bluuebunny/crossref_metadata_2025 huggingface dataset:
Hemosiderin in Leukocytes of Dogs with Immune‐Mediated Hemolytic Anemia
Authors: Gaunt Stephen D. , Baker Dale C.
Published: February 2009

Hemosiderin granules were identified in blood neutrophils and monocytes of three dogs. The brownish granules were 1 to 4 microns in diameter and stained positively for iron with Prussian blue stain. All three dogs had evidence of immune‐mediated hemolytic anemia and received whole blood transfusions prior to observation of hemosiderin. The mechanism of hemosiderin accumulation by blood leukocytes from these dogs was undetermined. Iron overload produced by administration of whole blood transfusions during immune‐mediated hemolytic anemia was implicated as a causative factor.
Potential Role of Serum Inhibitors of Erythropoiesis in the Anemia Associated With Infection, Renal Disease and Malignancy in the Dog
Authors: Weiss Douglas J.
Published: February 2009

Serum from dogs with anemia secondary to infection, malignancy and chronic renal disease was tested for the capacity to inhibit in vitro erythropoiesis. Serum inhibition was found in seven of 10 dogs with infectious diseases, four of 10 dogs with malignant diseases, and two of 10 dogs with chronic renal failure. The results were reproducible with at least three different marrow donor dogs. Serum inhibition did not correlate with decreased packed cell volumes. However, serum inhibition occurred in the two dogs with greatest azotemia suggesting that inhibitors of erythropoiesis are present only in advanced stages of renal failure.
Mean Platelet Volume Artifacts: The Effect of Anticoagulants and Temperature on Canine Platelets
Authors: Handagama Prem , Feldman Bernard , Kono Constance , Farver Thomas
Published: February 2009

Improper handling of specimens results in artifactually high Mean Platelet Volume (MPV) measurements limiting their usefulness as a clinical tool. MPV measurement and Scanning Electron Microscopy (SEM) were performed on split specimens collected from normal dogs using two anticoagulants and two temperatures over a period of 4 hours. Platelets exposed to EDTA and maintained at 4 °C (39.2 °F) exhibited the highest artifactual increase in MPV, while those exposed to citrate and maintained at 37°C (98.6 °F) exhibited minimal change. The increase in MPV was accompanied by platelet shape change from a smooth disc to an irregular sphere with filopodia. It is recommended that citrated specimens maintained at 37°C be used in all MPV measurements.
A Review of Feline Serum Protein Electrophoresis
Authors: Baker Rebecca J. , Valli Victor E.O.
Published: February 2009

A review of the current literature available on feline serum proteins is presented. Early studies concentrated on comparative aspects of species variations in the electro‐phoretic pattern. The feline electrophoreto‐gram was divided into five basic regions: albumin, alpha‐1, alpha‐2, beta and gamma. Different subdivisions of these areas were recognized depending on the support medium used. Current papers have compared the relative migration distances of each globulin peak to the migration of albumin. This “Rf” value enables reliable peak identification. To date, no data exists identifying the individual proteins responsible for the peaks in the alpha and beta regions. The only feline globulin to be studied is haptoglobin; however its precise location on the electrophoretic strip was not identified.
An Enzyme Immunoassay to Measure Canine Circulating Fibronectin
Authors: O'Neill Sharron L , Boothby John T. , Feldman Bernard F.
Published: February 2009

A competitive enzyme immunoassay has been used to detect and quantitate fibronectin in canine plasma. In this test, purified fibronectin, bound to microtiter plates, competes with plasma fibronectin for the conjugated antibody, rabbit‐anticanine, fibronec‐tin‐horseradish peroxidase. The assay could detect fibronectin in purified standards from 58 ng/ml to 580 μg/ml. The range of 1–100 μ g/ml was linear for plasma samples diluted 1:10, allowing samples with fibronectin concentrations from 10–1000 μg/ml to be easily measured by this method. The mean normal fibronectin concentration of 132 dogs, by this method, was determined to be 320 ± 74 μ g/ml.
Antithrombin III (ATIII) Activity in Plasmas from Normal and Diseased Horses, and in Normal Canine, Bovine and Human Plasmas
Authors: Johnstone I.B. , Petersen D. , Crane S.
Published: February 2009

Plasma Antithrombin III (ATIII) activity was quantitated in 24 clinically normal Standardbred/Thoroughbred horses using a clotting time technique. ATIII activity ranged from 80 to 106% of the pooled reference standard plasma, with a mean of 94%. Horses presenting with impaction or spasmotic colic (n = 17) had normal plasma ATIII activity, while 15 horses presenting with acute diarrhea/colitis had significantly lower plasma ATIII activity with a mean of only 74% of the reference plasma. Seven horses presenting with liver disease had significantly higher plasma ATIII activity with a range of 127 to 177% of the pooled reference plasma.

Fifty‐seven equine plasmas were retested using a rapid chromogenic substrate technique for quantitating plasma ATIII activity. A good correlation (r = +0.83) existed between clotting time and chromogenic determinations of ATIII.

