Quantitative buffy coat analysis for hematologic measurements of canine, feline, and equine blood samples and for detection of microfilaremia in dogs

A quantitative buffy coat (QBC) analysis was evaluated for 175 canine, 125 feline, and 125 equine blood samples. The method used centrifuged whole blood and yielded rapid results expressed as respective band lengths for RBC, granulocytes, nongranulocytes, and platelets. Simple regression analysis of band lengths and reference laboratory methods yielded correlation coefficients (r) ranging from 0.72 to 0.99. The PCV, granulocyte count, and total WBC count, as determined by the 2 methods, correlated well (r greater than or equal to 0.93 in all cases). Platelet and nongranulocyte counts were less well correlated. The QBC system provided a means of performing rapid hematologic screening. The principal problem encountered was poor separation of the RBC-granulocyte interface in 17% of canine samples, which interfered with measurement of band lengths. Evaluation of the QBC tube for detection of Dirofilaria immitis microfilaremia revealed 100% sensitivity to counts as low as 160 microfilariae/ml of whole blood.     

Thiopurine methyltransferase in red blood cells of dogs, cats, and horses

Our objective was to determine if thiopurine methyltransferase (TPMT), the enzyme important in the metabolism of azathioprine in human beings, is detectable in red blood cell lysates (RBCL) of healthy dogs, cats, and horses. Values for TPMT activity were determined from blood collected from 20 healthy dogs, cats, and horses. The TPMT activity in each animal's RBCL was determined using a radioenzymatic end point involving TPMT-facilitated metabolism of 6-mercaptopurine to 6-methylmercaptopurine (6-MMP). One unit of TPMT activity represents the formation of 1 nmol of 6-MMP per milliliter of packed red blood cells per hour of incubation at 37 degrees C. TPMT activity in RBCL was detectable in all species, with mean RBC values +/- standard deviation of 17.9 +/- 3.79 U/mL in dogs; 2.76 +/- 0.70 U/mL in cats; and 2.185 +/- 0.36 U/mL in horses. Values for TPMT in the 3 species were significantly (P < .05) different from one another. TPMT values for dogs were significantly higher than the other species, and TPMT values for cats were significantly higher than those for horses. We conclude that RBCL TPMT values are measurable in dogs. cats, and horses and that dogs have higher values than cats or horses. These findings are consistent with the lower tolerance for azathioprine in cats as compared with dogs. It remains to be determined whether RBCL TPMT values in these species correlate with TPMT activity in the liver, where most of the metabolization of azathioprine is believed to occur.     

Isolation of granulocytes and mononuclear cells from the blood of dogs, cats, horses and cattle

A simple discontinuous Percoll density-gradient technique was adapted for isolation of granulocytes and mononuclear cells from cats, dogs, horses and cattle. Separation was accomplished at low speeds using a standard tabletop centrifuge. Cell purity was 100% for both granulocytes and mononuclear cells and cell viability exceeded 95%. Percent recovery of leukocytes ranged from 69 to 83%.     

Scleral rupture in dogs, cats, and horses

OBJECTIVES: The aim of this retrospective study was to summarize the most frequent clinical signs, ultrasonographic, and histological findings accompanying scleral rupture as a result of blunt trauma in dogs, cats, and horses. ANIMALS STUDIED AND PROCEDURES: Thirty small animals and three horses diagnosed with scleral rupture resulting from blunt trauma. B-mode ultrasonography was performed on 20 animals. Histopathology was carried out on 18 enucleated globes. RESULTS: In small animals, 80% presented hyphema, 60% subconjunctival hemorrhage, and 53% eyelid and conjunctival swelling. In horses, 100% presented eyelid and conjunctival swelling, 67% hyphema, subconjunctival hemorrhage, and collapsed anterior chamber. Ultrasonographic findings were an area with ill-defined scleral margins (90%), echoic/hyperechoic contents in the anterior and posterior chamber (55%) and in the vitreous (80%). In small animals, scleral rupture location noted on gross examination was: at the posterior pole (4), close to the optic nerve (3), near the limbus (2), and in the dorsal aspect of the globe (1). In horses, the lesion was located at the limbus (3). In small animals, histopathology showed presence of hemorrhage in the anterior, posterior chamber, and vitreous (94%), retinal detachment (94%), choroidal edema and hemorrhages (88%), and choroidal detachment as a result of suprachoroidal hemorrhage (88%). The same lesions were found in the globes of two horses. In small animals, rupture location noted on histopathology was: at the posterior pole (8), close to the optic nerve (4), near the limbus (1), near the ciliary body (1). CONCLUSIONS: The most frequent clinical signs observed were hyphema, subconjunctival hemorrhage, and eyelid and conjunctival swelling. Ultrasonographic findings suggestive for scleral rupture were ill-defined scleral borders and/or echoic/hyperechoic material in the cavities of the globe. On histopathology, lesions severely altering the anatomy of the eye structures were: hemorrhage into the chambers of the globe, subretinal and suprachoroidal hemorrhage leading to retinal and choroidal detachment, respectively. In small animals, the most frequent locations for scleral rupture were the posterior pole and close to the optic nerve, whereas in horses it was the limbus.     

