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
计量
- 文章访问数: 832
- HTML全文浏览量: 273
- PDF下载量: 31
- 被引次数: 0
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.

下载: 全尺寸图片幻灯片

图 3 NP作用下蚯蚓的回避行为

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

下载: 全尺寸图片幻灯片

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).

下载: 全尺寸图片幻灯片

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.

下载: 全尺寸图片幻灯片

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

下载: 全尺寸图片幻灯片

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＞	-

下载: 导出CSV

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.

下载: 导出CSV

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.

下载: 导出CSV

参考文献(40)

[1]	ESPEJO R, VALTER K, SIMONA M, et al. Determination of nineteen 4-alkylphenol endocrine disrupters in Geneva municipal sewage wastewater [J]. Journal of Chromatography A, 2002, 976(1/2): 335−343.
[2]	张婷瑜, 张福金, 何江, 等. 壬基酚的土壤残留及其行为研究进展 [J]. 农业资源与环境学报, 2014, 31(2):118−125. ZHANG T Y, ZHANG F J, HE J, et al. Advances in nonylphenols residues and their behaviors in soil [J]. Journal of Agricultural Resources and Environment, 2014, 31(2): 118−125.(in Chinese)
[3]	黄超, 吴文铸, 单正军, 等. 壬基酚在土壤中的吸附和淋溶特性 [J]. 生态环境学报, 2015, 24(12):2062−2067. HUANG C, WU W Z, SHAN Z J, et al. Adsorption and leaching of nonylphenol in soils [J]. Ecology and Environmental Sciences, 2015, 24(12): 2062−2067.(in Chinese)
[4]	朱建林, 陈昱, 李昱辰, 等. 大豆异黄酮对壬基酚所致大鼠血清性激素紊乱及前列腺增生的改善作用 [J]. 福建医科大学学报, 2012, 46(4):231−234. doi: 10.3969/j.issn.1672-4194.2012.04.002 ZHU J L, CHEN Y, LI Y C, et al. Improvement effect of soy isoflavones(SI) on serum hormone disorders and benign prostatic hyperplasia(BPH) induced by nonylphenol(NP) in rat [J]. Journal of Fujian Medical University, 2012, 46(4): 231−234.(in Chinese) doi: 10.3969/j.issn.1672-4194.2012.04.002
[5]	SEIKE N, WANIBUCHI H, MORIMURA K, et al. Enhancement of lung carcinogenesis by nonylphenol and genistein in a F344 rat multiorgan carcinogenesis model [J]. Cancer Letters, 2003, 192(1): 25−36. doi: 10.1016/S0304-3835(02)00684-5
[6]	TANAKA T, KOHNO H, TANINO M, et al. Inhibitory effects of estrogenic compounds, 4-nonylphenol and genistein, on 7, 12-dimethylbenz[a]anthracene-induced ovarian carcinogenesis in rats [J]. Ecotoxicology and Environmental Safety, 2002, 52(1): 38−45. doi: 10.1006/eesa.2002.2159
[7]	CAI Q Y, HUANG H J, LYU H, et al. Occurrence of nonylphenol and nonylphenol monoethoxylate in soil and vegetables from vegetable farms in the Pearl River Delta, South China [J]. Archives of Environmental Contamination and Toxicology, 2012, 63(1): 22−28. doi: 10.1007/s00244-011-9741-4
[8]	WU A H, ARAKAWA K, STANCZYK F Z, et al. Tea and circulating estrogen levels in postmenopausal Chinese women in Singapore [J]. Carcinogenesis, 2005, 26(5): 976−980. doi: 10.1093/carcin/bgi028
