Enhancing liquid-chilled storage and cryopreservation capacities of ram spermatozoa by supplementing the diluent with different additives

Abstract

Objective: In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid 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°C 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 (4% glycerol, v/v) 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 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) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa.Conclusion: Melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.

Melatonin and selenium supplementation in extenders improves the post-thaw quality parameters of rat sperm

“…The plasma membrane is involved in protecting the physiology of sperm cytosol. In freezing, osmotic changes influence membrane integrity and sperm homeostasis by changing salt concentrations [ 35 ]. In the current study, although the PMI in the group containing 1 mM Se did not differ from that of the frozen control group, it had the highest amount compared to the other groups.…”

Chlorogenic Acid Improves Quality of Chilled Ram Sperm by Mitigating Oxidative Stress

“…When the balance of ROS production and elimination was disrupted, excessive ROS caused damage to sperm [ 22 , 34 ]. ROS can not only lead to DNA chain break or sister chromatid exchange, but also oxidize key enzymes in the methylation process, leading to DNA methylation [ 35 , 36 ].…”

Cryopreservation of ram semen: Manual versus programmable freezing and different lengths of equilibration

“…Cryopreservation of ram sperm is less successful compared with other farm animals (bulls and rabbits; Curry et al, 2000) due to its higher sensitivity to freezing–thawing procedures and low cryotolerance. Various attempts have been tried to improve post‐thaw sperm viability using, for example, different additives (Bohlool et al, 2015; Emamverdi et al, 2013; Forouzanfar et al, 2010; Küçük et al, 2014; Luna‐Orozco et al, 2019; Masoudi et al, 2016; Najafi et al, 2014; Sharafi et al, 2014; Toker et al, 2016) or antioxidants (Alipour et al, 2018; Câmara et al, 2011; Fang et al, 2020; Rateb et al, 2020) to semen extenders, as well as several modifications to cryoprotectant composition (Galarza et al, 2019; Jerez et al, 2016) or cryoprocedure processes (Ahmad et al, 2015; Šterbenc et al, 2019; Tibary & Manar, 2018). These attempts resulted in various cryopreservation success.…”