Carnivore semen cryopreservation procedures started with semen washing and centrifuging in culture media for seminal plasma removal and microorganisms elimination. The objective of this study was to perform coatis semen cryopreservation comparing the effects between two extenders Ham’s F-10 and M199 for washing and centrifugation before cryopreservation using Dilutris medium. Semen samples (n = 36) were collected by electroejaculation from six adult male coatis (Nasua nasua) between May and October of 2008 at the Universidade Federal de Mato Grosso Zoo. Sperm total motility (%), progressive sperm motility (0-5), plasma membrane integrity spermatozoa rates (%), and acrosome integrity (%) were analyzed. These fresh semen samples were divided in two fractions, diluted in 1 ml of Ham’s F-10 (Ham’s F-10, Nutricel S.A., Brazil) or M199 (M199, Nutricel S.A., Brazil) and centrifuged at 300 g for 10 min. The supernatant was discarded and pellets resuspended in 1 ml of Dilutris (Dilutris, Minitube®, Brazil), stored at 5ºC for 3 hours, transferred to 0.25 ml straws, placed in liquid nitrogen vapor for 20 min, and immersed in liquid nitrogen. The means/SD for fresh semen and cryopreserved semen using Ham’s F-10/Dilutris and M199/Dilutris were, respectively: 84.28 ± 11.57, 45.38 ± 27.26, and 44.61 ± 25.03 for total motility; 3.64 ± 1.44, 2.15 ± 1.14, and 2.07 ± 1.03 for progressive sperm motility; 92.76 ± 3.46, 84.69 ± 15.77, and 89.76 ± 13.97 for live spermatozoa rate; and 94.76 ± 2.89, 92.35 ± 4.73, and 90.58 ± 7.17 for acrosome integrity. No significant difference (P < 0.05) were observed between the values obtained from the Ham’s F-10/Dilutris or M199/Dilutris treatments. Both treatments demonstrated to be suitable for freezing semen from this species.