[9]	FUHRMAN B J, PFEIFFER R M, WU A H, et al. Green tea intake is associated with urinary estrogen profiles in Japanese-American women [J]. Nutrition Journal, 2013, 12: 25. doi: 10.1186/1475-2891-12-25
[10]	刘振兴, 柯浩, 郝乐, 等. 茶多酚对罗非鱼生长性能、抗氧化功能和非特异免疫指标的影响 [J]. 广东农业科学, 2012, 39(23):113−115. doi: 10.3969/j.issn.1004-874X.2012.23.037 LIU Z X, KE H, HAO L, et al. Effects of tea polyphenols on growth performance, antioxidant effect and non-specific immune indices of Oreochromis niloticus [J]. Guangdong Agricultural Sciences, 2012, 39(23): 113−115.(in Chinese) doi: 10.3969/j.issn.1004-874X.2012.23.037
[11]	汪小红, 武书庚, 崔耀明, 等. 茶多酚对蛋鸡生产性能、蛋品质和抗氧化能力的影响 [J]. 动物营养学报, 2017, 29(1):193−201. doi: 10.3969/j.issn.1006-267x.2017.01.022 WANG X H, WU S G, CUI Y M, et al. Effects of dietary tea polyphenols on performance, egg quality and antioxidant ability of laying hens [J]. Acta Zoonutrimenta Sinica, 2017, 29(1): 193−201.(in Chinese) doi: 10.3969/j.issn.1006-267x.2017.01.022
[12]	徐仁扣. 土壤酸化及其调控研究进展 [J]. 土壤, 2015, 47(2):238−244. XU R K. Research progresses in soil acidification and its control [J]. Soils, 2015, 47(2): 238−244.(in Chinese)
[13]	刘德鸿, 成杰民, 刘德辉. 蚯蚓对土壤中铜、镉形态及高丹草生物有效性的影响 [J]. 应用与环境生物学报, 2007, 13(2):209−214. doi: 10.3321/j.issn:1006-687X.2007.02.015 LIU D H, CHENG J M, LIU D H. Effect of earthworm on Cu and Cd forms and their availability to Sorghum bicolor × S. sudanense [J]. Chinese Journal of Applied & Environmental Biology, 2007, 13(2): 209−214.(in Chinese) doi: 10.3321/j.issn:1006-687X.2007.02.015
[14]	郭永灿, 王振中, 张友梅, 等. 重金属对蚯蚓的毒性毒理研究 [J]. 应用与环境生物学报, 1996, 2(2):132−140. doi: 10.3321/j.issn:1006-687X.1996.02.006 GUO Y C, WANG Z Z, ZHANG Y M, et al. Studies on toxicity and toxicology of heavymetals to earthworms in polluted soils [J]. Chinese Journal of Applied and Environmental Biology, 1996, 2(2): 132−140.(in Chinese) doi: 10.3321/j.issn:1006-687X.1996.02.006
[15]	戈峰, 刘向辉, 江炳缜. 蚯蚓对金属元素的富集作用分析 [J]. 农业环境保护, 2002, 21(1):16−18. GE F, LIU X H, JIANG B Z. Accumulation of several metals in earthworm (Eisenia foetida) [J]. Agro-Environmental Protection, 2002, 21(1): 16−18.(in Chinese)
[16]	肖能文, 刘向辉, 李薇, 等. 用蚯蚓溶酶体作为检测土壤污染的生物标志物 [J]. 应用生态学报, 2006, 17(3):516−519. doi: 10.3321/j.issn:1001-9332.2006.03.032 XIAO N W, LIU X H, LI W, et al. Lysosome of Eisenia fetida as biomarker of soil pollution [J]. Chinese Journal of Applied Ecology, 2006, 17(3): 516−519.(in Chinese) doi: 10.3321/j.issn:1001-9332.2006.03.032
[17]	马莉, 贾辉, 殷秀琴, 等. 蚯蚓在处理活性污泥过程中的生长繁殖 [J]. 应用与环境生物学报, 2013, 19(1):147−151. doi: 10.3724/SP.J.1145.2013.00147 MA L, JIA H, YIN X Q, et al. Growth and propagation of earthworm in activated sewage sludge treatment [J]. Chinese Journal of Applied and Environmental Biology, 2013, 19(1): 147−151.(in Chinese) doi: 10.3724/SP.J.1145.2013.00147
