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Asian-Australas J Anim Sci > Accepted Articles
DOI: https://doi.org/10.5713/ajas.19.0338    [Accepted] Published online October 21, 2019.
Enhancing liquid-chilled storage and cryopreservation capacities of ram spermatozoa by supplementing the diluent with different additives
Sherif A. Rateb1,*  , Marwa A. Khalifa1  , Ibrahim S. Abd El- Hamid1  , Hisham A. Shedeed1 
Animal and Poultry Production Division, Desert Research Center, Ministry of Agriculture and Land Reclamation, Egypt
Correspondence:  Sherif A. Rateb, Tel: +20-1097310306, Fax: +20-34480064, Email: rateb.drc@gmail.com
Received: 18 April 2019   • Revised: 8 September 2019   • Accepted: 30 September 2019
Abstract
Objective
In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 PUFA in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa.
Methods
In the first experiment, ejaculates (n=30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T0). The diluted specimens were stored at 4 oC for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T24, T48). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis (CASA) system.
Results
The results clarified superiority of both melatonin and omega-3 supplementation on maintaining (P<0.05) sperm properties, while reducing (P<0.05) livid peroxidase (LPO) reaction and enzymatic activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of sperm.
Conclusion
melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for artificial insemination (AI) or in vitro fertilization (IVF).
Keywords: Cryopreservation; Oxidative Stress; CASA; Ram Semen
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