Early in his investigations, Leeuwenhoek (1670s) deduced that spermatozoa were alive and an integral part of semen, rather than artifacts or parasites. He eventually observed spermatozoa in the semen of men, dogs, horses, birds, fishes, amphibians, molluscs, and many insects, and concluded that they must be a universal feature of male reproduction. The huge differences in sperm form among species have been discussed in relation to evolutionary changes dictated by the egg and its investments. Spallanzani (1800s) was the first scientist to develop successful methods for artificial insemination, first with amphibians and later with dogs. With these experiments, he showed that physical contact between intact spermatozoa and ova was necessary to achieve the fertilization. Some years later (1820s), Prévost and Dumas performed the defining experiment to identify correctly the function of spermatozoa in reproduction.