Abstract
Traditionally, egg yolk is a protective agent that is used to freeze semen in various species. However, the addition of egg yolk in extender risks the introduction of disease. Therefore, an alternative cryoprotective agent should be found to preserve ram semen. The aim of this study was to evaluate the effect of phosphatidylcholine (PC) as a protective agent in extender with or without egg yolk on semen characteristics and acrosome integrity of frozen then thawed Bapedi ram semen. Semen was collected from four mature Bapedi rams, in the Agricultural Research Council (ARC) Germplasm Conservation Programme, using an artificial vagina. Following collection, semen samples were randomly diluted into Tris-based extender (1: 2), with and without egg yolk, and supplemented with four concentrations of PC liposome (0 mg/ml), 0.25 mg/ml, 0.5 mg/ml and 0.75 mg/ml). Supplementation of PC liposome in extender with or without egg yolk did not improve the semen total motility (TM), progressive motility (PM) and rapid motility (RM) rate. The sperm cell membrane integrity in extender with or without egg yolk was not influenced by the supplementation of PC liposome after thawing (P >0.05). The addition of PC liposome to Tris-based extender with egg yolk had a similar result to control (Tris-based extender with egg yolk) on sperm cell acrosome integrity. In conclusion, supplementation of PC liposome to Tris-based extender without egg yolk had lower sperm cell viability and motility rates compared with the extender with egg yolk, regardless of concentration.Keywords: acrosome, cryoprotectant, liposome, membrane, motility
Highlights
Cryopreservation of semen is a useful technology for indefinite conservation of spermatozoa and preservation of genetic resources (Day & Stacey, 2007; Castro et al, 2016)
Freezing and thawing semen in extender with egg yolk resulted in better membrane integrity compared with semen that was frozen in extender without egg yolk (P
Integrity of the sperm cell membrane after freezing and thawing was not influenced by PC liposome supplementation of the extender (P >0.05)
Summary
Cryopreservation of semen is a useful technology for indefinite conservation of spermatozoa and preservation of genetic resources (Day & Stacey, 2007; Castro et al, 2016). Its use has been disputed because of the risk of introducing disease (Chelucci et al, 2015) and owing to inconsistency in its biochemical composition (Kulaksiz et al, 2010; Emamverdi et al, 2013). It may inhibit sperm metabolism and respiration, reduce motility, and cause difficulties during semen analysis (Amirat et al, 2004; Singh et al, 2012; Ugur et al, 2019).
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