Characteristics of high-quality Asian elephant (Elephas maximus) ejaculates and in vitro sperm quality after prolonged chilled storage and directional freezing

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doi:10.1071/RD12129

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Characteristics of high-quality Asian elephant (Elephas maximus) ejaculates and in vitro sperm quality after prolonged chilled storage and directional freezing

J. K. O’Brien ^A ^E, K. J. Steinman ^A, G. A. Montano ^A ^B, C. C. Love ^C, R. L. Saiers ^D and T. R. Robeck ^A

^A SeaWorld and Busch Gardens Reproductive Research Center, SeaWorld Parks and Entertainment, 2595 Ingraham St, San Diego, CA 92109, USA.
^B Department of Animal Science, Texas A&M University, College Station, TX 77843, USA.
^C Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843, USA.
^D Albuquerque Biological Park, Albuquerque, NM 87102, USA.
^E Corresponding author. Email: justine.obrien@seaworld.com

Reproduction, Fertility and Development - http://dx.doi.org/10.1071/RD12129
Submitted: 21 February 2012 Accepted: 2 July 2012 Published online: 15 August 2012





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Abstract

The in vitro quality of spermatozoa from one elephant (Elephas maximus) was examined after chilled storage and directional freezing (DF). High-quality, non-contaminated ejaculates (77.6 ± 6.0% progressive motility, 3.9 ± 1.5 µg creatinine mL^–1 raw semen, 2.7 ± 0.6% detached heads) were cryopreserved after 0 (0hStor), 12 (12hStor) and 24 h (24hStor) of chilled storage. At 0 h and 6 h post-thawing, total motility, plasma membrane integrity, acrosome integrity, mitochondrial activity and normal morphology were similar (P > 0.05) across treatments. In contrast, progressive motility, rapid velocity and several kinematic parameters were lower (P < 0.05) for 24Stor compared with 0hStor at 0 h post-thaw. By 6 h post-thaw, amplitude of lateral head displacement and velocity parameters (average pathway, straight-line and curvilinear velocity) were lower (P < 0.05) for 24hStor compared with 0hStor and 12hStor. DNA integrity was high and remained unchanged (P > 0.05) across all groups and processing stages (1.6 ± 0.6% of cells contained fragmented DNA). Results indicate that DF after up to 12 h of chilled storage results in a post-thaw sperm population of acceptable quality for artificial insemination. These findings have implications for the cryopreservation of sex-sorted spermatozoa, which typically undergo more than 12 h of chilled storage prior to sorting and preservation.

Additional keywords: CASA, DNA integrity, mitochondrial activity, SCSA, sperm cryopreservation.

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