TY - JOUR
T1 - Development of a technique for quantification of reticulocytes and assessment of erythrocyte regenerative capacity in birds
AU - Johns, Jennifer L.
AU - Shooshtari, Mahrokh P.
AU - Christopher, Mary M
PY - 2008/8
Y1 - 2008/8
N2 - 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°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 [p] = 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.
AB - 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°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 [p] = 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.
UR - http://www.scopus.com/inward/record.url?scp=49749103135&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=49749103135&partnerID=8YFLogxK
U2 - 10.2460/ajvr.69.8.1067
DO - 10.2460/ajvr.69.8.1067
M3 - Article
C2 - 18672972
AN - SCOPUS:49749103135
VL - 69
SP - 1067
EP - 1072
JO - American Journal of Veterinary Research
JF - American Journal of Veterinary Research
SN - 0002-9645
IS - 8
ER -