Atypical granulation in neutrophils of a domestic shorthair cat.

A 13-year-old male domestic short-hair cat presented for evaluation of labored breathing, hyporexia, and lethargy. Pertinent initial diagnostics yielded leukocytosis, characterized by neutrophilia and monocytosis. Numerous small, round, magenta granules were observed within all neutrophils in Wright-Giemsa-stained blood films on the day of presentation and the day thereafter. No other neutrophil morphologic abnormalities were present, making cytoplasmic toxicity highly unlikely. Hyperadrenocorticism was diagnosed based on the lack of suppression in a low-dose dexamethasone suppression test, and without other diagnostics, the cat was discharged on trilostane therapy. Neutrophil granules did not stain with Alcian blue pH 1.0, periodic acid-Schiff (PAS), PAS and Alcian blue pH 2.5, and toluidine blue. Electron microscopy identified no differences in the morphology of the secretory granules or other neutrophil features. Metabolic screening tests of the cat's urine did not identify a genetic metabolic disorder. However, serum α- and ß -hexosaminidase (HexA and HexB) activities were 4.3% and 0% of normal controls, respectively, which is supportive of GM2-gangliosidosis, that is, Sandhoff disorder. However, the historical, clinical, and electron microscopy findings did not provide evidence to confirm this genetic defect. To the author's knowledge, this is the first case of magenta-staining granules within neutrophils in a breed other than a Birman, Siamese, or Himalayan.

Hepatic insufficiency in two juvenile dogs with histoplasmosis.

A 9-month-old female intact toy poodle and a 1-year-old female intact Labrador retriever mix presented to separate teaching hospitals for chronic histories of malaise and clinicopathologic evidence of hepatic dysfunction. The signalment and clinical histories of these dogs prompted consideration of a congenital portosystemic shunt as a primary differential. However, microscopic evaluation of peritoneal effusion, pleural effusion, and peripheral blood samples from the dogs revealed round to ovoid yeast organisms morphologically most compatible with Histoplasma capsulatum. Additional testing confirmed histoplasmosis in each case. The poodle underwent a computed tomography (CT) study, which showed hepatomegaly with a spleno-gonadal shunt, pancreatic and gastric wall edema, and marked peritoneal effusion, findings compatible with portal hypertension and secondary acquired shunt formation. The dog was later humanely euthanized due to clinical deterioration, and on necropsy hepatic histoplasmosis was verified, with additional affected tissues comprising lungs and spleen. The Labrador Retriever mix responded clinically and clinicopathologically to antifungal therapy, though no abdominal imaging was performed to definitively exclude the possibility of a congenital portosystemic shunt. In retrospect, several features were more compatible with histoplasmosis than portosystemic shunt in these cases, including hyperbilirubinemia, effusion, and hepatomegaly. These findings serve as a reminder of the need to interpret serum biochemical findings in the context of the totality of the clinicopathologic data and imaging findings, as well as the diagnostic value of microscopy in the evaluation of hematologic and body cavity fluid samples.

Telangiectatic osteosarcoma in four dogs: Cytologic, histopathologic, cytochemical, and immunohistochemical findings.

Telangiectatic osteosarcoma is a rare variant of osteosarcoma histologically and clinically similar to hemangiosarcoma (HSA). This case series describes the imaging and cytologic features of four histologically confirmed telangiectatic osteosarcomas, including the use of cytochemical stains. Alkaline phosphatase (ALP) was applied to Wright-Giemsa-stained cytology slides, and Factor VIII immunohistochemistry was evaluated. Cytologic characteristics included atypical mesenchymal cells with evidence of acute and chronic hemorrhage. Telangiectatic osteosarcoma cases had positive ALP cytochemical staining, while control HSA cases were negative. Factor VIII immunohistochemistry was negative in telangiectatic osteosarcoma and positive in HSA. Cytologic diagnosis of telangiectatic osteosarcoma with positive ALP cytochemical staining can help differentiate this neoplasm from HSA.

