title: Cryopreservation of sperm from seven-band grouper, Epinephelus septemfasciatus.
abstract: In the present study, we examined methods for the cryopreservation of Epinephelus septemfasciatus spermatozoa. The percent motility, average path velocity, and linearity of movement (LIN) of fresh and corresponding post-thaw sperm were evaluated. Sperm motility was investigated using computer-assisted sperm analysis. Five percent dimethyl sulphoxide (Me₂SO) with 95% fetal bovine serum (FBS) was the most successful cryoprotectant diluent with a comparative post-thaw motility of 77.6±8.5%; 5% dimethyl formamide was also effective. Fetal bovine serum was significantly better as an extender when compared with artificial seminal plasma, glucose, and trehalose solution. Sperm tolerated a wide range of cooling rates (from 27.1 to 94.3 °C min⁻¹); however, the post-thaw motility of sperm cooled to -30 °C was significantly lower than that of other cooled temperatures (-40 to -70 °C). The velocity of post-thaw sperm was significantly lower than that of fresh sperm, although LIN remained the same. For effective cryopreservation of seven-band grouper sperm, samples should be diluted in 5% Me₂SO with 95% FBS and cooled to at least -40 °C before immersion in liquid nitrogen.
authors.affiliation: Department of Fisheries, Graduate School of Agriculture, Kinki University, Naka-Machi, Nara 631-8505, Japan.
authors.forename: Ivan Chong Chu
authors.initials: IC
authors.lastname: Koh
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Yokoi
authors.forename: Masaharu
authors.initials: M
authors.lastname: Tsuji
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Tsuchihashi
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ohta
mesh_terms: D000818:Animals; D017756:Aquaculture / Q000379:methods; D001492:Bass; D002417:Cattle; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004126:Dimethylformamide / Q000494:pharmacology; D005312:Fetal Blood / Q000502:physiology; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility; D013094:Spermatozoa / Q000187:drug effects
chemical_list: D004126:Dimethylformamide; D004121:Dimethyl Sulfoxide