Cryopreservation of saltwater crocodile (Crocodylus porosus) spermatozoa

Johnston, S. D., Qualischefski, E., Cooper, J., McLeod, R., Lever, J., Nixon, B., Anderson, A. L., Hobbs, R., Gosálvez, J., López-Fernández, C. and Keeley, T. (2017) Cryopreservation of saltwater crocodile (Crocodylus porosus) spermatozoa. Reproduction, Fertility and Development, . doi:10.1071/RD16511

Author Johnston, S. D.
Qualischefski, E.
Cooper, J.
McLeod, R.
Lever, J.
Nixon, B.
Anderson, A. L.
Hobbs, R.
Gosálvez, J.
López-Fernández, C.
Keeley, T.
Title Cryopreservation of saltwater crocodile (Crocodylus porosus) spermatozoa
Formatted title
Cryopreservation of saltwater crocodile (Crocodylus porosus) spermatozoa
Journal name Reproduction, Fertility and Development   Check publisher's open access policy
ISSN 1031-3613
Publication date 2017-03-30
Sub-type Article (original research)
DOI 10.1071/RD16511
Open Access Status Not yet assessed
Total pages 10
Place of publication Clayton, VIC, Australia
Publisher CSIRO Publishing
Collection year 2018
Language eng
Formatted abstract
The aim of the present study was to develop a protocol for the successful cryopreservation of Saltwater crocodile spermatozoa. Sperm cells were frozen above liquid nitrogen vapour in phosphate-buffered saline (PBS) containing either 0.3 M trehalose, 0.3 M raffinose or 0.3 M sucrose and compared with glycerol (0.3–2.7 M). Although the highest levels of mean post-thaw motility were observed following cryopreservation in 0.3 M trehalose (7.6%) and 0.3 M sucrose (7.3%), plasma membrane integrity (PI) was best following cryopreservation in 2.7 M glycerol (52.5%). A pilot study then assessed the cytotoxicity of glycerol and sucrose prior to cryopreservation and revealed no loss of survival when spermatozoa were diluted in 0.68 M glycerol or 0.2–0.3 M sucrose once cryoprotectants were washed out with PBS or Biggers, Whitten and Whittingham medium containing sperm capacitation agents (BWWCAP). A final study refined the combined use of permeating (0.68 or 1.35 M glycerol) and non-permeating (0.2 or 0.3 M sucrose) cryoprotectants. Spermatozoa were cryopreserved in liquid nitrogen vapour at rates of approximately -21°C min-1 (fast freeze) or -6.0°C min-1 (slow freeze). Post-thaw survival was highest with a combination of 0.2 M sucrose and 0.68 M glycerol and when these cryoprotectants were washed out with BWWCAP, regardless of whether spermatozoa were frozen using a fast (motility 14.2 ± 4.7%; PI 20.7 ± 2.0%) or slow (motility 12.0 ± 2.7%; PI 22 ± 4%) cryopreservation rate.
Keyword Glycerol
Non-permeating cryoprotectants
Permeating cryoprotectants
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ
Additional Notes Published online 30 March 2017

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
School of Agriculture and Food Sciences
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Created: Thu, 30 Mar 2017, 14:29:38 EST by Associate Professor Stephen Johnston on behalf of School of Agriculture and Food Sciences