EFFECT OF ROUTINE HANDLING AND TRANSPORTATION ON BLOOD LEUKOCYTE CONCENTRATIONS AND PLASMA CORTICOSTERONE IN CAPTIVE HISPANIOLAN AMAZON PARROTS ( AMAZONA VENTRALIS).

Increased glucocorticoids cause a characteristic stress leukogram in mammalian taxa. It is assumed that avians exhibit a similar response, but to date, there have been no controlled studies to correlate serial endogenous corticosterone levels to hematologic values. An established flock of 18 Hispaniolan Amazon parrots ( Amazona ventralis) was used as a model in a crossover study. The treatment group was subjected to the stress of transport, restraint, and common clinical procedures with serial blood samples collected at 20-min intervals for hematology and corticosterone levels; the control group was sampled at the same intervals. Longitudinal data analysis was performed with linear mixed modeling. For all hematologic analytes, the baseline value had a significant positive effect on subsequent values (all P < 0.001). The white blood cell, heterophil, and eosinophil counts and heterophil to lymphocyte ratio increased over time in the treatment group, whereas it remained stable in the control group ( P = 0.016, P < 0.001, P < 0.001, P = 0.02, respectively, for the time*treatment effect). Lymphocyte absolute counts decreased over time, although not significantly; the decrease was significant for the relative lymphocyte count in the treatment group. Monocytes and basophils were not significantly altered. The treatment group had a higher mean corticosterone level overall than the control group by approximately 60% ( P = 0.008). The mean corticosterone level also increased over time in both groups by three- to fourfold ( P < 0.001) by 20 min then plateaued. These results demonstrate that some significant hematologic changes may arise with routine handling and transportation of birds and should be accounted for in hematologic interpretation of cell counts.

Rhabdomyolysis and Artifactual Increase in Plasma Bicarbonate Concentration in an Amazon Parrot (Amazona species).

A 7-year-old male Amazon parrot housed outdoors presented with acute collapse, marked lethargy, and open-mouth breathing. The patient had stiffness of the pectoral muscles, and petechiation and ecchymosis noted around the eyes and beneath the mandible. Laboratory data revealed markedly increased aspartate aminotransferase, creatine kinase, and lactate dehydrogenase activity consistent with rhabdomyolysis, as well as markedly increased plasma bicarbonate concentration. Marked clinical improvement and resolution of laboratory abnormalities occurred with fluid therapy, administration of a nonsteroidal anti-inflammatory medication, and husbandry modifications, including indoor housing and dietary alteration. A spurious increase in bicarbonate measurement as documented in equine and bovine cases of rhabdomyolysis also occurred in this avian patient and must be considered for accurate interpretation of acid-base status in exotic species presenting with consistent clinical signs.

The effect of urea on refractometric total protein measurement in dogs and cats with azotemia.

BACKGROUND: While protein is the predominant solute measured in plasma or serum by a refractometer, nonprotein substances also contribute to the angle of refraction. There is debate in the current literature regarding which nonprotein substances cause factitiously high refractometric total protein measurements, as compared to the biuret assay. OBJECTIVES: The purpose of the study was to determine if the blood of azotemic animals, specifically with increased blood urea concentration, will have significantly higher refractometric total protein concentrations compared to the total protein concentrations measured by biuret assay. METHODS: A prospective case series was conducted by collecting data from azotemic (n = 26) and nonazotemic (n = 34) dogs and cats. In addition, an in vitro study was performed where urea was added to an enhanced electrolyte solution at increasing concentrations, and total protein was assessed by both the refractometer and spectrophotometer. Statistical analysis was performed to determine the effect of urea. RESULTS: The refractometric total protein measurement showed a positive bias when compared to the biuret protein measurement in both groups, but the bias was higher in the azotemic group vs the nonazotemic group. The mean difference in total protein measurements of the nonazotemic group (0.59 g/dL) was significantly less (P < .01) than the mean difference of the azotemic group (0.95 g/dL). The in vitro experiment revealed a positive bias with a proportional error. CONCLUSIONS: This study demonstrated that increasing concentrations of urea significantly increased the total protein concentration measured by the refractometer as compared to the biuret assay, both in vivo and in vitro.

Nannizziopsis guarroi infection in 2 Inland Bearded Dragons (Pogona vitticeps): clinical, cytologic, histologic, and ultrastructural aspects.

