doi:

DOI: 10.3724/SP.J.1035.2011.00325

Acta Hydrobiologica Sinica (水生生物学报) 2011/35:2 PP.325-332

EFFECT OF CRYOPRESERVATION AND SHORT-TERM STORAGE OF CHINESE GIANT SALAMANDER SPERM


Abstract:
Chinese giant salamander (Andrias davidianus) is a rare and endangered endemic amphibian caudata in China. Chinese giant salamander sperm was cryopreserved in liquid nitrogen or stored at 0—4℃ on a short-term basis. The objective was to establish reliable technologies for the conservation of this endangered species and for artificial assisted reproduction in Chinese giant salamander farms. The studies showed that the spermatozoa remained motility only 3-5h at the room temperature but 6d at 4℃ and 9d at 0℃. Besides, the viability of spermatozoa precipitous declined after adding a dilute solution. An improved cryopreservation method for Chinese giant salamander sperm was established according to the mature technologies for cryopreservation of fish. Cryopreseved sperm was diluted in buffered extenders (pH between 7 and 7.5) containing intracellular (DMSO). The concentration of intracellular cryoprotectants ranged from 5% to 25%. In this paper, ten to fifteen percents of the sperm were motile after thawing. The ultrastructures of Chinese giant salamander sperm before and after cryopreservation were observed by SEM and TEM. Many deformations were found in the cryopreserved sperm, such as swelling or breaking of the plasma membrane, disappearing of the nuclear envelope, fracture of the perforatorium and axoneme, degenerating of the undulating membrane and disappearing of the mitochondrial ridges. The results indicated that these damages might be the main reasons for the decline of the motility in the cryopreserved sperm. This study provided an important reference for the establishment of standardized procedures of the short-term storage and cryopresevation of Chinese giant salamander's sperm as well as the artificial breeding of Chinese giant salamander.

Key words:Chinese giant salamander,Sperm,Cryopreservation,Ultrastructural

ReleaseDate:2014-07-21 15:41:39



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