2，4，6-三氯酚慢性暴露对青鳉生长和生殖影响的研究
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摘要
2,4,6-三氯酚(2,4,6-TCP)是化工生产中的重要原料,可用作杀菌剂、防腐剂、脱叶剂,在有机合成、造纸和印染行业中得到使用广泛,目前在工业废水、污水处理厂排放废水、农药废水、垃圾渗滤液、饮用水和河水中都检测到它的存在,美国环境保护署(EPA)已经确定2,4,6-三氯酚为疑似致癌物。
目前,针对三氯酚的毒性研究大多集中于水生生物某一生理阶段上的急性和亚急性毒性,而它在整个世代暴露的慢性影响研究较少。在自然环境中,大多数情况下污染物对鱼类的影响可能作用于整个发育和繁殖过程,慢性毒性研究的结果将更接近于实际环境,有助于人们了解环境内分泌干扰物对鱼类生理生态的影响机制。
青(?)是发源于日本的一种小型鱼类,易于饲养,对水质的环境变化特别敏感。成熟青(?)个体小,体长3-4 cm,繁殖力强。20世纪初日本开始将青(?)用于毒理学实验研究,作为供试物种在生理学、生态学和内分泌学等领域被逐渐采用,研究涉及青(?)的培育、饲养管理和实验操作等方面,目前被广泛用于毒理学实验。
本文以日本青(?)(Oryzias latipes)为实验动物,研究了2,4,6-三氯酚对青(?)整个世代生长、生殖能力和子代质量的影响。取性成熟的青(?)鱼,暴露在不同浓度水平(0,10,30,100,100,10000μg/L)的2,4,6-TCP溶液中,以0.01％(v／v)DMSO为溶剂对照。暴露28 d后,收集鱼卵,统计分析每对产卵数、存活个数和受精率；选取一组清水恢复饲养,另一组继续暴露直到性成熟,分别观察胚胎发育、孵化天数、孵化率、畸形率等生长和生殖指标的变化；成鱼暴露5周后,取鱼样考察畸形率和死亡率。本文获得了如下研究成果：
1.总体来说,随着2,4,6-三氯酚暴露浓度的增加,青(?)的体长和体重均下降,但雄鱼在高浓度水平有回升现象；雌鱼的肝脏指数下降,而雄鱼在各浓度水平无明显影响。在低浓度组中,青(?)卵巢指数无显著变化；在高浓度组中,卵巢指数显著下降(p<0.05)。
2.随着2,4,6-三氯酚暴露浓度的增加,青(?)的每天产卵对数没有明显变化,但每条雌鱼的产卵量下降；鱼卵的受精率下降,特别是在1000μg/L浓度水平,存在显著性差异(p<0.05)；鱼卵的存活数在300和1000μgL浓度水平也显著下降(p<0.05)。
3.子代孵化率在清水饲养后其孵化天数没有明显变化。孵化率在300和1000μg/L浓度组中显著下降。畸形率在100、300和1000μg/L各浓度组中都显著增加。继续暴露的子代,在孵化天数上仍然没有显著的变化。孵化率和畸形率在100、300和1000μg/L浓度组中均有显著性上升。与在清水中饲养的鱼卵相比,孵化率下降,畸形率均上升。暴露后,幼鱼在不同程度上出现了大量的畸形,出生后不久就死亡。
2,4,6-trichlorophenol (2,4,6-TCP) is a kind of important chemical material which could be much used as a antiseptic, food preservative and defoliant in industries like organic synthesis, paper making, and priting and dyeing. It could be detected in drinking water, river water, landfill leachate, and wastewater from chemical factories, sewage plants, and agricultural chemicals, etc. So,2,4,6-TCP may be potentially harmful to the aquatic ecosystem. The EPA has claimed that 2,4,6-TCP is, to some extent, carcinogenic substance.