Silica and barium: Comparison of microscopic appearance and characterization of effects of silica on cytomorphology.

BACKGROUND: Silica from plastic red top sample collection tubes and barium cause recognized artifacts in slide preparations for microscopic examination. OBJECTIVES: The objectives of this study were to evaluate and directly compare the microscopic appearance of silica and barium particles and various slide preparation techniques (e.g., use of coverslips, oil immersion, and different stains). A secondary objective of this study was to evaluate the effects of silica particles on cellular morphology after mechanical trauma with cytocentrifugation. METHODS: Fluid samples (deionized water, pleural effusion, peritoneal effusion, cerebrospinal fluid, and urine) were collected and evaluated in silica- and non-silica-containing tubes. Barium was added to silica and non-silica samples. Direct and cytocentrifuge preparations were compared to evaluate the effect of silica particles on cellular morphology. Preparations were stained with Wright-Giemsa, rhodizonic acid disodium salt, Alizarin Red, Grocott's methenamine silver, and Prussian blue. RESULTS: Silica and barium particles were identifiable via light microscopy with and without polarized light, although silica particles diminished with immersion oil. Barium particles retained their structure and diminished less under oil. Cytoseal mounting medium for coverslip placement resulted in diminished refractility of silica and some barium particles. Silica particles with mechanical interaction during cytocentrifugation resulted in disrupted cellular morphology with many lysed cells. Silica and barium particles were negative for all special stains tested. CONCLUSIONS: Silica from plastic red top tubes adversely affects cell morphology in cytocentrifuge preparations, potentially affecting manual differential cell counts and compromising diagnostic interpretation. Samples intended for microscopic evaluation should not be collected in silica-containing tubes.

Cytochemical staining of leukocytes and platelets in the Florida manatee (Trichechus manatus latirostris): identification of a bilobed monocyte similar to other members of the Paenungulata.

Manatees (Antillean-, Amazonian, and African-) and dugongs belong to the Order Sirenia, and when combined with elephants and rock hyraxes, form the Paenungulata. A bilobed mononuclear cell has previously been identified in elephants and rock hyraxes, but not in manatees and dugongs, with cytochemical staining identifying these cells as bilobed monocytes in elephants. The objective of this study was to characterize leukocytes (white blood cells, WBC) and platelets in blood films of Florida manatees (Trichechus manatus latirostris; n = 8) using one routine hematological (Wright-Giemsa) and eight cytochemical stains: alkaline phosphatase (ALP), α-naphthyl butyrate esterase (ANBE), chloroacetate esterase (CAE), Luna, myeloperoxidase (MPx), periodic acid-Schiff (PAS), Sudan black B (SBB), and toluidine blue (TB). Heterophils and lymphocytes comprised most of the WBC, with low numbers of eosinophils, basophils, and monocytes. Additionally, 1-3% of the WBC were bilobed mononuclear cells. Bilobed mononuclear cell proportions were similar to rock hyraxes, but lower than elephants (approximate range 20-60%). Heterophils and eosinophils were positive for MPx, ALP, SBB, and PAS, with heterophils also being positive for CAE. Most of the lymphocytes were positive for ANBE and they were variably positive for CAE. Monocytes and bilobed mononuclear cells had similar cytochemical staining reactions (variably positive for all stains, except Luna and TB), supporting a monocytic origin, like elephants. Platelets were ANBE- and PAS-positive. Luna stain was useful for identifying eosinophils and TB was uninformative. This study provides new information on the morphological features and cytochemical staining characteristics of WBC and platelets and will aid in obtaining accurate hematological data of Florida manatees.

Successful multimodal treatment of extreme hypertriglyceridemia in a juvenile diabetic dog.

