Chlamydia trachomatis and Ureaplasma urealyticum have been implicated as causative organisms in infections involving the male and female urogenital tracts. Seminal fluid, anterior urethral swabs and first-voided urine specimens from men undergoing infertility evaluation, with and without pyosemia, or anterior urethritis were cultured for Chlamydia trachomatis and Ureaplasma urealyticum. The method used to isolate Chlamydia trachomatis involved cytochalasin-B treated McCoy cells, and NYC and A7 solid media were used for the isolation of Ureaplasma urealyticum. Isolation of Chlamydia trachomatis from seminal fluid has not been possible even in the face of pyosemia and the presence of Chlamydia trachomatis in urine and urethral swab material. The reasons for the inability to culture Chlamydia trachomatis will be explored.