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The IDEXX SediVue Dx (SediVue) is an automated, in-clinic urine sediment analyzer for veterinary patients. The bias between the results from manual microscopy and the SediVue is currently unknown.To assess the diagnostic accuracy of the SediVue, we aimed to determine the bias between the SediVue (index test) and manual microscopy (reference standard) for the quantification of RBCs and WBCs in urine.Urine remnant samples were collected from cats and dogs that contained RBCs (n = 462) and WBCs (n = 510). Retrospective analysis of results from urine sediment examinations using both manual microscopy (using a KOVA and DeciSlide system) and the SediVue (1.0.1.3) was performed. Bias was determined with Bland-Altman plots. SediVue-captured images from high-bias samples were reviewed, and biases were compared.The median bias for semi-quantitative RBC and WBC counts was determined for RBC and WBC counts. The cutoffs were RBC ≤ 5/HPF, 0.3; RBC 5.1-10/HPF, 10.1; RBC 10.1-20/HPF, 10.6; and RBC 20/HPF, 28.93; WBC ≤ 5/HPF, 0.1; WBC 5.1-10/HPF, 2.2; WBC 10.1-20/HPF, 9.4; and WBC 20/HPF, 26.6. High bias between the methods was identified in 98 samples (21.0%) with RBCs and 77 samples (15.7%) with WBCs. Reviewer-based enumeration of the SediVue-captured images decreased the percentage of samples with high bias to 17.3% for RBCs and to 11.4% for WBCs.Bias in the RBC and WBC counts between manual microscopy and the SediVue was unlikely to impact clinical interpretations in a majority of cases. Although reviewer enumeration of SediVue-captured images reduced observed bias, inherent differences between methodologies appeared to have a larger impact on the bias.

Objective To evaluate the proportion of blood samples diagnosed with reticulocytosis without anaemia in cats and dogs and report the aetiology and mortality rate of affected animals. Materials and methods Retrospective multicentre study including haematological examination of 3956 cats and 11,087 dogs admitted to seven German veterinary clinics (2012 to 2014). The proportion of blood samples with reticulocytosis without anaemia was calculated, and after exclusion of multiple measurements of the same animal, clinical data were evaluated. Animals with reticulocytosis without anaemia were classified as healthy or diseased, and diseased patients were assigned to 12 disease groups. Pretreatment (i.e. non-steroidal anti-inflammatory drugs, glucocorticoids, dipyrone) was recorded. Results The proportion of blood samples with reticulocytosis without anaemia was 3·1% (124/3956) in cats and 4·4% (492/11,087) in dogs. Overall, 1·8% (2/111) of cats and 1·5% (7/458) of dogs with reticulocytosis without anaemia were healthy. Blood loss/anaemia, cardiac/respiratory disorders, gastrointestinal disorders and inflammatory disorders as well as cancer were the most frequent underlying diseases. Pretreatment was noted in 39·5% (43/111) of cats and 42·4% (194/458) of dogs. The mortality rate was 37·8% (42/111) in cats and 29·7% (136/458) in dogs with reticulocytosis without anaemia; the median survival time in non-survivors was 1 day (range: 0 to 376 days in cats, 0 to 444 days in dogs). Clinical significance In both species, reticulocytosis without anaemia was observed in a low proportion of blood samples (dogs>cat). Though a bias towards sick animals is possible in our sample, reticulocytosis without anaemia was mainly seen in diseased animals and associated with a mortality rate of approximately one-third of patients.

Background Reticulocyte hemoglobin content provided by the Siemens ADVIA (CHr) is an established marker of iron deficiency. The IDEXX ProCyte Dx hematology analyzer now provides a similar variable, reticulocyte hemoglobin equivalent (RET-He). Objectives The objective was to evaluate RET-He and its diagnostic utility in dogs, and to calculate a cutoff value for diagnosing iron-deficient erythropoiesis (IDE). Furthermore, the prevalence of RET-He values below this cutoff value was established. Methods One hundred and seventy-one CBCs of healthy dogs were used to establish a RI. Stability of RET-He was evaluated by repeated measurements over 48 hours (n = 10). The 25-run coefficient of variation (CV) was calculated, and correlation and bias between measurements of RET-He and CHr were assessed (n = 190). A cutoff value for diagnosing IDE was calculated. The utility of RET-He in the detection of IDE was evaluated in 123 dogs. The prevalence of low RET-He values was assessed retrospectively in a multicenter study (2012-2014) under participation of 7 veterinary clinics. Results Reticulocyte hemoglobin equivalent with an RI of 22.2 to 28.6 pg was statistically stable over 48 hours (P = .10). The CV was 1.8%. A fair correlation (ρ = 0.74) between RET-He and CHr with a small bias of -0.6 pg was found. The cutoff value for diagnosing IDE was 20.9 pg (sensitivity: 85%; specificity: 99%). The prevalence of RET-He values below 20.9 pg was 10.3% (1084/10,553 dogs). Conclusions RET-He on the ProCyte Dx is a precise screening tool in dogs to detect iron-deficient erythropoiesis.

