An Anticoagulant for Transport of Bovine Blood

Various anticoagulant were assessed in order to develop a transport media for bovine blood which would improve leukocyte stability. The effects of time, disease in the donor and different personnel and laboratory equipment were also tested.

The transport media most effective for leukocyte stability was found to be Titriplex IV. This is a modification of Titriplex I by the inclusion of acetylsalicyclic acid to prevent cellular clumping and by the removal of methylene blue. Anticoagulant citrate dextroseformalin-ASA performed equally well for transport but cellular morphology was impaired.

Febrile or neoplastic disease in the donor animal did not alter the efficiency of the formalin containing anticoagulants.

     

Anticoagulant‐dependent in vitro hemagglutination in a cat

Abstract: A 17‐year‐old domestic shorthaired cat was presented to the Cornell University Hospital for Animals for recheck of hyperthyroidism previously treated with radioiodine. Marked agglutination was noted in a blood sample collected into EDTA for a CBC; no other clinical or hematologic evidence of hemolysis was observed and none developed despite persistent agglutination in additional EDTA samples collected over a 2‐month period. Blood drawn into heparin and sodium citrate tubes did not have grossly or microscopically detectable agglutination, unless EDTA was added. Plasma from the cat induced agglutination of washed RBCs from a control cat in the presence of EDTA but not in the presence of heparin. Flow cytometric analysis of samples created by mixing plasma from the patient with washed RBCs from a control cat showed immunoglobulin coating of the control RBCs, predominantly by IgM. These findings suggested an anticoagulant‐dependent antibody‐mediated mechanism for the agglutination. EDTA‐dependent hemagglutination has not been reported previously in cats, although rare cases have been described in humans. The phenomenon needs to be recognized as an in vitro occurrence in order to prevent erroneous diagnosis of immune‐mediated hemolytic anemia.     

Flow cytometric detection of activated platelets in the dog

Background: Platelet activation appears to play a role in a variety of canine thrombotic disorders. At present, tests for the detection of activated platelets are not used routinely in veterinary clinical laboratories. Objective: The purpose of this study was to develop a clinically applicable method to detect activated canine platelets. Methods: A flow cytometric assay was developed to detect activated platelets, platelet aggregates, and platelet microparticles in the dog. Blood was collected from healthy dogs using EDTA or sodium citrate as the anticoagulant, and platelet‐rich plasma was prepared. Platelets were activated by adding phorbol myristate acetate. In some experiments, platelets were fixed by incubation with 0.5% paraformaldehyde. In other experiments, platelets were stored for 4 or 24 hours at 4°C before analysis. Activated platelets were detected by measuring surface expression of P‐selectin and by determining the percentages of platelet aggregates and microparticles using forward‐angle vs side‐angle light scatter plots. Results were analyzed by using 2‐way ANOVA and the SchefféF‐test. Results: Platelets collected in EDTA had minimal expression of P‐selectin, whereas platelets collected in sodium citrate had greater median fluorescence intensity. Fixation with 0.5% paraformaldehyde before labeling platelets with anti‐P‐selectin did not affect antibody binding or the percentages of platelet aggregates and microparticles. Storage of platelet‐rich plasma at 4°C for 4 hours did not affect antibody binding or the percentages of platelet aggregates or microparticles. Activation of platelets ex vivo by addition of 10 ng/mL phorbol myristate acetate resulted in a large increase in expression of P‐selectin but only slight increases in platelet aggregates and microparticles. Conclusion: Determination of platelet P‐selectin expression and percentages of platelet aggregates and platelet microparticles may provide a clinically applicable means for detection of activated platelets in dogs. The capacity to use EDTA‐anticoagulated blood samples and to fix platelets for evaluation at a later time makes the test attractive as a routine diagnostic tool.     

Pseudothrombocytopenia secondary to the effects of EDTA in a dog

Pseudothrombocytopenia (PTCP) secondary to the effects of ethylenediaminetetraacetic acid (EDTA) has been noted in horses and pigs and should be considered in dogs with moderate thrombocytopenia and no clinical bleeding tendency. This type of pseudothrombocytopenia is not a pathological process by itself, but it can be clinically significant if diagnostics and medical treatments are initiated based on the reported thrombocytopenia. Platelet clumping occurs with EDTA-dependent PTCP, resulting in inaccurate hematology analyzer platelet concentrations. A nontraumatic venipuncture may be sufficient to obtain an accurate platelet count. However, rare cases in the dog may require blood drawn into a different anticoagulant, such as sodium citrate, to help discriminate a true thrombocytopenia from PTCP.     

