Using zona-free hamster eggs and salt-stored human eggs for assessing the fertilizing capacity of human spermatozoa, the effects of human seminal plasma on fertilizing capacity of human spermatozoa were investigated. The persistent presence of seminal plasma prevented sperm attachment to and penetration into the zona pellucida and vitellus. A series of experiments with zona-free hamster eggs revealed that, once the spermatozoa were preincubated in a seminal plasma-free environment known to induce the acrosome reaction, the seminal plasma no longer interfered with sperm-egg fusion. The native seminal plasma appears to interfere with both the acrosome reaction and vigorous motility of the spermatozoa, and this could be the reason fertilization fails when the plasma is consistently present around the spermatozoa. The fertilization-disturbing factor or factors in the seminal plasma appear to be macromolecular substances or substances associated with macromolecules.