Pooled normal canine, human and bovine plasmas had only 65, 62 and 79%, respectively, of the ATIII activity of the equine reference plasma.
Evaluation of Hepatic and Renal Function in Cats with Chediak‐Higashi Syndrome
Authors: Wardrop K. Jane , Dhein Cheryl R. , Prieur David J. , Meyers Kenneth M.
Published: February 2009

Cats with the Chediak‐Higashi Syndrome (CHS) have partial oculocutaneous albinism, a bleeding tendency, and enlarged cytoplasmic granules in many cell types including those in the liver and kidney. Hepatic and renal function was evaluated in six CHS and six age‐matched control cats to determine if the functions of these organs were compromised by the CHS trait. Serum concentrations of alanine aminotransferase, alkaline phosphatase, and total bilirubin were determined to assess the status of the liver. Sulfobromophthalein retention tests were also performed. Renal function was evaluated by determination of14C‐inulin clearance; blood urea nitrogen and serum creatinine concentrations; 24‐hour protein/creatinine ratios; percent clearance ratios of calcium, phosphorus, sodium, potassium and chloride; and urinalysis values. The CHS cats were not significantly (P>0.05) different from the control cats in any of the above tests. Use of a non‐parametric statistical test did reveal a mild difference (P = 0.047) in 24‐hour protein excretion between CHS and control cats. Complete blood counts were performed, and the packed cell volume and hemoglobin concentrations were significantly lower (P> 0.05) in the CHS cats than in the control cats.
Routine Hematology and Clinical Chemistry in Female Dogs with Benign and Malignant Mammary Tumors
Authors: Fayolled P. , Broulet V. , Braun J.P. , Rico A.G. , Magnol J.P. , Thouvenot J.P.
Published: February 2009

Routine hematological and serum biochemical screening was done in 61 female dogs with benign mammary tumors and 51 female dogs with malignant mammary tumors. Most parameters were not significantly different from age‐matched female controls; moreover, no significant difference could be observed between animals with benign and malignant tumors. It is concluded that routine hematology and biochemistry offer little diagnostic or prognostic benefit in female dogs with mammary tumors.
Direct Measurement of Progesterone in Plasma and Milk by a Simple Solid‐Phase Radioimmunoassay
Authors: Epelu‐Opiod Justin , Madejd Andrzej
Published: February 2009

A rapid method for determination of progesterone in bovine, goat and porcine plasma as well as in bovine milk was evaluated. The method employed was a solid‐phase 125***/ radioimmunoassay equipped with progesterone standards in human serum and called the Coat‐A‐Count procedure. The dilution curves of bovine plasma samples with high progesterone content were parallel with the standard curve based on human serum. The relation between measurements of progesterone levels in bovine plasma using the reference extraction method and the direct Coat‐A‐Count procedure was highly significant, resulting in the linear regression equation Y = 1.06x ‐ 0.04. In case of goat and porcine plasma, the direct method yielded higher results than the reference extraction method (Y = 1.37x + 1.38 and Y = 1.69x ‐ 6.47, respectively). Progesterone concentration in bovine whole milk was much higher when measured by the Coat‐A‐Count procedure than by the reference Farmos kit (Y = 1.59x + 1.51). However, when the same samples were assayed by a modified Coat‐A‐Count procedure, i.e. progesterone standards from Farmos kit, the values were more or less identical (Y = 0.88x – 0.21).
Effect of Age on Serum Chemistry Profile, Electrophoresis and Thyroid Hormones in Beagle Dogs Two Weeks to One Year of Age
Authors: Wolfordd S.T. , Schroer R.A. , Gohs F.X. , Gallo P.P. , Falk H.B. , Dente A.R.
Published: February 2009

hemistry tests were conducted on serum from young Beagle dogs in order to determine the effect of age on these parameters. Blood was collected at regular intervals from 25 normal Beagle puppies (13 males and 12 females) at ages ranging from 2 weeks to 12 months. Serum chemistry profiles, protein electrophoresis and radioimmunoassays for thyroxine and triiodothyronine were included.

Rapidly changing age related differences were observed in several parameters. Urea nitrogen, cholesterol, triglycerides, lactate dehydrogenase, thyroxine, glucose, gamma glutamyl transpeptidase, and total bilirubin values were elevated early in life, and decreased during the first 6 to 8 weeks, while alanine aminotransferase activity was low initially and increased during this period. Lactate dehydrogenase, thyroxine, gamma glutamyl transpeptidase, total bilirubin and alanine aminotransferase attained stability by 3 months, but the remaining parameters showed slight changes subsequently, gradually approaching adult values.

More gradual age related changes were observed in other parameters. These included alkaline phosphatase, inorganic phosphorus and calcium values, which were higher in younger dogs, and creatinine, aspartate aminotransferase and total protein values, which were lower in younger dogs. Creatinine and aspartate aminotransferase values were stable by approximately 6 months; alkaline phosphatase, inorganic phosphorus, calcium and total protein values continued to change gradually up to 1 year.
Automated Blood Cell Counting Systems: A Comparison of the Coulter S‐Plus IV, Ortho ELT‐8/DS, Ortho ELT‐8/WS, Technicon H‐1, and Sysmex E‐5,000
Authors: Tvedtend Harold W. , Wilkins Robert J.
Published: February 2009

Four automated blood cell counting systems were evaluated at the Michigan State University Veterinary Clinical Center (VCC) for their suitability for analyzing various animals' blood in a university teaching hospital laboratory. The instruments were compared to a Coulter Model S‐Senior (Coulter Electronics) which had been used for 8 years, and was to be replaced. The instruments were a Coulter Model S‐Plus IV, an Ortho ELT‐8/ds (Ortho Diagnostics Systems), a Technicon H‐1 (Technicon Instruments), and a Sysmex E‐5,000 (Toa Medical Electronics). Additionally, an Ortho ELT‐8/ws at a private laboratory (Cenvet) was compared to the Coulter S—Senior at the VCC.

