PARENT SESSION
Cryopreservation
524
SEMEN PARAMETERS AND CRYOPRESERVATION OF SPERMATOZOA OF ASSAMESE MACAQUE (Macaca assamensis).
Ji, Weizhi1,3, He, Xiechao1,3, Wang, Hong1,3, Bavister, Barry2,3, Li, Xilong1,3, 1 3 2
ABSTRACT- The Assamese macaque is a rare and endangered species found from northern India and Nepal to Vietnam. In China, this species is distributed only in southeast Tibet and southwest Yunnan. To our knowledge, there is no report on the semen characteristics of this species nor on cryopreservation of its spermatozoa. In the present study, spermatozoa were collected from 4 adult male monkeys using penile electroejaculation and semen parameters were analysed. Mean semen volume (ml), liquefaction (ml), concentration (X108/ml) and coagulum liquefaction rate (%) were 2.63±1.08, 1.13±0.38, 7.81±0.9 and 45.4±11.6, respectively (n=16). The semen was also cryopreserved using TTE extender (Sankai, 1994; 1.2% TES, buffer, 0.2% TRIS buffer, 2% Lactose, 2% Dextrose, 0.2% Raffinose, 5% Glycerol and 20% Egg yolk) and TEST extender (Tollner, 1990; 4.325% TES buffer, 1.0269% TRIS buffer, 1% Dextrose, 3% Glycerol and 30% Egg yolk). Sperm motility was used to evaluate post-thaw sperm viability. Samples were diluted with non-glycerolated TTE or TEST and equilibrated at 4 centigrade for 2h, then further diluted with equal volumes of TTE or TEST containing glycerol to reach final glycerol concentrations of 5% and 3% respectively, and frozen in 0.25ml in liquid nitrogen vapor for 10 min, finally placing them directly into liquid nitrogen. Semen was thawed by dropping straws directly into 37 centigrade water for 2 min. Post-thaw sperm motility in was 58% in TTE and 10% in TEST (n=10). After 2h, 4h, 6h, 8h and 10h incubation, post-thaw sperm motility in TTE was 38%, 31%, 21%, 17% and 2% vs. in fresh semen 59%, 54%, 30%, 11% and 4%, respectively. However, post-thaw sperm motility using TEST was only 5% after 2h incubation and 0% after 4h. We have demonstrated that TTE is a suitable extender for cryopreservation of spermatozoa of the Assamese macaque. This work was supported by the China National Science Foundation (39910117), the Chinese Academy of Sciences (KZ952-J1-109) and the Yunnan Nature Sciences Foundation (98C092M).
KEY WORDS: Assamese macaque, semen parameter, TTE, TEST