Infection with a novel derogenid trematode in a Flap-necked chameleon (Chamaeleo dilepis).

A 2-year-old female Flap-necked chameleon (Chamaeleo dilepis) was presented with a history of anorexia. On physical examination, three 11×4×2 mm, light tan-pink, raised, gelatinous, freely movable, poorly demarcated, raised nodules were identified within the oral cavity. An impression smear contained multiple, light blue to green structures exhibiting an 18×35 µm, elliptical-shaped anterior portion and a single 1×150 µm filament with a tapered end. Initial differential etiologies included trematode ova, flagellate protozoa, fungal sporangia, plant material, and sperm. Fungal culture yielded no growth. The structures described above did not stain with Gomori methenamine silver stain. A calcofluor white stain yielded positive fluorescence. The DNA was extracted from the impression smears and pan-trematode PCR was performed; products were directly sequenced and phylogenetic analysis revealed maximum similarity with a digenean trematode in the family Derogenidae that was distinct from other trematodes for which a reference sequence was available. To our knowledge, this is the first description of cytologic features of trematode ova from an oral lesion in a chameleon.

Effect of delayed serum separation and storage temperature on serum glucose concentration in horse, dog, alpaca, and sturgeon.

BACKGROUND: Although delays between blood sample collection and analysis are common in veterinary medicine, the effect of prolonged serum-clot contact time on serum glucose concentration is not well established and species differences have not been elucidated. OBJECTIVES: The objective was to investigate the effect of storage time and temperature on serum glucose concentration in stored whole blood samples from horse, dog, alpaca, and sturgeon. METHODS: Whole blood specimens were divided into 7 no-additive tubes and serum was separated from one sample within one hour, serving as the reference sample. The remaining samples were stored at 4°C and 25°C, then centrifuged and serum glucose measured by automated analysis at 2, 4, and 8 hours postcollection. Glucose concentrations were compared using linear mixed models. RESULTS: The decline in serum glucose concentration for all samples stored at 4°C was not statistically significant, except for the 8-hour samples from sturgeon and dog. At 25°C, serum glucose concentration was comparable to reference values at 2 hours in sturgeon and alpaca, but significantly lower at 4 and 8 hours in those species, and at all time points in equine and canine specimens, being most prominent after 8 hours of storage in canine specimens. CONCLUSIONS: Storage at 4°C limits serum glucose decline for at least 4 hours in all species tested and up to 8 hours in specimens of horse and alpaca. At 25°C, serum-clot contact time should not exceed 1 hour in equine and canine samples, and 2 hours in specimens from alpaca and sturgeon.

Erythrocyte dysplasia in peripheral blood smears from 5 thrombocytopenic dogs treated with vincristine sulfate.

Secondary dyserythropoiesis has been associated with vincristine administration in dogs. Evaluation of bone marrow aspirates for the presence of morphologic abnormalities in the erythroid lineage aids in the diagnosis. However, morphologic features of circulating erythroid precursors in these cases have not been described previously. The purpose of this report was to describe the cytologic features of dyserythropoiesis in peripheral blood and also bone marrow smears in a case series of dogs with immune-mediated thrombocytopenia (IMT) treated with vincristine sulfate. Nineteen dogs receiving vincristine for treatment of IMT were identified by retrospectively searching a computerized medical record system. There were 5 dogs that had dysplastic erythroid precursors in peripheral blood smears within 7 days of vincristine treatment. Two of those 5 dogs also had evidence for erythrodysplasia in modified Wright's-stained bone marrow smears obtained postvincristine administration. Morphologic changes included bizarre or inappropriate mitotic figures, abnormal nuclear configurations (fragmentation, elongation, indentation, and binucleation), atypical nuclear remnants (Howell-Jolly bodies), or nuclear and cytoplasmic asynchrony within the erythroid precursors. A brief review of the literature with discussion of the etiologies for dyserythropoiesis is provided. The dyserythropoiesis was clinically insignificant in all 5 cases and resolved. However, pathologists and clinicians should be aware of these potential findings to prevent misdiagnosis of other conditions.

Preliminary evaluation of a quantitative ethylene glycol test in dogs and cats.

Ethylene glycol (EG) toxicity is commonly encountered in dogs and cats. The purpose of the current study was to determine if the Catachem test kit (Catachem Inc., Oxford, Connecticut) could precisely and accurately detect the presence of EG added to serum and plasma from 6 dogs and 4 cats. Serum and plasma samples were spiked at various concentrations of EG (0, 20, 60, and 100 mg/dl) and analyzed using the Catachem kit. Twenty randomly selected samples were also submitted for gas chromatography-mass spectroscopy (GC-MS) analysis of EG concentration, which was considered the gold standard. Inter- and intra-assay coefficients of variation (CVs) were calculated. Bland-Altman analysis was performed to compare the Catachem results to the GC-MS analyses. Analysis of serum samples showed a bias of 8.48 mg/dl (95% limits of agreement: 17.8 to -0.9 mg/dl) while spiked plasma samples had a bias of 7.32 mg/dl (18.1 to -3.5 mg/dl). Intra-assay CV was 0.7%. Interassay CV ranged from 1.2% to 2.0%. For all samples, the Catachem kit read higher than GC-MS values and slightly overestimated in vitro concentrations. The Catachem test kit is an accurate quantitative test for EG in dogs and cats that may aid in timely recognition of EG exposure. Because of the positive bias in all samples, some pets may receive treatment unnecessarily. However, animals with blood EG concentrations at or above the published lethal serum or plasma concentration will be readily identified so that treatment may be initiated.

Comparison of manual polychromatophilic cell and automated reticulocyte quantification in evaluating regenerative response in anemic dogs.

BACKGROUND: Degree of polychromasia and reticulocyte counts are commonly used in veterinary medicine to evaluate response to anemia. The quantitative association between these 2 measurements has not been evaluated. OBJECTIVE: The aim of this retrospective study was to compare automated reticulocyte percents and counts with those of polychromatophilic cells on blood films from dogs. METHODS: Blood films and medical records from 30 clinically healthy dogs and 60 anemic dogs were evaluated. Manual percentage of polychromatophilic cells (MPP) was determined by counting cells in 10 1000× fields, and automated reticulocyte percentage (ARP) was measured using an ADVIA 120 hematology analyzer; absolute polychromatophilic cell and reticulocyte counts were then calculated. Degree of polychromasia, estimated as slight, mild, moderate, marked was documented. Percentages and absolute counts of polychromatophilic cells and reticulocytes were compared using least squares regression and Pearson's correlation coefficients. RESULTS: The reference interval for MPP in healthy dogs was 0.21-0.26%. Correlation between MPP and ARP was highest when all levels were considered (n = 60, r = .98, P < .0001), strong when ARP was > 5% (n = 20, r = .91, P < .0001), and poor when ARP was 1-2% (n = 20, r = .35, P < .0001). Results were similar for comparisons of absolute reticulocyte and polychromatophilic cell counts, with the best correlation found when all levels of counts were analyzed together (r = .96) and for automated reticulocyte counts > 150,000/µL (r = .94). Correlation of estimates of polychromasia with ARP was good (r = .83). CONCLUSION: In anemic dogs MPP can be used for assessment of regeneration, especially in dogs with higher levels of ARP. Standard quantification of reticulocytes is more accurate in dogs with lesser degrees of regeneration.