BACKGROUND Reticulocyte hemoglobin content (RET-He) is a diagnostic marker for iron deficiency (ID) in people and dogs. OBJECTIVES The aim of our study was to evaluate the clinical utility of RET-He in the diagnosis of different causes of iron-deficient erythropoiesis (IDE). METHODS Canine CBCs were separated into 2 groups according to RET-He values

C-reactive protein (CRP) is a major acute phase protein used to monitor response to treatment during surgical recovery. Depending on the anatomical problem, surgery type and technique, the level of CRP can change drastically. The aim of this study was to describe the changes in CRP and white blood cell (WBC) levels following surgery for medial patellar luxation in otherwise healthy dogs. Twenty-two dogs completed the study. CRP was measured using a commercially available dry chemistry slide on a commercially available in-clinic analyser. Analyses were performed using the Wilcoxon Rank Sum test and a mixed effects Poisson regression model. A significant change in CRP levels was found between the pre-anesthetic and 24 hr post-surgical timepoint with a median difference of 92.0 mg/dL (P < 0.001). Though a median drop in the CRP value of 13.9 mg/dL was observed between the 24 hr and 48 hr post-surgical time period, the result was not statistically significant (P = 0.456). Similarly, there was a significant increase in WBC between the pre-anesthetic and 24-hr post-surgical time point (P < 0.001) followed by a significant decrease in WBC between the 24 hr and 48-hr post-surgical time points (P = 0.015). In this study population, CRP levels were observed to aid in monitoring of the overall health of the dogs following surgery for medial patellar luxation. The results of this study suggest that both CRP and WBC values significantly increase by 24 hr but where CRP levels remain elevated through 48 hr post-surgery, WBC showed a significant drop between 24 and 48 hr. Further investigation into the length of time for both CRP and WBC to reach basal levels in this particular type of surgery would be of value to monitor recovery from surgery.

Background Microscopic evaluation of urine is inconsistently performed in veterinary clinics. The IDEXX SediVue Dx® Urine Sediment Analyzer (SediVue) recently was introduced for automated analysis of canine and feline urine and may facilitate performance of urinalyses in practice. Objective Compare the performance of the SediVue with manual microscopy for detecting clinically relevant numbers of cells and 2 crystal types. Samples Five-hundred thirty urine samples (82% canine, 18% feline). Methods For SediVue analysis (software versions [SW] 1.0.0.0 and 1.0.1.3), uncentrifuged urine was pipetted into a cartridge. Images were captured and processed using a convolutional neural network algorithm. For manual microscopy, urine was centrifuged to obtain sediment. To determine sensitivity and specificity of the SediVue compared with manual microscopy, thresholds were set at ≥5/high power field (hpf) for red blood cells (RBC) and white blood cells (WBC) and ≥1/hpf for squamous epithelial cells (sqEPI), non-squamous epithelial cells (nsEPI), struvite crystals (STR), and calcium oxalate dihydrate crystals (CaOx Di). Results The sensitivity of the SediVue (SW1.0.1.3) was 85%-90% for the detection of RBC, WBC, and STR; 75% for CaOx Di; 71% for nsEPI; and 33% for sqEPI. Specificity was 99% for sqEPI and CaOx Di; 87%-90% for RBC, WBC, and nsEPI; and 84% for STR. Compared to SW1.0.0.0, SW1.0.1.3 had increased sensitivity but decreased specificity. Performance was similar for canine versus feline and fresh versus stored urine samples. Conclusions and clinical importance The SediVue exhibits good agreement with manual microscopy for the detection of most formed elements evaluated, but improvement is needed for epithelial cells.

Background Centrifugation is the primary method used to perform urine sediment analyses, but evaluation of other methods is required to validate centrifugation. Objectives Non-urine materials were used to examine the repeatability (precision) and effectiveness (recovery) of four sediment methodologies on red blood cell (RBC) and white blood cell (WBC) counts. Methods Four urine sediment methods were compared using commercially available quality control material (QCM) and fresh canine RBCs in a diluent. Treatments included (a) 5 mL centrifugation at 390g for 5 minutes; (b) 1.5 mL centrifugation at 3900g for 45 seconds; (c) 60 µL of neat (unspun urine) in a microtiter well; and (d) 30 µL of neat on a slide with a coverslip. A within-run precision using QCM was followed by a one-run comparison test performed with a suspension of canine erythrocytes. RBC morphology was also examined. Results All results are listed in order of Methods A-D. Percent coefficients of variation (%CVs) for WBCs were 23.2%, 33.7%, 15.0%, and 27.2%. Red blood cells %CVs were 34.3%, 29.2%, 16.2%, and 24.4%. Average WBC counts in ten fields of view (FOV) ± 1 SD were 26.4 ± 6.1, 14.2 ± 4.8, 32.8 ± 4.9, and 1.6 ± 0.4. Average RBC counts in 10 fields of view (FOV) ± 1 SD were 45.3 ± 15.5, 23.9 ± 7.0, 38.4 ± 6.2, and 2.6 ± 0.6. The one-run comparison test reports average RBC counts per FOV at 55.2, 23.4, 92.8, and 13.8. The percentages of abnormal RBCs were 92.2%, 74.8%, 7.0%, and 55.1%. Conclusions Method C had the best reproducibility, a lower frequency of cell morphology abnormalities, and similar cellular counts to those of Methods A and B.

The purpose of this study was to determine the extent to which pretreatment prostaglandin E2 (PGE2) concentration and cyclooxygenase-2 (cox-2) expression could be used to predict the antitumor activity of cox inhibitor treatment in naturally occurring canine transitional cell carcinoma of the urinary bladder (TCC). Snap frozen tissues (to measure PGE2) and formalin-fixed TCC samples (for cox-2 immunohistochemistry) were obtained by cystoscopy or surgery. Complete tumor staging was performed before and after one month of treatment with the cox inhibitor, piroxicam (0.3 mg/kg q24 h po). The pretreatment PGE2 concentration ranged from 57 to 1624 ng/g of TCC tissue; n=18 dogs). Cox-2 immunoreactivity was observed in all TCC samples. There was no association between PGE2 concentration, cox-2 expression, and change in tumor volume with piroxicam treatment. In conclusion, cox-2 expression or PGE2 concentration alone, or the combination of the two was not useful in predicting response to piroxicam treatment in canine TCC.

