Research was conducted to develop an artificial insemination protocol optimizing the use of spermatozoa by turkey breeder hens. Large White turkey hens were inseminated on Days 14 and 17 postphotostimulation with 200 million spermatozoa from one male phenotype to fill the oviductal storage sites. Artificial inseminations were then performed weekly for 20 wk with different spermatozoa numbers of another male phenotype. Fertility and phenotype of each poult were determined at hatch to ascertain which insemination, initial or subsequent, was responsible for fertility. Inseminating weekly with 200 million viable spermatozoa cells resulted in better fertility but did not optimize the hen's utilization of spermatozoa from the initial inseminations. When fewer spermatozoa were inseminated weekly (50 million cells), more progeny were fertilized by spermatozoa already residing in the oviduct than would be expected. When the number of spermatozoa inseminated weekly was increased at intervals during a laying cycle, spermatozoa from the initial inseminations were utilized more efficiently, but fertility was depressed at times during the laying cycle. Gradually increasing weekly inseminated numbers of spermatozoa from 50 to 200 million viable cells/hen as the hens age results in nearly equivalent fertility to that resulting from insemination by 200 million cells each week. This represents a savings of 1.4 billion spermatozoa/hen over a 20-wk laying period.