title: Development of a technique for quantification of reticulocytes and assessment of erythrocyte regenerative capacity in birds.
abstract: 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.
  
  
  METHODS
  Blood samples from a red-tailed hawk and 31 ill birds.
  
  
  METHODS
  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
  Results indicated that quantification of ring-form reticulocytes provides an accurate assessment of erythrocyte regenerative capacity in birds.
authors.affiliation: Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Johns
authors.forename: Mahrokh P
authors.initials: MP
authors.lastname: Shooshtari
authors.forename: Mary M
authors.initials: MM
authors.lastname: Christopher
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000097:blood*; D002449:Cell Aggregation / Q000502:physiology; D004906:Erythrocyte Count; D004912:Erythrocytes / Q000166:cytology* / Q000473:pathology / Q000502:physiology*; D046428:Hawks / Q000097:blood*; D006403:Hematologic Tests / Q000662:veterinary; D012038:Regeneration; D012156:Reticulocytes / Q000166:cytology* / Q000473:pathology / Q000502:physiology