Cryopreservation of epididymal sperm collected postmortem in the Tasmanian devil (Sarcophilus harrisii)

Keeley, T., McGreevey, P. D. and O'Brien, J. K. (2012) Cryopreservation of epididymal sperm collected postmortem in the Tasmanian devil (Sarcophilus harrisii). Theriogenology, 78 2: 315-325. doi:10.1016/j.theriogenology.2012.02.005

Author Keeley, T.
McGreevey, P. D.
O'Brien, J. K.
Title Cryopreservation of epididymal sperm collected postmortem in the Tasmanian devil (Sarcophilus harrisii)
Formatted title
Cryopreservation of epididymal sperm collected postmortem in the Tasmanian devil (Sarcophilus harrisii)
Journal name Theriogenology   Check publisher's open access policy
ISSN 0093-691X
Publication date 2012
Sub-type Article (original research)
DOI 10.1016/j.theriogenology.2012.02.005
Open Access Status
Volume 78
Issue 2
Start page 315
End page 325
Total pages 11
Place of publication Philadelphia, PA, United States
Publisher Elsevier
Language eng
Formatted abstract
Effective sperm cryopreservation protocols are limited to a small number of marsupial species. In this study, postmortem gamete rescue (PMGR) epididymal sperm samples from Tasmanian devils (N = 34) euthanized due to the fatal Devil Facial Tumor Disease were used to develop long-term sperm storage techniques for the species. Cryoprotectant toxicity associated with equilibration of sperm samples in a TEST yolk diluent (TEST; 189 mM N-Tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid, 85 mM Trizma base [Tris], 11 mM glucose, 20% vol/vol egg yolk; pH 7.1, and 315.0 ± 5.0 mOsm/kg) with a final concentration of 0.06 M trehalose, or 4%, 10%, and 18% vol/vol of either glycerol or dimethyl sulfoxide (DMSO), was examined over 12 h at 15 °C. Trehalose supplementation resulted in an immediate decline (P < 0.05) of total motility. After 12 h, total motility was reduced (P < 0.05) in treatments containing 18% glycerol, and 10% and 18% dimethyl sulfoxide. The effects of final glycerol concentration (4% and 10%), glycerol equilibration duration (10 min 1 h, or 3 h) prefreeze, freezing rate and the addition of 0.10 M lactose or a combination of 0.10 M lactose and 0.11 M raffinose were assessed during three experiments on the cryopreservation of postmortem gamete rescue samples in TEST. In all experiments, motility and viability were reduced (P < 0.01 postthaw). Samples cryopreserved in TEST supplemented with lactose or lactose with raffinose using a fast freezing rate (-8 °C/min from 4 to -40 °C, then -65 °C/min until -165 °C) produced the highest (P < 0.05) postthaw motility (18.6 ± 5.5% and 16.9 ± 8.5%, respectively), which represented 35% to 48% retention of prefreeze motility. These results apparently were the best postthaw results of dasyurid sperm reported to date and will help lay the foundations for developing assisted reproductive technologies for marsupial species.
Keyword Cryopreservation
Tasmanian devil
Gamete rescue
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Agriculture and Food Sciences
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Citation counts: TR Web of Science Citation Count  Cited 3 times in Thomson Reuters Web of Science Article | Citations
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Created: Mon, 15 Dec 2014, 12:01:04 EST by Tamara Keeley on behalf of School of Agriculture and Food Sciences