Based on these evaluations, only the Sysmex E‐5,000 was considered unacceptable for the VCC laboratory. Certain advantages and disadvantages of the instruments are described in this article. The comparisons a‐mong instruments were not as consistent or repetitive as expected for a controlled experiment but did provide information likely useful to others considering using or purchasing an automated blood cell counting system for a veterinary laboratory.
Comparison of Various Staining Procedures in the Identification of Hepatozoon canis Gamonts
Authors: Mercer Susan H. , Craig Thomas M.
Published: February 2009

The staining characteristics of gamonts of Hepatozoon canis in peripheral blood smears were evaluated. Three stains, Diff‐Quik stain, Giemsa stain and a modification of the naphthol‐ASD‐chloroacetate esterase stain, were compared. The staining characteristics of the infected cell and the parasite were different in each stain used. The modified naphthol‐ASD‐chloroacetate esterase stain showed the most difference between the infected and noninfected cells, as well as a difference in staining characteristics of the parasite nucleus from those of the host nucleus. Although the parasite was identified with all stains, a definitive diagnosis was more easily obtained with the naphthol‐ASD‐chloroacetate esterase stain.
A Technique for Automated Quantification of Canine Glucocorticoid‐Induced Isoenzyme of Alkaline Phosphatase
Authors: Hoffmann Walter E. , Sanecki Robin K. , Dorner Joseph L.
Published: February 2009

A sensitive assay for the corticosteroid‐induced alkaline phosphatase isoenzyme (CAP), adaptable to most clinical chemistry autoanalyzers, is described and validated. This assay is based on the greater than 98% inhibition of liver alkaline phosphatase isoenzyme (LAP) activity with 4.2 mM levamisole, as compared to the 42% inhibition of CAP activity. Analysis of serum with total alkaline phosphatase (AP) activity within the reference range, resulted in a reference range of 0 to 19 U/L for CAP activity. Analysis of serum from 160 clinical patients with AP activity above the reference range, revealed 73% with increased CAP activity ranging from 20 to 7,000 U/L. The diagnostic significance of this increased CAP activity is discussed.
Studies on the Pathogenesis of the Erythrocyte Destruction Associated with the Anemia of Inflammatory Disease
Authors: Weiss Douglas J. , McClay Carolyn B.
Published: February 2009

Density‐separated feline RBC were analyzed for surface immunoglobulin and in vitro susceptibility to macrophage phagocytosis before and after introduction of a sterile abscess. Prior to abscess induction, least dense cells did not have detectable surface immunoglobulin G (IgG) as a direct antiglobulin reaction was negative. Two of four intermediate and three of four most dense cell fractions had detectable surface IgG as indicated by a positive direct antiglobulin reaction. On Days 3 and 5 after induction of sterile abscesses all three RBC fractions were positive and the size of the RBC clumps tended to increase. In vitro macrophage phagocytosis of all three RBC fractions increased significantly after abscess induction. The phagocytosis could be partially blocked by preincubation of macrophages with feline IgG.
The Intravenous Bile Acid Loading Test in the Goat: Determination of a Procedural Protocol and Reference Values
Authors: Hagemoser Wayne A.
Published: February 2009

An intravenous bile acid loading test was performed on 16 goats using various dosages and sampling protocols. A recently developed enzymatic bile acid assay provided an economical and technologically simple method for bile acid analysis. Intravenous injection of bile salts into goats was well tolerated. Clearance times were rapid.

A protocol for performance of the test was suggested and a practical reference range for T 1/2 clearance of 4 ± 1 minute was determined.

Further investigations of this test in the assessment of liver function in the ruminant appear warranted.
Isolation of Granulocytes and Mononuclear Cells from the Blood of Dogs, Cats, Horses and Cattle
Authors: Weiss Douglas J. , Kraemer Ruth , Schmit Kathleen
Published: February 2009

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%.
Effect of Desmopresssin in Normal Dogs and Dogs with von Willebrand's Disease
Authors: Giger Urs , Dodds W. Jean
Published: February 2009

Desmopressin acetate (DDAVPR), a synthetic analogue of vasopressin, was slowly administered intravenously to 12 healthy dogs of various breeds and 10 Doberman Pinschers with mild‐to‐moderate type I von Willebrand's disease at a dose of 0.3, 1.0 and 3.0 μg/kg body weight. Plasma von Willebrand factor: antigen was measured by an electroimmunoassay prior to and 30, 60, 90, 120 and 180 minutes after desmopressin infusion. Desmopressin induced only very modest and statistically insignificant increases in von Willebrand factor in both groups. We conclude that the response to desmopressin as measured by circulating von Willebrand factor is much less pronounced in healthy dogs and in Doberman Pinschers with von Willebrand's disease than in humans.
Idiopathic Dyserythropoiesis in a Dog
Authors: Weiss Douglas J. , Reidarson Thomas H.
Published: February 2009

Idiopathic dyserythropoiesis in a dog was characterized by chronic nonregenerative normocytic normochromic anemia, cellular marrow and abnormal morphology of erythroid precursors. Serum concentrations of Vitamin B12, folate and iron were inconsistent with secondary causes of dyserythropoiesis. The disorder appeared to be distinct from myelodysplastic syndromes described previously.
Hemolysis as a Factor in Clinical Chemistry and Hematology of the Dog
Authors: O'Neill Sharron L. , Feldman Bernard F.
Published: February 2009

Serum biochemical, hemostatic, and hematologic analytes were determined by four laboratories on dogserum or plasmas containing increasing amounts of hemolysate. From the results of testing interferographs were prepared to aid in decision making and to enhance visualization of the effects of hemolysis on the determination of the analytes. Hemolysis consistently interfered with the analysis of creatinine phosphokinase, lactic dehydrogenase, aspartate aminotransferase, lipase, and albumin, all of which appeared to increase with increasing hemolysis. The results of testing for the remainder of the biochemical, hemostatic, and hematologic analytes varied greatly among analyzers or methods, rarely in a predictable manner, indicating that each laboratory should evaluate the effects of hemolysis on each analyzer and for each method used, in order to make informed decisions on the use of hemolyzed, but irreplaceable, samples.
Cytologic Features of a Meningioma in a Dog
Authors: Altman David , Bolon Brad , Meyer D.J. , Homer Bruce
Published: February 2009