The purpose of this work was to determine cox-1 and cox-2 expression by immunohistochemistry in forms of naturally occurring canine cancer in order to identify animal systems for pre-clinical evaluation of cox inhibitors and cox-2 inhibitors in cancer. Canine lymphoma (LSA), prostatic carcinoma (PCA), osteosarcoma (OSA), oral melanoma (MEL), oral squamous cell carcinoma (SCC), oral fibrosarcoma (FSA), mammary carcinoma (MCA), and normal tissues were included. Cox-2 was expressed in epithelial tumors (17 of 26 SCC, 8 of 13 MCA, 5 of 9 PCA cases) and MEL (9 of 15 cases), but was generally absent in normal tissues. Cox-2 expression was minimal or absent in mesenchymal tumors and LSA. Cox-1 was expressed in normal epithelial tissues and in some osteoclast and osteoblast in bone, but was absent in normal lymph node. In conclusion, forms of canine cancer were identified for in vivo studies of the effects of cox inhibitors and selective cox-2 inhibitors on cancer.

A placebo-controlled experiment was performed to evaluate the effect of potassium bromide on the canine thyroid gland. Basal total thyroxine, free thyroxine, and basal thyrotropin serum concentrations were evaluated over a 6-month period in potassium bromide-treated and control dogs. A thyrotropin-releasing hormone stimulation test was also performed in all dogs at the beginning and conclusion of the study. Thyroid histopathology was compared between treated and control dogs at the end of the study. No difference was detected in any parameter between the two groups at the end of the study. A decline in thyroid hormone concentrations over the course of the study did occur in both groups of dogs. Potassium bromide does not appear to have a significant effect on canine thyroid function or morphology.

The purpose of this study was to determine if the presence of histopathologically tumor-free versus nontumor-free margins was prognostic for relapse or tumor-related death in dogs following surgical excision of single or multiple cutaneous mast cell tumors confined to the skin without evidence of metastasis to lymph nodes or other noncutaneous sites. Differences in tumor-related death or frequency of relapse between the two groups were not significant. Failure to achieve histopathological tumor-free margins frequently did not lead to local relapse. All tumor-related deaths occurred following local relapse. The lack of statistical support for an association between prognosis and histopathological tumor-free versus nontumor-free margins may be a result of small sample size.

Case Presentation A wild-caught male panther chameleon (Furcifer pardalis, previously known as Chamaleo pardalis) imported from Madagascar and of unknown age was presented to the Avian and Exotic Animal Clinic of Lafayette with a complaint of anorexia. The chameleon’s diet consisted primarily of crickets, which the owner occasionally would dust with calcium supplement. Although physically vigorous, the patient was thin (52 g), exhibited poor coloration (Figure 1A), and had pale oral mucous membranes. Physical examination revealed 4 multifocal, poorly defined small bulges underneath the skin (not shown). Two of the bulges were located behind the left forearm, and the other 2 were located cranial to the right rear limb. As part of the diagnostic workup, heparinized blood samples were collected and used to prepare several air-dried smears and wet preparations. Most blood smears were stained with a rapid Romanowsky stain (Diff-Quik, Dade Diagnostics, Aguada, Puerto Rico).The remaining blood smears were later stained with an automated Romanowsky stain (Hematek, Bayer Diagnostics, Elkhart, Ind) at the Purdue University Veterinary Clinical Pathology Laboratory (Figure 1B). Other laboratory tests (clinical chemistry, PCV, CBC) were not done because of the limited amount of blood available. (Continued on next page)

Objective—To evaluate the antitumor and toxic effects of treatment with doxorubicin combined with piroxicam or doxorubicin alone for multicentric lymphoma in dogs.Design—Nonrandomized clinical trial.Animals—75 dogs with multicentric lymphoma.Procedure—33 dogs were treated with doxorubicin (30 mg/m2, IV, q 21 d, for 3 doses) and piroxicam (0.3 mg/kg [0.14 mg/lb], PO, q 24 h); results were compared with a historical control group of 42 dogs treated with doxorubicin (30 mg/m2, IV, q 21 d, for 3 doses) alone.Results—The percentages of dogs that had remission with doxorubicin-piroxicam treatment (79%) or doxorubicin treatment alone (74%) were not significantly different. Median duration of first remission was 130 days with doxorubicin-piroxicam and 147 days with doxorubicin alone; these values were not significantly different. Severe toxicosis was observed in 22% of dogs treated with doxorubicin-piroxicam and 17% of dogs treated with doxorubicin alone.Conclusions and Clinical Relevance—Both treatment protocols were efficacious and well tolerated. The doxorubicin-piroxicam treatment was no more effective regarding response rate, remission duration, or survival duration, compared with the control group treated with doxorubicin alone. (J Am Vet Med Assoc2002;220:1813–1817)

A 4-month-old, intact female schipperke was presented for evaluation and treatment of subcutaneous (SC) emphysema. Radiographs revealed pneumomediastinum and SC emphysema. Sequential radiographs confirmed a worsening of the SC emphysema. Extensive, nonsurgical evaluation failed to reveal the source of the air within the mediastinum. Exploratory thoracotomy revealed an emphysematous right middle lung lobe. Lobectomy of the right middle lung lobe resolved both the pneumomediastinum and SC emphysema. Histopathological evaluation confirmed pulmonary emphysema. A variation of congenital pulmonary emphysema was considered in this case.

