Efficacy of short-term cold storage prior to cryopreservation of spermatozoa in a threatened lizard
Lachlan Campbell A , John Clulow A , Belinda Howe A , Rose Upton
A , Sean Doody A B and Simon Clulow C D E
+ Author Affiliations
- Author Affiliations
A School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
B Department of Biological Sciences, University of South Florida, St. Petersburg, FL 33701, USA.
C Institute for Applied Ecology, University of Canberra, Bruce, ACT 2617, Australia.
D Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, Australia.
E Corresponding author. Email: simon.clulow@canberra.edu.au
Reproduction, Fertility and Development 33(9) 555-561 https://doi.org/10.1071/RD20231
Submitted: 7 September 2020 Accepted: 21 January 2021 Published: 2 March 2021
Journal Compilation © CSIRO 2021 Open Access CC BY-NC
Abstract
Assisted reproductive technologies (ARTs) have a significant role to play in reptile conservation, yet are severely lacking. Previous attempts to cryopreserve spermatozoa in the threatened lizard Varanus panoptes achieved approximately 48% motile sperm post-thaw for samples frozen immediately after collection. However, the feasibility of extended cold storage before cryopreservation has not been tested. We held V. panoptes spermatozoa at either 25°C or 4°C for 8 days, assessing sperm motility at days 1, 2, 4 and 8. Subsamples were cryopreserved on days 1 and 4 following the previously reported protocol for this species. Percentage motility decreased rapidly at 25°C, but did not decrease significantly until 4 days after collection at 4°C, with >30% motility maintained after 8 days. There was no significant difference in post-thaw motility or viability of samples cryopreserved after 1 or 4 days storage at 4°C, yielding substantial results for both parameters (mean motility 23.8% and 28.1% and mean viability 50.1% and 57.5% after 1 and 4 days respectively). We demonstrate the capacity to extend sperm viability for up to 8 days in unfrozen samples and to produce acceptable post-thaw motility in samples frozen after 4 days of storage, contributing to the development of valuable ARTs for lizards and other reptiles.
Keywords: assisted reproductive technologies, conservation, genome storage, reptile, squamate.
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