Meningiomas are one of the most common nervous system tumors in dogs. They are mostly solitary, benign, intracranial tumors of the transitional type. Tentative diagnosis can usually be made on historical data, clinical and neurological signs, and radiographs, with an antemortem diagnosis confirmed by exploratory surgery and histologic evaluation of biopsy. The literature contains many reports of the gross, histological and ultrastructural morphology of canine meningiomas, but little on the cytologic features of these tumors. This report describes the cytologic appearance of an intracranial meningioma in a 9‐year‐old Miniature Schnauzer.
Evaluation of a Rapid Reagent Strip Test for Blood Glucose Determination in Diarrheic Calves
Authors: Sweeney Raymond W. , Divers Thomas J. , Zamos David T. , Spencer Pamela A.
Published: February 2009

Sixty‐three blood samples from 10 diarrheic calves were tested for glucose concentration by two methods. Plasma glucose concentration was measured by the conventional glucose‐6‐phosphate dehydrogenase method in the clinical laboratory, and the results compared to those obtained using a rapid reagent strip test for blood glucose concentration measurement.

The rapid reagent strip test result could not be used to make an accurate prediction of the actual plasma glucose concentration as determined by the conventional method, due to the wide variability in actual plasma glucose concentrations corresponding to each rapid test result.
Platelet Function in the Racing Thoroughbred: Implication for Exercise‐Induced Pulmonary Hemorrhage
Authors: Weiss Douglas J. , McClay Carolyn B. , Smith Clark M. , Rao Gundu H.R. , White James G.
Published: February 2009

Platelet function was evaluated in horses with exercise‐induced pulmonary hemorrhage (bleeder) and in control horses (nonbleeder). Platelet aggregation, secretion, and adhesion to rabbit aortic suben‐dothelium were similar for bleeders and nonbleeders. Platelets readily aggregated in response to ADP, thrombin, collagen, and arachidonic acid, but platelet secretion occurred only with high concentrations of thrombin. Platelets readily adhered to rabbit aortic subendothelium and tended to form large thrombi rather than platelet monolayers or aggregates. These data suggest that horses may be predisposed to thrombus formation and subsequent microvascular obstruction.
Evaluation of a Radioimmunoassay for Type III Procollagen Peptide in the Dog
Authors: Dimski Donna S. , Brooks Charles L , Johnson Susan E.
Published: February 2009

Type III procollagen peptide (P‐3‐P) is a serum marker for hepatic fibrosis in humans. The utility of a commercially available radioimmunoassay for P‐3‐P was evaluated in the dog. The specificity of the assay was assessed by polyacrylamide gel electrophoresis (PAGE) of canine serum and purified bovine P‐3‐P, followed by Western immunoblotting with rabbit anti‐P‐3‐P serum. The sensitivity was assessed by performing the radioimmunoassay on dilutions of sera from 22 dogs. Polyacrylamide gel electrophoresis of purified bovine P‐3‐P and sera from two dogs suspected of having elevated P‐3‐P concentrations revealed no homologous bands of staining. Western immunoblotting showed marked cross‐reactivity of the high antisera concentrations with several components of the serum proteins, but none corresponding to the purified P‐3‐P.

All tested sera from dogs had minimal competitive binding with radiolabeled P‐3‐P in the radioimmunoassay. Dilution curves of dog sera did not parallel either the standard curve or the dilution curve of a known test human serum. There were no statistically different P‐3‐P concentrations in any of the groups of dogs studied.

It was concluded that currently available radioimmunoassay kits for the measurement of P‐3‐P in the human are not applicable in the dog. Seemingly, the structure or metabolism of canine P‐3‐P may vary significantly from that of the bovine or human, limiting the sensitivity and specificity of this assay in the dog.
Cytochemical Staining Characteristics of Chicken Heterophils and Eosinophils
Authors: Andreasen Claire B. , Latimer Kenneth S.
Published: February 2009

Cytochemistry was used to differentiate chicken heterophils and eosinophils in blood smears and cytospin preparations of isolated leukocytes. Cytochemical staining also was performed to evaluate any alteration of cell staining characteristics after heterophils had been isolated using discontinuous Ficoll‐Hypaque gradients. Cytochemical staining reactions of heterophils and eosinophils were the same before and after discontinuous gradient cell separation. Characteristic positive reactions did not occur when chicken heterophils were stained with alkaline phosphatase, peroxidase, Sudan black B, acid phosphatase, alpha‐naphthyl acetate esterase, naphthol AS‐D chloroacetate esterase, periodic acid‐Schiff, or Luna's eosinophil granule techniques. Chicken eosinophils stained positively when peroxidase, Sudan black B, acid phosphatase, and Luna methods were performed. In highly cellular cytospin preparations, peroxidase reactions readily distinguished the few contaminating eosinophils (< 1%) from the heterophils.
Cerebrospinal Fluid Response Following Metrizamide Myelography in Normal Dogs: Effects of Routine Myelography and Postmyelographic Removal of Contrast Medium
Authors: Widmer William R. , Blevins William E. , Cantwell H. Dan , Cook James R. , DeNicola Dennis B.
Published: February 2009