Chrysosporium-related infections have been increasingly reported in reptiles over the last 2 decades. In this report, we describe clinical, cytologic, histopathologic, and ultrastructural aspects of Chrysosporium-related infection in 2 Inland Bearded Dragons (Pogona vitticeps). Case 1 was presented for an enlarging raised lesion over the left eye and multiple additional masses over the dorsum. Case 2 was submitted to necropsy by the referring veterinarian for suspected yellow fungus disease. Impression smears of the nodules in case 1 revealed granulomatous to pyogranulomatous inflammation and many septate, variably long, 4-10 µm wide, often undulated hyphae, and very rare conidia. Postmortem impression smears of the superficial lesions of case 2 contained large numbers of solitary conidia and arthroconidia and low numbers of hyphae with similar morphology to case 1. Histopathology of the 2 cases revealed severe, multifocal, chronic, ulcerative, nodular pyogranulomatous dermatitis, with myriad intralesional septate hyphae, and arthroconidia. Fungal culture and molecular sequencing in both cases indicated infection with Nannizziopsis guarroi.

Atypical fibrosarcoma in the skin of a Roborovski hamster (Phodopus roborovskii).

A 2.5-year-old intact male Roborovski hamster (Phodopus roborovskii) was presented with a large subcutaneous mass overlying the abdomen, affecting the animal's ambulation and access to different compartments of the cage through narrow tubing. Ultrasound examination delineated a well-circumscribed mass in the subcutis of the caudoventral abdominal region. The mass was surgically excised and on cytologic examination showed, in a background of blood, a small population of individually arranged oval to spindle-shaped cells that exhibited a moderate degree of anisokaryosis, coarsely stippled chromatin, one or more prominent nucleoli, and lightly basophilic well-defined cytoplasmic processes. Histologically, the mass was composed of interlacing streams and bundles of pleomorphic spindle cells (ganglion-like cells) with variable amounts of collagenous stroma. The neoplastic cells exhibited moderate features of malignancy. These cells stained intensely with vimentin, but not with any other markers, including antibodies to cytokeratin AE1/AE3, S100 protein, desmin, smooth muscle actin, synaptophysin, neurofilament, and androgen receptor. Based on histologic features, the mass was diagnosed as an atypical fibrosarcoma. This is the first report of an atypical fibrosarcoma in a Roborovski hamster and one of few reports of atypical fibrosarcoma in domesticated hamsters overall.

Idiopathic solitary cutaneous xanthoma in a dog.

A 6-year-old female spayed Boxer mix dog was presented with multiple cutaneous masses, one of which was determined to be a xanthoma. Fine-needle aspirates of this mass revealed large round cells that were consistent with macrophages. These macrophages had lightly basophilic cytoplasm that was filled with many clear circular spaces that varied in size. The nuclei of these cells displayed mild anisokaryosis with condensed chromatin and lacked prominent nucleoli. The cytologic interpretation was lipid-laden histiocytic inflammation most consistent with a cutaneous xanthoma, which was confirmed histologically. Mild hypertriglyceridemia and persistent moderate hypercholesterolemia were present. After ruling out other causes of hyperlipidemia, we concluded that the dog likely had idiopathic hyperlipidemia with secondary xanthoma formation.

Performance of a commercially available in-clinic ELISA for the detection of antibodies against Anaplasma phagocytophilum, Ehrlichia canis, and Borrelia burgdorferi and Dirofilaria immitis antigen in dogs.

OBJECTIVE: To evaluate the sensitivity and specificity of a commercially available in-clinic ELISA for detection of heartworm infection and tick-borne diseases in dogs. SAMPLE POPULATION: 846 serum, plasma, or blood samples obtained from dogs. PROCEDURES: Samples were evaluated via the in-clinic ELISA to detect antibodies against Anaplasma phagocytophilum, Ehrlichia canis, and Borrelia burgdorferi and Dirofilaria immitis (heartworm) antigen. True infection or immunologic status of samples was assessed by use of results of necropsy, an antigen assay for D immitis, and immunofluorescence assay or western blot analysis for antibodies against B burgdorferi, E canis, and A phagocytophilum. RESULTS: Sensitivity and specificity of the in-clinic ELISA for detection of heartworm antigen (99.2% and 100%, respectively), antibodies against B burgdorferi (98.8% and 100%, respectively), and antibodies against E canis (96.2% and 100%, respectively) were similar to results for a similar commercial ELISA. In samples obtained from dogs in the northeast and upper Midwest of the United States, sensitivity and specificity of the in-clinic ELISA for antibodies against Anaplasma spp were 99.1% and 100%, respectively, compared with results for an immunofluorescence assay. Samples from 2 dogs experimentally infected with the NY18 strain of A phagocytophilum were tested by use of the in-clinic ELISA, and antibodies against A phagocytophilum were detected by 8 days after inoculation. Antibodies against Anaplasma platys in experimentally infected dogs cross-reacted with the A phagocytophilum analyte. Coinfections were identified in several of the canine serum samples. CONCLUSIONS AND CLINICAL RELEVANCE: The commercially available in-clinic ELISA could be used by veterinarians to screen dogs for heartworm infection and for exposure to tick-borne pathogens.

