水-沉积物中花翅摇蚊标准化毒性测试体系研究
Study on Standardized Toxicity Test System of Chironomus striatipennis Kieffer in Water-sediment
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摘要: 花翅摇蚊（Chironomus striatipennis Kieffer）是一种分布广泛的本土底栖生物，在环境科学研究领域中应用较广泛，对于评价化学品对底栖生物的水-沉积物毒性具有重要意义。为了建立化学品对花翅摇蚊的水-沉积物毒性标准化测试体系，根据经济合作与发展组织（Organisation for Economic Co-operation and Development，OECD）颁布的《沉积物-水体中摇蚊生命周期毒性试验—水体加标法或沉积物加标法》（OECD No.233），研究了人工沉积物类型对花翅摇蚊羽化率和发育速率的影响以及受精卵孵化初期的生长密度对幼虫性别分化的影响。此外，选取典型的杀生剂敌草隆，通过加标上覆水法开展2代花翅摇蚊的生命周期毒性研究，对该系统能否满足标准方法的质控要求进行验证。结果显示，经平衡后的人工沉积物较未经平衡的沉积物更有利于花翅摇蚊的羽化和发育，平均羽化率为95.8%，平均发育速率（development rate）为0.1387 d-1。2组沉积物中花翅摇蚊的羽化率和发育速率的差异均具有统计学意义（Per=4.87×10-6，Pdr=0.015，P<0.05）。在其他条件一定的情况下，花翅摇蚊受精卵串孵化初期的生长密度对幼虫性别分化具有一定的影响，随着生长密度的增加，摇蚊幼虫的雌雄性别比率逐渐降低。低密度组、中密度组和高密度组雌雄性别比率分别为1.11、0.95和0.56，中、低密度组与高密度组之间具有显著性差异（P=0.002，P<0.01）。化学分析结果显示，敌草隆的检出限为0.0500 μg·mL-1，定量限为0.200 μg·mL-1，体系中的回收率范围为74.9%～94.3%。敌草隆对2代花翅摇蚊生命周期毒性研究结果表明，空白对照组2代花翅摇蚊的平均羽化率均为100%，其中完全羽化且存活的成蚊雌/雄性别比率分别为0.412/0.588（母代）和0.4/0.6（子一代）；母代对照组中花翅摇蚊繁殖力以及卵串的受精率分别为0.848和0.893，均满足标准方法的质控要求。花翅摇蚊标准化测试体系可满足其正常的生命周期需求，能够用于化学品对底栖生物的水-沉积物毒性评估。Abstract: Chironomus striatipennis Kieffer is a native benthic organism that widely distributed in China. This species has been extensively used in the research field of environmental science researches, which has a great significance for evaluating the water-sediment toxicity of chemicals towards benthic organisms. In order to establish the standardized water-sediment toxicity test system of Chironomus striatipennis Kieffer for chemical, according to the Organisation for Economic Co-operation and Development testing guideline No.233, namely Sediment-Water Chironomid Life-Cycle Toxicity Test Using Spiked Water or Spiked Sediment. The effects of different types of artificial sediments on the emergence rate and development rate of Chironomus striatipennis Kieffer was compared. The relationship between the sex differentiation of larvae and the growth density at the early hatching stage was studied as well. In addition, diuron, a typical biocide, was selected to study the life-cycle toxicity of two generations of Chironomids by using spiked water, to verify whether the system could meet the quality control requirements of the standard method. The results showed that the balanced artificial sediments were more favorable to the emergence and development of Chironomus striatipennis Kieffer compared with the artificial sediments without balance. The average emergence rate of Chironomus striatipennis Kieffer is as high as 95.8% and the average development rate is 0.1387 d-1. The difference between the two groups of standardized artificial sediments were statistically significant (Per=4.87×10-6, Pdr=0.015, P<0.05). Under certain conditions, the growth density at the early hatching stage had a certain effect on the sex differentiation of the larvae. With the increasing of the growth density, the sex ratio (♀:♂) of the larvae decreased gradually. The sex ratio of the low-density group, medium density group and high-density group was 1.11, 0.95 and 0.56, respectively. There was a significant difference between the high-density group and the other two groups (P=0.002, P<0.01). The chemical analysis results showed that the detection limit of diuron was 0.0500 μg·mL-1, and the quantitative limit was 0.200 μg·mL-1. The range of the recovery rate of the diuron in the test system was 74.9%~94.3%. The results of life-cycle toxicity of two generation Chironomus striatipennis Kieffer induced by diuron showed that the average emergence rate of the two generations both was 100% in the blank control group. The sex ratios in the blank control group of fully emerged and alive adults (♀:♂) were 0.412/0.588 (P generation) and 0.4/0.6 (F1 generation), respectively. The reproduction and the fertilization rate of eggs spawned by P generation in the blank control group were 0.848 and 0.893, respectively, which met the quality control requirements of the standard method. Therefore, the standardized test system can satisfy the normal life cycle requirement of Chironomus striatipennis Kieffer and can be used to assess the water-sediment toxicity of chemicals to sediment-dwelling organisms.
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