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Abstract

OBJECTIVE

To measure plasma and tissue activities of alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase (AST), creatine kinase, and γ-glutamyltransferase in 2 snake species.

ANIMALS

6 banded water snakes (Nerodia fasciata) and 6 diamondback water snakes (Nerodia rhombifer).

PROCEDURES

Blood was collected via the ventral tail vein to measure plasma enzyme activities. Animals were then euthanized, and samples of 9 tissues were collected from each snake: skeletal muscle, cardiac muscle, liver, spleen, lung, kidney, testicle, pancreas, and gallbladder. Tissues were frozen for 30 days, then homogenized and processed. Supernatants were collected and analyzed within 24 hours of processing. A linear mixed model was used to determine differences in enzyme activity between tissues and species and assess interactions between tissues and species.

RESULTS

Activities of all enzymes were found to differ significantly among tissues. There were also significant differences between species for all enzyme activities, except AST activity. The kidney had the highest alanine aminotransferase and γ-glutamyltransferase activities. Alkaline phosphatase activity was significantly highest in liver and kidney tissues than in other tissue. Creatine kinase activity was highest in skeletal muscle, followed by cardiac muscle and kidney. AST activity was present in all tissues evaluated, but was highest in liver, kidney, and cardiac muscle in both species.

CLINICAL RELEVANCE

Results reinforced the importance of characterizing the origin of tissue enzymes in reptiles to improve our understanding of biochemistry results and highlighted the differences that can exist in tissue enzyme activities between closely related species.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To develop a testing algorithm that incorporates multiple assays to evaluate host cellular and humoral immunity and antigen detection concerning Mycobacterium tuberculosis complex (MTBC) infection in captive nonhuman primates.

ANIMALS

Cohorts of captive-bred and wild-caught macaques from 5 different geographic regions.

PROCEDURES

Macaques were tested for MTBC infection by use of a γ interferon tuberculosis (GIFT) assay, an interferon-γ release assay, and other assays. In the first 2 cohorts (n = 15 and 181), initial validation of the GIFT assay was performed by use of experimentally infected and unexposed control macaques. In the next 3 cohorts (n = 59, 42, and 11), results were obtained for opportunistically collected samples from macaques exposed during spontaneous outbreaks.

RESULTS

Sensitivity and specificity of the GIFT assay in the control cohorts were 100% and 97%, respectively, and were variable but enhanced by incorporating results from multiple assays in spontaneous outbreaks.

CLINICAL RELEVANCE

The detection and management of MTBC infection in captive nonhuman primate populations is an ongoing challenge, especially with animal imports and transfers. Despite standardized practices of initial quarantine with regular intradermal tuberculin skin testing, spontaneous outbreaks continue to be reported. Since infection encompasses a range of disease manifestations over time, a testing algorithm that incorporates multiple assays, such as the GIFT assay, to evaluate host cellular and humoral immunity in addition to agent detection is needed. Testing a combination of samples from controlled studies and spontaneous outbreaks of MTBC infection in nonhuman primates would advance the development and validation of a functional algorithm that incorporates promising tools such as the GIFT assay.

Open access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate the effect of 1-Desamino-8-d-arginine vasopressin (DDAVP; desmopressin acetate) on platelet aggregation in healthy dogs receiving aspirin or clopidogrel.

ANIMALS

7 healthy staff-owned dogs.

PROCEDURES

In this randomized double-blinded crossover study, impedance aggregometry was performed on samples of lithium-heparinized whole blood samples from dogs before (T0) treatment with aspirin (1 mg/kg, PO, q 24 h for 4 days; ASP group) or clopidogrel (1 mg/kg, PO, q 24 h for 4 days; CLP group) and then before (T1) and after (T2) treatment with DDAVP (0.3 µg/kg, IV, once). There was a 14-day washout period before the crossover component. Aggregometry was performed with 4 different assays, each of which involved a different agonist reagent to stimulate platelet function: ADP, thrombin receptor activating peptide-6, arachidonic acid, or collagen type 1.

RESULTS

Median results for platelet aggregometry with agonist reagents ADP, arachidonic acid, or thrombin receptor activating peptide-6 significantly decreased between T0 and T1 for the CLP group; however, no meaningful difference in platelet aggregation was detected in the ASP group. Results for platelet aggregometry did not differ substantially between T1 and T2 regardless of treatment group or assay.