Background Greyhounds have several hematologic variables that are outside of the respective reference intervals of other dog breeds. In addition, increases in HCT, total protein and HGB concentration, and RBC and WBC counts occur immediately after exercise; these values return to resting values within a few hour after racing. Objective This study evaluated the effects of exercise on the concentration of reticulocytes in circulating blood in racing Greyhounds. We hypothesized that reticulocyte numbers are significantly increased immediately after a race, and return to baseline within one to 2 h postrace. Methods Fifty actively racing Greyhounds at the Wheeling Island Racetrack and Casino were included in the study. Samples were collected by jugular venipuncture one day prior to racing at the kennel (resting), immediately after racing, and one to 2 h after the race (recovery). Reticulocyte counts were determined with an IDEXX ProCyte Dx Hematology Analyzer (IDEXX Laboratories, Inc., Westbrook, ME, USA). Due to a nonparametric distribution, the results were statistically compared using the Friedman test. Results Reticulocyte concentrations were significantly different among the 3 sample collection times (P

A 7to 11-week-old female rabbit from a commercial rabbit farm with a history of diarrhea and sudden death was submitted alive to the Animal Disease Diagnostic Laboratory at Purdue University for euthanasia and necropsy. On presentation, the rabbit was thin with reduced fat stores and muscle wasting. The rabbit weighed 2 lb 4 oz. The hair coat was rough, and fecal material was adherent to hair on the perineum. Primary gross pathologic changes were observed in the small intestines and liver. The small intestines were distended and filled with gray-green semisolid ingesta. Undigested food pellets were present in the colon. The liver had multiple 1to 3-mm–diameter, slightly raised, discrete to coalescing, yellow-white nodules scattered throughout the parenchyma. Many of these nodules contained yellow-white caseous material. The remaining parenchyma was severely congested and edematous. Air-dried scrapings and impression smears of the nodules were prepared and stained with an automated Wright’s stain (Hematek, Bayer Diagnostics, Elkhart, Ind; Figure 1).

The purpose of this study was to determine the PGE2 concentration in naturally-occurring cancer in pet dogs and in canine cancer cell lines in order to identify specific types of canine cancer with high PGE2 production which could serve as preclinical models to evaluate anticancer strategies targeting PGE2. PGE2 concentrations were measured by enzyme immunoassay in canine melanoma, soft tissue sarcoma, transitional cell carcinoma, osteosarcoma, and prostatic carcinoma cell lines; in 80 canine tumor tissue samples including oral melanoma (MEL), oral squamous cell carcinoma (SCC), transitional cell carcinoma of the urinary bladder (TCC), lymphoma (LSA), mammary carcinoma (MCA), osteosarcoma (OSA), prostatic carcinoma (PCA); and in corresponding normal organ tissues. High concentrations of PGE(2)(range 400-3300 pg/10(4)cells) were present in cell culture medium from the transitional cell carcinoma, prostatic carcinoma, and osteosarcoma cell lines. PGE2 concentrations in tumor tissues were elevated (tumor PGE2 concentration>mean+2X sd PGE(2)concentration of normal organ tissue) in 21/22 TCC, 5/6 PCA, 7/10 SCC, 5/10 MEL, 3/8 MCA, 4/15 OSA, and 0/9 LSA. Results of this study will help guide future investigations of anticancer therapies that target cyclooxygenase and PGE2.

Purpose: More than 12,000 people are expected to die from invasive transitional cell carcinoma (TCC) of the urinary bladder each year in the United States, indicating that more effective therapy is needed. Drugs inhibiting cyclooxygenase (cox) have recently been found to have chemopreventive and antitumor activity and may potentiate the effects of chemotherapy. The purpose of this study was to determine whether cisplatin combined with the cox-inhibitor piroxicam would induce remission more frequently than cisplatin alone in a relevant animal model of human invasive TCC. Methods: Pet dogs with naturally occurring, histopathologically confirmed, measurable TCC of the urinary bladder were randomized to receive cisplatin (60 mg/m2 i.v. every 21 days) or cisplatin (same dosage) combined with piroxicam (0.3 mg/kg orally every 24 h). Complete staging was performed prior to and at 6-week intervals during therapy. Results: After eight dogs had been evaluated in each treatment group, a significant difference in remission rate was noted (Fisher's Exact test, P < 0.004). Tumor responses in the cisplatin/piroxicam group included two complete remissions (CR), four partial remissions (PR), two stable disease (SD), and no progressive disease (PD). Tumor responses to cisplatin alone in eight dogs were no CR, no PR, four SD, and four PD. Six additional dogs were treated with cisplatin/piroxicam, and in total 10 of 14 dogs had remission (two CR, eight PR). Renal toxicity of cisplatin/piroxicam was frequent and dose limiting. Conclusions: Cisplatin/piroxicam induced remission more frequently than cisplatin alone in a canine model of human invasive TCC. Strategies to reduce renal toxicity need to be developed prior to evaluation of cisplatin/piroxicam in humans or general use of this treatment in pet dogs.

