Abstract
The aim of our study was to examine the effects of crocin (0.5 (C0.5), 1 (C1) and 1.5 (C1.5) mM) and naringenin (50 (N50), 100 (N100) and 150 (N150) μM) in cryopreservation extender for freezing rooster semen. Sperm motility, viability, abnormalities, membrane functionality, active mitochondria, apoptosis status, lipid peroxidation (LP), GPX, SOD, TAC, the mRNA expression of pro-apoptotic (CASPASE 3) and anti-apoptotic (Bcl-2) genes, fertile eggs, hatched eggs and hatching rate were investigated following freeze-thawing. C1 and N100 resulted in higher (P < 0.05) total motility and progressive motility in comparison to the control group. The C1 and N100 groups improved viability, membrane functionality and reduced lipid peroxidation. We found higher values for active mitochondria with C1 and N100 compared to control group. The C1 and N100 groups showed lower percentages of early apoptosis when compared with control group. Also, C1 and N100 had higher TAC, compared to the control group. The mRNA expressions of BCL-2 in the C1 and N100 groups were significantly higher than that of other treatments. The expression of CASPASES 3 was significantly reduced in C1 and N100 group (P < 0.05) when compared to control group. Significantly higher percentages of fertile eggs, hatched eggs and hatching rate were observed in C1 and N100 compared to the control group. In conclusion, crocin at 1 mM and naringenin at 100 μM seem to improve the post-thawing rooster semen quality, fertility and could protect the sperm by reducing the pro-apoptotic (CASPASE 3) and increasing anti-apoptotic (Bcl-2) genes.
Highlights
Despite its utilization over 70 years ago [1], cryopreservation of poultry sperm leads to low fertility, which limits its application in genetic stock preservation [2]
C1 and N100 resulted in higher (P < 0.05) sperm total motility (TM) (74.43±0.79 and 71.21±1.47, respectively) compared to the control group (61.06±1.64)
The results showed that viability of sperm in C1 and N100 groups were improved compared with control
Summary
Despite its utilization over 70 years ago [1], cryopreservation of poultry sperm leads to low fertility, which limits its application in genetic stock preservation [2]. Effect of crocin and naringenin supplementation in cryopreservation medium on post-thawed sperm quality harmful effect on sperm which decreases sperm viability and motility [3]. Avian sperm are susceptible to oxidative stress [4], though reactive oxygen species (ROS), in physiological quantities, are necessary for important sperm events leading to successful fertilization [5]. Oxidative stress disturbs motility and mitochondrial activity in sperm, induces lipid peroxidation of the membrane [6], and DNA fragmentation [7]. Adding antioxidant compounds to the freezing medium is known as one of the ways to reduce the harmful effects of excessive ROS on sperm fertility after thawing because they block or inhibit oxidative stress. Antioxidants provide positive effects on semen and improve sperm parameters such as motility and membrane integrity [8,9,10]
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