In vitro effects of cyclooxygenase inhibitors in whole blood of horses, dogs, and cats.

OBJECTIVE: To determine potency and selectivity of nonsteroidal anti-inflammatory drugs (NSAID) and cyclooxygenase- (COX-) specific inhibitors in whole blood from horses, dogs, and cats. SAMPLE POPULATION: Blood samples from 30 healthy horses, 48 healthy dogs, and 9 healthy cats. PROCEDURE: Activities of COX-1 and COX-2 were determined by measuring coagulation-induced thromboxane and lipopolysaccharide-induced prostaglandin E2 concentrations, respectively, in whole blood with and without the addition of various concentrations of phenylbutazone, flunixin meglumine, ketoprofen, diclofenac, indomethacin, meloxicam, carprofen, 5-bromo-2[4-fluorophenyl]-3-14-methylsulfonylphenyl]-thiophene (DuP 697), 5,5-dimethyl-3-(3-fluorophenyl)-4-(4-methylsulphonyl) phenyl-2(5H)-furan one (DFU), 3-(3,4-difluorophenyl)-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone (MF-tricyclic), and celecoxib. Potency of each test compound was determined by calculating the concentration that resulted in inhibition of 50% of COX activity (IC50). Selectivity was determined by calculating the ratio of IC50 for COX-1 to IC50 for COX-2 (COX-1/COX-2 ratio). RESULTS: The novel compound DFU was the most selective COX-2 inhibitor in equine, canine, and feline blood; COX-1/COX-2 ratios were 775, 74, and 69, respectively. Carprofen was the weakest inhibitor of COX-2, compared with the other COX-2 selective inhibitors, and did not inhibit COX-2 activity in equine blood. In contrast, NSAID such as phenylbutazone and flunixin meglumine were more potent inhibitors of COX-1 than COX-2 in canine and equine blood. CONCLUSIONS AND CLINICAL RELEVANCE: The novel COX-2 inhibitor DFU was more potent and selective in canine, equine, and feline blood, compared with phenylbutazone, flunixin meglumine, and carprofen. Compounds that specifically inhibit COX-2 may result in a lower incidence of adverse effects, compared with NSAID, when administered at therapeutic dosages to horses, dogs, and cats.     

Epidemiologic analysis of oral and pharyngeal cancer in dogs, cats, horses, and cattle.

Four hundred sixty-nine oral-pharyngeal malignancies diagnosed in dogs, cats, horses, and cattle and submitted to the Viterinary Medical Data Program between March 1, 1964, and Dec 31, 1974, were analyzed. Of these cases, 84% were in dogs. The most frequent oral-pharyngeal cancer in dogs was melanoma; in cats and horses, it was squamous cell carcinoma. In dogs, the risk of developing melanoma increased more with age than did the risk of developing squamous cell carcinoma and fibrosarcoma. Male dogs had significantly greater risk of developing fibrosarcomas and melanomas than did female dogs. The German Shorthaired Pointer, Weimaraner, Golden Retriever, Boxer, and Cocker Spaniel breeds had significantly higher risk and Dachshunds and Beagles had significantly lower risk, as compared with all breeds combined. There was no significant difference between observed and expected numbers of tonsillar carcinomas diagnosed at veterinary colleges located in small urban areas (less than 50,000 persons) as compared with large urban populations (greater than 500,000).     

