Preliminary evaluation of a gel tube agglutination major cross-match method in dogs.

BACKGROUND: A major cross-match gel tube test is available for use in dogs yet has not been clinically evaluated. OBJECTIVES: This study compared cross-match results obtained using the gel tube and the standard tube methods for canine samples. METHODS: Study 1 included 107 canine sample donor-recipient pairings cross-match tested with the RapidVet-H method gel tube test and compared results with the standard tube method. Additionally, 120 pairings using pooled sera containing anti-canine erythrocyte antibody at various concentrations were tested with leftover blood from a hospital population to assess sensitivity and specificity of the gel tube method in comparison with the standard method. RESULTS: The gel tube method had a good relative specificity of 96.1% in detecting lack of agglutination (compatibility) compared to the standard tube method. Agreement between the 2 methods was moderate. Nine of 107 pairings showed agglutination/incompatibility on either test, too few to allow reliable calculation of relative sensitivity. Fifty percent of the gel tube method results were difficult to interpret due to sample spreading in the reaction and/or negative control tubes. CONCLUSIONS: The RapidVet-H method agreed with the standard cross-match method on compatible samples, but detected incompatibility in some sample pairs that were compatible with the standard method. Evaluation using larger numbers of incompatible pairings is needed to assess diagnostic utility. The gel tube method results were difficult to categorize due to sample spreading. Weak agglutination reactions or other factors such as centrifuge model may be responsible.

Comparison of wet-mount, Wright-Giemsa and Gram-stained urine sediment for predicting bacteriuria in dogs and cats.

This study assessed the standard urinalysis technique and sediment stain techniques as predictors of bacterial culture results for canine and feline urine. Canine (n = 111) and feline (n = 79) urine samples were evaluated using unstained wet-mount and air-dried Gram and Wright-Giemsa stained sediment; results were compared to aerobic bacterial culture. Eleven canine and 7 feline urine samples were culture positive. Unstained wet-mount and stained sediment had sensitivities of 89% and 83% and specificities of 91% and 99%, respectively. The specificity of using either stain was higher (P < 0.01) than wet-mount examination for detecting bacteriuria. There were significant differences among 3 technologists in detecting true positives (P < 0.01). Association of sediment and culture results used 112 canine and 81 feline samples. There was a negative association (P < 0.01) between lipid detection and wet-mount identification of bacteria.

Comparaison de sédiments d'urine à l'état frais, avec la coloration Wright-Giemsa et la coloration de Gram pour la prédiction de la bactériurie chez les chiens et les chats. Cette étude a évalué la technique d'analyse urinaire standard et des techniques de coloration du sédiment d'urine comme prédicteurs des résultats de la culture bactérienne. Les échantillons d'urine canine (n = 111) et féline (n = 79) ont été évalués en utilisant des sédiments à l'état frais et des sédiments séchés à l'air avec coloration de Gram et de Wright-Giemsa; les résultats ont été comparés à une culture bactérienne aérobie. Onze échantillons d'urine canine et 7 échantillons d'urine féline ont obtenu des résultats positifs pour la culture. Le sédiment à l'état frais non coloré et le sédiment coloré présentaient des sensibilités de 89 % et de 83 % et des spécificités de 91 % et de 99 %, respectivement. La spécificité de l'utilisation de l'une ou l'autre de la coloration était supérieure (P < 0,01) à celle de l'examen à l'état frais pour la détection de la bactériurie. Il y avait une différence significative entre les technologues pour la détection des vrais positifs (P < 0,01). L'association des résultats des sédiments et des cultures a utilisé 112 échantillons canins et 81 échantillons félins. Il y avait une association négative (P < 0,01) entre la présence de lipide et l'examen à l'état frais des bactéries.(Traduit par Isabelle Vallières).

Comparison of white and red blood cell estimates in urine sediment with hemocytometer and automated counts in dogs and cats.

BACKGROUND: Therapeutic decisions regarding urinalysis are commonly based on the presence of white and red blood cells. Traditionally, numbers per high-power field are estimated using wet-mount microscopic examination. This technique is not standardized and counts are likely prone to inaccuracy. In addition, differentiation of leukocyte types is not possible. OBJECTIVES: The aims of this study were to (1) compare WBC and RBC estimates using wet-mount examination with counts obtained using a hemocytometer, (2) assess if a hematology automated analyzer (Sysmex ST-2000iV/XT) provides reliable WBC and RBC counts in urine comparable to hemocytometer counts, and (3) evaluate air-dried Wright-Giemsa-stained urine drop sediment preparations for the determination of differential leukocyte counts. METHODS: WBC and RBC counts were obtained by performing wet-mount estimates, manual hemocytometer counts, and Sysmex automated counts on 219 canine and feline urine samples. Results were correlated using Spearman rank correlation. Air-dried Wright-Giemsa stained sediment drop preparations (n = 215) were examined for differential counts of leukocytes. RESULTS: A low but significant association was found between WBC estimates on wet-mount examination and hemocytometer counts (rho = 0.37, P < .01). There was a high and significant association when RBC counts were compared between wet-mount and hemocytometer evaluation (rho = 0.7, P < .01). There was very high and significant interassay correlation between Sysmex data from duplicate samples for what the analyzer classified as WBC (rho = 0.97, P < .01) and RBC (rho = 0.94, P < .01). Low correlations were found between the Sysmex RBC counts and both wet-mount estimates and hemocytometer RBC counts (rho = 0.43, P < .01 and rho = 0.39, P < .01, respectively). Cell preservation in the air-dried sediment preparations was so poor that differential counts could not be performed. CONCLUSION: WBC and RBC estimates on wet-mount examination agreed with hemocytometer counts and are therefore considered adequate. The Sysmex ST-2000iV/XT did not provide reliable cell counts under the conditions used.

