Kunming Institute of Zoology, Chinese Academy of Sciences
Abstract
Cryopreservation of sperm from Neolissochilus benasi was studied in
2011. The effects of various cryoprotectants of different
concentrations, dilution ratios of milt to extender, storage volume and
thawing temperature on motility of post-thawing of spermatozoa were
examined to optimize cryopreservation procedures. Semen was stored in
liquid nitrogen in 1.8 mL cryovial for 24 h, and the intensity of sperm
motility was measured before and after cryopreservation. Post-thawing
motility of frozen sperm obtained with cryoprotectants 10% MeOH or 15%
EG were higher than for others. The most effective dilution ratio of
milt to extender is 1:7. The maximal storage volume is 60 μL of
1.8 mL cryovial and the optimal sperm equilibration period in the
extender D-15+10% MeOH was between 10-60 min. Thawing was optimal in a
37 °C water bath. When fresh sperm motility is (62.33±2.05)%,
this cryopreservation protocol resulted in frozen-thawed semen with
20%-30% motile. The overall effect is not ideal, and cannot achieve
extensive application. Different breeding management of different
ground protection may have contributed to this result. Therefore, it is
necessary to reduce stress capture induced in management of parent fish
and provide suitable forming conditions. In the ex situ conservation of
rare fish the broodstocks management of males is as important as that
for females and the key to obtaining high quality larval fish