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Summary Experiment 1: Percentages of spermatozoa were scored viable or nonviable (non-eosinophilic or eosinophilic) in an ejaculate from each of ten stallions stained with Hancock's stain, i.e., eosin Y-nigrosin (EN) or a mixture of ethidium bromide (EB) and acridine orange (AO). When spermatozoa were stained immediately after seminal collection, no significant difference (P>0.05) was detected in percentage of spermatozoa scored as viable with EN or EB-AO (55 ± 3.4 vs 52 ± 5.2), respectively. Similarly, when aliquots from each ejaculate were subjected to conditions expected to drastically reduce viability, percentages of spermatozoa scored viable with EN were not different (P>0.05) from those obtained using EB-AO stain. Thus, EN appeared to provide a practical means for rapidly estimating percentages of viable spermatozoa in equine ejaculates. Experiment 2: Six ejaculates from each of six stallions were stained with EN and evaluated by two experienced technicians. To determine effects of region of slide and number of spermatozoa evaluated on percentages of spermatozoa scored either normal or abnormal, and eosinophilic or non-eosinophilic, each slide was divided into three regions and 100, 200 and 300 spermatozoa evaluated. When spermatozoa were scored abnormal, abnormalities were sub-classified into specific anomalies of the head, mid-piece or tail. In addition, data were analyzed to determine variation between technicians evaluating the spermatozoa, and percentage of spermatozoa scored eosinophilic or non-eosinophilic after storage for 0, 7 or 28 days after slide preparation. Region within slide, number of spermatozoa evaluated, technician or storage had no significant effects (P>0.05) on percentage of spermatozoa classified as normal, abnormal, eosinophilic or non-eosinophilic. However, there was a difference (P |