Effects of toxic and non‐toxic blue mussel meal on health and product quality of laying hens

Blue mussels (Mytilus edulis) are a potential protein source in poultry feeds because of their high methionine and cystein content. However, mussels may temporarily be contaminated with the toxin, okadaic acid. The aim of this small scale study was to evaluate the effects on animal health and morphology of the digestive tract of laying hens fed three different diets; a commercial feed and diets containing 15% normal or 15% toxic mussel meal, i.e. a concentration of 198.6 μg toxin per kg feed. Twelve laying hens were divided into six groups and fed the experimental diets for 8 weeks. Animal health, production performance and egg quality were recorded. At sacrifice, tissue samples were prepared for histological evaluation using light microscopy. Okadaic acid did not have any adverse effects on animal health or production parameters and no histological changes indicating disturbances in the digestive tract was observed. Internal egg quality was improved as eggs from hens fed either of the two mussel meal diets had an increased yolk colour. These results show that mussel meal functions as a novel protein source for laying hens and those toxic mussels at this level may be included in the feed without negative effects on parameters evaluated in this study.     

Evaluation of a citrate-based anticoagulant with platelet inhibitory activity for feline blood cell Counts

Abstract: Aggregation of feline platelets in vitro results in difficulty assessing platelet number. A citrate-based anticoagulant containing the platelet inhibitors theophylline, adenosine, and dipyridamole (CTAD; Diatube-H, Becton Dickinson, Oxford, UK) has been developed for use in human platelet studies and heparin assays. To evaluate the efficacy of CTAD in reducing platelet aggregation in feline blood samples, aliquots of blood from 51 cats were anticoagulated with EDTA, CTAD, and for 12 samples, citrate solution. Samples preserved in CTAD had significantly higher (P ≤ .001) platelet counts, as determined by an impedance counter, hemacy-tometer, and smear estimation, than samples preserved in EDTA. In addition, subjective assessment of blood smears showed significantly fewer platelet aggregates (P<.001) in CTAD-treated samples compared with EDTA samples. Although values were similar, automated platelet counts and smear estimates of platelet number were significantly higher (P < .05) and platelet aggregation was significantly less (P < .05) in CTAD samples than in citrate samples. These results suggest that the platelet inhibitory activity of CTAD reduced feline platelet aggregation. Automated total WBC counts in CTAD samples were significantly lower (P<.001) than automated counts in EDTA samples but were similar to manual WBC counts in EDTA samples. Differences in both platelet and WBC counts between CTAD and EDTA or citrate samples were clinically relevant. Mean platelet volume and MCV were significantly lower (P< .05) in CTAD samples than in EDTA samples. No effect was seen on cell morphology or staining characteristics. The anticoagulant CTAD offers an advantage over both EDTA and citrate for feline hematologic analysis, by decreasing pseudothrombocytopenia and pseudoleukocytosis.     

Chemical form of dietary l‐Carnitine affects plasma but not tissue Carnitine concentrations in male Sprague–Dawley rats

In Experiment 1, rats (n = 54) were randomly assigned to control or one of the four sources of l ‐Carnitine supplemented at either 100 or 200 μmol/kg/day and were allowed to acclimate for 14 days. Following a 12‐h fast, plasma samples were obtained at 0, 5, 10, 15, 30, 60, 120, 240, 480 and 720 min after l ‐Carnitine feeding and assayed for free l ‐Carnitine concentration. Plasma‐free l ‐Carnitine levels were affected by time after treatment intake (p < 0.0001) and l ‐Carnitine source (p < 0.0001). The time × source interaction was not statistically significant (p = 0.99). In Experiment 2, rats (n = 54) were randomly assigned to control or one of the four sources of l ‐Carnitine at either 100 or 200 μmol/kg/day and were acclimated as in experiment 1. Rats were sacrificed 120 min after feeding. Samples of liver and skeletal muscle were obtained and assayed for free l ‐Carnitine concentration. Neither skeletal muscle (p = 0.44) or liver (p = 0.59) tissue concentrations of l ‐Carnitine were affected by any l ‐Carnitine source as compared with the control. We conclude that some differences exist in plasma concentrations of free l ‐Carnitine following ingestion of different chemical forms of l ‐Carnitine. It is unclear if these differences in the circulating concentration of free l ‐Carnitine translate into any physiological differences for the animal. In this study, chemical form of l ‐Carnitine had no effect on skeletal muscle or liver tissue concentrations of l ‐Carnitine in young male Wistar rats.     