OBJECTIVE: To describe the therapeutic protocol used to normalize severe hypertriglyceridemia in a dog. CASE SUMMARY: A 7-month-old, 1.2-kg female Pomeranian presented with acute polyuria, polydipsia, and ocular discoloration. Diagnoses included diabetic ketosis, severe hypertriglyceridemia (>225 mmol/L [>20,000 mg/dl]), lipemia retinalis, and bilateral uveitis. The triglyceride concentration was near normal within 2 days of initiating treatment with fenofibrate, regular insulin constant rate infusion (CRI), manual therapeutic plasma exchange (TPE), and a low-fat diet. All clinical signs resolved. The dog has had no relapse of hypertriglyceridemia at the time of writing the manuscript, 6 months later, with continued treatment of diabetes mellitus. NEW OR UNIQUE INFORMATION PROVIDED: This is the first case report documenting the combination of fenofibrate, insulin CRI, and manual TPE for treatment of severe hyperlipidemia in a dog. Detailed protocols for manual TPE and a novel insulin CRI are provided. A discussion of multiple spurious biochemical and hematologic errors associated with the severe hypertriglyceridemia is also provided.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Indications and outcome associated with positive-pressure ventilation in dogs and cats: 127 cases.

OBJECTIVES: To determine the indications and outcomes of positive-pressure ventilation (PPV) and identify factors associated with successful weaning. DESIGN: Retrospective study from October 2009 to September 2013. SETTING: University teaching hospital. ANIMALS: One hundred and eleven dogs and 16 cats. MEASUREMENTS AND MAIN RESULTS: Medical records were retrospectively reviewed; signalment, indication for PPV, patient characteristics, blood gas, and ventilator variables during PPV, duration of PPV, and outcome were recorded. Dogs were most commonly ventilated for pneumonia (36/111; 32%) and cats for multiple pulmonary diseases (8/16; 50%). The median duration of PPV for all animals was 25.7 h (range, 0.1-957 h). Long-term PPV (≥24 h) was performed in 53% of cases. No differences were noted in successful weaning rates between cases ventilated for pulmonary etiologies (23/99; 23%) versus nonpulmonary etiologies (9/28; 32%). Overall, 32 of 127 (25%; 30 dogs, 2 cats) animals were successfully weaned from PPV and 28 of 127 (22%; 26 dogs, 2 cats) survived to hospital discharge. Long-term ventilation had a higher likelihood of successful weaning (26/67 [39%] vs 6/60 [10%], P = 0.0002) and higher rates of survival to discharge (23/67 [34%] vs 5/60 [8%], P = 0.0005) than short-term ventilation. Animals with higher Pao2 /Fio2 and Spo2 /Fio2 and lower APPLE and SOFA scores on day 1 of PPV were more likely to be weaned (P < 0.03). CONCLUSIONS: The outcome of PPV appears to be most heavily determined by the underlying disease process and no clear improvement in outcome could be demonstrated in this study, despite advances in veterinary critical care and ventilator management strategies since previous studies. Dogs and cats receiving PPV for more than 24 h in this study had a higher likelihood of a positive outcome. Several indices of oxygenation and illness severity at the onset of PPV were predictive of outcome and maybe useful when considering prognosis of these cases.

Complications associated with long-term positive-pressure ventilation in dogs and cats: 67 cases.

OBJECTIVES: To determine the complications associated with positive-pressure ventilation (PPV) in dogs and cats. DESIGN: Retrospective study from October 2009 to September 2013. SETTING: University Teaching Hospital. ANIMALS: Fifty-eight dogs and 9 cats. MEASUREMENTS AND MAIN RESULTS: Medical records were retrospectively reviewed; signalment, complications associated with PPV, duration of PPV, and outcome were recorded. Complications most commonly recorded during PPV included hypothermia 41/67 (61%), hypotension 39/67 (58%), cardiac arrhythmias 33/67 (49%), a positive fluid balance 31/67 (46%), oral lesions 25/67 (37%), and corneal ulcerations 24/67 (36%). A definition of ventilator-associated events (VAE) extrapolated from the Center of Disease Control's criteria was applied to 21 cases that received PPV for at least 4 days in this study. Ventilator-associated conditions occurred in 5 of 21 (24%) of cases with infection-related ventilator-associated conditions and ventilator-associated pneumonia identified in 3 of 21 (14%) cases. CONCLUSIONS: Complications are common and diverse in dogs and cats receiving long-term PPV and emphasizes the importance of intensive, continuous patient monitoring and appropriate nursing care protocols. Many of the complications identified could be serious without intervention and suggests that appropriate equipment alarms could improve patient safety. Development of veterinary specific surveillance tools such as the VAE criteria would aid future investigations and allow for effective multicenter studies.

