What is your diagnosis? Blood smear from an injured red-tailed hawk.

An injured juvenile red-tailed hawk (Buteo jamaicensis) was evaluated at the Veterinary Medical Teaching Hospital at the University of California, Davis. The hawk was quiet, alert, and emaciated, and had a closed comminuted, mid-diaphyseal ulnar fracture. CBC results included heterophilia with a left shift, monocytosis, and increased plasma fibrinogen concentration. The blood smear included rare heterophils containing small, dark blue inclusions approximately 1-2 mum in diameter that ranged from round to coccobacillary in shape and formed variably shaped aggregates; the morphology of the inclusions was suspicious for Chlamydophila or Ehrlichia spp. pathogens. The hawk died, and histopathologic examination of tissues obtained at necropsy found severe multifocal histiocytic and heterophilic splenitis in addition to chronic hepatitis, myocarditis and epicarditis, meningoencephalitis, and airsacculitis. Using immunohistochemistry the presence of Chlamydia/Chlamydophila spp. antigen within multiple tissues was confirmed. Chlamydophila psittaci DNA was demonstrated in whole blood and fresh splenic tissue via real-time PCR. Direct fluorescent antibody staining of air-dried blood smears was positive in rare leukocytes for Chlamydia/Chlamydophila spp. antigen, and immunocytochemical staining of blood smears for Chlamydia/Chlamydophila spp. antigen was focally positive in rare heterophils. These findings may represent the first reported diagnosis of natural avian C. psittaci infection by visualization of organisms in peripheral blood heterophils. Immunocytochemical evaluation of blood smears was valuable in confirming the diagnosis and may be a useful antemortem test to discriminate between bacteria and other inclusions within heterophils.

Development of a technique for quantification of reticulocytes and assessment of erythrocyte regenerative capacity in birds.

OBJECTIVE: To develop a reticulocyte classification scheme, optimize an avian reticulocyte staining protocol, and compare the percentages of reticulocyte types with polychromatophil percentage in blood samples from birds. SAMPLE POPULATION: Blood samples from a red-tailed hawk and 31 ill birds. PROCEDURES: A single blood sample obtained from a red-tailed hawk (Buteo jamaicensis) was used to optimize the staining protocol. For optimization of the staining protocol, 4 dilutions of whole blood with new methylene blue stain and 4 incubation times were evaluated. From samples submitted for avian CBCs, EDTA-anticoagulated whole blood samples from 31 ill birds were randomly selected and examined to compare polychromatophil and reticulocyte percentages. Reticulocyte staining was performed in all samples by use of a 1:3 (whole blood to new methylene blue) dilution with incubation for 10 minutes at room temperature (approx 22 degrees C); reticulocytes were assessed as a percentage of 1,000 RBCs by 2 independent observers. In Wright-Giemsa-stained blood smears, a polychromatophil percentage was similarly determined. RESULTS: 4 avian reticulocyte types were defined: ring-form reticulocytes, aggregate reticulocytes, and 2 subcategories of punctate reticulocytes. A reticulocyte-staining protocol was optimized. Interobserver and intraobserver variations in assessment of reticulocyte and polychromatophil percentages were not significant. A strong positive correlation (Spearman coefficient of rank correlation [rho] = 0.978) was identified between the percentage of polychromatophils and the percentage of ring-form reticulocytes. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that quantification of ring-form reticulocytes provides an accurate assessment of erythrocyte regenerative capacity in birds.

Assessment of erythrocyte morphologic abnormalities in mallards with experimentally induced zinc toxicosis.

OBJECTIVE: To describe and quantify morphologic abnormalities in RBCs of mallards (Anas platyrhynchos) with experimentally induced zinc toxicosis. ANIMALS: 120 female mallards. PROCEDURE: Farm-raised mallards (6 to 8 months old) were used in the study. On day 0, 60 ducks received shot pellets orally by gavage (mean dose of zinc, 0.97 g); another 60 ducks underwent the same procedure without administration of pellets. On day 15, Romanowsky-stained blood smears were prepared from 53 control and 45 zinc-treated ducks (smears were examined retrospectively). In each smear, 200 RBCs were examined and numbers of erythrocytes with abnormal size, shape, or color were expressed as a percentage. Results were compared with PCV values and zinc dose. RESULTS: Mean PCV value was lower in all zinc-treated ducks, compared with control ducks, and was lower in zinc-treated ducks that died or were euthanatized before day 30 because of severe clinical disease, compared with those that survived. Zinc-treated ducks that survived had a high percentage of polychromatophilic RBCs, and those that that died before day 30 had high percentages of hypochromic RBCs, fusiform RBCs, and RBC nuclear abnormalities. There was no correlation between PCV value or RBC morphologic abnormalities and dose of zinc. CONCLUSIONS AND CLINICAL RELEVANCE: In ducks with severe zinc-induced hemolysis, findings indicated that functional iron deficiency may impair the effectiveness of the erythroid regenerative response and contribute to death. Erythrocyte nuclear abnormalities were consistent with mild dyserythropoiesis. These findings may be applicable to effects of other metal toxicoses and regenerative anemias in birds.