壬基酚对赤子爱胜蚯蚓（<i>Eisenia foetida</i>）的生态胁迫及茶多酚的缓解作用

陈建华; 张碧尧; 李丹佩; 陈世红; 谢艳颖; 王国红

doi:10.19303/j.issn.1008-0384.2020.11.014

壬基酚对赤子爱胜蚯蚓（Eisenia foetida）的生态胁迫及茶多酚的缓解作用

doi: 10.19303/j.issn.1008-0384.2020.11.014

福建师范大学生命科学学院，福建　福州　350117

基金项目: 福建省自然科学基金项目（2015J01125）；福建省自然科学基金引导性项目（2015N0027）

详细信息

作者简介:
陈建华（1973−），女，博士，讲师，研究方向：遗传毒理（E-mail：jhchen@fjnu.edu.cn）

通讯作者:
王国红（1967−），女，博士，教授，研究方向：动物生态（E-mail：guohongw@fjnu.edu.cn）

中图分类号: Q 89
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出版历程
- 收稿日期: 2020-04-08
- 修回日期: 2020-07-14
- 网络出版日期: 2020-11-13
- 刊出日期: 2020-11-30

Ecotoxicity of Nonylphenol on Eisenia foetida and Detoxification by Tea Polyphenols

College of Life Sciences, Fujian Normal University, Fuzhou, Fujian　350117, China

摘要

摘要: 目的研究壬基酚（NP）对赤子爱胜蚯蚓（Eisenia foetida）的生态胁迫效应，并探讨茶多酚对NP引发蚯蚓生态胁迫的缓解作用。方法采用自然土壤法研究壬基酚（NP）对赤子爱胜蚯蚓的生态胁迫效应，通过在NP基础液中添加不同质量分数的茶多酚，探讨茶多酚对NP胁迫下蚯蚓生活状态的影响、致死毒性、回避行为及生长抑制的缓解作用，同时采用HE组织切片染色法初步探讨NP毒性的作用机理。结果 400 mg·kg⁻¹ NP对蚯蚓具有强烈的致死毒性，100～200 mg·kg⁻¹NP对蚯蚓具有一定的致死毒性，其致死毒性与染毒时间、染毒量呈正相关；NP为200 mg·kg⁻¹时对蚯蚓生长抑制率达34%，具有显著差异；通过组织切片试验证实，NP胁迫下，蚯蚓空泡化腺细胞减少，环肌层变薄，角质层脱落，表皮层增厚；蚯蚓对12.5～200 mg·kg⁻¹的NP具有回避反应。在壬基酚基础液中添加70～700 μg·kg⁻¹的茶多酚后，蚯蚓致死率和生长抑制率均有显著下降。0.7～7.0 μg·kg⁻¹的茶多酚能明显缓解蚯蚓的回避反应。与NP对照组相比，添加0.7～700.0 μg·kg⁻¹茶多酚后蚯蚓体壁环肌层厚度增加，角质层恢复光滑完整。结论 100～400 mg·kg⁻¹的NP对赤子爱胜蚯蚓具有生态毒性作用，0.7～700.0 μg·kg⁻¹的茶多酚对NP胁迫蚯蚓生态毒性具有缓解作用。
- 壬基酚 /
- 蚯蚓 /
- 茶多酚 /
- 生态毒性 /
- 缓解作用
Abstract: Objective Ecotoxicity imposed by nonylphenol (NP) on Eisenia foetida and potential detoxification effect of tea polyphenols were studied. Method Adding different concentrations of tea polyphenols into NP base solution, the relief of tea polyphenols (TP) on the life state, lethal toxicity, avoidance behavior and growth inhibition induced by NP were observed in E. foetida, and the toxic mechanism of NP was studied by HE staining. Result At a concentration in soil ranging between 100-200 mg·kg⁻¹, NP exerted varied degrees of toxicity to the earthworms with lethality directly correlated with the concentration as well as exposure time. The growth of earthworms was retarded by 34% at 200 mg NP·kg⁻¹, and a high mortality rate observed when the concentration reached 400 mg NP·kg⁻¹. The stained tissues of the earthworms under NP stress showed decreased vacuolated gland cells, thinned circular muscles, bared cuticles, and thickened epidermis. Toward the soil with NP in the range of 12.5-200 mg·kg⁻¹, the earthworms displayed a movement avoidance. Upon the addition of tea polyphenols at the concentrations of 70-700 μg·kg⁻¹ in the NP solution, the lethality and growth retardation of the earthworms declined significantly. A tea polyphenols addition of 0.7-7 μg·kg⁻¹ in the NP solution significantly lessened the avoiding response of the earthworms. With 0.7-700 μg·kg⁻¹ of added tea polyphenols, the thickness of the muscular layer of the earthworm body wall increased and the stratum corneum became smooth again. Conclusion NP at a concentration in the range from 100 mg·kg⁻¹ to 400 mg·kg⁻¹ in soil could be toxic or lethal to E. foetida. Whereas, the presence of tea polyphenols at a level between 0.7 μg·kg⁻¹ and 700 μg·kg⁻¹ in the NP solution that was blended in the soil could significantly mitigate the adverse effect induced by NP.
- Nonylphenol /
- Eisenia foetida /
- tea polyphenol /
- ecotoxicity /
- detoxification effect

HTML全文

图 1 壬基酚对蚯蚓生长的抑制

注：不同小写字母表示各质量浓度处理组间存在显著差异（P＜0.05）

Figure 1. Growth retardation of E. foetida caused by NP

Note: Data with different lowercase letters indicate significant differences between treatments (P＜0.05).