Objective—To evaluate expression of cyclooxygenase (COX)-1 and COX-2 in the urinary bladder epithelium of clinically normal dogs and in transitional cell carcinoma cells of dogs. Animals—21 dogs with transitional cell carcinoma of the urinary bladder and 8 dogs with clinically normal urinary bladders. Procedure—COX-1 and COX-2 were evaluated by use of isoform-specific antibodies with standard immunohistochemical methods. Results—COX-1, but not COX-2, was constitutively expressed in normal urinary bladder epithelium; however, COX-2 was expressed in neoplastic epithelium in primary tumors and in metastatic lesions of all 21 dogs and in new proliferating blood vessels in 3 dogs. Also, COX-1 was expressed in the neoplastic cells. Conclusions and Clinical Relevance—Lack of expression of COX-2 in normal bladder epithelium and its substantial expression in transitional cell carcinoma cells suggest that this isoform may be involved in tumor cell growth. Inhibition of COX-2 is a likely mechanism of the antineoplastic effects of non steroidal antiinflammatory drugs. (Am J Vet Res 2000;61:478–481)

Sensitivity and specificity were determined for the cytologic detection of malignant tumors in canine and feline body cavity effusions. In a prospective study, 424 body cavity effusions from dogs and cats were collected and evaluated, including 70 pleural and 163 peritoneal effusions from dogs, and 77 pleural and 114 peritoneal effusions from cats. Final diagnoses were confirmed in 339 of the 424 cases by clinical follow-up, necropsy, and in the case of malignant tumors, Histopathology. Malignant tumors were found in 18% of canine and 25% of feline body cavity effusions. Approximately one-half of tumors in both dogs and cats were carcinomas. Discrete cell tumors accounted for 56% of feline neoplastic effusions. The sensitivity of cytologic evaluation for the detection of malignant tumors in body cavity effusions was 64% for dogs and 61% for cats. Specificity was 99% for canine and 100% for feline effusions. Sensitivity and specificity were comparable to those obtained with cytologic evaluation of human samples.

Styrene is pneumotoxic in mice. It is metabolized by pulmonary microsomes of both mouse and rat to styrene oxide (SO), presumed to be the toxic metabolite of styrene, and known to be genotoxic. To determine which pulmonary cell types are responsible for styrene metabolism, and which cytochromes P450 are associated with the bioactivation of styrene, we isolated enriched fractions of mouse and rat Clara and type II cells in order to determine the rate of styrene metabolism, with and without chemical inhibitors. Mouse Clara cells readily metabolized styrene to SO. Diethyldithiocarbamate, a CYP2E1 inhibitor, caused less inhibition of SO formation in Clara cells isolated from mice than previously found with pulmonary microsomes. As in microsomes, 5-phenyl-1-pentyne, a CYP2F2 inhibitor, inhibited the formation of both enantiomers. alpha-Naphthoflavone, a CYP1A inhibitor, did not inhibit SO formation in Clara cells. alpha-Methylbenzylaminobenzotriazole, a CYP2B inhibitor, exhibited minimal inhibition of SO production at 10 microM and less at 1 microM. The microsomal and isolated cell studies indicate that CYP2E1 and CYP2F2 are the primary cytochromes P450 involved in pulmonary styrene metabolism. Styrene metabolizing activity was much greater in Clara cells than in type II pneumocytes, which demonstrated essentially no activity. Styrene-metabolizing activity was several-fold higher in the mouse than in rat Clara cells. The more pneumotoxic and genotoxic form, R-SO, was preferentially formed in mice, and S-SO was preferentially formed in rats. These findings indicate the importance of Clara cells in styrene metabolism and suggest that differences in metabolism may be responsible for the greater susceptibility of the mouse to styrene-induced toxicity.

Asian pox was diagnosed in three sanderlings (Calidris alba) on Sanibel Island, Florida (USA) in February 1997. All three cases had large tumor-like lesions which contributed significantly to their mortality. Poxvirus infection was confirmed by cytology, histopathology, and electron microscopy. This is the first report of poxvirus infection in sanderlings.

Glomerulonephritis has been associated with exogenous glucocorticoid administration and spontaneous hyperadrenocorticism in the dog. The purpose of this study was to determine the effects of long-term glucocorticoid therapy on urine protein:creatinine ratios (UP/Cs) and renal morphology. Nine young-adult male dogs were determined to be healthy and have normal renal function as assessed by physical examination, CBC, serum biochemistry analysis, Knott's test for Dirofilaria immitis, urinalysis, urine culture, urine protein electrophoresis, endogenous creatinine clearance, 24-hour urinary protein excretion, and UP/C. Prednisone was administered to each dog at a dosage of 2.2 mg/kg PO bid for 42 days. Urinalysis and UP/C were performed on days 0, 7, 14, 21, 28, and 42 of treatment. Mean UP/C on day 0 was 0.29 +/- 0.10. Mean UP/C increased progressively to a maximum of 1.27 +/- 1.02 on day 28. Mean UP/C on day 42 decreased slightly (0.92 +/- 0.56) but remained significantly increased above baseline. The most consistent renal light microscopic finding on necropsy examination was generalized hypercellular glomerular tufts, suggestive of mesangial cell proliferation. Four dogs also had occasional adhesions of glomerular tufts to Bowman's capsule, accompanied by thickening of the capsule. Direct immunofluorescence for immunoglobulin deposition was negative in all dogs. Electron microscopy, evaluated in 7 dogs, was characterized by occasional mild segmental thickening of basement membranes, fusion of visceral cell foot processes, and glomerular adhesions. The results of this study indicate that long-term administration of glucocorticoids results in significant proteinuria and glomerular changes in the dog.

