- Title
- Cryopreservation of saltwater crocodile (Crocodylus porosus) spermatozoa
- Creator
- Johnston, S. D.; Qualischefski, E.; Keeley, T.; Cooper, J.; McLeod, R.; Lever, J.; Nixon, B.; Anderson, A. L.; Hobbs, R.; Gosálvez, J.; López-Fernández, C.
- Relation
- Reproduction, Fertility and Development Vol. 29, Issue 11, p. 2235-2244
- Publisher Link
- http://dx.doi.org/10.1071/RD16511
- Publisher
- C S I R O Publishing
- Resource Type
- journal article
- Date
- 2017
- Description
- 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.
- Subject
- glycerol; non-permeating cryoprotectants; permeating cryoprotectants; raffinose; sucrose; trehalose
- Identifier
- http://hdl.handle.net/1959.13/1350731
- Identifier
- uon:30601
- Identifier
- ISSN:1031-3613
- Language
- eng
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