CONCLUSIONS AND CLINICAL RELEVANCE

Findings suggested that administration of DDAVP may have no effect on platelet aggregation (measured with platelet aggregometry) in healthy dogs treated with clopidogrel. Because no inhibition of platelet aggregation was detected for dogs in the ASP group, no conclusion could be made regarding the effects of DDAVP administered to dogs treated with aspirin.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To compare serum concentrations of histones and inflammatory markers in dogs with acute pancreatitis and healthy control dogs, investigate associations of these variables with coagulation test results and survival (vs nonsurvival) to hospital discharge, and examine the prognostic utility of clinical findings and routine laboratory and coagulation tests in affected dogs.

ANIMALS

36 dogs.

PROCEDURES

Dogs with findings consistent with acute pancreatitis (n = 29) and healthy control dogs (7) were enrolled in a prospective, observational study. Serum concentrations of histones, interleukin (IL)-6, and tumor-necrosis factor-α were assessed for all dogs. Clinical (including ultrasonographic) findings, relevant history, routine laboratory and coagulation test results, and outcomes were recorded for dogs with pancreatitis. Variables were assessed to determine an association with outcome for affected dogs and hospitalization time for survivors; histone concentrations and markers of inflammation were compared among survivors, nonsurvivors, and controls. Correlation between quantitative variables was investigated.

RESULTS

Serum histone and IL-6 concentrations did not differentiate survivors (n = 23) from nonsurvivors (6); IL-6 concentrations in affected dogs were correlated with 1,2-o-dilauryl-rac-glycero glutaric acid-(6′-methylresorufin) ester lipase activity (r S = 0.436) and hospitalization time (r S = 0.528). Pancreatitis-associated peritoneal fluid, obtundation, and jaundice were more common, and serum bilirubin concentration, serum alanine aminotransferase and aspartate aminotransferase activities, and prothrombin and activated partial thromboplastin times were greater in nonsurvivors than in survivors. Thromboelastometric changes consistent with hypercoagulability were detected in survivors; hypocoagulability was detected in 2 nonsurvivors.

CONCLUSIONS AND CLINICAL RELEVANCE

Serum histone concentrations were not associated with presence of acute pancreatitis or outcome for affected dogs. Further research is needed to investigate the clinical and therapeutic implications of hypocoagulability, hepatocellular injury, and pancreatitis-associated peritoneal fluid in affected dogs. (Am J Vet Res 2021;82:701–711)

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To assess agreement between 2 benchtop blood gas analyzers developed by 1 manufacturer (BGA 1 and BGA 2 [a newer model with reduced maintenance requirements]) and a reference chemistry analyzer for measurement of electrolyte (sodium, chloride, and potassium) in blood samples from dogs.

ANIMALS

17 healthy staff- and student-owned dogs and 23 client-owned dogs admitted to an emergency and intensive care service.

PROCEDURES

Blood collected by venipuncture was placed in lithium heparin–containing tubes. Aliquots were analyzed immediately with each BGA. Samples were centrifuged, and plasma was analyzed with the reference analyzer. Results for each BGA were compared with results for the reference analyzer by Passing-Bablok regression analysis. Percentage differences between BGA and reference analyzer results were compared with published guidelines for total allowable error.

RESULTS

Proportional bias was detected for measurement of chloride concentration (slope, 0.7; 95% CI, 0.7 to 0.8), and constant positive bias was detected for measurement of chloride (y-intercept, 34, mmol/L; 95% CI, 16.9 to 38 mmol/L) and potassium (y-intercept, 0.1 mmol/L; 95% CI, 0.1 to 0.2 mmol/L) concentrations with BGA 1. There was no significant bias for measurement of potassium or chloride concentration with BGA 2 or sodium concentration with either BGA. Differences from the reference analyzer result exceeded total allowable error guidelines for ≥ 1 sample/analyte/BGA, but median observed measurement differences between each BGA and the reference analyzer did not.