Evaluation of pulse oximetry as a surrogate for PaO2 in awake dogs breathing room air and anesthetized dogs on mechanical ventilation.

OBJECTIVE: To evaluate the ability of arterial hemoglobin oxygen saturation measurement via pulse oximetry (SpO2 ) to serve as a surrogate for PaO2 in dogs. DESIGN: Two-part study: prospective observational and retrospective components. SETTING: University teaching hospital. ANIMALS: Ninety-two dogs breathing room air prospectively enrolled on a convenience basis. Retrospective evaluation of 1,033 paired SpO2 and PaO2 measurements from 62 dogs on mechanical ventilation. INTERVENTIONS: Dogs with concurrent SpO2 and PaO2 measured on room air had a data sheet completed with blood gas analysis. SpO2 , PaO2 , and FiO2 values were collected from medical records of dogs on mechanical ventilation. MEASUREMENTS AND MAIN RESULTS: Predicted PaO2 was calculated from SpO2 using the dog oxyhemoglobin dissociation curve. The correlation coefficient between measured and predicted PaO2 was 0.49 (P < 0.0001) in room air dogs and 0.74 (P < 0.0001) in ventilated dogs. In room air dogs, Bland-Altman analysis between measured minus predicted PaO2 versus the average showed a mean bias of -6.0 mm Hg (95% limit of agreement, -35 to 23 mm Hg). The correlation coefficient between PaO2 /FiO2 and SpO2 /FiO2 ratios was 0.76 (P < 0.0001). After combining data sets, receiver operating characteristic curve analysis showed the optimal cutoff value for detecting hypoxemia (PaO2  < 80 mm Hg) was an SpO2 of 95%, with sensitivity and specificity of 77.8% and 89.5%, respectively. Using this cutoff, 6.9% of SpO2 readings failed to detect hypoxemia, whereas 7.2% predicted hypoxemia that was not present. CONCLUSIONS: The SpO2 was not clinically suitable as a surrogate for PaO2 , though it performed better in mechanically ventilated dogs. As sensitivity for the detection of hypoxemia was poor, pulse oximetry does not appear to be an acceptable screening test. The SpO2 /FiO2 ratio may have value for evaluation of anesthetized dogs on supplemental oxygen. Arterial blood gas analysis remains ideal for assessment of oxygenation.

Severe hypernatremia and transient azotemia in a cat following inadvertent intravenous administration of a commercial polyethylene glycol solution.

OBJECTIVE: To describe the clinical signs, clinicopathologic abnormalities, treatment, and outcome after IV administration of polyethylene glycol 3350 (PEG3350) in a cat. CASE SUMMARY: A cat was inadvertently administered 6 g/kg of PEG3350 in electrolyte solution, IV, resulting in severe hypernatremia (203 mmol/L), diffuse encephalopathy, hemolysis, and moderate azotemia. The hemolysis and acute kidney injury observed immediately following PEG3350 administration resolved with supportive care. Administration of IV and oral electrolyte-free water slowly corrected the hypernatremia and the neurologic signs subsequently improved. Complete resolution of clinical signs was documented one month following hospital discharge. The PEG3350 concentrations in serum, plasma, and urine samples confirmed toxic exposure to PEG3350. Efficacy of treatment was evident by decreasing concentrations of PEG3350 in serum after the first 24 hours of treatment. Renal elimination of PEG3350 was significant and PEG3350 was still detected in the urine 17 days after exposure. NEW INFORMATION PROVIDED: This is the first report to describe the clinical signs and clinicopathologic abnormalities in a cat intoxicated with IV PEG3350. Potential pathophysiologic mechanisms are discussed, and the successful supportive medical treatment is outlined.

