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Supplementation of Underfed Twin-Bearing Ewes with Herbal Vitamins C and E: Impacts on Birth Weight, Postnatal Growth, and Pre-Weaning Survival of the Lambs
Open AccessArticle

Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function

Clinic for Reproduction and Large Animals, Veterinary Faculty, University of Ljubljana, Gerbičeva 60, 1000 Ljubljana, Slovenia
School of Veterinary Medicine, Clinical Sciences & Advanced Medicine, University of Pennsylvania, Philadelphia, PA 19143, USA
Division of Gynaecology and Obstetrics, University Medical Centre Ljubljana, Zaloška cesta 2, 1000 Ljubljana, Slovenia
Author to whom correspondence should be addressed.
Animals 2020, 10(4), 653;
Received: 3 February 2020 / Revised: 5 April 2020 / Accepted: 7 April 2020 / Published: 9 April 2020
(This article belongs to the Section Animal Reproduction)
Soy lecithin and sucrose were used at different concentrations to develop and compare different vitrification methods for the cryopreservation of canine semen. All of the sperm quality characteristics were effectively preserved after devitrification when vitrification extenders containing soy lecithin at 1% and 0.25 M sucrose were used. The results suggest that vitrification is effective, fast, and simple for cryopreservation of canine semen. Furthermore, the use of soy lecithin in lieu of animal proteins (e.g., egg yolk) facilitates semen shipping to countries with strict import requirements.
A challenge in freezing semen for short and long-term availability is avoiding damage to intact spermatozoa caused by the freezing process. Vitrification protocols provide better results through less manipulation of semen and shorter freezing time compared to slow freezing techniques. Our research was aimed at improving vitrification methods for canine semen. Semen quality was determined in 20 ejaculates after collection. Each ejaculate was divided into eight aliquots, each with a different extender. The control extender contained TRIS, citric acid, fructose, and antibiotics. Soy lecithin and sucrose were added to the control extender at different concentrations to make up the test extenders and final concentration of 50 × 106 spermatozoa/mL. From each group, a 33 µL (1.65 × 106 spermatozoa) suspension of spermatozoa was dropped directly into liquid nitrogen and devitrified at least one week later and evaluated as before. Soy lecithin at 1% and 0.25 M sucrose added to the base vitrification media effectively preserved all sperm qualities. Our results demonstrate the effectiveness of our methods. Vitrification media containing sucrose and soy lecithin cause a minimal decline in quality of canine semen after devitrification. Furthermore, extenders used in our research did not contain egg yolk, which was replaced by soy lecithin, thus allowing for ease of shipping to other countries with strict requirements. View Full-Text
Keywords: Vitrification; soy lecithin; sucrose; semen preservation; canine Vitrification; soy lecithin; sucrose; semen preservation; canine
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Zakošek Pipan, M.; Casal, M.L.; Šterbenc, N.; Virant Klun, I.; Mrkun, J. Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function. Animals 2020, 10, 653.

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