Evaluation of peptide- and recombinant protein-based assays for detection of anti-Ehrlichia ewingii antibodies in experimentally and naturally infected dogs. [corrected]

OBJECTIVE: To evaluate microtiter-plate format ELISAs constructed by use of different diagnostic targets derived from the Ehrlichia ewingii p28 outer membrane protein for detection of E ewingii antibodies in experimentally and naturally infected dogs. SAMPLE POPULATION: Serum samples from 87 kenneled dogs, 9 dogs experimentally infected with anti-E ewingii, and 180 potentially naturally exposed dogs from Missouri. PROCEDURES: The capacities of the synthetic peptide and truncated recombinant protein to function as detection reagents in ELISAs were compared by use of PCR assay, western blot analysis, and a full-length recombinant protein ELISA. Diagnostic targets included an E ewingii synthetic peptide (EESP) and 2 recombinant proteins: a full-length E ewingii outer membrane protein (EEp28) and a truncated E ewingii outer membrane protein (EETp28) RESULTS: A subset of Ehrlichia canis-positive samples cross-reacted in the EEp28 ELISA; none were reactive in the EESP and EETp28 ELISAs. The EESP- and EETp28-based ELISAs detected E ewingii seroconversion at approximately the same time after infection as the EEp28 ELISAs. In afield population, each of the ELISAs identified the same 35 samples as reactive and 27 samples as nonreactive. Anaplasma and E can is peptides used in a commercially available ELISA platform did not detect anti-E ewingii antibodies in experimentally infected dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The EESP and EETp28 ELISAs were suitable for specifically detecting anti-E ewingii antibodies in experimentally and naturally infected dogs.

Malignant B-cell lymphoma with Mott cell differentiation in a ferret (Mustela putorius furo).

A 3.5-year-old, male, neutered ferret (Mustela putorius furo) was presented with a 3-day history of lethargy and anorexia. Splenic aspirates revealed high numbers of intermediate-sized lymphocytes and Mott cells interpreted as lymphoma with Mott cells. The ferret was euthanized because of a poor clinical prognosis. Postmortem examination revealed markedly enlarged spleen and lymph nodes, with multifocal white nodules in the liver parenchyma. Histologically, the spleen had multifocal large nodules composed of neoplastic lymphocytes with frequent Mott cells. Similar neoplastic cells were present in the sections of liver, lymph nodes, and bone marrow. These cells were cluster of differentiation (CD)3-negative, CD79alpha-positive, and lambda light-chain-positive. Electron microscopy revealed that the cytoplasm of the neoplastic Mott cells had increased, disorganized, dilated, rough endoplasmic reticulum containing electron-dense immunoglobulin. On the basis of cytologic, histopathologic, immunohistochemical, and electron microscopic findings, a malignant B-cell lymphoma with Mott cell differentiation was diagnosed.

Reference intervals of plasma calcium, phosphorus, and magnesium for African grey parrots (Psittacus erithacus) and Hispaniolan parrots (Amazona ventralis).

