论文部分内容阅读
光唇鱼(Acrossocheilus fasciatus)是我国溪流性特色经济鱼类之一,因生境变化及捕捞过度等原因,导致种质资源衰退,因此需要开展种质资源保护及种质细胞保存研究,精子冷冻保存是种质细胞保存的有效方式之一。本研究,从人工养殖的性成熟雄性光唇鱼采集精液,以0.25 m L麦细管为冻存管,开展稀释液及其稀释比例、抗冻剂种类及其浓度、平衡时间、两步降温法降温高度以及解冻温度等对该种鱼精子超低温冻存效果影响的研究,并通过酶活性检测对冻精质量进行初步评价。结果表明,在稀释液为D-15液、稀释比为1∶5、抗冻剂为10%二甲基亚砜(dimethyl sulfoxide,DMSO)、平衡时间为30 min,距离液氮面3.5 cm处降温5 min后投入液氮中保存以及37℃解冻的精子效果较佳,冻精解冻后激活率达(35.33±2.52)%,但与鲜精激活率(87.67±3.06)%相比仍差异显著(P<0.05);鲜精总ATP酶、琥珀酸脱氢酶(succinate dehydrogenase,SDH)及乳酸脱氢酶(lactate dehydrogenase,LDH)活性分别为(9.31±0.17)U/m L、(30.33±5.69)U/m L和(7 454.84±252.42)U/L,冻精总ATP酶、SDH和LDH活性分别为(7.23±1.08)U/m L、(17.67±6.03)U/m L和(2 172.48±209.62)U/L,两者差异亦显著(P<0.05)。本研究取得了光唇鱼精子冷冻保存实验的初步成功,为光唇鱼精子冷冻保存库建立提供了一定的技术基础,但有关技术参数还有待于进一步探索优化,以提高冻精质量。
Acrossocheilus fasciatus is one of the characteristic stream fish in China. Due to habitat changes and overfishing, Acrossocheilus fasciatus causes the decline of germplasm resources. Therefore, it is necessary to carry out germplasm conservation and germplasm conservation research and sperm cryopreservation It is one of the effective ways to preserve germplasm cells. In this study, sperm were collected from artificially cultured, male, mature malar lip fish, and 0.25 m L of straw was stored as a cryogenic tube. Dilutions and dilutions were carried out. The type and concentration of antifreeze, balance time, Method of cooling height and thawing temperature on the cryopreservation of fish sperm cryogenic effects of the study, and through the enzyme activity test for the initial evaluation of the quality of sperm. The results showed that when the dilution was D-15 solution, the dilution ratio was 1: 5, the antifreeze agent was 10% dimethyl sulfoxide (DMSO), the equilibrium time was 30 min and the liquid nitrogen surface was 3.5 cm After cooling for 5 min, the sperm stored in liquid nitrogen and thawed at 37 ℃ had a better effect. After thawing, the activation rate of frozen sperm was (35.33 ± 2.52)%, but still significantly different from that of fresh sperm activation (87.67 ± 3.06)% (P <0.05). The activities of total ATPase, succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) in fresh semen were (9.31 ± 0.17) U / m L, (7.29 ± 1.08) U / m L, (17.67 ± 6.03) U / m L and ((7454.84 ± 252.42) U / 2 172.48 ± 209.62) U / L, the difference was also significant (P <0.05). In this study, the preliminary successes of cryopreservation of spermatophytes were obtained, which provided some technical basis for the establishment of sperm cryopreservation of lip lip fish. However, the technical parameters need to be further explored and optimized to improve the quality of frozen sperm.