Application of the slope‐ratio growth assay technique to estimate tryptophan availability in soybean meal fed to young rats

The slope‐ratio assay for rat was used to determine whether tryptophan (Trp) availability in soybean meal (SBM) is affected by the presence of other amino acids (AAs). In a preliminary study, rats were fed graded levels of Trp‐supplemented diets to establish the Trp concentration range over which the weight gain response was linear. This range was found to be from 0.04% to 0.12% Trp. Subsequently, rats were fed basal (0.045% Trp) or Trp‐supplemented diets from three different sources: l ‐Trp alone, SBM, or l ‐Trp mixed with other AAs to reflect AA levels in the test SBM (AA‐mix). Weight gain in rats increased linearly with supplemental Trp intake (p < .05) for all Trp sources. Compared to the slope achieved with l ‐Trp alone, the estimated availability of Trp in SBM was 84.4%, while for the AA‐mix it was 93.4%. It is evident that the 6.6% reduction in l ‐Trp availability in AA‐mix is due to metabolic costs derived from excess levels of other AAs beside Trp, given that the absorption of crystalline l ‐Trp in the small intestine is 100%. In conclusion, the Trp availability of SBM was estimated to be around 90.4% (i.e., 84.4/93.4 × 100) after correcting for the effects of the other AAs in SBM.     

Effect of dosage and application mode of l‐carnitine on plasma lipid and egg‐yolk cholesterol of turkeys,hatchability of eggs and post‐hatch growth of their offsprings

The effect of dosage and application mode of l ‐carnitine on plasma lipid and egg‐yolk cholesterol of breeder turkeys, hatchability of eggs and post‐hatch growth response was investigated using 180 breeder hens. The hens were assigned to six dietary treatments in a 2 × 3 factorial arrangements of two application modes of l ‐carnitine (diet and drinking water) supplemented at 0, 50 and 100 ppm (mg/kg or mg/l) levels, respectively. Each treatment was replicated five times with six hens per replicate. Dietary inclusion of 50 ppm l ‐carnitine showed the lowest (p < 0.01) plasma total cholesterol (TC) and low‐density lipoprotein concentration (LDL). Breeder hens offered 50 ppm l ‐carnitine with no regard to application mode recorded the highest (p < 0.01) plasma high‐density lipoprotein (HDL). Hens offered 50 and 100 ppm l ‐carnitine irrespective of application mode also showed reduced (p < 0.01) egg‐yolk TC concentration at 32 weeks of age. Dietary supplementation of 50 ppm l ‐carnitine for breeder turkeys recorded the lowest (p < 0.01) egg‐yolk triglyceride (TG) at 40 weeks of age. Hens offered 50 ppm l ‐carnitine irrespective of application mode recorded the highest (p < 0.05) hen‐day egg production. Incidence of dead‐in‐shell also reduced (p < 0.05) with increasing dosage of l ‐carnitine. Dietary supplementation of 50 ppm and oral application in drinking water of 100 ppm l ‐carnitine for breeder turkeys resulted in highest (p < 0.05) egg fertility. Offsprings from breeder hens fed diets supplemented with l ‐carnitine recorded no post‐hatch mortality. Highest (p < 0.05) post‐hatch final live weight and weight gain was obtained with poults obtained from hens fed diet supplemented with 50 ppm l ‐carnitine. In conclusion, dietary supplementation of 50 ppm l ‐carnitine for turkey hens showed improved serum lipid profile, egg fertility, reduced dead‐in‐shell, egg‐yolk cholesterol and resulted in improved post‐hatch growth performance.     