The effect of metrizamide myelography on 90‐minute postmyelographic cerebrospinal fluid (CSF) samples was evaluated in a paired crossover study in 16 normal dogs. Each dog received a routine cervical myelogram (nonwithdrawal myelography) and a myelogram followed by contrast medium removal via aspiration from the subarachnoid space (withdrawal myelogram). Following nonwithdrawal myelography, the CSF was characterized by mild inflammation with a mixed pleocytosis and increased protein concentration. Compared with the nonwithdrawal CSF samples, the postmyelographic CSF of the withdrawal dogs had a more severe Inflammatory response with significant increases (p < 0.05) in absolute numbers of neutrophils, monocytoid cells, eosinophils, lymphocytes, and protein concentration. The withdrawal procedure may have contributed an additional mechanical effect on the leptomeninges producing the more severe inflammatory response in the withdrawal dogs. Although seizure data are not reported here, postmyelographic seizures were more frequent following non‐withdrawal myelography as compared with withdrawal myelography (p < 0.05), suggesting a decrease in metrizamide‐induced neurotoxicity for the withdrawal dogs.
Lipoprotein Electrophoresis Differentiation of Chylous and Nonchylous Pleural Effusions in Dogs and Cats and Its Correlation with Pleural Effusion Triglyceride Concentration
Authors: Waddle Jerry R. , Giger Urs
Published: February 2009

Pleural effusions from 23 dogs and 25 cats hospitalized between November 1986 and April 1987 at the Veterinary Hospital of the University of Pennsylvania (VHUP) were classified as chylous (chylomicrons present) or nonchylous (chylomicrons absent) by the presence or absence, respectively, of a chylomicron band on lipoprotein electrophoresis gels of the effusions. Triglyceride concentrations, cholesterol concentrations, and cholesterol/triglyceride (C/T) ratios were compared between the chylous and nonchylous groups in each species. Cholesterol concentrations were not significantly different between chylous and nonchylous effusions in both dogs and cats. Cholesterol/triglyceride ratios of less than 1 were present in all dogs and cats with chylous effusions; however, 12% of dogs with nonchylous effusions and 50% of cats with nonchylous effusions also had a C/T ratio less than 1. Triglyceride concentrations accurately classified all effusions as chylous or nonchylous in both dogs and cats. Pleural effusions with triglyceride concentrations greater than 100 mg/dl were chylous in all cases, whereas, effusions with triglyceride concentrations less than 100 mg/dl were nonchylous in all cases.
Determination of Uric Acid in Canine Serum and Urine By High Performance Liquid Chromatography
Authors: Felice Lawrence J. , Dombrovskis Dzintra , Lafond Elizabeth , Bartges Joseph , Osborne Carl A.
Published: February 2009

A reproducible high performance liquid chromatography (HPLC) method was developed for analysis of uric acid in canine serum and urine. The method consists of precipitating serum proteins with phosphotungstic acid prior to HPLC analysis. Urine is analyzed after dilution with buffer. Chromatography is performed on a reversed‐phase C‐18 column with UV detection at 292 nm. Sensitivity of the method will allow reproducible measurement of uric acid at concentrations of 0.05 mg/dl in serum and 0.1 mg/dl in urine. The HPLC method has been used to quantify hundreds of canine serum and urine samples. The method is superior to UV absorption or colorimetric methods because its lower limit of detection allows measurement of uric acid at concentrations found in canine serum and urine.
Experimental Transmission of Granulocytic Ehrlichial Organisms in Dogs
Authors: Stockham S.L. , Tyler J.W. , Schmidt D.A. , Curtis K.S.
Published: February 2009

Canine granulocytic ehrlichial organisms were transmitted from an infected dog from Missouri to two male, 10‐month‐old dogs by an intravenous injection of whole blood. Physical or behavioral abnormalities were not detected during the 98 days of evaluation other than a mild pyrexia from Day 18 to 20. Ehrlichial morulae were found in blood granulocytes of Dog 1 from Day 13 to 44 and of Dog 2 from Day 14 to 34 with the peak rickettsemia occurring on Day 16 for both dogs. By Day 21 after inoculation, both dogs had positive titers to Ehrlichia canis. The highest titers for both dogs were found 63 days after inoculation, after which the titers decreased. Most of the hematologic abnormalities (i.e., neutropenia, lymphocytosis, thrombocytopenia) and fever occurred between 18 and 24 days after inoculation. The pathologic bases of these abnormalities were not investigated but their concurrent presence suggested an association with the dogs' immunologic responses to the granulocytic ehrlichial agent. Results from the study indicated that the canine granulocytic ehrlichial agent of Missouri may produce subclinical infections and suggested that dogs may be able to clear the organism without antimicrobial therapy.
Serum 5'Nucleotidase Activity in Rats: A Method for Automated Analysis and Criteria for Interpretation
Authors: Carakostas Michael C. , Power Richard J. , Banerjee Asit K.
Published: February 2009

A manual kit for determining serum 5'nucleotidase (5'NT, EC 3.1.3.5) activity was adapted for use with rat samples on a large discrete clinical chemistry analyzer. The precision of the method was good (within‐run C.V. = 2.14%; between‐run C.V. = 5.5%). A comparison of the new automated method with a manual and semi‐automated method gave regression statistics of y = 1.18x − 3.66 (Sy·x = 4.54), and y = 0.733x + 1.97 (Sy·x = 1.69), respectively. Temperature conversion factors provided by the kit manufacturer for human samples were determined to be inaccurate for converting results from rat samples. Analysis of components contributing to normal variation in rat serum 5'NT activity showed age and sex to be major factors. Increased serum 5'NT activity was observed in female rats when compared to male rats beginning at about 5 to 6 weeks of age. An analysis of variance of serum 5'NT, alkaline phosphatase, and GGT activities observed over a 9‐week period in normal rats suggests several advantages for 5'NT as a predictor of biliary lesions in rats.
Quantitative Evaluation of Echinocytes in the Dog
Authors: Weiss Douglas J. , Kristensen Annemarie , Papenfuss Nancy , McClay Carolyn B.
Published: February 2009