CONCLUSIONS AND CLINICAL RELEVANCE

Good agreement with reference analyzer results was found for measurement of the selected electrolyte concentrations in canine blood samples with each BGA.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To determine effects of PCV on blood glucose (BG) concentration measurements obtained with a human portable blood glucometer (HPBG) and a veterinary portable blood glucometer (VPBG) on canine (cVPBG) and feline (fVPBG) settings (test methods) when used in rabbits and to develop correction formulas to mitigate effects of PCV on such measurements.

SAMPLE

48 resuspended blood samples with known PVCs (range, 0% [plasma] to 92% [plasma and packed RBCs]) from 6 healthy research rabbits (experimental sample set) and 252 historic measurements of BG concentration and PCV in 84 client-owned rabbits evaluated at a veterinary hospital (validation data set).

PROCEDURES

Duplicate measurements of BG concentration with each test method and of PCV were obtained for each sample in the experimental sample set, and the mean results for each variable for each test method and sample were compared with results from a clinical laboratory analyzer (reference method) used to determine the true BG concentration for each sample. Mean ± SD differences in measurements between the reference and test methods were calculated. Linear regression and modified Clarke error grid analysis were used to develop correction formulas for the test methods given known PCVs, and these formulas were evaluated on the validation data set with linear regression and a modified Clarke error grid.

RESULTS

Blood glucose concentrations were falsely low for cVPBG and fVPBG used on samples with PCV < 31% and were falsely high for all test methods used on samples with PCV > 43%. Compared with original measurements, formula-corrected measurements overall had better agreement with reference method measurements for the experimental sample set; however, only the formula-corrected HPBG measurements had improved agreement for the validation data set.

CONCLUSIONS AND CLINICAL RELEVANCE

Findings indicated that, in rabbits, HPBG measurements had improved accuracy with the use of the correction formula HPBG measurement of BG concentration + ([0.75 × PCV] − 15); however, the correction formulas did not improve the accuracy of VPBG measurements, and we believe that neither the cVPBG nor fVPBG should be used in rabbits.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To assess the effect of packed RBC (pRBC) transfusion on thromboelastographic (TEG) tracings in dogs with naturally occurring anemia.

ANIMALS

22 clinically anemic dogs that received a pRBC transfusion.

PROCEDURES

For each dog, a blood sample was collected before and within 3 hours after completion of the pRBC transfusion for a CBC, nonactivated TEG analysis, and measurement of blood viscosity. Wilcoxon signed rank tests were used to compare CBC, viscosity, and TEG variables between pretransfusion and posttransfusion blood samples. Multivariable linear regression was used to assess the effects of pretransfusion-posttransfusion changes in Hct, WBC count, and platelet count on changes in TEG variables.

RESULTS

Median posttransfusion Hct (21%; range, 13% to 34%) was significantly greater than the median pretransfusion Hct (12.5%; range, 7% to 29%). Packed RBC transfusion was associated with a median increase in Hct of 6.2% (range, 1.2% to 13%). Maximum amplitude significantly decreased from 74.9 to 73.8 mm and clot strength significantly decreased from 14,906 to 14,119 dynes/s after pRBC transfusion. Blood viscosity significantly increased, whereas platelet and WBC counts significantly decreased after transfusion. Multivariable linear regression revealed that pretransfusion-posttransfusion changes in Hct, WBC count, and platelet count were not associated with changes in TEG variables.

CONCLUSIONS AND CLINICAL RELEVANCE

Results indicated that pRBC transfusion had only small effects on the TEG tracings of hemodynamically stable dogs. Therefore, large changes in TEG tracings following pRBC transfusion are unlikely to be the result of the transfusion and should be investigated further.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To develop and validate a sandwich ELISA for the measurement of α1-proteinase inhibitor (α1-PI) concentrations in serum and fecal samples obtained from common marmosets (Callithrix jacchus).

SAMPLE Leftover serum (n = 42) and fecal (23) samples submitted for diagnostic testing; paired serum and fecal samples obtained from 30 common marmosets at 2 research colonies.

PROCEDURES A sandwich ELISA was developed and analytically validated by determining the lower limit of detection, linearity, accuracy, precision, and reproducibility. Reference intervals for α1-PI concentrations in serum and feces of common marmosets were calculated.

