title: Cryopreservation of sperm from Atlantic halibut (Hippoglossus hippoglossus, L.) for commercial application.
abstract: Development of Atlantic halibut (Hippoglossus hippoglossus) aquaculture will be enhanced with cryopreservation of halibut sperm by ensuring a reliable supply of sperm of desired quality and quantity. To assist in its commercial application, the cryopreservation of large volumes of halibut sperm was investigated. Three cryoprotectants were compared: dimethyl sulfoxide (DMSO), polyethylene glycol (PG) and glycerol (GLY) at two concentrations (10% or 15%). Two salt solutions, Hanks' balanced salt solution (HBSS) and 0.1M KHCO(3) with 0.125 M sucrose solution (KS) were tested as diluents. Both factors were examined in 1.6 mL volumes. A cryopreservation volume of 4 mL and a low dilution ratio (1:1) were examined separately. Based on motility and fertilization rate, 10% and 15% DMSO diluted with HBSS or KS solution proved to be effective extenders with mean fertilization rates ranging from 52.2 ± 27.2% to 65.8 ± 26.1%; none of which were significantly different from that of the control. Four other extenders, 10% PG or 10% GLY with HBSS or KS, resulted in significantly lower fertilization rates. Use of a 4 mL cryopreservation volume did not exhibit a significant effect on fertilization rate or motility of post-thawed sperm compared to a 1.6 mL volume (P>0.05); while the use of a dilution ratio of one part sperm with three parts cryopreservation solution (1:3 v/v with sperm concentration of 0.51± 0.11×10(10)cells/ml) had a significantly better preservation effect than using a ratio of 1:1 with sperm concentration of 1.02 ± 0.21×10(10)cells/ml (P<0.05). From these results, an optimized protocol for the cryopreservation of Atlantic halibut sperm using a volume as large as 4mL has been established.
authors.affiliation: Yellowsea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, China. dingfuhong@hotmail.com
authors.forename: Fuhong
authors.initials: F
authors.lastname: Ding
authors.forename: Santosh P
authors.initials: SP
authors.lastname: Lall
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Jilin
authors.initials: J
authors.lastname: Lei
authors.forename: Melissa
authors.initials: M
authors.lastname: Rommens
authors.forename: Joyce E
authors.initials: JE
authors.lastname: Milley
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D005432:Flounder; D008297:Male; D012662:Semen Preservation / Q000379:methods; D013094:Spermatozoa
chemical_list: D003451:Cryoprotective Agents