Effects of zinc-L-carnosine and vitamin E on aspirin-induced gastroduodenal injury in dogs

Background: Nonsteroidal anti‐inflammatory drugs frequently cause gastrointestinal (GI) injury. Zinc‐l ‐carnosine has antioxidant, anti‐inflammatory, mucosal protective, and healing properties in rodent models and in some human studies of GI injury. Hypothesis: The combination of zinc‐l ‐carnosine and vitamin E attenuates aspirin‐induced gastroduodenal mucosal injury. Animals: Eighteen healthy random‐source Foxhound dogs. Methods: In this randomized, double‐blinded, placebo‐controlled study dogs were treated with placebo (n = 6; 0X group), 30 mg/30 IU (n = 6; 1X group), or 60 mg/60 IU (n = 6; 2X group) zinc‐l ‐carnosine/vitamin E orally every 12 hours for 35 days. Between Day 7 and 35, GI mucosal lesions were induced with aspirin (25 mg/kg PO q8h). Mucosal injury lesions (hemorrhage, erosion, and ulcer) were assessed by gastroduodenoscopy on Days 14, 21, and 35 with a 12‐point scoring scale. Results: At baseline (Day ?1) gastroscopy scores were not significantly different between groups (mean ± SD: 0X, 4.4 ± 0.8; group 1X, 4.4 ± 0.6; group 2X, 4.2 ± 0.3; P= .55). Gastroscopy scores increased significantly in all groups between Day ?1 and Days 14, 21, and 35 (P < .0001). On Day 35, gastroscopy scores were 29.2 ± 5.2 (0X), 27.3 ± 3.7 (1X), and 28.6 ± 3.3 (2X). Mean gastroscopy scores were not significantly different among treatment groups on any of the days (P= .61). Conclusions and Clinical Importance: Administration of the combination of zinc‐l ‐carnosine and vitamin E at 1X or 2X dosing did not attenuate aspirin‐induced gastroduodenal mucosal injury.     

Cytologically atypical anal sac adenocarcinoma in a dog

A 10-year-old intact female Shetland Sheepdog with tenesmus had a subcutaneous mass at the left ventral aspect of the anus. On cytologic examination, 2 types of cells were observed. Most of the cells were oval to polygonal and had elliptical or elongate nuclei and a moderate amount of pale to basophilic cytoplasm. The remaining cells had round to oval nuclei and pale to basophilic cytoplasm. Cells of both types were loosely adhered to each other and were arranged in rosette-like structures. Both neoplastic cell types had fine homogenous chromatin and either a small indistinct nucleolus or no visible nucleolus. Mild anisokaryosis and anisocytosis were observed. Histologically, the mass consists of glandular structures formed by cuboidal cells admixed with bundles of spindle cells. Eosinophilic PAS- and Alcian blue-positive secretory material was found in the center of some glandular structures. Both neoplastic cell types had positive staining with paradoxical concanavalin A and expressed cytokeratin, but not vimentin, S-100, α-smooth muscle actin, or desmin. Based on location and histologic and immunohistochemical features, the final diagnosis was adenocarcinoma of the apocrine gland of the anal sac, which should be included as a cytologic differential diagnosis when spindle cells and typical epithelial cells are observed in masses in the region of the anal sac of dogs.     

Glycomolecule Modifications in the Seminiferous Epithelial Cells and in the Acrosome of Post‐testicular Spermatozoa in the Alpaca

A lectin histochemical investigation of the seminiferous epithelium and acrosomes of spermatozoa present in the efferent ductules and epididymal regions was carried out in the alpaca. The histochemical characterization was performed using a battery of different lectins: Con‐A, UEA‐I, LTA, WGA, GSA‐IB4, SBA, PNA, ECA, DBA, MAL‐II and SNA. Sialidase digestion and deglycosilation pre‐treatments were also employed. The cytoplasm of the Sertoli cells contained N‐linked oligosaccharides with α‐d ‐Man/α‐d ‐Glc and GlcNAc and O‐linked glycans with α‐l ‐Fuc, β‐GalNAc, β‐d ‐Gal‐(1‐4)‐d ‐GlcNAc, α–Gal and Neu5Acα2,6α‐GalNAc moieties whereas β‐d ‐Gal‐(1‐3)‐d ‐GalNAc residues were included in both O‐ and N‐glycoproteins. Spermatogonia expressed α‐d ‐Man/α‐d ‐Glc residues included in N‐glycoproteins and α‐Fuc in O‐glycoproteins. Spermatocytes contained the N‐glycoproteins residues α‐d ‐Man/α‐d ‐Glc and GlcNAc and the O‐glycoproteins residues α‐l ‐Fuc, β‐d ‐Gal‐(1‐4)‐d ‐GlcNAc, α–Gal, β‐GalNAc, Neu5Acα2,6α‐GalNAc and Neu5Acα2,6β‐d ‐Gal‐(1‐3)‐d ‐GalNAc. The results of the present study show differences in the presence and distribution of lectin reactive sites throughout the acrosomal development in the alpaca. In particular, Fuc moieties were found only during the Golgi‐phase of spermatids, α‐Gal were found in the acrosome of Golgi‐ and cap‐phase spermatids, sialic‐acid/α‐GalNAc sequence was revealed during the cap‐phase and elongated spermatids, and α‐d ‐Man/α‐d ‐Glc and GlcNAc were detected only in the acrosomes of elongated spermatids. Finally, β‐GalNAc, β‐d ‐Gal‐(1‐3)‐d ‐GalNAc and β‐d ‐Gal‐(1‐4)‐d ‐GlcNAc were added to acrosomal glycoproteins in the early stages of spermatogenesis and remained unchanged during the later phases. Differences in the carbohydrate expression were also demonstrated on the sperm acrosomes during passage through the post‐testicular ducts.     