Echinocytes were evaluated quantitatively in both healthy and sick dogs and compared with red blood cell (RBC) morphology on Wright's‐stained blood smears. Of 68 samples submitted from sick dogs, 32 had high echinocyte numbers. Two diseases, glomerulonephritis and lymphosarcoma, accounted for 15 of the 32 cases. In only four instances were echinocytes reported from blood smear evaluation and not found to be high by quantitative studies. We conclude that nonartifactual echinocytosis occurs in the dog and that it may be associated with specific diseases.
Anti‐A Isoagglutinins in Two Blood Type B Cats Are IgG and IgM
Authors: Wilkerson Melinda J. , Meyers Kenneth M. , Wardrop K. Jane
Published: February 2009

Sera from two blood type B cats had strong isoagglutinating and isohemolyzing titers against blood type A erythrocytes. In order to determine the class of the immunoglobulins, sera from the cats were pooled, ammonium sulfate precipitated, and gel filtered using sepharose 6B to separate the immunoglobulins by molecular size. The immunoglobulin concentrate separated into two fractions. The initial part of the first fraction was shown in an ELISA to contain IgM and to be devoid of IgG by immunoelectrophoresis and to have agglutinating activity (a titer of 1:4 with a protein concentration of 0.8 mg/ml). Treating this fraction with the reducing agent dithiothreitol (DTT) eliminated agglutinating activity. The latter portion of the second column fraction was shown to contain IgG by immunoelectrophoresis, and to be devoid of IgM by ELISA. Agglutinating activity was also present in the second fraction (a titer of 1:2 with a protein concentration of 1.9 mg/ml); hemagglutinating activity was not decreased by DTT treatment. These studies show that the predominant anti‐A isoagglutinating activity in pooled sera from two blood type B cats is IgM and that some isoagglutinin activity can be associated with IgG.
Detection of Antiplatelet Antibody: Comparison of Platelet Immunofluorescence, Agglutination, and Immunoinjury Tests Using Rabbit Antiequine Platelet Serum
Authors: Jain Nemi C. , Dhawedkar Ram G. , Vegad J.L. , Kono Constance S.
Published: February 2009

Immunofluorescence, tube agglutination, and platelet factor‐3 immunoinjury tests for detecting antiplatelet antibody were compared using a heterologous system of equine platelets and rabbit antiequine platelet serum. Platelet immunofluorescence tests were performed using paraformaldehyde‐fixed platelets in suspension as well as in air‐dried smears on glass slides (solid phase). Bright homogeneous, membranous, specific fluorescence was seen in both assays with anti‐immunoglobulin G (IgG) and protein G fluorescein isothiocynate conjugates (FITC‐conjugates). Protein A conjugate gave nonspecific fluorescence irrespective of normal or antiserum treatment. Anti‐IgG and protein G conjugates in suspension immunofluorescence tests with the same antiserum yielded antibody titers of 1:1024 and 1:128, respectively. Similarly, respective titers of 1:512 and 1:64 were obtained with solid phase immunoassay. Platelet suspension assay was slightly better than the solid phase assay. These observations indicated that anti‐IgG was more sensitive than protein G in detecting antiplatelet antibody by fluorescence microscopy, while protein A was ineffective because of its nonspecificity. Chloroquine treatment of platelets failed to reduce the nonspecific fluorescence. Platelet agglutination and platelet factor‐3 tests were relatively less sensitive to detect equine antiplatelet antibody.
Epidemiologic Survey of Thrombocytopenia in Dogs: A Report on 987 Cases
Authors: Grindem Carol B. , Breitschwerdt Edward B. , Corbett Wayne T. , Jans Heather E.
Published: February 2009

Thrombocytopenia was documented in 987 of 18,910 (5.2%) dogs admitted to North Carolina State University, College of Veterinary Medicine, Veterinary Teaching Hospital, between 1983 and 1989. Classifying thrombocytopenic dogs by etiologic groups revealed the following proportionate ratios: 5% (48/987) immune‐mediated thrombocytopenia; 13% (130/987) neoplasia‐associated thrombocytopenia; 23% (224/987) inflammatory***linfectious thrombocytopenia; and 59% (585/987) miscellaneous thrombocytopenia. Dogs with immune‐mediated thrombocytopenia had significantly (P < 0.05) lower platelet counts (mean 36,760 ± 50,288/μl) than dogs in the other three groups, and Doberman Pinschers were overrepresented in all groups except the immune‐mediated thrombocytopenic group. We conclude that thrombocytopenia is a prevalent and potentially important diagnostic finding in a variety of disease states.
Inherited Erythrocyte Pyruvate Kinase Deficiency in a Beagle Dog
Authors: Giger Urs , Mason Gail D. , Wang Ping
Published: February 2009

A 1‐year‐old, female Beagle dog with minimal exercise intolerance was found to have a persistent, severe, and highly regenerative anemia, splenomegaly, and progressive osteosclerosis. Despite near‐normal in vitro erythrocyte pyruvate kinase (PK) activity, the authors diagnosed PK deficiency by demonstrating a glycolytic block at the PK step, the lack of normal R‐type PK isoenzyme, and the presence of M2‐type PK in the animal's erythrocytes. The dam had half‐normal erythrocyte PK activity, which supports an autosomal recessive mode of inheritance. We conclude from our studies that close similarities exist between erythrocyte PK deficiency in Beagle, Basenji, and West Highland White Terrier dogs and that this form of PK deficiency may be more widespread than previously thought.
Using the Correct Regression Analysis Technique in Method Comparison Studies
Authors: Carakostas Michael C. , Green John W.
Published: February 2009