Effects of in vitro hemodilution of canine blood on platelet function analysis using the PFA-100.

BACKGROUND: The platelet function analyzer (PFA)-100 is a point-of-care instrument previously evaluated in humans and dogs. In both species, artificially prolonged platelet closure time (CT) occurs with anemia. Reliability of the analyzer in dogs becomes a concern when the HCT is between 0.25 and 0.35 L/L. OBJECTIVE: The objective of this study was to further define the level of HCT at which CT is prolonged, using in vitro diluted canine blood. METHODS: Citrated whole blood samples were collected from 22 healthy dogs. Initial HCT was determined and autologous platelet-rich plasma was added to samples to achieve HCTs of 0.33, 0.30, and 0.27 L/L. CT was determined in duplicate on the PFA-100 using collagen/adenosine-5'-diphosphate cartridges. RESULTS: Compared with the initial CT in samples with HCT 0.39-0.54 L/L (CT mean+/-SD=57.8+/-5.75 seconds), significantly prolonged CTs were found in hemodiluted samples with HCT 0.33 L/L (61.1+/-4.64 seconds), 0.30 L/L (64.3+/-6.79 seconds), and 0.27 L/L (70.8+/-7.90 seconds) (P=0.029; repeated measures ANOVA). CONCLUSION: Although statistical differences were found, further studies are needed to determine the clinical significance of the mild prolongation in CT associated with mild anemia. Until then, dogs with HCTs slightly <0.35 L/L should be evaluated cautiously for platelet dysfunction using the PFA-100.

Evaluation of platelet function in dogs with cardiac disease using the PFA-100 platelet function analyzer.

BACKGROUND: Cardiac disease has the potential to alter platelet function in dogs. Evaluation of platelet function using the PFA-100 analyzer in dogs of multiple breeds and with a broad range of cardiac conditions would help clarify the effect of cardiac disease on platelets. OBJECTIVES: The objective of this study was to assess differences in closure time (CT) in dogs with cardiac disease associated with murmurs, when compared with that of healthy dogs. METHODS: Thirty-nine dogs with cardiac murmurs and turbulent blood flow as determined echocardiographically were included in the study. The dogs represented 23 different breeds. Dogs with murmurs were further divided into those with atrioventricular valvular insufficiency (n=23) and subaortic stenosis (n=9). Fifty-eight clinically healthy dogs were used as controls. CTs were determined in duplicate on a PFA-100 analyzer using collagen/ADP cartridges. RESULTS: Compared with CTs in the control group (mean+/-SD, 57.6+/-5.9 seconds; median, 56.5 seconds; reference interval, 48.0-77.0 seconds), dogs with valvular insufficiency (mean+/-SD, 81.9+/-26.3 seconds; median, 78.0 seconds; range, 52.5-187 seconds), subaortic stenosis (71.4+/-16.5 seconds; median, 66.0 seconds; range, 51.5-95.0 seconds), and all dogs with murmurs combined (79.6+/-24.1 seconds; median, 74.0 seconds; range, 48.0-187 seconds) had significantly prolonged CTs (P<.01). CONCLUSIONS: The PFA-100 analyzer is useful in detecting platelet function defects in dogs with cardiac murmurs, most notably those caused by mitral and/or tricuspid valvular insufficiency or subaortic stenosis. The form of turbulent blood flow does not appear to be an important factor in platelet hypofunction in these forms of cardiac disease.

Evaluation of ionized and total serum magnesium concentrations in hyperthyroid cats.

Hyperthyroidism can increase the renal excretion of magnesium and thus cause hypomagnesemia in various species. Anaerobically collected blood samples from 15 hyperthyroid and 40 normal, healthy cats were analyzed with an ion-selective electrode analyzer and a serum biochemical analyzer. There was no significant difference in ionized or total serum magnesium concentration between the 2 groups, but there was a significant difference (P = 0.004) in the ratio of ionized to total serum magnesium concentrations between the healthy cats and the hyperthyroid cats with thyroxine (T4) concentrations at or above the median. There was a significant correlation (r = 0.894, P = 0.000) between the ionized and total magnesium concentrations in the hyperthyroid cats. The hyperthyroid cats had a significantly lower (P = 0.003) total serum protein concentration than the healthy cats. A significant negative correlation (r = -0.670, P = 0.006) was detected between the ionized magnesium and logarithmically transformed total T4 concentrations in the hyperthyroid cats, which suggests that the severity of hyperthyroidism may contribute to a decrease in the ionized magnesium concentration.

