title: In vitro sperm characterization and development of a sperm cryopreservation method using directional solidification in the killer whale (Orcinus orca).
abstract: Research was conducted to characterize seminal traits and to develop a sperm cryopreservation method using directional freezing (DF) for the killer whale (Orcinus orca). Experiments evaluated effects of: (i) freezing rate (SLOW, MED, FAST) by diluent (BF5F, Biladyl®, EYC) in 0.5 mL straws; and (ii) freezing method (straw or DF) by glycerol (3, 6, or 9% final concentration, v:v) on in vitro sperm quality. Fresh ejaculates (n = 161) were (mean ± SD) 7.8 ± 7.4 mL at 740 × 10(6) sperm/mL with 92.2 ± 6.3% total motility (TM), 85.4 ± 6.9% progressive motility (PM), 89.6 ± 9.0% viability and 89.8 ± 9.2% acrosome integrity. Samples frozen using straws by the MED or SLOW method were improved (P < 0.05) over FAST across all diluents. At 3 h post thaw (PT), TM, PM, Rapid motility (RM), VAP, VCL, ALH and viability for 3% and 6% glycerol were improved (P < 0.05) over 9% glycerol. Directional freezing samples at 0 h and 3 h PT, at all glycerol concentrations, displayed higher (P < 0.001) TM, PM, RM, VAP, VSL, VCL and viability /intact acrosomes (PI/FITC-PNA) than straw. These data provided the first information on ejaculate characteristics and the development of a semen cryopreservation method using DF in the killer whale.
authors.affiliation: Sea World and Busch Gardens Reproductive Research Center, SeaWorld Parks & Entertainment, San Diego, CA 92109, USA. Todd.Robeck@SeaWorld.com
authors.forename: T R
authors.initials: TR
authors.lastname: Robeck
authors.forename: S A
authors.initials: SA
authors.lastname: Gearhart
authors.forename: K J
authors.initials: KJ
authors.lastname: Steinman
authors.forename: E
authors.initials: E
authors.lastname: Katsumata
authors.forename: J D
authors.initials: JD
authors.lastname: Loureiro
authors.forename: J K
authors.initials: JK
authors.lastname: O'Brien
mesh_terms: D000177:Acrosome / Q000502:physiology; D000818:Animals; D002470:Cell Survival; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods / Q000662:veterinary*; D008297:Male; D055101:Semen Analysis / Q000379:methods / Q000662:veterinary*; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility; D050685:Whale, Killer