An Improved Protocol for Hematology Staining With Pallid Sturgeon

Hematological staining is useful for the diagnosis of disease and stress; however, methods are not yet standardized within Acipenseridae.  Capitalizing on a previous study using pallid sturgeon (Scaphirhynchus albus), variations of two staining protocols were tested to assess whether peripheral blood cell types could be better differentiated and identified.  Either Wright-Giemsa (WG), or Sudan Black B (SBB) counterstained with methyl green (MG), were applied to blood smears.  The experimental treatments differed by staining time and by counterstaining method and time.  Sodium heparinized blood (1:9) was used (n=1 sturgeon), and each treatment group was comprised of five replicate slides (total = 85). Ranked endpoints included the ability to differentiate erythrocytes from leukocytes and the clarity of intracellular structures.  Results showed that a 10 minute WG staining period was better than the previously used 10-15 seconds.  For SBB, staining for 90 minutes (not 30) followed by counterstaining with MG for 15 minutes (not 5) was better.  Overall, the SBB with MG was more effective than WG for cell differentiation and identification.  These improved blood staining protocols for use with pallid sturgeon are likely to be useful with other Acipenseriformes.