Abstract
ABSTRACT The aim of this study was to evaluate the addition of vitamin C, reduced glutathione and the association thereof to the bovine semen cryopreservation extender. The ejaculate from nine bulls were divided into four fractions, each corresponding to a treatment, namely: control group-semen diluted with Tris-yolk extender; vitamin C group-semen diluted in Tris-yolk extender supplemented with vitamin C (2.5mmol/mL); glutathione group-semen diluted in Tris-yolk extender supplemented with reduced glutathione (2.5mmol/mL) and associated group-semen diluted in Tris-yolk extender supplemented with vitamin C (1.25mmol/mL) and reduced glutathione (1.25mmol/mL). Afterwards, the semen was packed into French straws and submitted to cryopreservation using automated equipment. After cryopreservation, the semen was thawed and evaluated considering sperm motility, morphology, plasma membrane, acrosome, mitochondrial potential and oxidative stress, as well as the thermo resistance test. Extender’s supplementation with the association of vitamin C and reduced glutathione showed benefic effects on sperm motility and preservation of plasma and acrosomal membranes during semen cryopreservation, being also the group that showed higher values of reactive oxygen species. Thus, the association of both antioxidants contributed to the preservation of sperm cells in every analyzed characteristic, suggesting its use on bovine semen cryopreservation.
Highlights
The use of cryopreserved semen allows the choice of best breeders that meet farm needs, being indispensable in artificial insemination, and in vivo and in vitro embryo transfer programs (Leite et al, 2011)
When sperm motility was assessed after the resistance test (RTT) test, within each post thawing time, there was no statistical difference between treatments in relation to the CON group (P> 0.05). (Table 1)
Reduced glutathione and vitamin C, when single added to the extender, did not affect the progressive motility after cryopreservation, contrasting with what was found by Zhao et al (2015) in bovine semen supplemented with vitamin C, as well as Ansari et al (2012) in buffalo semen, Oliveira et al (2013) in equine semen and Ogata et al (2015) in canine semen supplemented with reduced glutathione
Summary
The use of cryopreserved semen allows the choice of best breeders that meet farm needs, being indispensable in artificial insemination, and in vivo and in vitro embryo transfer programs (Leite et al, 2011). Semen cryopreservation is a process that causes great stress to the spermatozoa, which can occur during cooling, freezing and thawing, diminishing sperm quality compared to fresh semen (Maia and Bicudo, 2009). Oxidative stress occurs when there is an imbalance between the production of reactive oxygen species (ROS) and the protective action of the antioxidant system, responsible for its neutralization and removal (Jedrzejowska et al, 2013). All radicals and non-radicals from oxygen are considered ROS, which has high electron reactivity and instability. The ROS can react with a great number of compounds, acting as donors or receivers of electron (Agarwal et al, 2005), and they are considered the main prompters of damage to living organisms (Bernard and Krause, 2007). Main ROS includes oxygen ions, free radicals and peroxide, and are naturally formed as sub-products of oxygen metabolism and play an important role in cellular signalization (Paparella et al, 2015)
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