Calcium (Ca), phosphorus (P), and magnesium (Mg) are important elements for body homeostasis in several diseases associated with imbalances in the plasma concentration of these ions. This is the first published report of reference intervals for Mg in association with Ca and P levels for psittacine species. One milliliter of blood was collected from 26 Hispaniolan parrots (Amazona ventralis) and 24 African grey parrots (Psittacus erithacus). The plasma concentrations of Ca, P, and Mg were determined for each sample. Statistical analyses were performed including all data (analysis 1) and after exclusion of the subjects with Ca > or = 14.00 mg/dl (3.5 mmol) (analysis 2). The data from analysis 1 have a narrower interval than that observed in analysis 2. Following the normality test (Shapiro-Wilk, alpha = 0.05), the univariate and mean procedures were run. For the reference intervals, the lower and upper values were used, after elimination of the outliers calculated by Blom scores from the ranked variables. The analysis 1 references for the Hispaniolans were Ca = 8.80-10.40 mg/dl (2.20-2.60 mmol/L), P = 1.80-4.40 mg/dl (0.58-1.42 mmol/L), Mg = 1.80-3.10 mg/dl (0.74-1.27 mmol/L), and Ca:P ratio = 2.62-5.39; for the African greys analysis 1 references were Ca = 8.20-20.20 mg/dl (2.05-5.05 mmol/L), P = 2.50-5.90 mg/dl (0.81-1.91 mmol/L), Mg = 2.10-3.40 mg/dl (0.82-1.4 mmol/L), and Ca:P ratio = 1.81-3.77. The analysis 2 references for the Hispaniolans were Ca = 8.80-10.30 mg/dl (2.20-2.58 mmol/L), P = 1.80-3.80 mg/dl (0.58-1.23 mmol/L), Mg = 1.90-3.00 mg/dl (0.82-1.07 mmol/L), Ca:P ratio = 2.62-5.39; for the African greys analysis 2 references were Ca = 1.07 mmol/L), Ca:P ratio = 1.67-3.50. The results of this study are important for evaluating Mg concentrations in relation to the Ca and P parameters in psittacines. This information will be particularly helpful for veterinarians evaluating the hypocalcemic syndrome in African grey parrots and other disease processes associated with Ca, P, and Mg physiologic imbalances.

Comparison of hematologic values in blood samples with lithium heparin or dipotassium ethylenediaminetetraacetic acid anticoagulants in Hispaniolan Amazon parrots (Amazona ventralis).

Blood samples were collected from 20 Hispaniolan Amazon parrots (Amazona ventralis) and were divided into tubes that contained dipotassium ethylenediaminetetraacetic acid (K2EDTA) and lithium heparin. Complete blood cell counts were determined in each sample within 2 hours of collection. The level of agreement in results was moderate for plasma protein, packed cell volume (PCV), and leukocyte, monocyte, and lymphocyte counts between the anticoagulants. Plasma protein and PCV values were significantly lower in samples with lithium heparin than in those with K2EDTA, whereas lymphocyte numbers were significantly higher in lithium heparin samples than in K2EDTA samples. The level of agreement was good for the other cell types (heterophils, eosinophils, and basophils) when comparing the different anticoagulants. The poor level of agreement between anticoagulants with the increase in thrombocyte clumping in lithium heparin samples indicates that the use of lithium heparin as anticoagulant may affect thrombocyte count. No negative effects on morphology and staining of blood cells were apparent in smears from heparin samples compared with K2EDTA samples. Within the different values compared, the limits of agreement are small enough to be confident that lithium heparin can be used for routine CBC counts in a clinical setting. The use of the same anticoagulant should be recommended to follow trends within the same patient, especially when considering plasma protein concentration, PCV, and lymphocyte count.

Clinical hematology practices at veterinary teaching hospitals and private diagnostic laboratories.

The clinical hematology practices utilized at veterinary teaching hospitals and private veterinary diagnostic laboratories were surveyed using a questionnaire. The hematology caseload at private diagnostic laboratories was larger, and comprised predominantly of canine and feline submissions. The Coulter S Plus IV and Serono Baker 9000 were the hematology analyzers used most frequently at veterinary medical laboratories. The Abbott Cell-Dyn 3500, a multispecies analyzer capable of leukocyte differential counting, was utilized more by private laboratories. Commercial hematology control reagents were used at all laboratories; teaching hospital laboratories more often used reagents supplied by the manufacturer of the analyzer. A greater percentage of private diagnostic laboratories participated in the external quality assurance programs offered by Veterinary Laboratory Association and College of American Pathologists. While private diagnostic laboratories retained the EDTA blood specimens longer after initial testing, the teaching hospital laboratories retained blood smears and complete blood count reports longer. The complete blood count reports at veterinary teaching laboratories more often included red blood cell volume distribution width, mean platelet volume, manual hematocrit, plasma protein, and leukocyte differentials as absolute concentrations. The laboratory practices utilized by these veterinary medical laboratories were generally similar, and differences were attributed to divergent emphasis on economic accountability and clinical investigation.