Abstract
The fertility rate in dairy cattle with cryopreserved semen significantly depends on the reduction of the toxic effects on the sperms induced during cryopreservation. DNA integrity in sperm is vital for the precise transmission of genetic information and therefore the production of high yielders and maintenance of good health in future generations. The reliable and consistent assessment of sperm motility can be succeeded by Computer Assisted Semen Analyzer (CASA). Ten ejaculates were collected from Frisian bulls kept at Centre of excellence for bovine genetics (CEBG) Renala Khurd District Okara. The deleterious effects of three cryoprotectants glycerol, dimethylsulfoxide (DMSO) and ethylene glycol as a constituent of tris-egg yolk-citrate extender were compared on diluted post thawed cryopreserved Frisian bull semen. The quality of semen in terms of viability/motility/progressive velocity was determined with computerized external real image optical system (CEROS) and sperm DNA fragmentation was quantified with comet assay using single cell gel electrophoresis (SCGE) technique. Glycerol was the least followed by DMSO and ethylene glycol was the most toxic cryoprotectant both for semen quality and sperm DNA fragmentation. The present study suggests that sperm DNA fragmentation is an associated feature of semen cryopreservation resulting into cell death (non-motile sperms) and glycerol as a cryoprotectant constituent of tris-egg yolk citrate extender offers a better protection for storage of cryopreserved Frisian bull semen in liquid nitrogen.
Highlights
The fertility rate in animal production mainly depends upon the quality of semen
Though the whole process of cryopreservation right from collection of semen to thawing and insemination cause varying degree of semen quality deterioration, the present study targeted the Cryoprotective agents (CPAs) used in extenders keeping rest of the factors standardized
The present study provides persuasive support for the genotoxic effects of CPAs glycerol, DMSO and ethylene glycol as a part of tris-egg yolk-citrate extender during the cryopreservation process of Frisian bull semen
Summary
The fertility rate in animal production mainly depends upon the quality of semen. In dairy cattle, the extensive usage of artificial insemination (AI) has ensued in increased selection intensity, resultantly achieving enhanced productivity (Naveen et al, 2014). There are different approaches in practice to evaluate the DNA disintegration, such as the orthodox agarose gel electrophoresis but a more current technique is single cell gel electrophoresis (Anderson and Laubenthal, 2013, Soudabeh et al, 2016) This technique was first presented in 1984 and has developed intoa renowned method toidentify DNA fragmentations in a range of cell types, including sperm cells (Steele et al, 2000).Commonly this assay is termed as Comet assay, since broken and intact DNA as a whole looks like a comet and is a simple, quick, optical and sensitive technique for detecting primary DNA damage at the single cell level (Singh et al, 1989). The objective of present study was to evaluate the proficiency of Glycerol, DMSO and Ethylene glycolas CPA of tris egg yolk citrate extender by assessing semen quality descent / sperm DNA fragmentation and to find out any correlation between these two phenomenon
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