Quantitative genetic aspects of coat color in horses

The aim of this study was to estimate genetic parameters for coat color in horses. Besides defining coat color classes (gray, chestnut, bay, and black), the phenotypes were also measured quantitatively according to standardized international procedures (Commission Internationale de l'Eclairage L*, a*, b*), where L* describes lightness, a* describes color saturation from red to green, and b* describes color saturation from yellow to blue. The total color saturation was derived from a* and b* and referred to as Chroma. A total of 294 horses from the breeds Lipizzan, Nonius, Arabian Pure Bred, Shagya Arabian, and Gidran were measured at neck, shoulder, and belly. Heritabilities (within and between breeds or color classes) and repeatabilities were estimated using REML from univariate animal models defined separately for gray and nongray horses. For gray horses, the estimated within-breed heritabilities for L* ranged from 0.45 to 0.49 and for a*, b*, and Chroma from 0.09 to 0.52, indicating moderate polygenic effect. For nongray horses, between-color class heritabilities were high (0.70 to 0.85) and within-color class heritabilities were negligible (except for L* measured on neck and belly, 0.21 and 0.34, respectively). Additionally, the importance of L* was described by the relation with the total melanin content of horse coat hair; for gray and nongray horses, a strong negative linear relationship was detected (P < 0.01). The spectrometric measures and the results of this study demonstrate a possible approach to the estimation of the polygenic component involved in coat color inheritance.     

Conjunctival fungal flora in horses, cattle, dogs, and cats

Conjunctival swab specimens were obtained from both eyes of 43 horses, 25 cows, 50 dogs, and 25 cats without keratitis or other ophthalmologic problems. Fungi were isolated from 95% of the horses, 100% of the cows, 22% of the dogs, and 40% of the cats. Aspergillus spp were isolated from 56% of the horses, 12% of the cows, 8% of the cats, and none of the dogs. Penicillium spp and Cladosporium spp were isolated ubiquitously. Collectively, 28 species from 209 isolants were identified.     

Lymphoid nodules in skin biopsies from dogs, cats, and horses with nonneoplastic dermatoses

In a retrospective histopathologic study of nonneoplastic dermatoses, lymphoid nodules were found in 0.3% of 3,408 canine, 5.1% of 469 feline, and 4.5% of 325 equine skin biopsies. In all 3 species, the majority of cases wherein lymphoid nodules were found were diseases of presumed immune-mediated nature. In cats and horses, the majority of cases were also diseases characterized by tissue eosinophilia.     

An evaluation of pulse oximeters in dogs, cats and horses

Objective Evaluation of five pulse oximeters in dogs, cats and horses with sensors placed at five sites and hemoglobin saturation at three plateaus. Study design Prospective randomized multispecies experimental trial. Animals Five healthy dogs, cats and horses. Methods Animals were anesthetized and instrumented with ECG leads and arterial catheters. Five pulse oximeters (Nellcor Puritan Bennett‐395, NPB‐190, NPB‐290, NPB‐40 and Surgi‐Vet V3304) with sensors at five sites were studied in a 5 × 5 Latin square design. Ten readings (SpO₂) were taken at each of three hemoglobin saturation plateaus (98, 85 and 72%) in each animal. Arterial samples were drawn concurrently and hemoglobin saturation was measured with a co‐oximeter. Accuracy of saturation measurements was calculated as the root mean squared difference (RMSD), a composite of bias and precision, for each model tested in each species. Results Accuracy varied widely. In dogs, the RMSD for the NPB‐395, NPB‐190, NPB‐290, NPB‐40 and V3304 were 2.7, 2.2, 2.4, 1.7 and 2.7% respectively. Failure to produce readings for the NPB‐395, NPB‐190, NPB‐290, NPB‐40 and V3304 were 0, 0, 0.7, 0, and 20%, respectively. The Pearson correlation coefficients for the tongue, toe, ear, lip and prepuce or vulva were 0.95, 0.97, 0.69, 0.87 and 0.95, respectively. In horses, the RMSD for the NPB‐395, NPB‐190, NPB‐290, NPB‐40 and V3304 were 3.1, 3.0, 4.7, 3.3 and 2.1%, respectively while rates of failure to produce readings were 10, 21, 0, 17 and 60%, respectively. The Pearson correlation coefficients for the tongue, nostril, ear, lip and prepuce or vulva were 0.98, 0.94, 0.88, 0.93 and 0.94, respectively. In cats, the RMSD for all data for the NPB‐395, NPB‐190, NPB‐290, NPB‐40 and V3304 were 5.9, 5.6, 7.9, 7.9 and 10.7%, respectively while failure rates were 0, 0.7, 0, 20 and 32%, respectively. The correlation coefficients for the tongue, rear paw, ear, lip and front paw were 0.54, 0.79,.0.64, 0.49 and 0.57, respectively. For saturations above 90% in cats, the RMSD for the NPB‐395, NPB‐190, NPB‐290, NPB‐40 and V3304 were 2.6, 4.4, 4.0, 3.5 and 4.8%, respectively, while failure rates were 0, 1.7, 0, 25 and 43%, respectively. Conclusions and clinical relevance Accuracy and failure rates (failure to produce a reading) varied widely from model to model and from species to species. Generally, among the models tested in the clinically relevant range (90–100%) RMSD ranged from 2–5% while failure rates were highest in the V3304.     