l‐carnitine and fat type in the maternal diet during gestation and lactation modify the fatty acid composition and expression of lipid metabolism‐related genes in piglets

This study examined the influence of adding different amounts of maternal dietary l ‐carnitine and two fat types on fatty acid (FA) composition and the expression of lipid metabolism‐related genes in piglets. The experiment was designed as a 2 × 2 factorial with two fat types (3.5% soyabean oil, SO, and 3.5% fish oil, FO) and two levels of l ‐carnitine (0 and 100 mg/kg) added to the sows' diets. A higher proportion of n‐3 polyunsaturated fatty acids (PUFA) and a lower ratio of n‐6/n‐3 PUFA in sow milk and piglet tissues were observed in the FO groups than in the SO groups. Adding l ‐carnitine increased the proportion of C16:1 in sow milk and decreased n‐3 PUFA in piglet subcutaneous fat. Hepatic peroxisome proliferator‐activated receptor α (PPAR‐α) was more abundantly expressed in piglets from the FO groups than from the SO groups (p < 0.05), whereas stearoyl‐CoA‐desaturase (SCD), sterol regulatory element binding protein‐1 (SREBP1) and ?6‐desaturase (D6D) genes were less expressed in the FO groups compared with piglets from the SO groups. The expression of fatty acid synthase (FAS) genes was decreased in the SO groups with l ‐carnitine compared to that of the other dietary treatments. No differences among dietary treatments were observed with regard to the expression of acetyl‐CoA carboxylase (ACC). In conclusion, FO and l ‐carnitine supplementation in sows affect FA composition and hepatic gene expression in piglets.     

Evaluation of EDTA hematology tubes for collection of blood samples for tests of secondary hemostasis in dogs

OBJECTIVE: To evaluate the use of EDTA tubes for collection of blood samples for assays of secondary hemostasis in dogs. ANIMALS: 108 dogs of various ages, breeds, and sexes (19 healthy and 89 with abnormalities of secondary hemostasis). PROCEDURES: Blood samples were collected via cephalic venipuncture and transferred to sodium citrate tubes and EDTA tubes. Plasma was harvested from each type of tube for assays of concentrations of fibrinogen and D-dimer as well as prothrombin time, activated partial thromboplastin time, and antithrombin activity. Intra-assay and interassay precision and correlation coefficients for all hemostatic tests were calculated for each type of plasma sample. The effect of storage conditions on assay results for the 2 types of plasma samples was also evaluated. RESULTS: Results of hemostatic tests were highly correlated between citrated and EDTA-treated plasma samples. Intra-assay imprecision for all hemostatic tests with the exception of D-dimer concentration was < 10% for both citrated and EDTA-treated plasma samples; interassay imprecision was higher for EDTA-treated versus citrated plasma samples. Storage of plasma samples for 1 hour did not result in significantly different assay results for either type of plasma sample, but storage for 2 hours significantly affected values for EDTA-treated plasma samples. CONCLUSIONS AND CLINICAL RELEVANCE: Although evaluation of the sensitivity and specificity of hemostatic tests that use EDTA-treated plasma samples is required, EDTA may be a suitable alternative to sodium citrate as an anticoagulant for use in hemostatic testing in conditions in which tests could be performed within 1 hour after sample collection.     

