Optimizing the conventional method of sperm freezing in liquid nitrogen vapour for Wallachian sheep conservation program

Filipp Georgijevic Savvulidi; Martin Ptacek; Anezka Malkova; Jakub Beranek; Ludek Stadnik

doi:10.17221/226/2020-CJAS

Optimizing the conventional method of sperm freezing in liquid nitrogen vapour for Wallachian sheep conservation program

Filipp Georgijevic Savvulidi , Martin Ptacek , Anezka Malkova, Jakub Beranek, Ludek Stadnik

https://doi.org/10.17221/226/2020-CJASCitation:

Savvulidi F.G., Ptacek M., Malkova A., Beranek J., Stadnik L. (2021): Optimizing the conventional method of sperm freezing in liquid nitrogen vapour for Wallachian sheep conservation program. Czech J. Anim. Sci., 66: 55–64.

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The aim of the present study was to optimize the conventional method of sperm freezing in liquid nitrogen (LN₂) vapour for successful cryopreservation of Wallachian ram sperm, the genetic resources of the Czech Republic. Sperm in straws were frozen using the conventional freezing method via a static exposure of sperm doses to LN₂ vapour, or by four different modified freezing methods. Under modified freezing, straws were frozen by a discontinuous, time-dependent decremental change in the distance between the straws and the surface of LN₂. The viability of sperm was evaluated by flow cytometry after sperm equilibration, and immediately after thawing. Besides the observed inter-sire and daily variation, the obtained results suggest the methodological weakness of the conventional freezing method via the static exposure of sperm doses to LN₂ vapour. With the use of the optimized freezing procedure, all parameters of thawed sperm were significantly (P < 0.05) improved in comparison with the conventional method: percentage of thawed sperm viability increased up to 48.3%, percentage of sperm with plasma membrane damage after thawing decreased to 6.58%, percentage of sperm with acrosome damage decreased to 24.4%, and percentage of sperm with deteriorated mitochondrial activity decreased to 6.28%. In conclusion, our results suggest that an optimized freezing procedure should be routinely used instead of the conventional method to cryopreserve Wallachian ram sperm.

Keywords:

cryopreservation; flow cytometry; multicolour analysis; sperm doses; static exposure

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Czech Academy of Agricultural Sciences

Optimizing the conventional method of sperm freezing in liquid nitrogen vapour for Wallachian sheep conservation program

Filipp Georgijevic Savvulidi , Martin Ptacek , Anezka Malkova, Jakub Beranek, Ludek Stadnik

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