下载: 全尺寸图片幻灯片

图 2 NP对蚯蚓体壁显微结构的影响（14 d）（400×）

注：A：清水对照组；B：乙醇对照组；C：25 mg·kg^>−1NP组；D～E：50 mg·kg^>−1NP组；F～G：100 mg·kg^>−1NP组；H～I：200 mg·kg^>−1NP组。a:表皮层；b:环肌层；c:腺细胞空泡化；d:角质层；e：箭头所示角质层脱落后裸露的柱状上皮细胞。

Figure 2. Effects of NP on microstructure of E. foetida body wall (14 d) (400×)

Note: A: Normal body wall under water control; B: Ethanol control; C: 25 mg·kg⁻¹ NP treatment; D and E: 50 mg·kg⁻¹ NP treatment; F and G: 100 mg·kg⁻¹ NP treatment; H and I: 200 mg·kg⁻¹ NP treatment. a: Epidermis; b: Circular muscle; c: Vacuolated gland cells; d: Cuticle layer; e: Arrow shows columnar epithelial cells with bare cuticles.

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图 3 NP作用下蚯蚓的回避行为

Figure 3. Avoidance behavior of E. foetida in presence of NP

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图 4 茶多酚对壬基酚所致生长毒性的缓解

注：不同小写字母表示各质量浓度处理组间存在显著差异（P＜0.05）

Figure 4. Effect of tea polyphenols on growth of E. foetida under NP stress

Note: Date with different lowercase letters indicate significant differences between treatments (P＜0.05).

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图 5 茶多酚对NP胁迫下蚯蚓体壁损伤的缓解作用（400×）

注：A：NP对照组；B: 0.7 μg kg^-1茶多酚组；C:700 μg·kg^-1茶多酚组。a: 角质层；b: 空泡化腺细胞；c:环肌层

Figure 5. Effect of tea polyphenols on body wall injury of E. foetida under NP stress (400×)

Note：A: NP control; B: 0.7 μg·kg⁻¹ tea polyphenols treatment; c: 700 μg·kg⁻¹ tea polyphenols treatment. a: Epidermis; b: Vacuolated secretory cells; c: Circular muscle.

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图 6 茶多酚对NP胁迫下蚯蚓回避的缓解

Figure 6. Effect of tea polyphenols on avoidance behavior of E. foetida under NP stress

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表 1 有害物质毒性分级标准^[26]

Table 1. Standards for toxicity classification of hazardous substances

毒性等级 Toxicity level	剧毒 Poisonous	高毒 High toxic	中毒 poisoning	低毒 low-toxic	微毒 slight toxicity
LC₅₀范围 LC₅₀ Range/（mg·kg⁻¹）	≤0.1	0.1～≤1.0	1.0 ～≤10.0	10.0＞	-

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表 2 NP对蚯蚓的毒性分析

Table 2. Toxicity induced by NP on E. foetida

暴露时间 Exposure time/d	LC₅₀/（mg·kg⁻¹）	LC₅₀95% 置信区间95% Confidence interval	毒力回归式 Toxicity regression	相关系数r² Correlation coefficient
7	292.817	274.058～312.860	Y=0.281X−28.913	0.914
10	214.355	184.714～284.752	Y=0.303X−21.739	0.991
14	141.421	124.32～160.875	Y=0.299X−6.087	0.886
注：Y代表蚯蚓的慢性致死率，X代表NP质量分数。 Note: Y represents chronic lethality on earthworms; and X, NP concentration in soil.

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表 3 茶多酚对壬基酚胁迫下蚯蚓致死率

Table 3. Mortality of E. foetida under NP stress in presence of tea polyphenolsy　　　　　　　　　　　（单位/%）

处理 Treatment	第10 d Day 10	第14 d Day 14
100 mg·kg⁻¹NP（CK）	25.00±21.21	90.00±14.14
0.7 μg·kg⁻¹茶多酚 0.7 μg·kg⁻¹tea polyphenols	5.00±7.07	40.00±14.14*
7 μg·kg⁻¹茶多酚 7 μg·kg⁻¹tea polyphenols	0.00±0.00	0.00±0.00**
70 μg·kg⁻¹茶多酚 70 μg·kg⁻¹tea polyphenols	0.00±0.00	0.00±0.00**
700 μg·kg⁻¹茶多酚 700 μg·kg⁻¹ tea polyphenols	0.00±0.00	0.00±0.00**
注：代表与NP对照相比差异显著（P＜0.05），代表与NP对照相比差异极显著（P＜0.01）。 Note: represents significant difference compared with NP control at P＜0.05; ** extremely significant difference compared with NP control at P＜0.01.

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