Johns Jennifer L, Shooshtari Mahrokh P, Christopher Mary M
Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
Am J Vet Res. 2008 Aug;69(8):1067-72. doi: 10.2460/ajvr.69.8.1067.
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.
Blood samples from a red-tailed hawk and 31 ill birds.
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.
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.
Results indicated that quantification of ring-form reticulocytes provides an accurate assessment of erythrocyte regenerative capacity in birds.
制定一种网织红细胞分类方案，优化禽类网织红细胞染色方案，并比较禽类血液样本中网织红细胞类型的百分比与多染性红细胞百分比。
一只红尾鹰和31只患病禽类的血液样本。
从一只红尾鹰（赤肩鵟）采集的一份血液样本用于优化染色方案。为优化染色方案，评估了用新亚甲蓝染液对全血进行的4种稀释度以及4种孵育时间。从提交进行禽类全血细胞计数的样本中，随机选取31只患病禽类的乙二胺四乙酸抗凝全血样本进行检查，以比较多染性红细胞和网织红细胞的百分比。所有样本均采用1:3（全血与新亚甲蓝）稀释，在室温（约22摄氏度）下孵育10分钟进行网织红细胞染色；由2名独立观察者将网织红细胞评估为每1000个红细胞中的百分比。在瑞氏-吉姆萨染色的血涂片上，同样确定多染性红细胞百分比。
定义了4种禽类网织红细胞类型：环形网织红细胞、聚集性网织红细胞以及点状网织红细胞的2个亚类。优化了网织红细胞染色方案。观察者间和观察者内对网织红细胞和多染性红细胞百分比评估的差异不显著。多染性红细胞百分比与环形网织红细胞百分比之间存在强正相关（斯皮尔曼等级相关系数[rho]=0.978）。
结果表明，环形网织红细胞的定量可为禽类红细胞再生能力提供准确评估。
