Medycyna Wet. 64 (4b), 608-612, 2008
Demianowicz W., Giżejewski Z., Kubiak D., Kowalski R., Glogowski J.
Improvement of the cryopreservation of red deer (Cervus elaphus L.) semen collected with an artificial vagina
Cryopreservation of red deer sperm is essential for establishing the biodiversity of this species. The aim of the study was to test four extenders and two freezing methods on the cryosurvival of red deer spermatozoa. Semen collected with an artificial vagina from 4 stags was diluted with compared extenders: A (citrate-fructose-egg yolk-glicerol), B (Tris-fructose-citric acid-egg yolk-glicerol), C (Triladyl® with egg yolk) and D (Bioxcell®). Sperm, loaded into 0.25 ml straws, was frozen in nitrogen vapor (method L) and in the Minicool 40 PC (method M) cryogenic unit. After thawing the motility of spermatozoa was evaluated subjectively. Viability was assessed using nigrosin-eosin (N-E test), SYBR-14 with propidium iodide (L/D test) and hypoosmotic swelling test (test HOS) to detect membrane integrity. The best post-thaw motility was obtained with the use of extender D (43.0 ± 14.2%). Similarly, more viable spermatozoa (p < 0.05) at N-E and L/D tests (42.4 ± 11.3% and 39.9 ± 14.0% respectively) were preserved in extender D. The lowest results in those tests were received in extender A. The percentage of HOST-responsive spermatozoa was higher (p < 0.05) in method M than in L (19.8 ± 9.8 % vs 14.3 ± 7.6 %), independently of the utilized extenders. In conclusion, the use of extender D in combination with freezing method M significantly improved freezability of red deer spermatozoa.
Keywords: red deer, cryopreservation, sperm