Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. s...Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. sauliae, which is a potential preparation for its artificial reproduction and further research, in this study a protocoi for the cryopreservation of veliger larvae of trumpet shell was optimized. Through a two-step cryopreservation procedure, four kinds of cryoprotectants （ethylene glycol, 1, 2-propanediol, dimethyl sulfoxide and glycerol） were employed at three concentrations （1.0, 1.5 and 2.0molL^-1） respectively and survival rates of larvae were determined after a storage of lh. The larvae frozen with these four cryoprotectants after 1 h storage were cultured, and then survival rates were determined at 24, 72 and 120 h after thawing. Dimethyl sulfoxide at a concentration of 1.5 molL^-1 showed the best protective effect in all experiments （p〈0.05）. And survival rates of larvae frozen with dimethyl sulfoxide were determined after 1, 7 and 15 d of storage. The survival rates of larvae frozen with 1.5 molL^-1 dimethyl sulfoxide after 1 h, 1 d, 7 d and 15 d of storage were 80.77% ±7.51%, 80.34%±11.28%, 83.10%±9.14% and 77.23%±6.22% respectively. No significant differences in survival rates of larvae frozen with dimethyl sulfoxide were observed after various storage periods （p〉0.05）.展开更多
Although cryopreservation of embryos has been used in most terrestrial animals, the application of this technique has not been succeeded for aquatic animals. In this study, the authors investigate the effect of differ...Although cryopreservation of embryos has been used in most terrestrial animals, the application of this technique has not been succeeded for aquatic animals. In this study, the authors investigate the effect of different combinations of sucrose （SUC, C12H22OH） and cryoprotectants （CPAs） on the survival of the catfish embryos （Pangasidae hypophthalmus） at low temperatures （4, 0 and -20 ℃） for short-term storage. For this aim, embryos with somites and optic cups were exposed to different combinations of sucrose with methanol （SUC ＋ MeOH）, 1.2-propylene glycol （SUC ＋ PROH） and ethylene glycol （SUC ＋ EG） at four concentrations ratios： （1） 0.5 M SUC ＋ 0.5 M CPA; （2） 1 M SUC ＋ 0.5 M CPA; （3） 0.5 M SUC ＋ 1 M CPA; （4） 1 M SUC ＋ 1 M CPA for 40 min at 4, 0 and -20 ℃. Embryos kept in water at room temperature （RT）, 4, 0 and -20℃ were used as controls. The survival rate was expressed as a percentage of hatched embryos per total embryos treated. The results showed that the hatching rate declined significantly when embryos were stored in water at 0 ℃ and -20℃. For embryos at 0 ℃ storage, the highest survival rate （87.78%） was obtained with 1 M SUC ＋ 1 M MeOH combination while at -20 ℃, only embryos in the combined treatments of 0.5 M SUC ＋ 1 MEG and 0.5 M SUC ＋ 1 M PROH reached the hatching stage （40% and 83.33%, respectively）. In conclusion, the results showed that the catfish embryos are sensitive to sub-zero temperatures and the combined treatment of 0.5 M sucrose and 1 M propylene glycol can be used to protect catfish embryos from damages caused by low temperature （0 ℃ and -20 ℃）.展开更多
This study was to research the cryopreservation effect of plant antifieeze proteins(AFPs) on day 7 porcine expanded and hatched blastocysts that were frozen in 1.5 M glycerol by conventional slow freezing method. The ...This study was to research the cryopreservation effect of plant antifieeze proteins(AFPs) on day 7 porcine expanded and hatched blastocysts that were frozen in 1.5 M glycerol by conventional slow freezing method. The developmental rates of porcine embryos frozen with 0.5 mg/ml AFPs in freeze medium and cultured for 12 and 24 hours in vitro are 25% and 0%respectively. With the concentration of AFPs increased to 5 mg/ml, the corresponding values became 80% and 40%. The hatched rates for porcine embryos frozen with 0.5 mg/ml and 5 mg/ml of AFPs and cultured for 24 hours in vitro are 0% and 20% respectively..The developmental and hatched rate of the contnrol are all 0%(0/4). These results indicate that the survival rates of porcine expanded and hatched blastocyst can be improved by supplementing freeze medium with plant AFPs.展开更多