A 15-year-old, castrated male domestic shorthair was presented for a recurrent, perineal apocrine gland adenocarcinoma. A right adrenal mass was identified on abdominal radiography and ultrasonography performed as routine staging prior to surgical excision of the perineal tumor. An ultrasonographic-guided biopsy of the adrenal mass was performed and a pheochromocytoma was diagnosed upon histopathology. The perineal and adrenal tumors were removed surgically. The cat developed fatal thromboembolic disease following surgery and was euthanized. This is the third reported case of phenochromocytoma in a cat and is unusual in that two rare tumors were identified in one animal.

An extensive, laryngeal tumor was identified in a nine-year-old, spayed female, mixed-breed dog. Clinical staging of the tumor included computed tomography. Six days prior to surgery, a percutaneous gastrostomy tube was placed under endoscopic guidance. Surgical treatment included total laryngectomy and permanent tracheostomy. The histologic diagnosis of the tumor was rhabdomyosarcoma. There were no major postoperative complications, and there have been no signs of local recurrence, metastatic disease, or long-term complications associated with the surgical procedure during an 18-month follow-up period.

These studies were designed to evaluate the importance of bile duct obstruction in the pathogenesis of alpha-naphthylisothiocyanate (ANIT)-induced biliary epithelial cell (BEC) hyperplasia in rats. Hepatobiliary function and morphology were evaluated in adult male Sprague-Dawley rats 16, 24, 48, 72, 120, and 168 hr after a single oral dose of ANIT (0, 25, 75, or 150 mg/kg). After 75 or 150 mg/kg ANIT, multifocal bile duct obstruction was observed at 48 and 72 hr and preceded BEC hyperplasia which occurred at 120 and 168 hr. BEC proliferation, reflected by 5-bromo-2'-deoxyuridine (BrdU) incorporation, occurred at doses and at time points coinciding with BEC necrosis and/or bile duct obstruction. In contrast, 25 mg/kg ANIT produced minimal BEC damage and no evidence of bile duct obstruction or BEC hyperplasia. In a separate experiment, BEC proliferation was evaluated following bile duct ligation or ANIT treatment (150 mg/kg). The onset and peak of BEC proliferation occurred 24 and 48 hr, respectively, following bile duct obstruction resulting from either ligation or ANIT treatment. Furthermore, BEC proliferation occurred at all levels of the biliary tree in both bile duct-ligated and ANIT-treated rats. These data indicate that (a) dose-response curves for ANIT-induced bile duct obstruction and BEC hyperplasia are similar; (b) ANIT-induced BEC proliferation and bile duct obstruction precedes BEC hyperplasia; (c) BEC proliferation occurred at doses/timepoints associated with BEC damage and bile duct obstruction; and (d) once ANIT-induced bile duct obstruction occurs, the spatial and temporal aspects of BEC proliferation are comparable to those following biliary obstruction induced by bile duct ligation. Collectively, these data suggest that ANIT-induced BEC hyperplasia is secondary to intrahepatic bile duct obstruction.

Tumor-associated glycoprotein 72 (TAG-72) is a large, high molecular weight, mucinlike antigen that is expressed in a wide variety of human carcinomas. Three different TAG-72 monoclonal antibodies (MAbs), designated B72.3, CC49, and CC83, were applied to the following archived samples from the dog: 1) 51 transitional cell carcinomas of the urinary bladder, 2) 15 hyperplastic/inflamed urinary bladders, and 3) eight normal urinary bladders. Immunoreactivity was detected with an avidin-biotin complex immunoperoxidase method. Fifty-three percent (27/51) of transitional cell carcinomas were positive (≥ 5% staining) for MAb B72.3. MAb B72.3 staining of these transitional cell carcinomas did not statistically correlate with any of the examined features of malignancy, including histologic grade, clinical stage, DNA ploidy, or presence of vascular/lymphatic invasion. In regard to the staining of transitional cell carcinoma by the other two TAG-72 antibodies, 53% (27/ 51) of the samples were positive for MAb CC83 and 63% (32/51) were positive for MAb CC49. The finding that similar populations of neoplastic urothelial cells in serial sections from the same neoplasm stained with all three TAG-72 antibodies supports the hypothesis that an antigen similar to TAG-72 was expressed in canine transitional cell carcinoma. None of the normal urinary bladders nor any of the hyperplastic/inflamed urinary bladders stained with any of the three TAG-72 antibodies tested. The results of these studies demonstrated that the staining of canine transitional cell carcinoma with all three TAG-72 antibodies was specific for neoplastic urothelial cells and that an antigen similar to TAG-72 was expressed.

Background Automated in-house diagnostic analyzers, most commonly used for hematologic and biochemical analysis, are typically calibrated, and then control materials are used to confirm the quality of results. Although this approach provides indirect knowledge that the system is performing correctly, it does not provide direct knowledge of system performance between control runs. Objectives The objectives of this study were to apply analysis of weighted moving averages to assess performance of hematology analyzers using animal patient samples from dogs, cats, and horses as they were analyzed and apply correction factors to mitigate instrument-driven biases when they developed. Methods A set of algorithms was developed and applied to sequential batches of 20 samples. Repeated samples within a batch and large populations of samples with similar abnormalities were excluded. Data for 6 hematologic variables were grouped into batches of weighted moving averages; data were analyzed with control chart rules, a gradient descent algorithm, and fuzzy logic to define and apply adjustments. Results A total of 102 hematology analyzers that had developed biases in RBC count, HCT, hemoglobin (HGB) concentration, MCV, MCH, and MCHC were evaluated. Following analysis, all variables except HGB concentration required adjustment, with RBC counts requiring only slight change and MCV requiring the greatest change. Adjustments were validated by comparing PCVs with the original and adjusted HCT values. Conclusions The proposed system provides feedback control to minimize system bias for RBC count, HCT, HGB concentration, MCV, MCH, and MCHC. Fundamental assumptions must be met for the approach to assure proper functionality.

