The Effect of Canthaxanthin on the Quality of Frozen Ram Spermatozoa

Abstract

The addition of antioxidants to semen cryopreservation extenders has been employed for combating oxidative damage. This work aimed to evaluate the addition of carotenoid canthaxanthin to a cryopreservation extender of ram semen. Three breeder rams were used and, after semen collection, with 48-hour intervals between collection, the samples were included in the pool formation (n = 6). The experimental groups comprised 0 (control), 0.1, 1, 10, and 25 μM of canthaxanthin. After thawing (37°C/30 s) and incubation at 37°C for 2 hours, semen aliquots from each group were evaluated for sperm kinetics (CASA), the integrity of the plasma and acrosomal membranes (iPAM), intracellular reactive oxygen species (ROS) production, and lipid peroxidation (LPO) by flow cytometry associated with the image. The control group and canthaxanthin 1 μM after incubation at 37°C for 2 hours showed increases of curvilinear velocity and amplitude of lateral head displacement with decreases of linearity, straightness, and wobble (p < 0.05), which were not observed for the canthaxanthin 10 and 25 μM. The supplementation of a Tris-egg yolk extender with canthaxanthin had no effect on the iPAM, intracellular ROS production in viable spermatozoa, or LPO. In conclusion, supplementation with 10 and 25 μM of canthaxanthin in a Tris-egg yolk extender used for ram semen cryopreservation is able to protect ovine sperm from kinetic changes after incubation at 37°C for 2 hours post-thawing.

Bacterioruberin extracts from a genetically modified hyperpigmented Haloferax volcanii strain: antioxidant activity and bioactive properties on sperm cells

“…Our results are consistent with those reported by Souza et al . () which showed a clear positive effect on ram sperm motility after 2 h post‐thawing with similar concentrations of canthaxanthin supplemented to the freezing medium. Similar to our study, Sapanidou et al .…”

“…; Souza et al . ). Even under conditions of induced oxidative stress where the effect of a ROS scavenger should be more evident, the reports in sperm cells are not consistent among different authors who did (Domínguez‐Rebolledo et al .…”

“…; Souza et al . ). Controls were performed with the corresponding amount of ethanol (0·125% (v/v), 0·35% (v/v), 0·75% (v/v) or 1% (v/v) in PBS respectively) or the maximum concentration of DMSO (1% (v/v) in PBS).…”

“…Thawing was carried out by dropping the semen straws in a water bath at 37°C for 30 s. A motile sperm population was selected by a glass wool filtration and diluted to 10 9 10 6 cell per ml in phosphate-buffered saline, pH 7Á4 (PBS) to avoid extender and seminal plasma effect. Sperm suspension (in PBS medium) was incubated with different Bctr extract or bcar concentrations (2Á5, 7, 15 and 20 lmol l À1 ) for at least 120 min (Fernandez-Santos et al 2007;Sapanidou et al 2015;Souza et al 2017). Controls were performed with the corresponding amount of ethanol (0Á125% (v/v), 0Á35%…”

“…Vitamin A and retinol promoted the production of motile sperm during in vitro maturation (Arkoun et al 2015;Dumont et al 2016). On the other hand, the addition of carotenoids (lycopene, canthaxanthin, lutein) to freezing extenders exhibited significant ROS-trapping and antioxidant properties preventing oxidative damage to frozen-thawed bovine (Tuncer et al 2014;Tvrda et al 2017), ram (Souza et al 2017), boar (Varo-Ghiuru et al 2015 and fish sperm (Liu et al 2015) among others. Crocin applied to semen after thawing was an effective ROS scavenger and improved motility in bovine (Sapanidou et al 2015) and red deer sperm (Dom ınguez-Rebolledo et al 2010).…”