1. Four methods of semen collecting that involved interruption of mating in two breeding ostrich pairs were tested: an artificial vagina was tested without promising results; the funnel method, in which a funnel was placed under the phallus of the tested male immediately after mating allowing semen drips to be collected; the vacuum method, using a turkey semen collector, inserted into the seminal canal; and the tube method, conducted by placing a test tube inside the seminal canal, allowing semen to enter by gravity. 2. For the funnel, vacuum and tube methods, respectively, average semen volume was 0.1 ± 0.02, 1.12 ± 0.22, and 0.58 ± 0.13 ml, sperm concentration was 0.66 ± 0.14, 2.35 ± 0.26, and 2.13 ± 0. 27 × 109 cells/ml, and percentage of abnormal cells was 5.82 ± 1.79%, 4.68 ± 1.19%, and 7.09 ± 1.72%. 3. Semen characteristics varied throughout the reproductive season reaching peak concentration in June-July. 4. The vacuum method proved to be the most efficient and was a low stress, restraint-free method for collecting ostrich semen.