Effect of Different Levels of 2, 4 Dinitrophenol and Luteolin on Semen Quality of Rooster during Freezing-Thawing Process

During sperm freezing, the production of reactive oxygen species and the reduction of antioxidant activity cause damage to sperm. Most of the stress is due to the high production of reactive oxygen species and the impermeability of antioxidants to mitochondria. In this study, the effects of four levels of a combination of 2, 4- dinitrophenol which is a targeted mitochondrial antioxidant and Luteolin antioxidants, which are among the strongest flavonoid compounds with antioxidant and antimicrobial properties, were evaluated in lecithin-based Lake Extender.

For this purpose, semen samples from 15 roosters were collected by dorso-abdominal massage method. Semen samples are mixed together after initial evaluation after diluting and adding different levels, treatment 1 (0.5 nanomolar 2, 4- dinitrophenol + 0.75 micromolar Luteolin), treatment 2 (0.5 nanomolar 2, 4- dinitrophenol + 1 micromolar Luteolin), treatment 3 (0.75 nanomolar 2, 4- dinitrophenol + 0.75 micromolar Luteolin) and treatment 4 (0.75 nanomolar 2, 4- dinitrophenol + 1 micromolar Luteolin), of antioxidants, samples were frozen by nitrogen vapor. After thawing, the samples were evaluated for sperm performance parameters.

The results of the present study showed that the addition of treatments 2 and 3 to the leak diluent during freezing and thawing of rooster sperm significantly increased the parameters of total motility and progressive movement and also treatment 3 significantly increased sperm velocity in a straight line and the criterion of direct sperm motility. Also, treatments 2 and 3 significantly increased survival compared to the control group and reduced the amount of semen malondialdehyde and sperm with abnormal morphology during freezing and thawing (p<0.05). Also, treatment 3 was able to increase membrane integrity. Plasma increase compared to the control group (p<0.05).

The results of the present study show that the combined use of these two antioxidants has increased the properties of both antioxidants and improved the quality and quantity of sperm.