At present, research work on 2,4,6-TCP mostly lies on the acute and subacute researches on a certain physiological stage of the aquatic animals, while there has been less research work on the chronic influence researches exposed in the whole generation. In the natural environment and in most cases, the pollutants may be influential on the whole development and breeding process. And the results of the chronic research will be much closer to the actual environment, which contributes to people's comprehension of the physiological and ecological effect mechanism that the endocrine disruptors effect on the fish.
medaka which is a kind of small fish originated from Japan, is easy to raise and very sensitive to the changes of water quality. The mature medaka is small with a full-length of 3 to 4 cm, and has strong reproductive capacity. In the beginning of 19~(th) century in Japan, the research on medaka has been started, which has been commonly used in physiology, ecology, endocrinology, and alike. Also, there have been systematic researches on breeding, feeding management, experiment operations, and so on. Nowadays, the medaka is widely used in toxicity test.
With the medaka as the model organism, this dissertation researches into the influence of 2,4,6-TCP on the whole generation's development, reproductive performance, and the filial-generation's quality of the medaka.
Take 20 pairs, each set, of medaka of sexual maturity and respectively expose them in the 2,4,6-TCP (0,10,30,100,100,10000μg/L),0.01%(v/v) DMSO of different concentration solvent as comparison. After 28 d of exposure, collect the roes and take statistics; choose one set and breed them in clear water, and another set still in the solvent until they get sexually mature; observe the embryos respectively; after the 5-week exposure of the mature fish, take some samples and make analysis.
After the decline of the length and weight of the male, the dose-effect relationship takes on as the strength of the solvent intensifies. For the female, the length and weight declines in light concentration solvent as the concentration goes up, and picks up in high concentration solvent. The liver index of the male lowers with the solvent strength's increasing, while female's liver index doesn't take on significant changes in each solvent set of different concentration. The ovary index doesn't change markedly in sets of light concentration solvent; but it declines notably (p< 0.05) in sets of high concentration solvent.
For medaka in the solvent, the number of laying-egg medaka doesn't make notable difference, but the egg laying amount of the female goes down, while, about the filial-generation's hatchability, the number of hatching days is of no marked changes during the feeding in the clear water. The hatching rate doesn't change significantly in solvent sets of light concentration, but goes down notably in solvent sets of high concentration (p< 0.05). The teratological rate begins to increase significantly in solvent sets of relatively high concentration (p< 0.05).
目前,针对三氯酚的毒性研究大多集中于水生生物某一生理阶段上的急性和亚急性毒性,而它在整个世代暴露的慢性影响研究较少。在自然环境中,大多数情况下污染物对鱼类的影响可能作用于整个发育和繁殖过程,慢性毒性研究的结果将更接近于实际环境,有助于人们了解环境内分泌干扰物对鱼类生理生态的影响机制。
青(?)是发源于日本的一种小型鱼类,易于饲养,对水质的环境变化特别敏感。成熟青(?)个体小,体长3-4 cm,繁殖力强。20世纪初日本开始将青(?)用于毒理学实验研究,作为供试物种在生理学、生态学和内分泌学等领域被逐渐采用,研究涉及青(?)的培育、饲养管理和实验操作等方面,目前被广泛用于毒理学实验。
本文以日本青(?)(Oryzias latipes)为实验动物,研究了2,4,6-三氯酚对青(?)整个世代生长、生殖能力和子代质量的影响。取性成熟的青(?)鱼,暴露在不同浓度水平(0,10,30,100,100,10000μg/L)的2,4,6-TCP溶液中,以0.01％(v／v)DMSO为溶剂对照。暴露28 d后,收集鱼卵,统计分析每对产卵数、存活个数和受精率；选取一组清水恢复饲养,另一组继续暴露直到性成熟,分别观察胚胎发育、孵化天数、孵化率、畸形率等生长和生殖指标的变化；成鱼暴露5周后,取鱼样考察畸形率和死亡率。本文获得了如下研究成果：
1.总体来说,随着2,4,6-三氯酚暴露浓度的增加,青(?)的体长和体重均下降,但雄鱼在高浓度水平有回升现象；雌鱼的肝脏指数下降,而雄鱼在各浓度水平无明显影响。在低浓度组中,青(?)卵巢指数无显著变化；在高浓度组中,卵巢指数显著下降(p<0.05)。
2.随着2,4,6-三氯酚暴露浓度的增加,青(?)的每天产卵对数没有明显变化,但每条雌鱼的产卵量下降；鱼卵的受精率下降,特别是在1000μg/L浓度水平,存在显著性差异(p<0.05)；鱼卵的存活数在300和1000μgL浓度水平也显著下降(p<0.05)。
