Shelley A. Burton

Liver Histopathology and Liver and Serum Alanine Aminotransferase and Alkaline Phosphatase Activities in Epileptic Dogs Receiving Phenobarbital

Phenobarbital (PB) therapy is frequently associated with elevated serum alanine aminotransferase (ALT) and alkaline phosphatase (AP) activities in dogs without clinical signs of liver disease. The goal of this study was to determine if increased serum ALT and AP activities in clinically healthy PB-treated epileptic dogs are due to hepatic enzyme induction or to subclinical liver injury. Liver biopsies were obtained from 12 PB-treated dogs without clinical signs of liver disease but with elevated serum ALT and/or AP activities or both. Liver biopsies were obtained from eight healthy control dogs not receiving PB. Biopsies were evaluated histopathologically (all dogs) and liver homogenates were assayed for ALT (all dogs) and AP (six treated dogs, all controls) activities. As a positive control, liver cytochrome P4502B, an enzyme known to be induced by PB, was measured by benzyloxyresorufin-O-dealkylase activity and immunoblotting (five treated dogs, all controls). Serum AP isoenzyme analyses were performed. Results showed that ALT and AP activities in liver homogenates were not increased in treated dogs compared with controls, whereas the positive control for induction, CYP2B, was dramatically increased in treated dogs. Histopathological examination of liver biopsies revealed more severe and frequent abnormalities in treated dogs compared to controls, but similar types of abnormalities were found in both groups. Serum AP isoenzyme analyses in treated dogs demonstrated increased corticosteroid-induced and liver isoenzyme activities compared to controls. Results do not support induction of ALT or AP in the liver as the cause of elevated serum activities of these enzymes due to PB.

Babesia (Theileria) annae in a red fox (Vulpes vulpes) from Prince Edward Island, Canada.

A 4–6-mo-old female red fox (Vulpes vulpes) was presented to the Atlantic Veterinary College (AVC) Teaching Hospital, Prince Edward Island, Canada. On presentation, the fox was weak and had pale mucous membranes. A complete blood count and a serum biochemistry profile were performed. Blood smear examination revealed low numbers of erythrocytes containing centrally to paracentrally located, single, rarely multiple, approximately 1×2 μm, oval to round organisms with morphology similar to Babesia microti. Polymerase chain reaction testing and DNA sequencing of the Babesia species 18S rRNA gene were performed on DNA extracted from whole blood. Results were positive for a Babesia microti–like parasite genetically identical to Babesia (Theileria) annae. The fox was euthanized due to poor prognosis for recovery. Necropsy examination revealed multifocal to locally extensive subacute nonsuppurative meningoencephalitis, an eosinophilic bronchopneumonia, a moderate diffuse vacuolar hepatopathy, and lesions associated with blunt trauma to the left abdominal region. This is the first reported case of a red fox in Canada infected with a piroplasm. It remains uncertain whether the presence of this hemoparasite in this fox was pathogenic or an incidental finding. The potential for competent vectors of Babesia species on Prince Edward Island, the potential for this Babesia microti–like parasite to infect other wild and domestic canids, and the significance of this parasite to the health of infected individuals are yet to be determined.

Association between excess body weight and urine protein concentration in healthy dogs

BACKGROUND Markedly overweight people can develop progressive proteinuria and kidney failure secondary to obesity-related glomerulopathy (ORG). Glomerular lesions in dogs with experimentally induced obesity are similar to those in people with ORG. OBJECTIVES The aim of this study was to evaluate if urine protein and albumin excretion is greater in overweight and obese dogs than in dogs of ideal body condition. METHODS Client-owned dogs were screened for underlying health conditions. These dogs were assigned a body condition score (BCS) using a 9-point scoring system. Dogs with a BCS of ≥ 6 were classified as being overweight/obese, and dogs with a BCS of 4 or 5 were classified as being of ideal body weight. The urine protein:creatinine ratio (UPC) and urine albumin:creatinine ratio (UAC) were then determined, and compared between 20 overweight/obese dogs and 22 ideal body weight control dogs. RESULTS Median UPC (0.04 [range, 0.01-0.14; interquartile range, 0.07]) and UAC (0.41 [0-10.39; 3.21]) of overweight/obese dogs were not significantly different from median UPC (0.04 [0.01-0.32; 0.07]) and UAC (0.18 [0-7.04; 1.75]) in ideal body weight dogs. CONCLUSIONS Clinicopathologic abnormalities consistent with ORG were absent from overweight/obese dogs in this study.

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

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

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

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

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

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

What is your diagnosis? Blood smear from a cat

A 16-year-old spayed female Domestic Shorthaired cat was presented for a recent episode of vomiting. The cat had been diagnosed with hyperthyroidism 4 months prior to presentation, at which time oral methimazole therapy had been initiated. The cat had been doing well clinically. Whole blood was submitted in EDTA and red top tubes for hematologic analysis on a Sysmex XT-2000iv (Sysmex Corporation, Kobe, Japan) and biochemical analysis on a Roche COBAS (Roche Diagnostics Corp., Indianapolis, IN, USA). Evaluation of CBC data revealed a mild neutrophilia (segmented neutrophils 13.7 9 10/L, reference interval [RI] 2.2–9.5 9 10/L) with a mild left shift (band neutrophils 0.3 9 10/L, RI 0.0–0.1 9 10/L). Inclusions were noted in the cytoplasm of ~65% of neutrophils (Figure 1). Significant changes in serum biochemistry included moderate azotemia (urea 15.9 mmol/L, RI 6.4–11.8 mmol/L, creatinine 318 umol/L, RI 67–157 lmol/L) and mildly increased activities of alanine aminotransferase (163 U/L, RI 34–90 U/L), aspartate transaminase (180 U/L, RI 11–44 U/L), and lipase (293 U/L, RI 16–139 U/L). Figure 1. Neutrophil with cytoplasmic inclusions in a blood smear from a cat. Wright–Giemsa.

Pathology in practice. Neutrophilic and histiocytic inflammation with intracellular bacteria (consistent with R equi).

Palpation of the limb did not elicit signs of pain, and no other abnormalities were noted during physical examination. Results of a CBC and serum biochemical analyses were within reference limits. A test for circulating FeLV antigen and anti-FIV antibodies yielded negative results. Hair on the lower portion of the right limb was clipped, and the area was cleaned with chlorhexidine gluconate. Fusidic acid cream was applied topically