RESULTS The standard curve was generated for concentrations between 1 and 100 ng/mL. Mean ± SD observed-to-expected ratio for serial dilutions of serum and fecal samples was 117.1 ± 5.6% (range, 112.2% to 123.0%) and 106.1 ± 19.7% (range, 82.6% to 130.2%), respectively. Mean observed-to-expected ratio for spiking recovery of serum and fecal samples was 102.9 ± 12.1% (range, 86.8% to 115.8%) and 97.9 ± 19.0% (range, 83.0% to 125.1%), respectively. Reference interval for serum concentrations of α1-PI was 1,254 to 1,813 μg/mL, for 3-day mean fecal concentrations was 11.5 to 42.2 μg/g of feces, and for 3-day maximum fecal concentrations was 13.2 to 51.2 μg/g of feces.

CONCLUSIONS AND CLINICAL RELEVANCE The ELISA was linear, accurate, precise, and reproducible for quantification of α1-PI concentrations in serum and feces of common marmosets. However, the ELISA had limited linearity and accuracy for spiking recovery of fecal samples.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To determine reference intervals for total nucleated cell count, total protein concentration, pH, RBC count, and percentages of neutrophils, lymphocytes, and large mononuclear cells in synovial fluid samples (SFSs) obtained from the carpal and tarsal joints of healthy swine.

ANIMALS 54 healthy commercial finisher pigs that had no evidence of lameness or gross joint swelling.

PROCEDURES Each pig was anesthetized, and SFSs were collected from 1 carpal and 1 tarsal joint for fluid analysis, cytologic evaluation, bacterial culture, and PCR analyses for common swine joint pathogens. Each pig was euthanized after SFS collection, and synovial tissue samples were collected for histologic assessment. If necessary, postmortem SFSs were collected.

RESULTS Overall, 37 of 50 tarsal and 46 of 53 carpal SFSs met inclusion criteria of sufficient volume, no gross blood contamination, and negative results of bacterial culture and PCR analyses, and were from joints with histologically normal synovial tissues. For the carpal and tarsal joints, upper reference limits were as follows: total nucleated cell count, 3,281 cells/μL and 2,368 cells/μL, respectively; total protein concentration, 3.6 g/dL and 3.6 g/dL, respectively; pH, 7.2 and 7.0, respectively; RBC count, 0.8 × 106 cells/μL and 0.1 × 106 cells/μL, respectively; and percentage of neutrophils, 46.5% and 33.7%, respectively; percentage of lymphocytes, 40.6% and 56.3%, respectively; and percentage of large mononuclear cells, 92.0% and 95.3%, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE Results have provided reference intervals for selected variables in SFSs obtained from the carpal and the tarsal joints of healthy swine, which should be useful in diagnostic investigations of swine lameness and arthritis.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To investigate effects of storage duration and temperature on biochemical analytes in plasma from red-eared sliders (Trachemys scripta elegans).

ANIMALS 8 red-eared sliders.

PROCEDURES Blood samples were collected. Plasma was harvested and analyzed at room temperature (approx 23°C; time = 1 hour) and then fractioned into 0.1-mL aliquots that were stored at room temperature or were refrigerated (4°C) or frozen (−20°C). Biochemical analysis of stored samples was performed at 4 (room temperature), 8 (4°C), 24 (4°C), 48 (4° and −20°C), and 72 (−20°C) hours and at 7 days (−20°C). For each time point for each storage temperature, bias was calculated by subtracting values from the value obtained at 1 hour. Bias was modeled by use of a linear mixed model.

RESULTS Storage temperature had a significant effect on several plasma biochemical analytes. In general, aspartate aminotransferase activity and uric acid, total protein, and potassium concentrations increased after storage at 4° and −20°C. Differences in values after storage were mostly within the acceptable range for allowable total error, except for calcium and potassium concentrations for samples stored at −20°C. Both storage temperatures increased variability of measurement results. Results for samples stored at room temperature for 4 hours did not differ significantly from values at 1 hour. Results differed significantly between refrigerated and frozen samples stored for 48 hours.

CONCLUSIONS AND CLINICAL RELEVANCE Short-term storage conditions influenced results for some biochemical analytes. These effects should be considered when performing biochemical analyses of plasma samples obtained from red-eared sliders.

Restricted access
in American Journal of Veterinary Research