[18]	刘向辉, 戈峰, 徐张红, 等. 亚硒酸钠对蚯蚓的毒性及蚓体富硒作用的研究 [J]. 应用与环境生物学报, 2001, 7(5):457−460. doi: 10.3321/j.issn:1006-687X.2001.05.011 LIU X H, GE F, XU Z H, et al. The toxicity of sodium selenite to earthworm and selenium-accumulating effect of earthworm [J]. Chinese Journal of Applied and Environmental Biology, 2001, 7(5): 457−460.(in Chinese) doi: 10.3321/j.issn:1006-687X.2001.05.011
[19]	戈峰, 刘向辉, 潘卫东, 等. 蚯蚓在德兴铜矿废弃地生态恢复中的作用 [J]. 生态学报, 2001, 21(11):1790−1795. doi: 10.3321/j.issn:1000-0933.2001.11.007 GE F, LIU X H, PAN W D, et al. The role of earthworm in the ecological restoration of mining wasteland of Dexing Copper Mine in China [J]. Acta Ecologica Sinica, 2001, 21(11): 1790−1795.(in Chinese) doi: 10.3321/j.issn:1000-0933.2001.11.007
[20]	郑丽萍, 林玉锁, 冯艳红, 等. 氯丹和灭蚁灵污染场地土壤对陆生生物的毒性效应 [J]. 应用与环境生物学报, 2012, 18(1):93−99. doi: 10.3724/SP.J.1145.2012.00093 ZHENG L P, LIN Y S, FENG Y H, et al. Toxic effects of chlordane and mirex-contaminated soil on terrestrial organisms [J]. Chinese Journal of Applied and Environmental Biology, 2012, 18(1): 93−99.(in Chinese) doi: 10.3724/SP.J.1145.2012.00093
[21]	张伟, 郑彬, 马静静, 等. 土壤B[a]P多次叠加污染的生物有效性及对蚯蚓体腔细胞染色体和溶酶体的毒性效应 [J]. 应用与环境生物学报, 2014, 20(6):1020−1026. ZHANG W, ZHENG B, MA J J, et al. Effect of multiple Benzo(a)Pyrene addition on its bioavailability and coelomocyte lysosomes and chromosomes in earthworms [J]. Chinese Journal of Applied and Environmental Biology, 2014, 20(6): 1020−1026.(in Chinese)
[22]	CASEY F X M, ŠIMŮNEK J, LEE J, et al. Sorption, mobility, and transformation of estrogenic hormones in natural soil [J]. Journal of Environmental Quality, 2005, 34(4): 1372−1379. doi: 10.2134/jeq2004.0290
[23]	翟洪艳, 于泳, 孙红文. 壬基酚在海河沉积物中的耗氧和厌氧降解 [J]. 环境化学, 2007, 26(6):725−729. doi: 10.3321/j.issn:0254-6108.2007.06.001 ZHAI H Y, YU Y, SUN H W. Aerobic and anaerobic biodegradation of nonylphenol in Haihe sediments [J]. Environmental Chemistry, 2007, 26(6): 725−729.(in Chinese) doi: 10.3321/j.issn:0254-6108.2007.06.001
[24]	王艳平. 高风险农药助剂在土壤中的残留特征及环境行为初探[D]. 北京: 中国农业科学院, 2011. WANG Y P. Exploratory studies on the residue and environmental behavior of high-risk pesticide adjuvants in soils[D]. Beijing: Chinese Academy of Agricultural Sciences, 2011. (in Chinese)
[25]	成云峰, 刘鹏, 薛章荣. 农药敌草快的生态毒性研究 [J]. 科技通报, 2015, 31(11):128−130. doi: 10.3969/j.issn.1001-7119.2015.11.026 CHENG Y F, LIU P, XUE Z R. Study on environment toxicity of 10% Diquat AS [J]. Bulletin of Science and Technology, 2015, 31(11): 128−130.(in Chinese) doi: 10.3969/j.issn.1001-7119.2015.11.026
[26]	陈春, 周启星, 刘潇威, 等. 多环麝香对蚯蚓的急性和亚急性毒性效应 [J]. 生态毒理学报, 2012, 7(4):401−407. CHEN C, ZHOU Q X, LIU X W, et al. Acute and sub-acute toxicological effects of polycyclic musks on earthworm, Eisenia fetida [J]. Asian Journal of Ecotoxicolog, 2012, 7(4): 401−407.(in Chinese)
[27]	徐冬梅, 文岳中, 李立, 等. PFOS对蚯蚓急性毒性和回避行为的影响 [J]. 应用生态学报, 2011, 22(1):215−220. XU D M, WEN Y Z, LI L, et al. Effects of perfluorooctane sulfonate on acute lethality and avoidance behavior of earthworm [J]. Chinese Journal of Applied Ecology, 2011, 22(1): 215−220.(in Chinese)