Regression analysis frequently is used to evaluate a new clinical laboratory method. Results from the new method are compared to results from an existing more established method. If measurement error exists in the established method, then least squares may not be an appropriate statistical method to use for the regression analysis. An errors variable regression analysis model was used to evaluate data from five method comparison studies. Results were compared to least squares analyses performed on the same data. When significant measurement error existed in the “reference” method, the errors variable analysis provided a less blased estimate of the regression statistics, which differed markedly from the least squares results. Inappropriate use of least squares in method comparison studies can lead to erroneous conclusions about the relationship of a new analytical method to an existing method.
Urine Protein Determination in Dogs and Cats: Comparison of Dipstick and Sulfasalicylic Acid Procedures
Authors: Moore Frances M. , Susan L. , Brown Lynne
Published: February 2009

Protein levels in urine specimens from 91 dogs and 65 cats were evaluated by sulfasalicylic acid precipitation (SSA) and dipstick methods. The dipstick frequently yielded reactions for protein that were greater than the level of protein indicated by SSA (i.e., false positive reactions), although no false negative reactions for protein were noted. All urine specimens with protein levels equal to or greater than 100 mg/dl by SSA had dipstick results of 3 +. Results of this study suggest that dipstick analysis for urine protein is an adequate screening procedure for the selection of urines for quantitative analysis of protein and creatinine to assess proteinuria.
Hematology Analyzer Comparison: Ortho ELT‐8/ds vs. Baker 9000 for Healthy Dogs, Mice, and Rats
Authors: Weingand Kurt W. , Odioso Leslie W. , Dameron Gregory W. , Laytart Melanie J. , Stitzel Katherine A.
Published: February 2009

A Baker 9000 hematology analyzer (electronic impedance) was purchased to replace an Ortho ELT‐8/ds analyzer (laser optics) due to discontinued technical support. An analytical comparison of hemograms from healthy dogs, rats, and mice was made from paired disodium ethylenediamine tetra‐acetate anticoagulated blood samples. Both instruments were calibrated with human blood products, and the ELT‐8/ds hematocrit (HCT) was calibrated to a spun packed cell volume (PCV) for each species. For Beagle dogs (n = 49), Baker 9000 mean platelet (PLT) counts had a negative bias of −89 × 103/μl when compared to ELT‐8/ds values. Mean ± standard error manual PLT counts compared well with Baker 9000 values for dogs (n = 10): 369 ± 28 vs. 367 ± 27 × 103/μl; r = 0.93. For CD‐1 mice (n = 44), Baker 9000 mean white blood cell (WBC) counts had positive biases of 1.1 × 103/μl when compared to ELT‐8/ds and 0.5 × 103/μl when compared to hemacytometer counts. Diluted microsamples using the predilution mode on the Baker 9000 compared well with undiluted samples for mice. For Sprague‐Dawley rats (n = 70), Baker 9000 mean WBC, red blood cell (RBC), and PLT counts had absolute biases of 0.8 × 103/μl, −1.09 × 106/μl, and −357 × 103/μl, respectively, when compared to ELT‐8/ds values. Baker 9000 RBC, WBC, and PLT counts from rats compared well with reference hemacytometer counts. The Baker 9000 HCT determination for rats had an absolute negative bias of 6% when compared to the ELT‐8/ds values or spun PCV. The Baker 9000 required whole blood calibration to PCV for accurate determination of HCT for rats. The biases between analyzers may be due to inherent physical differences between the analytical methods and/or the calibration techniques.
Analysis of Serum Protein Concentrations After Severe Thermal Injury in a Dog
Authors: Kern Margaret R. , Stockham Steven L. , Coates Joan R.
Published: February 2009

Serial serum electrophoreses and routine serum protein assays were used to assess changes in serum protein concentrations after severe thermal injury in a dog. Electrophoretic patterns during the month of evaluation were consistent with protein‐losing dermatopathy (thermal burn) and inflammatory dysproteinemias.
Development of a Teaching Laboratory Aid for Instruction of Fine Needle Aspiration Biopsy Cytology Technique
Authors: Simpson R. Mark , Meuten Donald J.
Published: February 2009

Specialized training is required to obtain adequate cytology smears by needle aspiration biopsy. An educational method that would provide opportunity for numerous trainees to acquire needle aspiration biopsy and cytodiagnostic skills, and which would not result in the interruption of teaching hospital patient care nor result in the use or sacrifice of animals from laboratory animal resources was sought. An aspiration biopsy cytology teaching aid was developed by placing portions of fresh organ tissue from routine submissions to the necropsy laboratory into a specimen container partially filled with chilled normal saline solution. Specimens were stored immersed in saline at 4°C for up to 48 hours. Prior to the cytopathology teaching laboratory period, the specimen container was removed from storage and covered with a latex diaphragm. Instructors responsible for teaching cytodiagnostic techniques used this ex vivo aspiration biopsy cytology device for instruction of fine needle aspiration biopsy, specimen processing, and microscopy. In the teaching laboratory, puncture of the diaphragm and aspiration of a tissue sample with hypodermic needle and syringe was instructed to, and practiced by, numerous trainees simultaneously using clinical operative technique without the need for live animals. Trainees prepared and stained cytology smears as well as evaluated and interpreted specimens in a single, realistic exercise. Such training experience may improve ability of trainees to procure diagnostic‐quality biopsy specimens for cytodiagnosis of lesions from clinic patients.
Cytochemical Staining Characteristics of Lymph Nodes From Normal and Lymphoma‐Affected Dogs
Authors: Raskin Rose E. , Nipper Marlin N.
Published: February 2009

