasma and muscle total SDM concentration among individual fish ( n = 25). Results of this study indicate that plasma total SDM concentration may be used to identify samples containing violative SDM muscle residue. No fish contained total SDM muscle residues greater than the FDA tolerance (0.1 μg/g) by 48 hours following the final dose.     

An evaluation of the copper sequestrated during clotting in cattle: is it just caeruloplasmin?

In cattle, sequestration of copper (Cu) occurs during the clotting process so that serum Cu concentrations are markedly lower than plasma Cu. It has been suggested that all of the Cu lost during clotting is caeruloplasmin (CP). This study used paired samples from 125 cattle to assess whether this assumption was correct. The regression equations for plasma CP activity against plasma Cu concentration and serum CP against serum Cu had significantly different intercepts suggesting that at zero CP activity the amount of Cu remaining was dependent on sample type. Furthermore, the difference between serum and plasma Cu was unrelated to Cu status, whereas the difference between serum and plasma CP was related to Cu status. The regression equation for the loss of CP activity against change in Cu concentration had an intercept that was different from zero, indicating that a reduction in Cu concentration could occur even if CP activity was unaffected by clotting.     

Pharmacokinetics of difloxacin after intravenous, intramuscular, and intragastric administration to horses

OBJECTIVE: To study the pharmacokinetics of difloxacin (5 mg/kg) following IV, IM, and intragastric (IG) administration to healthy horses. ANIMALS: 6 healthy mature horses. PROCEDURES: A crossover study design with 3 phases was used (15-day washout periods between treatments). An injectable formulation of difloxacin (5%) was administered IV and IM in single doses (5 mg/kg); for IG administration, an oral solution was prepared and administered via nasogastric tube. Blood samples were collected before and at intervals after each administration. A high-performance liquid chromatography assay with fluorescence detection was used to determine plasma difloxacin concentrations. Pharmacokinetic parameters of difloxacin were analyzed. Plasma creatine kinase activity was monitored to assess tissue damage. RESULTS: Difloxacin plasma concentration versus time data after IV administration were best described by a 2-compartment open model. The disposition of difloxacin following IM or IG administration was best described by a 1-compartment model. Mean half-life for difloxacin administered IV, IM, and IG was 2.66, 5.72, and 10.75 hours, respectively. Clearance after IV administration was 0.28 L/kg.h. After IM administration, the absolute mean +/- SD bioavailability was 95.81 +/- 3.11% and maximum plasma concentration (Cmax) was 1.48 +/- 0.12 mg/L. After IG administration, the absolute bioavailability was 68.62 +/- 10.60% and Cmax was 0.732 +/- 0.05 mg/L. At 12 hours after IM administration, plasma creatine kinase activity had increased 7-fold, compared with the preinjection value. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that difloxacin is likely to be effective for treating susceptible bacterial infections in horses.     

Artifactual changes in PCV, hemoglobin concentration, and cell counts in bovine, caprine, and porcine blood stored at room and refrigerator temperatures

BACKGROUND: Blood samples collected from farm animals for hematology testing may not reach the laboratory or be examined immediately upon collection, and in some cases may need to be transported for hours before reaching a laboratory. OBJECTIVE: The objective of this study was to investigate the artifactual changes that may occur in PCV, hemoglobin (Hgb) concentration, and cell counts in bovine, caprine, and porcine blood samples stored at room (30 degrees C) or refrigerator (5 degrees C) temperature. METHODS: Baseline values for PCV, Hgb concentration, and RBC and WBC counts were determined immediately after blood collection from 36 cattle, 32 goats, and 48 pigs using manual techniques. Blood samples were split into 2 aliquots and stored at 30 degrees C or 5 degrees C. Hematologic analyses were carried out at specified intervals during 120 hours of storage. Results were analyzed by repeated measure ANOVA; results at different temperatures were compared by paired t-tests. RESULTS: Compared to baseline values, there were no significant changes in Hgb concentration, RBC count, or WBC count in samples from cattle; in Hgb concentration and RBC count in samples from goats; and in Hgb concentration and WBC count in samples from pigs throughout the 120 hours of storage at both 30 degrees C and 5 degrees C. Significant changes (P <.05) from baseline occurred in PCV after 14 hours of storage at 30 degrees C and after 19 hours of storage at 5 degrees C in cattle and goats; and after 10 hours of storage at 30 degrees C and 14 hours of storage at 5 degrees C in pigs. Significant changes also were observed in Hgb concentration at 96 hours at 30 degrees C and 5 degrees C, and in RBC counts at 48 hours at 30 degrees C and 96 hours at 5 degrees C in porcine samples; and in total WBC counts at 120 hours at 30 degrees C and 5 degrees C in caprine samples. Artifactual changes were more pronounced in the samples stored at 30 degrees C. CONCLUSIONS: At both 30 degrees C and 5 degrees C, blood samples from cattle and goats can be stored for up to 12 hours, while blood samples from pigs can be stored for up to 8 hours without any significant changes in PCV. Blood samples from all 3 species can be stored for more than 24 hours without significant changes in Hgb concentration, RBC count, and total WBC count.