Effects of positive end-expiratory pressure and recruitment maneuvers in a ventilator-induced injury mouse model.

BACKGROUND: Positive-pressure mechanical ventilation is an essential therapeutic intervention, yet it causes the clinical syndrome known as ventilator-induced lung injury. Various lung protective mechanical ventilation strategies have attempted to reduce or prevent ventilator-induced lung injury but few modalities have proven effective. A model that isolates the contribution of mechanical ventilation on the development of acute lung injury is needed to better understand biologic mechanisms that lead to ventilator-induced lung injury. OBJECTIVES: To evaluate the effects of positive end-expiratory pressure and recruitment maneuvers in reducing lung injury in a ventilator-induced lung injury murine model in short- and longer-term ventilation. METHODS: 5-12 week-old female BALB/c mice (n = 85) were anesthetized, placed on mechanical ventilation for either 2 hrs or 4 hrs with either low tidal volume (8 ml/kg) or high tidal volume (15 ml/kg) with or without positive end-expiratory pressure and recruitment maneuvers. RESULTS: Alteration of the alveolar-capillary barrier was noted at 2 hrs of high tidal volume ventilation. Standardized histology scores, influx of bronchoalveolar lavage albumin, proinflammatory cytokines, and absolute neutrophils were significantly higher in the high-tidal volume ventilation group at 4 hours of ventilation. Application of positive end-expiratory pressure resulted in significantly decreased standardized histology scores and bronchoalveolar absolute neutrophil counts at low- and high-tidal volume ventilation, respectively. Recruitment maneuvers were essential to maintain pulmonary compliance at both 2 and 4 hrs of ventilation. CONCLUSIONS: Signs of ventilator-induced lung injury are evident soon after high tidal volume ventilation (as early as 2 hours) and lung injury worsens with longer-term ventilation (4 hrs). Application of positive end-expiratory pressure and recruitment maneuvers are protective against worsening VILI across all time points. Dynamic compliance can be used guide the frequency of recruitment maneuvers to help ameloriate ventilator-induced lung injury.

Injectable Anesthesia for Mice: Combined Effects of Dexmedetomidine, Tiletamine-Zolazepam, and Butorphanol.

Anesthetic protocols for murine models are varied within the literature and medetomidine has been implicated in the development of urethral plugs in male mice. Our objective was to evaluate the combination of butorphanol, dexmedetomidine, and tiletamine-zolazepam. A secondary objective was to identify which class of agent was associated with urethral obstructions in male mice. BALB/c male (n = 13) and female (n = 23) mice were assigned to dexmedetomidine and tiletamine-zolazepam with or without butorphanol or to single agent dexmedetomidine or tiletamine-zolazepam. Anesthesia was achieved in 58% (14/24) of mice without butorphanol and in 100% (24/24) of mice with butorphanol. The combination of dexmedetomidine (0.2 mg/kg), tiletamine-zolazepam (40 mg/kg), and butorphanol (3 mg/kg) resulted in an induction and anesthetic duration of 12 and 143 minutes, respectively. Urethral obstructions occurred in 66% (25/38) of trials in male mice that received dexmedetomidine with a mortality rate of 38% (5/13). Tiletamine-zolazepam, when used alone, resulted in a 0% (0/21) incidence of urethral obstructions. Combination use of dexmedetomidine, tiletamine-zolazepam, and butorphanol results in a longer and more reliable duration of anesthesia than the use of dexmedetomidine and tiletamine-zolazepam alone. Dexmedetomidine is not recommended for use in nonterminal procedures in male mice due to the high incidence of urethral obstructions and resultant high mortality rate.