文摘Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. sauliae, which is a potential preparation for its artificial reproduction and further research, in this study a protocoi for the cryopreservation of veliger larvae of trumpet shell was optimized. Through a two-step cryopreservation procedure, four kinds of cryoprotectants （ethylene glycol, 1, 2-propanediol, dimethyl sulfoxide and glycerol） were employed at three concentrations （1.0, 1.5 and 2.0molL^-1） respectively and survival rates of larvae were determined after a storage of lh. The larvae frozen with these four cryoprotectants after 1 h storage were cultured, and then survival rates were determined at 24, 72 and 120 h after thawing. Dimethyl sulfoxide at a concentration of 1.5 molL^-1 showed the best protective effect in all experiments （p〈0.05）. And survival rates of larvae frozen with dimethyl sulfoxide were determined after 1, 7 and 15 d of storage. The survival rates of larvae frozen with 1.5 molL^-1 dimethyl sulfoxide after 1 h, 1 d, 7 d and 15 d of storage were 80.77% ±7.51%, 80.34%±11.28%, 83.10%±9.14% and 77.23%±6.22% respectively. No significant differences in survival rates of larvae frozen with dimethyl sulfoxide were observed after various storage periods （p〉0.05）.
文摘Although cryopreservation of embryos has been used in most terrestrial animals, the application of this technique has not been succeeded for aquatic animals. In this study, the authors investigate the effect of different combinations of sucrose （SUC, C12H22OH） and cryoprotectants （CPAs） on the survival of the catfish embryos （Pangasidae hypophthalmus） at low temperatures （4, 0 and -20 ℃） for short-term storage. For this aim, embryos with somites and optic cups were exposed to different combinations of sucrose with methanol （SUC ＋ MeOH）, 1.2-propylene glycol （SUC ＋ PROH） and ethylene glycol （SUC ＋ EG） at four concentrations ratios： （1） 0.5 M SUC ＋ 0.5 M CPA; （2） 1 M SUC ＋ 0.5 M CPA; （3） 0.5 M SUC ＋ 1 M CPA; （4） 1 M SUC ＋ 1 M CPA for 40 min at 4, 0 and -20 ℃. Embryos kept in water at room temperature （RT）, 4, 0 and -20℃ were used as controls. The survival rate was expressed as a percentage of hatched embryos per total embryos treated. The results showed that the hatching rate declined significantly when embryos were stored in water at 0 ℃ and -20℃. For embryos at 0 ℃ storage, the highest survival rate （87.78%） was obtained with 1 M SUC ＋ 1 M MeOH combination while at -20 ℃, only embryos in the combined treatments of 0.5 M SUC ＋ 1 MEG and 0.5 M SUC ＋ 1 M PROH reached the hatching stage （40% and 83.33%, respectively）. In conclusion, the results showed that the catfish embryos are sensitive to sub-zero temperatures and the combined treatment of 0.5 M sucrose and 1 M propylene glycol can be used to protect catfish embryos from damages caused by low temperature （0 ℃ and -20 ℃）.
文摘This study was to research the cryopreservation effect of plant antifieeze proteins(AFPs) on day 7 porcine expanded and hatched blastocysts that were frozen in 1.5 M glycerol by conventional slow freezing method. The developmental rates of porcine embryos frozen with 0.5 mg/ml AFPs in freeze medium and cultured for 12 and 24 hours in vitro are 25% and 0%respectively. With the concentration of AFPs increased to 5 mg/ml, the corresponding values became 80% and 40%. The hatched rates for porcine embryos frozen with 0.5 mg/ml and 5 mg/ml of AFPs and cultured for 24 hours in vitro are 0% and 20% respectively..The developmental and hatched rate of the contnrol are all 0%(0/4). These results indicate that the survival rates of porcine expanded and hatched blastocyst can be improved by supplementing freeze medium with plant AFPs.
