The Effect of using Different Levels of Luteolin on Reducing Malondialdehyde Content and Increasing the Motility and Viability of Ross 308 Rooster Sperm during Cryopreservation and Thawing Process

Oxidative stress is an important factor in poor sperm function that causes morphological changes and oxidative damage. Various studies have shown that antioxidants can reduce oxidative damage during freezing. In fact, antioxidants reduce, eliminate, and inactivate free radicals. Many compounds, such as some plant extracts, some industrial chemicals, and some amino acids, can act as active antioxidants and neutralize free radicals. luteolin is an important member of flavonoids which is present in glycosylated form in plants and studies have shown that luteolin has antioxidant properties. The aim of this study was to reduce lipid peroxidation and minimize oxidative stress by adding different levels of luteolin in the freezing medium.

Semen samples were collected from 15 roosters at 28 weeks of age. Sperm collection from roosters was performed by back-abdominal massage method. To eliminate individual effects after initial evaluations, the samples were mixed together and used for cryopreservation. After extending the samples and adding the levels of 0 (control), 0.25, 0.5, 0.75, 0.1 and 1.25 μM luteolin to the base extender (lake extender), sperm were added at 1:20 ratio. The test samples were cooled to 4 ° C and then frozen in liquid nitrogen. The samples were thawed at 37 ° C and then the motility parameters were measured by CASA computer system, survival by eosin nigrosine test, plasma membrane integrity by host test, morphology by Hancock test and the amount of produced malondialdehyde. Then the data were analyzed by completely randomized design.

The results showed that the addition of 0.75, 1 and 1.25 μM levels significantly increased the total motility and the levels of 0.75 and 1 increased the progressive mobility and VAP. In addition, 1 μM of luteolin caused a significant increase in VSL parameter (p <0.05). The results showed that levels of 0.75 and 1 μM significantly increased membrane integrity, and also level of 0.75 μM of luteolin decreased malondialdehyde content and sperm percentage with abnormal morphology (p <0.05).

The results of this study showed that the addition of luteolin in the extender during freezing and thawing can have a positive effect on improving the quality of rooster sperm and subsequently increase the total and progressive motility after freezing and thawing and also the level of 0.75 μM could be the best performance among different levels added to the extender.