The effects of different anticoagulants on routine canine plasma biochemistry

The effects of heparin, ethylenediaminetetraacetic acid (EDTA), sodium citrate and sodium fluoride/potassium oxalate on plasma biochemistry results in dogs were studied and compared with serum. Blood specimens from 10 apparently clinical healthy dogs were collected and placed in different tubes containing each anticoagulant tested. Differences in albumin, acetylcholinesterase, ionized calcium and potassium were found between serum and heparinized plasma. Most metabolites and enzymes did not show any variation, but significant decreases in electrolytes, alkaline phosphatase, acetylcholinesterase, bile acids, fructosamine and albumin were found when EDTA was used. Sodium citrate produced a 10-15% decrease in most metabolites and enzymes, possibly due to a sample dilution effect. Sodium fluoride/potassium oxalate produced haemolysis which may have influenced changes in some biochemical parameters.     

Increased plasma leptin through l‐carnitine supplementation is associated with an enhanced glucose tolerance in healthy ponies

In this study 0 or 4 g of l ‐carnitine was supplemented for 7 days in a cross‐over design of six healthy ponies to modulate glucose metabolism and leptin production. At the end of each period, serial blood samples were taken to measure glucose and insulin response, leptin, triglyceride (TG), non‐esterified fatty acids (NEFA) and creatine phosphokinase. l ‐carnitine supplementation was associated with a decrease in postprandial plasma glucose and insulin concentration, indicating an enhanced glucose tolerance. In contrast, postprandial plasma leptin concentration was increased when l ‐carnitine was supplemented. Yet, this increase in leptin concentration was not preceded by an increase in insulin concentration, suggesting that other factors apart from plasma insulin concentration could influence plasma leptin concentration. Although NEFA and TG were not significantly influenced by l ‐carnitine supplementation under these experimental conditions, further research must clarify whether net TG synthesis might be responsible for this increase in leptin.     

Comparison of aspiration and nonaspiration techniques for obtaining cytologic samples from the canine and feline spleen

Background: Ultrasound‐guided fine needle aspiration of the spleen is commonly used in the diagnostic evaluation of veterinary patients. Techniques using suction delivered through a 6–20‐cm³ syringe are the most commonly described means of obtaining cytologic samples of the spleen. Comparison studies of various human lesions have shown nonaspiration techniques to produce equal or superior cytologic specimens with less blood than specimens obtained using aspiration techniques. Objective: The purpose of this study was to compare the quality of splenic cytology specimens obtained using aspiration and nonaspiration techniques. Methods: Client‐owned dogs (n=24) and cats (n=7) receiving an abdominal ultrasound at the University of Tennessee College of Veterinary Medicine were enrolled in the study between January and June 2005. Samples were obtained from patients with and without sonographic splenic abnormalities. Two clinical pathologists, working independently and blinded to the method of sample collection, graded the cytologic specimens using a subjective scoring system for cellularity, amount of blood, and preservation of cellular morphology. Results: Agreement between the 2 independent observers was good. Direct comparison of the 2 techniques showed that samples obtained by the nonaspiration method had higher cellularity (P=.0002), less blood (P=.0023), and similar cell morphology (P=1.0000) compared with samples obtained by the aspiration method. Conclusion: These results suggest the nonaspiration technique is a superior method for obtaining a high‐quality cytologic specimen from the canine and feline spleen.     

Effects of 2 concentrations of sodium citrate on coagulation test results, von Willebrand factor concentration, and platelet function in dogs

BACKGROUND: Blood collection tubes containing 3.2% (0.109 M) sodium citrate, instead of 3.8% (0.129 M) sodium citrate, have recently become available in the United States. These tubes are visually indistinguishable from the traditional 3.8% sodium citrate tubes, except for wording on the label. Consequently, samples for hemostatic evaluation are frequently collected in tubes containing the lower concentration of sodium citrate. HYPOTHESIS: Results of hemostasis assays are different in samples collected in 3.2% versus 3.8% sodium citrate. ANIMALS: Twenty healthy dogs. METHODS: This study aimed at determining whether results of standard coagulation tests, von Willebrand factor concentration (vWF:Ag), and platelet function with the platelet function analyzer PFA-100a were affected by the different concentrations of sodium citrate. Blood samples were collected in tubes containing either 3.2% or 3.8% sodium citrate concentrations and processed routinely for coagulation assays (one-stage prothrombin time [OSPT], activated partial thromboplastin time [aPTT], fibrinogen concentration, and platelet count), vWF:Ag, and platelet function assays with a PFA-100. RESULTS: There was no significant difference between samples collected in 3.2% versus those collected in 3.8% sodium citrate for OSPT, aPTT, fibrinogen concentration, platelet count, or vWF:Ag. The closure times with collagen/adenosine diphosphate were significantly shorter (66 +/- 8.1 versus 74.8 +/- 9.7 seconds; P < .0001) with the 3.2% than with 3.8% sodium citrate concentration, and the hematocrit was significantly higher (47.9 +/- 5.6 versus 46.0 +/- 4.7 seconds; P = .03) in samples collected in 3.2% than in those collected in 3.8% sodium citrate. CONCLUSIONS AND CLINICAL IMPORTANCE: There is no clinically relevant effect of collection of blood into 3.2% or 3.8% sodium citrate.     

