AUTOMATED MORPHOMETRIC ANALYSIS OF STALLION SPERMATOZOA

Tissue variation in microscope slides made for spermatozoon analysis and variation introduced by the subjective techniques used to analyze these slides reduce the statistical power of studies that seek to use spermatozoon morphology to predict fertility. A simple specimen preparation method was developed to standardize stallion spermatozoon morphologic smears, and a new, automated spermatozoa morphometry instrument was used to objectively analyze the efficacy of the specimen preparation technique. The method achieved a standard spermatozoon concentration and reduced field-to-field variation in the number of spermatozoa analyzed. Metric measurements of spermatozoon head dimensions from clinically normal, fertile stallions revealed small, but highly significant, differences between stallions. The variation in metric measurements between replicate slides within stallions was small, indicating that replicate slide analysis probably is not necessary for clinically normal stallions. Coefficients of variation were generally < 11% for metric measurements between stallions, and were < 4% within stallions. This study revealed that a high degree of statistical power can be achieved when using these new, standardized specimen preparation and objective analysis techniques. Such power makes possible the detection of subtle differences between clinically normal stallions, and may facilitate accurate detection of abnormal fertility (ie, subfertility) in stallions.