The aim of this study is comparing the Sperm Chromatin condensation status between the first and second ejaculate using Aniline Blue Stain. Also each ejaculate was subjected to semen analysis including the semen volume, sperm count, and sperm motility. The results showed no statistically significant difference in the sperm chromatin condensation stained with Aniline Blue stain between the first and second ejaculates in all patients. Although the volume of the second ejaculate was decreased significantly than the first one, there was no significant difference in semen parameters (sperm concentration, percentage of total sperm motility, and motility grades (a+b)) between first and second ejaculates generally. However some patients showed improvement in semen parameters in the second ejaculate as compared with the first one.