Our study aimed at addressing the question “Do spermatozoa biometric measures prove its fertility potential?”. Therefore, we carried out a comparative study of the buffalo bull’s fertility potential between Italian and Egyptian breeds based on morphometric analysis of frozen-thawed semen and in vitro fertilization. Semen was evaluated post-thawing for plasma membrane integrity, morphology, and motility. Sperm morphology was quantified using computer-assisted evaluation. The cleavage rates, morula, and blastocyst morphologies were recorded for the in vitro assessment of the fertilizing capacity. Our results show substantial differences in motility, viability, abnormalities, plasma membrane (P