Cryopreservation of Asian Elephant Semen (Elephas Maximus) in Conjunction with the Use of Spermac Stain to Evaluate Acrosome Status in Both Asian And African Elephants (Loxodonta Africana)


Wendy K. Kiso

Date of Graduation

Fall 2004


Master of Natural and Applied Science in Agriculture


College of Agriculture

Committee Chair

Dennis Schmitt


Semen cryopreservation has become an important research priority for the endangered Asian elephant (Elephas maximus) population. The ability to cryopreserve Asian elephant semen will enhance the success of artificial insemination and have several implications for long-term storage of genome resource banks. This research discusses the comparisons for long-term storage of genome resource banks. This research discusses the comparison of two egg-yolk based extenders (TriBuff extender and Trehalose extender), cryoprotectants (DMSO, glycerol, trehalose), and other constituents (cholesterol, vitamin E, EDTA, Equex STM paste) on post-thaw improvements after cryopreservation of Asian elephant semen. Semen was collected from four Asian elephant bulls (n=24). Ejaculates of adequate quality were divided into separate aliquots to test the various treatments. Only treatments showing adequate motility after cooling were eventually frozen using liquid nitrogen. Motility and acrosome status were assessed during pre-freeze and post-thaw evaluations. There were no significant differences between treatments in motility (P > 0.11) and acrosome status (P > 0.19). This research also evaluated the use of Spermac® stain to assess acrosome status in fresh semen collected from both Asian and African elephants. Six Asian elephants (n=21) and two African elephants (n=5) were utilized for this portion of the study. Freshly collected semen was aliquoted and prepared for Spermac® stain analysis. Spermac® stain effectively assessed acrosome status in both Asian and African elephants, exhibiting a good indicator of cryopreservation success. Asian elephant semen demonstrated a significantly higher percentage of acrosome damage (P < 0.01) compared to African elephant semen.


semen, cryopreservation, Asian elephant, African elephant, Spermac®stain

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© Wendy K. Kiso