中国科学院海洋研究所机构知识库

        近年来，有害藻华在我国近岸海域频繁暴发，对海洋浮游动物以及生态系统产生了不利影响。本研究以日本虎斑猛水蚤（Tigriopus japonicus）为实验生物，选用成体存活率、无节幼体至桡足幼体（N-C）存活率、无节幼体至成体（N-A）存活率、无节幼体至桡足幼体（N-C）发育时间、桡足幼体至成体（C-A）发育时间、雌体12 d内产卵次数、雌体12 d内产卵量以及毒素积累为指标，以青岛大扁藻（Tetraselmis helgolandica var. tsingtaoensis）为对照藻，评价了两株米氏凯伦藻（Karenia mikimotoi）（福建苏澳株和莆田株）、东海原甲藻（Prorocentrum donghaiense）、抑食金球藻（Aureococus anophagefferens）和链状亚历山大藻（Alexandrium catenella）四种典型有毒有害藻的生物毒性。研究结果如下：

         两株米氏凯伦藻均显著影响日本虎斑猛水蚤无节幼体的存活、发育和雌体的繁殖。实验藻为米氏凯伦藻（莆田株）时，在较低藻细胞密度（1.0×10³ cells/mL）时，实验第5 d无节幼体存活率为46.7%，第25 d低于20%；在较高藻细胞密度（3.0×10⁴ cells/mL）时，实验第5 d的存活率即为0，结果表明米氏凯伦藻（莆田株）能显著影响日本虎斑猛水蚤无节幼体的存活，并严重抑制发育和繁殖。与莆田株相比，苏澳株对日本虎斑猛水蚤的生物毒性相对较低。实验藻为米氏凯伦藻（苏澳株）时，在较高藻细胞密度（3×10⁴ cells/mL）时，N-C存活率、N-A存活率、雌体12 d内产卵次数和产卵量均低于对照组，存在潜在的不利影响，虽然无显著差异，N-C发育时间和C-A发育时间显著长于对照组，表明米氏凯伦藻（苏澳株）能够影响日本虎斑猛水蚤无节幼体的存活、发育和雌体的繁殖。

        东海原甲藻和抑食金球藻对日本虎斑猛水蚤的生长无显著性影响。当两种藻细胞密度为1.0×10⁵ cells/mL和1.0×10⁷ cells/mL时，5 d后成体的存活率均为100%，而且日本虎斑猛水蚤体内分别检测到了含量较高的甲藻特征色素多甲藻素（Peri）和抑食金球藻特征色素19’-丁酰氧基岩藻黄素（But-fuco），表明日本虎斑猛水蚤能够摄食这两种藻。实验藻为东海原甲藻时，在藻细胞密度为1.0×10⁵ cells/mL和5.0×10⁴ cells/mL时，N-C存活率、N-A存活率、N-C发育时间和C-A发育时间与对照组无显著差异，雌体12 d内产卵次数和产卵量分别显著高于对照组，表明东海原甲藻对日本虎斑猛水蚤无节幼体的存活、发育和雌体的繁殖没有不利影响。实验藻为抑食金球藻时，在较高藻细胞密度（1.0×10⁷ cells/mL）时，实验组N-C存活率、N-A存活率、N-C发育时间，C-A发育时间、雌体12 d内的产卵次数和产卵量均与对照组无差异，表明抑食金球藻不会影响日本虎斑猛水蚤无节幼体的存活、发育和雌体繁殖。

        链状亚历山大藻对日本虎斑猛水蚤的生长无显著性影响，但体内会积累麻痹性贝类毒素。暴露于链状亚历山大藻中，5 d后成体的存活率均为100%，日本虎斑猛水蚤体内检测到了含量较高的甲藻特征色素多甲藻素，表明日本虎斑猛水蚤能够摄食这种藻。在藻细胞密度为1.0×10⁴ cells/mL和5.0×10³ cells/mL时，N-C存活率、N-A存活率、C-A发育时间和雌体12 d内的产卵次数与对照组无差异，而N-C发育时间显著短于对照组，雌体12 d内的产卵量显著高于对照组，表明链状亚历山大藻对日本虎斑猛水蚤无节幼体的存活、发育和雌体繁殖没有不利影响。毒素检测结果表明，摄食链状亚历山大藻（密度为2.0×10³ cells/mL）48 h后每个日本虎斑猛水蚤体内所积累的毒素含量为598.2 pmol/ind。

