title: Sperm of the giant grouper: cryopreservation, physiological and morphological analysis and application in hybridizations with red-spotted grouper.
abstract: In order to develop excellent germplasm resources for giant grouper (Epinephelus lanceolatus), cryopreservation of giant grouper sperm was examined in the present study. Firstly, 13 kinds of sperm dilution (ELS1-3, EM1-2, TS-2, MPRS, ELRS0-6) were prepared with physiological salt, sucrose, glucose and fetal bovine serum. The physiological parameters of ELRS3 (ratio of fast motion, ratio of slow motion, time of fast motion, time of slow motion, lifespan and motility) and ELS3 (sperm ratio of slow motion, time of slow motion and motility) were significantly higher than those of the other dilutions (P < 0.05). Secondly, after adding 15% DMSO and 10% FBS to ELRS3 and ELS3, most physiological parameters of frozen sperm were also significantly higher than the other gradients (P < 0.05), and sperm motility was as high as 63.68 ± 4.16% to74.75 ± 12.71% (fresh sperm motility, 80.70 ± 1.37% to 80.71 ± 1.49%). Mixed with the above dilutions, a final volume of 105 ml semen was cryopreserved. Finally, the sperm of giant grouper cryopreserved with cryoprotectants (ELRS3 + 15% DMSO + 10% FBS) was used for electron-microscopic observation and crossbreeding with red-spotted groupers (Epinephelus akaara). The electron-microscopic observation revealed that part of the frozen-thawed sperm was cryodamaged, e.g., flagellum fracturing and mitochondria falling out, while the ultrastructure of sperm membrane, mitochondria and flagellum remained intact. Also, the fertilization and hatchability rates of giant grouper frozen sperm and red-spotted grouper eggs were as high as 94.56% and 75.56%, respectively. Thus, a technique for cryopreservation of giant grouper sperm was successfully developed and applied to crossbreeding with red-spotted grouper eggs.
authors.affiliation: Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Tian
authors.forename: Jing
authors.initials: J
authors.lastname: Jiang
authors.forename: Na
authors.initials: N
authors.lastname: Wang
authors.forename: Wenshan
authors.initials: W
authors.lastname: Qi
authors.forename: Jieming
authors.initials: J
authors.lastname: Zhai
authors.forename: Bo
authors.initials: B
authors.lastname: Li
authors.forename: You
authors.initials: Y
authors.lastname: Liang
authors.forename: Youming
authors.initials: Y
authors.lastname: Chen
authors.forename: Chuanjun
authors.initials: C
authors.lastname: Yang
authors.forename: Songlin
authors.initials: S
authors.lastname: Chen
mesh_terms: D000818:Animals; D017756:Aquaculture; D001492:Bass / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D002681:China; D003433:Crosses, Genetic; D015925:Cryopreservation / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000009:adverse effects; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D008297:Male; D008855:Microscopy, Electron, Scanning / Q000662:veterinary; D046529:Microscopy, Electron, Transmission / Q000662:veterinary; D010062:Ovulation Induction / Q000662:veterinary; D012662:Semen Preservation / Q000009:adverse effects / Q000662:veterinary*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D016019:Survival Analysis
chemical_list: D003451:Cryoprotective Agents