title: Vitrification of Sperm from Marine Fishes: Effect on Motility and Membrane Integrity.
abstract: Our goal was to develop a standardized approach for sperm vitrification of marine fishes that can be applied generally in aquatic species. The objectives were to: 1) estimate acute toxicity of cryoprotectants over a range of concentrations; 2) evaluate the properties of vitrification solutions (VS); 3) evaluate different thawing solutions, and 4) evaluate sperm quality after thawing by examination of motility and membrane integrity. Sperm were collected from red snapper (Lutjanus campechanus), spotted seatrout (Cynoscion nebulosus), and red drum (Sciaenops ocellatus). A total of 29 combinations of cryoprotectants were evaluated for toxicity and glass formation. Samples were loaded onto 10-µL polystyrene loops and plunged into liquid nitrogen. There was a significant difference (P< 0.05) in post-thaw motility among VS and among species when using the same VS. The sperm in VS of 15% DMSO + 15% ethylene glycol + 10% glycerol + 1% X-1000™ + 1% Z-1000™ had an average post-thaw motility of 58% and membrane integrity of 19% for spotted seatrout, 38% and 9% for red snapper, and 30% and 19% for red drum. Adaptations by marine fish to high osmotic pressures could explain the survival in the high cryoprotectant concentrations. Vitrification offers an alternative to conventional cryopreservation.
authors.affiliation: Aquaculture Research Station, Louisiana Agricultural Experiment Station, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
authors.forename: Rafael
authors.initials: R
authors.lastname: Cuevas-Uribe
authors.affiliation: Louisiana Universities Marine Consortium, 8124 Highway 56, Chauvin, LA 70344, USA.
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Chesney
authors.affiliation: Aquaculture Research Station, Louisiana Agricultural Experiment Station, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Daly
authors.affiliation: Aquaculture Research Station, Louisiana Agricultural Experiment Station, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
authors.forename: Terrence R
authors.initials: TR
authors.lastname: Tiersch
keywords: cryopreservation; red drum; red snapper; sperm vitrification; spotted seatrout