Novel droplet vitrification combined with fish antifreeze protein type III enhances cryoprotection of semen in wild endangered Persian sturgeon Acipenser persicus (Borodin, 1897)

作者：Abed Elmdoust Amirreza; Farahmand Hamid^*; Mojazi Amiri Bagher; Rafiee Gholamreza; Rahimi Ruhollah

来源：Aquaculture Research, 2015, 46(10): 2392-2397.

DOI：10.1111/are.12397

• Droplet vitrification
• Wild Persion sturgeon semen
• Fish antifreeze protein type III

In this study, the efficiency of a novel droplet vitrification technique along with different doses of fish antifreeze protein (AFP) type III on Persian sturgeon thawed spermatozoa quality (motility duration and motility percentage) was investigated. Semen of seven male individuals was pooled in equal volumes and diluted with 4 degrees C Tris-Hcl (100 mM), pH=8 extenders containing 0, 5, 10, 15 mu M of AFP type III in a ratio of 1: 1 (semen/extenders). Treated semen was dropped into liquid nitrogen. Solidified droplets were stored for 2, 60 and 120 days and thawed by plunging them into a tube containing 5 mL Tris-Hcl (100 mM), pH=8 with 1% BSA at 37 degrees C. Motility duration in all treatments had no significant difference comparing to fresh sperm (P>0.05), but their motility percentage was significantly lower. Treatment with 10 mu M of AFP had significantly higher motility percentage (16.11 +/- 0.5%) comparing to other treatments (P<0.05). There was no significant difference between 0, 5, 15 mu M of antifreeze protein treatments (P>0.05), suggesting that antifreeze protein effectiveness are highly dose dependent, and dose of 10 mu M is appropriate in Persian sturgeon spermatozoa droplet vitrification. Besides, the present technique obtained higher quality of spermatozoa comparing to its analogue techniques.

• 出版日期2015-10