        以上结果，四种不同类型的有毒有害藻对日本虎斑猛水蚤的危害途径和效应明显不同，日本虎斑猛水蚤适合作为评价有毒有害藻的测试生物，反映有毒有害藻的不同生物特征。因此，利用日本虎斑猛水蚤，结合其他生物的毒性评价研究，所得结果有利于更全面的了解有毒有害藻对整个生态系统浮游动物不同组分的影响。日本虎斑猛水蚤具有个体较小、雌雄异形、较高的繁殖力、较短的生命周期和易于在实验室培养等特点，利用无节幼体（N-C和N-A）的存活率、无节幼体（N-C和C-A）的发育时间、雌体12 d内产卵次数和产卵量，可以作为评价有毒有害良好的指标。

         In recent years, frequent outbreaks of harmful algae blooms in Chinese coastal waters have adversely affected marine zooplankton and ecosystems. In this study, the biological toxicity of four typical toxic and harmful algae, including two strains of Karenia mikimotoi (Fujian Suao and Putian), Prorocentrum donghaiense, Aureococus anophagefferens and Alexandrium catenella , were evaluated by using Tigriopus japonicus as the experimental organism, adult survival, napolii to copepod (N-C) survival rate, napolii to adult (N-A) survival rate, napolii to copepod (N-C) development time, copepod to adult (C-A) development time, number of spawns by females within 12 days, spawn production by females within 12 days and toxin accumulation as index, and Tetraselmis helgolandica var. tsingtaoensis as the control algae. The result of the study are as follows:

         Both K. mikimotoi strains significantly affected the survival of napulii, growth, development and female reproduction in T. japonicus. When the experimental algae was the Putian strain, the survival rate of nauplii was 46.7% on the 5th day and less than 20% on the 25th day at lower algal cell density (1.0×10³ cells/mL), and 0 on the 5th day when fed with 3.0×10⁴ cells/mL K. mikimotoi. Compared with the Putian strain, the suao strain had relatively low toxicity to T. japonicus. At a higher algal cell density (3.0×10⁴ cells/mL), N-C survival rate, N-A survival rate, number of spawns by females and spawn production by females within 12 days were lower than those of the control group, and there was a potential negative effect, although there was no significant difference, and the N-C development time and C-A development time were significantly longer than those of the control group, indicating that K. mikimotoi Suao strain could affect the survival, development of napulii and reproduction of female in T. japonicus.

        P. donghaiense and A. anophagefferens had no significant effect on the growth of T. japonicus. When fed with P. donghaiense at 1.0×10⁵ cells/mL or A. anophagefferens at 1.0×10⁷ cells/mL, the survival rate of adults T. japonicus after 5 days was 100%, and the dinoflagellates characteristic pigment polydinocophytin (Peri) and A. anophagefferens characteristic pigment 19'-butyroxyfucoxanthin (But-fuco) were detected in T. japonicus, indicating that T. japonicus was able to feed on these two algae. When the experimental algae were P. donghaiense, there was no significant difference in N-C survival rate, N-A survival rate, N-C development time and C-A development time at algal cell density of 1.0×10⁵ cells/mL and 5.0×10⁴ cells/mL, and number of spawns and spawn production by females within 12 days were significantly higher than those in the control group, indicating that P. donghaiense had no adverse effect on the survival, development of nauplii and the reproduction of females in T. japonicus. When T. japonicus was fed with A. anophagefferens, at the high algal cell density of 1×10⁷ cells/mL, there was no difference in N-C survival rate, N-A survival rate, N-C development time, C-A development time, number of spawns and spawn production by females within 12 days, indicating that A. anophagefferens could not affect the survival, development of nauplii and female reproduction in T. japonicus.