Determination of the number of mast cells in lymph node, bone marrow, and buffy coat cytologic specimens from dogs.

Cytologic samples of popliteal lymph node, proximal femoral bone marrow, and the buffy coat fraction of blood were obtained from 56 dogs. The number of mast cells on 1 slide of each sample was determined by microscopic examination. Eleven of 46 slides of lymph node aspirate contained mast cells (range, 1 to 16; mean, 6.4; median, 5 mast cells/slide). Fifty-one bone marrow aspirate slides were evaluated. Two of these contained a single mast cell. None of the 53 buffy coat smear slides examined contained any mast cells. These results indicated that in clinically normal dogs, a few to several mast cells may be encountered in smears of lymph node aspirate, mast cells are rare in smears of bone marrow aspirate, and mast cells are absent from smears of buffy coat.     

Antimicrobial drug susceptibility of bacteria isolated from disease processes in cattle, horses, dogs and cats

Results are presented of antimicrobial disc diffusion susceptibility testing on commonly isolated bacterial pathogens made at the Ontario Veterinary College Diagnostic Bacteriology Laboratory in 1981 and 1982. Nearly 2 000 isolates from horses, cattle, dogs and cats were tested. Comparison of resistance in the same bacterial species isolated from different animal species showed significant differences between some of the same antibiotics.     

Effects of platelet clumping on platelet concentrations measured by use of impedance or buffy coat analysis in dogs.

OBJECTIVE: To determine whether platelet clumps are homogeneously distributed in blood samples, and whether platelet concentrations (PC) obtained by use of impedance and buffy coat analysis can be considered minimum values when platelet clumps are present. DESIGN: Prospective study. SAMPLE POPULATION: 50 blood samples obtained from 30 dogs. PROCEDURE: 10 blood samples containing platelet clumps were used and 10 smears were made from each sample; amount of platelet clumping was graded for all 100 smears. Blood from each of 20 healthy dogs was divided between 2 EDTA tubes before and after platelet clumping was induced by adenosine diphosphate (ADP). The PC for each ADP-treated and untreated sample were measured, using impedance and quantitative buffy coat analyzers. RESULTS: Platelet clumps were evident in all 100 blood smears, but the amount of clumping varied considerably within some samples. Using the impedance analyzer, the PC of ADP-treated samples were significantly lower and never higher than the PC of untreated samples. Using the buffy coat analyzer, some ADP-treated samples had increased PC; however, significant differences were not detected between treated and untreated samples. CONCLUSIONS AND CLINICAL RELEVANCE: Platelet clumping was not homogeneous within blood samples. When platelet clumps were identified by direct examination of blood smears, the PC detected by use of the impedance analyzer could be considered minimum values. In contrast, the PC detected by use of the buffy coat analyzer were sometimes increased. Useful information can be obtained by measuring PC in blood with platelet clumps; values obtained by use of impedance can be considered minimums, and values obtained by use of buffy coat analysis may be either minimum values or reasonable estimates of PC.     

Blood vitamin and choline concentrations in healthy domestic cats, dogs, and horses

Blood concentrations of thiamin, biotin, nicotinates, pantothenates, folates, riboflavin, vitamins A, B6, B12, C, E, beta-carotene and choline were analyzed in healthy animals (23 horses, 25 dogs, and 29 cats). B-Complex vitamins and choline also were analyzed in the liver of the dogs and cats. Vitamin concentrations in the blood and livers of dogs were similar; however, blood vitamin A and beta-carotene concentrations were lower in the cat than in the dog. Horses had a higher B12 blood concentration than did the dogs and cats. These data can be useful for detecting overt and hidden vitamin deficits in these species due to various conditions.     

Prevalence of antibodies to Toxoplasma gondii in cats, dogs and horses in Sweden

Samples of serum or plasma taken during 1986 and 1987 from 244 pet cats, 303 dogs and 219 horses, randomly selected among animals referred to the Animal Clinics of the Swedish University of Agricultural Sciences, were screened by enzyme-linked immunosorbent assay (ELISA) for antibodies to Toxoplasma gondii. 42% of cats, 23% of dogs and 1% of horses examined were found seropositive.