Canine complete blood counts: a comparison of four in‐office instruments with the ADVIA 120 and manual differential counts

Background: With more use of bench‐top in‐office hematology analyzers, the accuracy of reported values is increasingly important. Instruments use varied methods for cell counting and differentiation, and blood smears may not always be examined. Objective: The purpose of this study was to compare canine CBC results using 4 bench‐top instruments (Hemavet 950, Heska CBC‐Diff, IDEXX LaserCyte, and IDEXX VetAutoread) with ADVIA 120 and manual leukocyte counts. Methods: EDTA‐anticoagulated canine blood samples (n=100) were analyzed on each instrument. Manual differentials were based on 100‐cell counts. Linear regression, difference plots, paired t‐tests, and estimation of diagnostic equivalence were used to analyze results. Results: Correlations of HCT, WBC, and platelet counts were very good to excellent between all in‐office instruments and the ADVIA 120, but results varied in accuracy (comparability). Hemavet 950 and Heska CBC‐Diff results compared best with ADVIA results and manual leukocyte differentials. HCT and platelet counts on the IDEXX VetAutoread compared well with those from the ADVIA. Except for neutrophil counts, leukocyte differentials from all instruments compared poorly with ADVIA and manual counts. Reticulocyte counts on the LaserCyte and VetAutoread compared poorly with those from the ADVIA. Conclusions: The Hemavet 950 and Heska CBC‐Diff performed best of the 4 analyzers we compared. HCT, WBC, and platelet counts on the LaserCyte had minimally sufficient comparability for diagnostic use. Except for neutrophils (granulocytes), leukocyte differential counts were unreliable on all in‐office analyzers. Instruments with a 5‐part leukocyte differential provided no added benefit over a 3‐part differential. Assessment of erythrocyte regeneration on the LaserCyte and VetAutoread was unreliable compared with the ADVIA 120.     

Feline platelet counting with prostaglandin E1 on the Sysmex XT‐2000iV

Background: The large size of many feline platelets and the high frequency of platelet aggregation often results in falsely low platelet counts in this species. A combination of optical platelet counting to detect even large platelets and the use of prostaglandin E1 (PGE1) to inhibit platelet clumping may increase the accuracy of feline platelet counting. Objective: The objective of this study was to compare platelet counts in feline whole blood samples with and without the addition of PGE1 and using different analytical methods in a clinical setting. Methods: Platelet counts were determined in 10 feline patients in a referral veterinary hospital using 2 sample types (EDTA, EDTA with PGE1) and 2 methods of analysis (optical counting [PLT‐O] and impedance counting [PLT‐I]) on the Sysmex XT 2000 iV analyzer. Results: All PGE1–PLT‐O samples had platelet counts of >200 × 10⁹/L. Mean platelet count using PGE1–PLT‐O (410,256±178 × 10⁹/L) was significantly higher (P<.03) compared with PGE1–PLT‐I (256±113 × 10⁹/L), EDTA–PLT‐O (238±107 × 10⁹/L), and EDTA–PLT‐I (142±84 × 10⁹/L) methods. Depending on the method, platelet counts in 2 to 7 of 10 cats were <200 × 10⁹/L when PGE1‐PLT‐O was not used. A slightly increased platelet count in response to treatment of a feline patient with thrombocytopenia would have been missed without use of PGE1–PLT‐O. Conclusions: Using PLT‐O analysis on EDTA samples containing PGE1 provides higher, and therefore likely more accurate, feline platelet counts in a clinical setting.