Efficacy of short-term cold storage prior to cryopreservation of spermatozoa in a threatened lizard

Lachlan Campbell, John Clulow, Belinda Howe, Rose Upton, Sean Doody, Simon Clulow*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Number of pages7
JournalReproduction, Fertility and Development
Early online date2 Mar 2021
Publication statusE-pub ahead of print - 2 Mar 2021

Bibliographical note

Publisher Copyright:
© 2021 CSIRO.

Copyright 2021 Elsevier B.V., All rights reserved.


  • assisted reproductive technologies
  • conservation
  • genome storage
  • reptile
  • squamate

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