animals Article Effects of Cryoprotective Medium Composition, Dilution Ratio, and Freezing Rates on Spotted Halibut (Verasper variegatus) Sperm Cryopreservation Irfan Zidni 1 , Yun Ho Lee 1, Jung Yeol Park 1, Hyo Bin Lee 1, Jun Wook Hur 2 and Han Kyu Lim 1,* 1 Department of Marine and Fisheries Resources, Mokpo National University, Mokpo 58554, Korea;
[email protected] (I.Z.);
[email protected] (Y.H.L.);
[email protected] (J.Y.P.);
[email protected] (H.B.L.) 2 Faculty of Marine Applied Biosciences, Kunsan National University, 558 Daehak-ro, Gunsan, Jeonbuk 54150, Korea;
[email protected] * Correspondence:
[email protected]; Tel.: +82-61-450-2395; Fax: +82-61-452-8875 Received: 12 October 2020; Accepted: 16 November 2020; Published: 19 November 2020 Simple Summary: The spotted halibut, Verasper variegatus, is a popular fish species occurring naturally in the East China Sea and coastal areas of Korea and Japan. However, when reared in captivity, male and female spotted halibut do not usually mature synchronously. Maintaining production of this commercial fish in hatcheries through sperm cryopreservation is important. This study investigated the effect of several factors for successful cryopreservation of fish sperm including cryoprotective agents (CPAs), diluents, dilution ratios (Milt: CPA + diluents), and freezing rates. The observed factors significantly affected movable sperm ratio (MSR), sperm activity index (SAI), survival rate, and DNA damage after cryopreservation. In the present study, the mixture of 15% dimethyl sulfoxide (DMSO) + 300 mM sucrose with a dilution ratio lower than 1:2 and a freezing rate slower than 5 C/min provided the best treatment and reduced DNA damage.