Semen samples from 15 male American Kestrels (Falco sparverius) were frozen in dimethyl sulfoxide (DMSO). The semen was thawed 1-14 mo later and used to inseminate six females during three breeding seasons. Kestrels inseminated with thawed semen containing 4% DMSO produced only infertile eggs (N = 14). Kestrels inseminated with thawed semen containing 6%, 8%, or 10% DMSO produced fertile eggs (N = 14) and live chicks (N = 6). Progressive motility of spermatozoa in thawed semen containing 10% DMSO was less (44 ? 6%) than in thawed semen containing 6% (62 ? 10%) or 8% (61 ? 1%) DMSO.