温度突变和非离子氨胁迫对淡水养殖凡纳滨对虾cyt-C和caspase-3的影响  

Effects of abrupt change in temperature and non-ionic ammonia on the cyt-C and caspase-3 of Litopenaeus vannamei in freshwater

作　　者：贾旭颖[1,2] 张丹[1] 路允良[1] 王芳[1] 董双林[1] 

机构地区：[1]中国海洋大学教育部海水养殖重点实验室,山东青岛266003 [2]广西海洋研究所广西海洋生物技术重点实验室,广西北海536000

出　　处：《中国水产科学》2014年第4期700-710,共11页Journal of Fishery Sciences of China

基　　金：国家重点基础研究发展规划项目(2009CB118706);广西海洋生物技术重点实验室开放课题(GLMBT-201301)

摘　　要：采用酶联免疫吸附(ELISA)法,研究了温度突变(16℃←22℃→28℃)和非离子氨胁迫(0.1 mg·L–1←0→0.5 mg·L–1)后,淡水养殖凡纳滨对虾(Litopenaeus vannamei)细胞色素C(cyt-C)含量和天冬氨酸–半胱氨酸蛋白酶-3(caspase-3)活性的变化规律,并与海水养殖条件下的相关指标进行了比较。结果显示:(1)温度突变5 d,淡水和海水养殖凡纳滨对虾血淋巴和肝胰腺cyt-C含量均显著高于突变前的水平(P<0.05)。(2)温度突变后,淡水养殖对虾血淋巴caspase-3活性显著升高(P<0.05),而海水养殖对虾血淋巴caspase-3活性在低温突变后无显著变化(P>0.05),在高温突变后显著升高(P<0.05);淡水养殖对虾肝胰腺caspase-3活性在低温突变后显著升高(P<0.05),在高温突变后呈无规则波动,海水养殖对虾肝胰腺caspase-3活性在温度突变后无显著变化(P>0.05)。(3)淡水和海水养殖对虾血淋巴和肝胰腺cyt-C在0.1 mg·L–1非离子氨胁迫后显著升高(P<0.05),而在0.5 mg·L–1非离子氨胁迫后则呈现先升高后下降的趋势。(4)0.1 mg·L–1非离子氨胁迫后,淡水和海水养殖对虾血淋巴和肝胰腺caspase-3活性均显著升高(P<0.05);0.5 mg·L–1非离子氨胁迫后,淡水和海水养殖对虾血淋巴caspase-3活性先升高后显著降低,而肝胰腺caspase-3活性则持续升高。实验结果表明,温度和非离子氨胁迫对淡水养殖凡纳滨对虾cyt-C和caspase-3均有显著影响;与海水养殖条件相比,淡水养殖凡纳滨对虾cyt-C和caspase-3受低温突变的影响更加显著,而两种养殖条件对虾对非离子氨胁迫的响应规律基本相似。To investigate the effect of temperature variation and non-ionic ammonia stresses on the apoptosis of Lito-penaeus vannamei cultured in freshwater, changes of cyt-C content and caspase-3 activity were determined by en-zyme-linked immunosorbent assay （ELISA） methods after acute temperature variation （16℃←22℃→28℃） and non-ionic ammonia （NH3–N） stress （0.1 mg·L–1←0→0.5 mg·L–1）, and compared with those of seawater shrimps. It was found that：（1） Thermal stresses significantly increased the cyt-C content in heamolymph and hepatopancreas of both freshwater and seawater shrimps within 5 d （P〈0.05）;three-way ANOVA analysis results showed that culture condition, thermal stress and exposure time all had a significant impact on cyt-C content in shrimps （P〈0.05） although their inter-action was not significant （P〉0.05）. （2） In freshwater shrimps, the caspase-3 activity in heamolymph and hepatopan-creas changed significantly after thermal stresses, and increased significantly at 5 d compared to that before experiment （0 d） （P〈0.05）, except the heamolymph after hyperthermal stress, which was different to those in seawater shrimps;culture condition, thermal stress and exposure time all had a significant impact on caspase-3 activity of shrimps and the interaction among them was significant （P〈0.05）. （3） Under the stress of 0.1 mg·L–1 concentration of NH3–N, the cyt-C content of heamolymph and hepatopancreas increased significantly in both freshwater and seawater groups （P〈0.05）, while at 0.5 mg·L–1 concentration, cyt-C content in freshwater and seawater shrimps showed a similar change trend：cyt-C content increased significantly within 2–3 d and then decreased to initial levels;NH3–N stress and exposure time significantly affected cyt-C content but only cyt-C content in hepatopancreas was significantly affected by culture con-dition and interaction among them was significant（P〈0.05）. （4） Change trend of caspase-3 acti

关 键 词：凡纳滨对虾 盐度 温度突变 非离子氨 CYT-C CASPASE-3 

分 类 号：S96[农业科学—水产养殖]