         A. catenella had no significant effect on the growth of T. japonicus, but paralytic shellfish toxins can accumulate in the body. The survival rate of adults after 5 days of exposure to A. catenella was 100%, and a high level of pigment polydinoflagellin, a characteristic pigment of dinoflagellates, was detected in T. japonicus, suggesting that T. japonicus is capable of ingesting this algae. At the algal cell density of 1.0×10⁴ cells/mL and 5.0×10³ cells/mL, there was no difference in N-C survival rate, N-A survival rate, C-A development time and number of spawns by females within 12 days in females compared with the control group, while the development time of N-C was significantly shorter than that in the control group, and spawn production by females within 12 days was significantly higher than that in the control group, indicating that A. catenella had no adverse effect on the survival, development of napulii and reproduction of female in T. japonicus. However, the results of toxin detection showed that the toxin content accumulated in each T. japonicus was 598.2 pmol/ind after ingestion of A. catenella (density of 2.0×10³ cells/mL) for 48h.

        The above results showed that the harmful pathways and effects of four different types of toxic and harmful algae on T. japonicus were obviously different. T. japonicus is suitable as a test organism for the evaluation of toxic and harmful algae, reflecting the different characteristics of toxic and harmful algae. Therefore, the results obtained by using T. japonicus and the toxicity evaluation studies of other organisms are helpful for a more comprehensive understanding of the effects of toxic and harmful algae on different components of zooplankton in the whole ecosystem. The survival rate of nauplii (N-C and N-A), the development time of nauplii (N-C and C-A), the number of spawns and spawn production by females within 12 days and spawn production can be used as indicators to evaluate toxicity and harmful algae.

第1章 绪论      1

1.1 有害藻华危害及日本虎斑猛水蚤概述      1

1.1.1 有害藻华概述及对浮游动物的主要危害方式      1

1.1.2 日本虎斑猛水蚤生物学特征及毒理学应用   3

1.2 四种有毒有害藻概述  6

1.2.1 米氏凯伦藻概述      6

1.2.2 东海原甲藻概述      7

1.2.3 抑食金球藻概述      8

1.2.4 链状亚历山大藻概述      10

1.3 研究目的及意义  10

第2章 两株米氏凯伦藻对日本虎斑猛水蚤摄食、存活、发育和繁殖的影响    13

2.1 材料与方法  13

2.1.1 实验生物   13

2.1.2 实验方法   13

2.1.3 统计分析   17

2.2 结果      17

2.2.1 米氏凯伦藻（苏澳株）对日本虎斑猛水蚤摄食的影响      17

2.2.2 两株米氏凯伦藻对日本虎斑猛水蚤存活的影响   18

2.2.3 两株米氏凯伦藻对日本虎斑猛水蚤发育的影响   21

2.2.4 两株米氏凯伦藻对日本虎斑猛水蚤繁殖的影响   23

2.3 讨论      25

2.4 小结      26

第3章 日本虎斑猛水蚤在东海原甲藻和抑食金球藻中的摄食、存活、发育和繁殖    27

3.1 材料与方法  27

3.1.1 实验生物   27

3.1.2 实验方法   27

3.1.3 统计分析   28

3.2 结果      29

3.2.1 日本虎斑猛水蚤在东海原甲藻和抑食金球藻中的摄食      29

3.2.2 日本虎斑猛水蚤在东海原甲藻和抑食金球藻中的存活      31

3.2.3 日本虎斑猛水蚤在东海原甲藻和抑食金球藻中的发育      35

3.2.4 日本虎斑猛水蚤在东海原甲藻和抑食金球藻中的繁殖      39

3.3 讨论      43

3.4 小结      44

第4章 日本虎斑猛水蚤在链状亚历山大藻中的生长繁殖和毒素累积       45

4.1 材料与方法  45

4.1.1 实验生物   45

4.1.2 实验方法   45

4.1.3 统计分析   46

4.2 结果      46

4.2.1 日本虎斑猛水蚤在链状亚历山大藻中的摄食      46

4.2.2 日本虎斑猛水蚤在链状亚历山大藻中的存活      48

4.2.3 日本虎斑猛水蚤在链状亚历山大藻中的发育      49

4.2.4 日本虎斑猛水蚤在链状亚历山大藻中的繁殖      51

4.2.5 日本虎斑猛水蚤对麻痹性贝类毒素的累积   53

4.3 讨论      54

4.4 小结      56

第5章 结论与展望   57

5.1  总结    57

5.2  结论    57

5.3  展望    57

参考文献     59

致  谢     67

作者简历及攻读学位期间发表的学术论文与其他相关学术成果     69