Cryopreservation of paddlefish Polyodon spathula milt

Abstract

A practical procedure for cryopreserving milt of paddlefish Polyodon spathula was developed to obtain thawed spermatozoa that would fertilize eggs and permit hatching of normal larvae. Milt was mixed with a cryoprotectant medium containing DMSO (2.4 M) in a ratio of 3:1(milt:medium; final concentration of DMSO 0.6 M). stored in 5.0-mL freezing straws, and frozen in dry ice (15 min) and then in liquid nitrogen. A total of three replicates were made; the milt of a different male was used in each replicate. Motility of the thawed spermatozoa decreased to 50%-25% as compared to 100% motility of the fresh (control) spermatozoa. Hatching of paddlefish (16.3 ± 2.2%) from eggs fertilized with thawed spermatozoa was significantly lower (P ≤ 0.01) than the hatch rate (90.8 ± 2.5%) for the control. It was suggested that an increase in viable motile spermatozoa to egg would result in better fertilization and hatching of paddlefish.