3.子代孵化率在清水饲养后其孵化天数没有明显变化。孵化率在300和1000μg/L浓度组中显著下降。畸形率在100、300和1000μg/L各浓度组中都显著增加。继续暴露的子代,在孵化天数上仍然没有显著的变化。孵化率和畸形率在100、300和1000μg/L浓度组中均有显著性上升。与在清水中饲养的鱼卵相比,孵化率下降,畸形率均上升。暴露后,幼鱼在不同程度上出现了大量的畸形,出生后不久就死亡。
2,4,6-trichlorophenol (2,4,6-TCP) is a kind of important chemical material which could be much used as a antiseptic, food preservative and defoliant in industries like organic synthesis, paper making, and priting and dyeing. It could be detected in drinking water, river water, landfill leachate, and wastewater from chemical factories, sewage plants, and agricultural chemicals, etc. So,2,4,6-TCP may be potentially harmful to the aquatic ecosystem. The EPA has claimed that 2,4,6-TCP is, to some extent, carcinogenic substance.
At present, research work on 2,4,6-TCP mostly lies on the acute and subacute researches on a certain physiological stage of the aquatic animals, while there has been less research work on the chronic influence researches exposed in the whole generation. In the natural environment and in most cases, the pollutants may be influential on the whole development and breeding process. And the results of the chronic research will be much closer to the actual environment, which contributes to people's comprehension of the physiological and ecological effect mechanism that the endocrine disruptors effect on the fish.
medaka which is a kind of small fish originated from Japan, is easy to raise and very sensitive to the changes of water quality. The mature medaka is small with a full-length of 3 to 4 cm, and has strong reproductive capacity. In the beginning of 19~(th) century in Japan, the research on medaka has been started, which has been commonly used in physiology, ecology, endocrinology, and alike. Also, there have been systematic researches on breeding, feeding management, experiment operations, and so on. Nowadays, the medaka is widely used in toxicity test.
With the medaka as the model organism, this dissertation researches into the influence of 2,4,6-TCP on the whole generation's development, reproductive performance, and the filial-generation's quality of the medaka.
Take 20 pairs, each set, of medaka of sexual maturity and respectively expose them in the 2,4,6-TCP (0,10,30,100,100,10000μg/L),0.01%(v/v) DMSO of different concentration solvent as comparison. After 28 d of exposure, collect the roes and take statistics; choose one set and breed them in clear water, and another set still in the solvent until they get sexually mature; observe the embryos respectively; after the 5-week exposure of the mature fish, take some samples and make analysis.
After the decline of the length and weight of the male, the dose-effect relationship takes on as the strength of the solvent intensifies. For the female, the length and weight declines in light concentration solvent as the concentration goes up, and picks up in high concentration solvent. The liver index of the male lowers with the solvent strength's increasing, while female's liver index doesn't take on significant changes in each solvent set of different concentration. The ovary index doesn't change markedly in sets of light concentration solvent; but it declines notably (p< 0.05) in sets of high concentration solvent.
For medaka in the solvent, the number of laying-egg medaka doesn't make notable difference, but the egg laying amount of the female goes down, while, about the filial-generation's hatchability, the number of hatching days is of no marked changes during the feeding in the clear water. The hatching rate doesn't change significantly in solvent sets of light concentration, but goes down notably in solvent sets of high concentration (p< 0.05). The teratological rate begins to increase significantly in solvent sets of relatively high concentration (p< 0.05).
引文
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