Sperm of the giant grouper: cryopreservation, physiological and morphological analysis and application in hybridizations with red-spotted grouper

Yongsheng TIAN; Jing JIANG; Na WANG; Wenshan QI; Jieming ZHAI; Bo LI; You LIANG; Youming CHEN; Chuanjun YANG; Songlin CHEN

doi:10.1262/jrd.2014-087

Original Articles

Sperm of the giant grouper: cryopreservation, physiological and morphological analysis and application in hybridizations with red-spotted grouper

Yongsheng TIAN, Jing JIANG, Na WANG, Wenshan QI, Jieming ZHAI, Bo LI, You LIANG, Youming CHEN, Chuanjun YANG, Songlin CHEN

Author information

Yongsheng TIAN
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
Jing JIANG
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
Na WANG
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
Wenshan QI
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
Jieming ZHAI
Ming Bo Aquatic Co. Ltd. Laizhou 261400, China
Bo LI
Ming Bo Aquatic Co. Ltd. Laizhou 261400, China
You LIANG
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
Youming CHEN
Hainan Blue Ocean Marine Science and Technology Breeding Company, Hainan Sanya 572426, China
Chuanjun YANG
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
Songlin CHEN
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China

Keywords: Cryopreservation, Crossbreeding, Epinephelus lanceolatus, Physiological characteristics, Sperm

JOURNAL FREE ACCESS

2015 Volume 61 Issue 4 Pages 333-339

DOI https://doi.org/10.1262/jrd.2014-087

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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.

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