Flow cytometric analysis of DNA ploidy was performed on 51 formalin-fixed, paraffin-embedded samples of canine transitional cell carcinoma of the urinary bladder. The DNA ploidy data obtained were compared to several clinicopathologic features. In addition, the DNA ploidy of 15 hyperplastic/inflamed and 8 normal canine urinary bladders was measured. Forty-three of the 51 neoplastic samples had interpretable DNA histograms. DNA aneuploidy was found in 34/43 (79%) of the transitional cell carcinoma samples. Of the 34 aneuploid neoplasms, 16 (47%) were hyperdiploid, 17 (50%) were tetraploid, and 1 (3%) was hypertetra-ploid. No significant correlation was found between the presence of DNA aneuploidy and the growth pattern. histologic grade, clinical stage, or individual morphologic features of this neoplasm. Additionally, the DNA ploidy was not related to the sex, age, or survival time of dogs with transitional cell carcinoma. All of the normal and all but one of the hyperplastic/inflamed urinary bladders were diploid. The results from this study demonstrated that DNA ploidy can be measured from paraffin-embedded canine samples by flow cytometry, a majority of the canine transitional cell carcinomas were aneuploid, and a significant correlation did not exist between the DNA ploidy and specific clinicopathologic features of this neoplasm.

The complete blood count is one of the basic building blocks of the minimum database in veterinary medicine. Over the past 20 years, there has been a tremendous advancement in the technology of hematology analyzers and their availability to the general practitioner. There are 4 basic methodologies that can be used to generate data for a complete blood count: manual methods, quantitative buffy coat analysis, automated impedance analysis, and flow cytometric analysis. This article will review the principles of these methodologies, discuss some of their advantages and disadvantages, and describe some of the hematology analyzers that are available for the in-house veterinary laboratory.

Natural killer (NK) cells spontaneously lyse a variety of tumor cells in vitro, and are believed to play an important role in host resistance to tumor growth and metastasis in vivo. As part of our work in comparative oncology, we have designed and validated a canine NK cell assay. Of several lymphocyte isolation techniques evaluated, sedimentation of whole blood through a two-step Ficoll/Hypaque gradient (sp. gr. 1.066/1.119) followed by plastic adherence of monocytes resulted in the most pure lymphocyte population (> 95% lymphocytes). Of four cell lines evaluated as targets in the NK assay, a canine thyroid adenocarcinoma (CTAC) cell line was determined to be most sensitive, and a lymphoblastoid (CT45-S) cell line was determined to be most resistant to NK lysis. A 15 h effector-target incubation period using these targets resulted in reproducible measurement of cell specific lytic activity. Passage of canine lymphocytes through nylon wool columns did not result in a significant increase in NK activity. A final sedimentation of purified lymphocytes through a 45/50% Percoll gradient concentrated NK activity into a single band of lymphocytes. Lymphocytes forming conjugates with CTAC target cells were 5.5-6.5 microns in diameter, and were characterized by a reniform nucleus and varying numbers of electron-dense cytoplasmic granules.

Evaluation of respiratory tract disease is a challenge for several reasons: no serum biochemical or hematologic tests that localize injury to the respiratory system are available, imaging techniques do not usually lead to etiologic diagnoses, and excisional biopsies are often very difficult to obtain from respiratory lesions. For these reasons, specific diagnosis of respiratory tract disease often resides in cytologic evaluation. This article reviews the various cytologic collection techniques that yield high-quality specimens from the upper and lower respiratory tract. Cytologic features of the normal respiratory tract as well as common respiratory disorders are described and illustrated.

It is unclear whether the pneumotoxicity observed with bromobenzene (BB) in phenobarbital-induced rats is related to BB bioactivation in lung, liver or both. To help differentiate pulmonary from hepatic bioactivation, BB was administered alone and in combination with p-xylene, which inhibits pulmonary but induces hepatic cytochromes P450. Exposure to p-xylene alone (3400 ppm for 4 hr) produced no changes in bronchoalveolar lavage fluid (BALF) measurements (γ-glutamyl transpeptidase, lactate dehydrogenase, protein, white blood cell count) or serum sorbitol dehydrogenase. p-Xylene increased hepatic microsomal benzyloxy- (BROD), pentoxy- (PROD), and ethoxy- (EROD) resorufin O-dealkylase activities but decreased pulmonary microsomal BROD and PROD. Immunoblot analysis revealed an induction of hepatic but not pulmonary microsomal P450IIB apoprotein. When rats were exposed to p-xylene (2800 ppm) or room air for 4 hr, treated 12 hr later with BB (0.5 ml/kg, ip) or corn oil, and killed after 12 hr, p-xylene increased hepatic P450IIB (27-fold) concomittant with a similar increase in BROD activity. p-Xylene also increased hepatic P450IA apoprotein (3.4-fold) with a complimentary increase in EROD activity. p-Xylene potentiated BB-induced hepatotoxicity. In pulmonary microsomes p-xylene and BB each produced similar decreases in both EROD and BROD activities. The combination of p-xylene and BB had an additive effect on pulmonary P450IA1 reduction. BALF analysis and histopathology revealed no pneumotoxicity with any treatment. p-Xylene potentiation of BB-induced hepatotoxicity without pneumotoxicity suggests that the liver does not produce metalites of BB which are directly involved in pulmonary damage.