Validation of the Nova CRT8 for the measurement of ionized magnesium in feline serum.

BACKGROUND: Evaluation of serum magnesium (Mg) concentration is becoming important in human and veterinary critical care medicine. An ion-selective electrode can measure the physiologically active ionized fraction. OBJECTIVES: The purpose of this study was to validate an ion-specific electrode analyzer and assay for measuring ionized Mg in feline serum and to determine a reference interval for this analyte in cats. METHODS: Venous blood samples were collected anaerobically from clinically healthy cats, and the serum was used to validate the analyzer and assay. This included investigating the stability of samples stored at different temperatures, intra- and interassay precision, linearity, analytical sensitivity, and potential interferences from bilirubin, lipemia, hemoglobin, or serum separator tubes. A reference interval was calculated. RESULTS: Serum samples evaluated for ionized Mg concentrations can be stored at 20 degrees C for < or =24 hours, at 4 degrees C for < or =72 hours, and at 20 degrees C for < or =4 weeks, when samples are minimally exposed to air. Intra- and interassay precisions had coefficients of variation (CVs) of 1.23% and 2.02%, respectively. There was good linearity using serum (r = .998; y = -0.0057 + 1.0256x) and manufacturer-supplied aqueous solutions and quality control materials (r = .999; y = 0.0110 + 0.9213x). Apparent analytical sensitivity was at least 0.015 mmol/L. Mean recovery was good for ionized Mg in samples with 1+ icterus (104%), 4+ lipemia (99.3%) and 1-4+ hemolysis (98.6%). There was no significant difference (P = .52) in ionized Mg concentrations in serum collected in tubes containing no additives compared with serum collected in glass separator tubes. The serum ionized Mg reference interval was 0.47-0.63 mmol/L (n = 40). CONCLUSIONS: The Nova CRT8 analyzer and assay provide a precise and reliable method of measuring ionized Mg concentration in feline serum. Strict adherence to sampling techniques, handling, and storage are necessary for reliable results.

Evaluation of an automated, homogeneous enzyme immunoassay for serum thyroxine measurement in dog and cat serum.

A homogenous enzyme immunoassay (EIA) for measurement of serum thyroxine (T4) concentration was evaluated for use with canine and feline serum. The EIA method was linear from 0 to 150 nmol T4/L for human serum, 0 to 94 nmol T4/L for feline serum and 10 to 60 nmol T4/L for canine serum. Intra- and interassay precision studies yielded coefficients of variation

Agarose gel electrophoresis of alkaline phosphatase isoenzymes in the serum of hyperthyroid cats.

Cats with hyperthyroidism [(increased serum thyroxine (T(4))] commonly have increased serum alkaline phosphatase (ALP) activity in addition to other serum biochemical abnormalities. Serum biochemical profiles were obtained from 10 hyperthyroid cats which had increased serum ALP. Agarose gel electrophoresis of serum from these cats was performed and stained for alkaline phosphatase activity. Alkaline phosphatase activity was calculated for each of the separate bands obtained, and the results were compared to those of tissue extracts, serum from normal cats, and serum from normothyroid cats with increased serum ALP activity. The hyperthyroid cats had increased ALP activity in bands corresponding to isoenzymes originating in the liver, bone, and an unidentified tissue source.

Stability of sorbitol dehydrogenase activity in bovine and equine sera.

Serum sorbitol dehydrogenase (SDH) activities in 10 cows and nine horses were measured using an automated clinical analyzer. The serum samples were divided into aliquots that were stored at room temperature (21 degrees C), refrigerated (0-5 degrees C), or frozen (-30 degrees C). The stability of the SDH activity was monitored at various intervals. SDH activity in bovine sera remained stable for at least 5 hours at room temperature, 24 hours refrigerated, and 72 hours frozen without any significant (p < 0.05) differences from the initial serum values. In equine sera, SDH activity remained stable for at least 5 hours at room temperature and 48 hours frozen. The activity of the refrigerated equine sera was stable for at least 5 hours but less than 24 hours. An evaluation of fresh bovine serum and heparinized plasma samples indicated that there was no significant difference (p < 0.05) between the two sampling methods and that either may be employed for automated measurement of SDH activity following the established protocol. Sample type comparison indicated that there was a small but statistically significant (p < 0.05) difference between the results obtained comparing fresh serum and heparinized plasma samples for the horse. A reference range for Holstein cows was established using sera from 71 clinically healthy cattle (mean -/+ 2 SD = 32 -/+ 26 U/L).