Frozen sections and imprint smears were used to evaluate the presence and pattern of cytochemical staining reactions in the B‐ and T‐cell regions of lymph nodes from normal dogs and dogs with lymphoma. Staining procedures evaluated included peroxidase (PER), Sudan black B (SBB), naphthol AS‐D chloroacetate esterase (CAE), α‐naphthyl butyrate esterase (NBE), acid phosphatase (ACP), and leukocyte alkaline phosphatase (LAP). In normal lymph nodes, macrophages and some lymphocytes within the interfollicular (T‐cell) region and medulla stained positive with ACP and NBE. Smaller numbers of macrophages also occurred sporadically within the germinal follicles. Cells positive for PER, SBB, and CAE were scattered infrequently throughout all regions of the normal lymph node, consistent with granulocytes and mast cells. The LAP stained cells were predominantly and prominently located within the mantle zone of secondary follicles and to a much lesser extent within the germinal centers, compatible with B‐cell lymphocytes derived from follicular center cells. Of the 12 dogs with lymphoma, 7 cases (4 immunoblastic, 2 large noncleaved, 1 small noncleaved) stained diffusely positive with LAP; 4 cases (all lymphoblastic) had numerous focally positive lymphocytes using ACP and NBE; and 1 case (immunoblastic) did not stain positive with any of the cytochemical reactions. Cytochemical staining of canine lymph nodes with NBE, ACP, and LAP proved useful in distinguishing between B‐ or T‐cell regions and detecting different cell types of canine lymphoma.
Inflammatory Disorders of Bone Marrow
Authors: Weiss Douglas J. , Greig Barbara , Aird Betsy , Geor Raymond J.
Published: February 2009

Inflammatory lesions in bone marrow, observed during a 2 year period at the College of Veterinary Medicine, University of Minnesota, were reviewed. Of 24 bone marrow specimens with evidence of inflammation, six were classified as acute inflammation, nine as fibrinous inflammation, five as chronic inflammation/hyperplasia, three as granulomatous inflammation and one as nodular lymphoid hyperplasia. Acute inflammation commonly accompanied bacterial sepsis. Two patterns of acute inflammation were identified. One pattern consisted of multifocal microabscesses. The other pattern of acute inflammation consisted of perivascular infiltrates of neutrophils, fibrin, edema, and hemorrhage. The most common disorder associated with fibrinous inflammation was disseminated intravascular coagulopathy. Chronic inflammation was difficult to differentiate from chronic immune stimulation. Discrete granulomas were identified in the marrow of animals with systemic mycotic disease, idiopathic systemic granulomatous disease, and serous atrophy of fat. This study indicates that a broad variety of inflammatory disorders occur in animal bone marrow and that these disorders can be classified based on general categories of inflammation described in other tissues.
Detection of Intravaginal Spermatozoa After Natural Mating in the Bitch
Authors: Whitacre M.D. , Yates D.J. , VanCamp S.D. , Meuten D.J.
Published: February 2009

Mismating is one of the most frequently presented reproductive problems presented to veterinarians. Confirming whether or not a mismating has occurred is necessary to determine if therapy will be instituted. Detection of spermatozoa in the vagina is irrefutable evidence of copulation; however, absence of sperm in a vaginal cytology cannot rule out coitus. In order to improve detection of spermatozoa post‐coitus, a prospective study was initiated utilizing natural breedings of purebred beagles.
Effects of Storage Time on Chemistry Results from Canine Whole Blood, Heparinized Whole Blood, Serum and Heparinized Plasma
Authors: Thoresen Stein Istre , Havre Gustav N. , Morberg Hanne , Mowinckel Petter
Published: February 2009

The stability and storage characteristics of 24 blood constituents from dogs including nine enzymes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, lipase), 15 metabolites and minerals (albumin, bile acids, bilirubin, calcium, cholesterol, creatinine, fructosamine, glucose, magnesium, phosphate, potassium, protein, sodium, triglycerides, urea) were studied. Conditions studied included storing of nonanticoagulated and heparinized whole blood for 3 days (Part A), and storing of serum and heparinized plasma for 3 days (Part B). The storage temperature for both studies was + 4°C from day 0 to day 1, and + 20°C, from day 1 to day 2 and 3.

Eight of 24 analytes showed no significant differences (p > 0.05) for three days in whole blood. However, the stability of all 24 analytes greatly improved by storing serum or heparinized plasma compared to nonanticoagulated or heparinized whole blood. In stored serum or heparinized plasma, 20 of 24 analytes showed no significant differences (p < 0.05) for 3 days.

Nine of 24 analytes showed significant differences (p < 0.05) between serum and heparinized plasma, where CK, LDH, GGT, and potassium showed differences of possible clinical importance.

This study strongly supports the practice of separating serum/plasma from clot/cells as promptly as possible to achieve improved stability for most analytes under test.
Bone Marrow Contamination of Canine Cerebrospinal Fluid
Authors: Christopher Mary M.
Published: February 2009

Bone marrow cells were identified in cytocentrifuge preparations of cerebrospinal fluid (CSF) obtained by lumbar punctures from two dogs. CSF analyses were characterized by normal or increased total cell counts, normal or increased total protein concentration and the presence of erythrocytes. Hematopoietic cells present included both erythroid and myeloid precursors and, in one case, an erythroblastic island was seen. Peripheral blood smear examination confirmed that the hematopoietic cells observed were not the result of blood contamination. Neither case was associated with a difficult tap procedure, nor with a specific disease process. Contamination may have resulted from actual marrow penetration or from extramedullary hematopoiesis in the dura mater. Accurate interpretation of CSF cytology requires the consideration of possible bone marrow contamination when hematopoietic cells are observed.