A retrospective study was conducted between two groups of dogs with histopathologically diagnosed multicentric malignant lymphoma to determine if treatment with either short-term or continuous chemotherapy resulted in a significant difference in first-remission length or survival time. One group was treated with single agent, short-term (three cycles) of doxorubicin. Dogs obtaining complete remission while receiving doxorubicin were given no further chemotherapy. The other group received combination agent, long-term chemotherapy consisting of cyclophosphamide, vincristine sulfate, and prednisone (COP). Dogs obtaining complete remission on COP by the end of 6 weeks were given maintenance chemotherapy of cyclophosphamide, prednisone and methotrexate. One hundred and five dogs were treated. Thirty-eight dogs received doxorubicin and 67 received COP. All dogs were evaluated at 6 weeks for response to chemotherapy and followed until death. No significant differences were observed in first-remission length or survival time when comparing dogs treated with either short-term doxorubicin or long-term COP (P greater than 0.05). Sex, weight, age, clinical stage, performance status, histopathologic cell type, and grade were not significant factors for determining the responsiveness to either chemotherapy protocol. However, within either treatment group, significant differences in first-remission length were observed in dogs evaluated histopathologically by the Keil and NCI working formulation and in survival time when evaluated by performance status (P less than 0.05).

Cytosine arabinoside (ara-C) is a component of many protocols for the treatment of CNS (central nervous system) leukemia and lymphoma in humans and dogs. It is also used for the prophylaxis of CNS metastasis in acute lymphoblastic leukemia. Although ara-C enters the cerebrospinal fluid (CSF) of human cancer patients after i.v. administration, it is unclear whether a similar CNS distribution occurs in humans whose blood-brain barrier has not been compromised by invasive disease. No information on the penetration of ara-C into the CSF in dogs is available. We studied the plasma and CSF pharmacokinetics of 600 mg/m2 ara-C in ten healthy male dogs after its administration as a rapid i.v. bolus (six dogs) or as a 12-h i.v. infusion (four dogs). Ara-C concentration in blood and CSF samples was determined by high-performance liquid chromatography (HPLC). After an i.v. bolus of ara-C, the mean plasma distribution half-life was 7.1 +/- 4.5 min and the mean elimination half-life was 69 +/- 28 min. The mean plasma clearance was 227 +/- 125 ml min-1 m-2. The peak concentration of ara-C in the CSF was 29 +/- 11 microM, which occurred at 57 +/- 13 min after the ara-C bolus. The CSF elimination half-life was 113 +/- 26 min. During a 12-h infusion of ara-C (50 mg m-2 h-1), the plasma steady-state concentration was 14.1 +/- 4.2 microM, the CSF steady-state concentration was 8.3 +/- 1.1 microM, and the CSF: plasma ratio was 0.62 +/- 0.14. The plasma elimination half-life was 64 +/- 19 min and the plasma clearance was 214 +/- 69 ml min-1 m-2. The CSF elimination half-life was 165 +/- 28 min. No clinically significant toxicity was observed over a 21-day period following drug administration in either of the treatment groups. Our data indicate that ara-C crosses the blood-brain barrier in normal dogs and that i.v. administration of this drug has potential as a treatment modality for neoplasia involving the CNS.

Bronchoalveolar lavage is a diagnostic procedure used to obtain specimens representative of disease processes involving the deep lung. Saline is instilled into an airway in sufficient volumes to bathe the alveoli dependent on that airway. The saline is retrieved by suction along with cellular and acellular material lining the epithelial surfaces of the lung. Cytologic and microbiologic evaluation of the fluid can be used to characterize pulmonary diseases in the dog and cat.

Bronchoalveolar lavage fluid analysis has gained popularity as a rapid in vivo screen to evaluate the toxicity of both systemic and inhaled pneumotoxicants and is used in addition to the more commonly evaluated pathologic changes. This study evaluated gamma-glutamyltranspeptidase (GGT) in the bronchoalveolar lavage fluid (BALF) along with the more commonly measured enzyme, lactate dehydrogenase (LDH), as a useful indicator of acute lung injury from systematically administered pneumotoxicants. Adult male rats were injected ip with 2, 3, or 3.5 mg/kg body weight of alpha-naphthylthiourea (ANTU) or 5, 10, or 20 mg/kg of 4-ipomeanol, and measurements were made 8 or 24 hr postdose, respectively. ANTU, which selectively damages pulmonary endothelial cells, caused extensive pleural effusions with striking increases in BALF protein and white blood cell (WBC) content. 4-Ipomeanol, which selectively damages nonciliated bronchiolar Clara cells, caused dose dependent increases in both GGT and LDH activities in the BALF with GGT being increased at all doses tested. BALF protein content was also increased in the 4-ipomeanol-treated groups, but this change was not dose dependent. Analysis of GGT in BALF was a sensitive method to assess cytotoxicity associated with 4-ipomeanol-induced injury but was less useful in monitoring pulmonary endothelial cell damage induced by ANTU. Measurements of BALF protein and WBC content proved to be better in assessing injury by agents such as ANTU that primarily affect vascular permeability.