[28]	王彦华, 俞卫华, 杨立之, 等. 22种常用除草剂对蚯蚓(Eisenia fetida)的急性毒性 [J]. 生态毒理学报, 2012, 7(3):317−325. WANG Y H, YU W H, YANG L Z, et al. Acute toxicity of twenty-two commonly used herbicides to earthworm (Eisenia fetida) [J]. Asian Journal of Ecotoxicolog, 2012, 7(3): 317−325.(in Chinese)
[29]	SHAN J, WANG T, LI C L, et al. Bioaccumulation and bound-residue formation of a branched 4-nonylphenol isomer in the geophagous Earthworm Metaphire guillelmiin a rice paddy soil [J]. Environmental Science & Technology, 2010, 44(12): 4558−4563.
[30]	智勇, 单正军, 卜元卿, 等. 壬基酚异构体在威廉腔环蚓(Metaphire guillelmi)体内的蓄积特征及对其生长的影响 [J]. 生态与农村环境学报, 2015, 31(6):935−941. doi: 10.11934/j.issn.1673-4831.2015.06.021 ZHI Y, SHAN Z J, BU Y Q, et al. Bioaccumulation of nonylphenol isomer in Metaphire guillelmi and its influence on growth [J]. Journal of Ecology and Rural Environment, 2015, 31(6): 935−941.(in Chinese) doi: 10.11934/j.issn.1673-4831.2015.06.021
[31]	ZHOU S P, DUAN C Q, FU H, et al. Toxicity assessment for chlorpyrifos-contaminated soil with three different earthworm test methods [J]. Journal of Environmental Sciences, 2007, 19(7): 854−858. doi: 10.1016/S1001-0742(07)60142-9
[32]	LUKKARI T, HAIMI J. Avoidance of Cu-and Zn-contaminated soil by three ecologically different earthworm species [J]. Ecotoxicology and Environmental Safety, 2005, 62(1): 35−41. doi: 10.1016/j.ecoenv.2004.11.012
[33]	XIAO N W, JING B B, GE F, et al. The fate of herbicide acetochlor and its toxicity to Eisenia fetida under laboratory conditions [J]. Chemosphere, 2006, 62(8): 1366−1373. doi: 10.1016/j.chemosphere.2005.07.043
[34]	黄若男, 范军杰, 涂宏志, 等. 咪唑氯盐离子液体对蚯蚓急性毒性及体重影响研究 [J]. 环境科学, 2013, 34(4):1380−1385. HUANG R N, FAN J J, TU H Z, et al. Effects of imidazolium chloride ionic liquids on the acute toxicity and weight of earthworm [J]. Environmental Science, 2013, 34(4): 1380−1385.(in Chinese)
[35]	MOTTIER L, ALVAREZ L, CEBALLOS L, et al. Drug transport mechanisms in helminth parasites: Passive diffusion of benzimidazole anthelmintics [J]. Experimental Parasitology, 2006, 113(1): 49−57. doi: 10.1016/j.exppara.2005.12.004
[36]	高玉红, 孙振钧, 孙新胜, 等. 兽药阿苯哒唑对蚯蚓皮肤和肠道超显微结构的影响 [J]. 环境科学学报, 2008, 28(12):2578−2582. doi: 10.3321/j.issn:0253-2468.2008.12.028 GAO Y H, SUN Z J, SUN X S, et al. Effects of albendazole on the ultrastructure of the skin and intestinalepithelium of the earthworm [J]. Acta Scientiae Circumstantiae, 2008, 28(12): 2578−2582.(in Chinese) doi: 10.3321/j.issn:0253-2468.2008.12.028
[37]	李辉龙, 雷震, 吴尔苗, 等. 1, 2, 4-三氯苯对蚯蚓生长和表皮及肠道超微结构的影响 [J]. 生态毒理学报, 2011, 6(3):247−254. LI H L, LEI Z, WU E M, et al. Effects of 1, 2, 4-trichlorobenzene on growth and ultrastructure of skin and intestinal epithelium of earthworm [J]. Asian Journal of Ecotoxicolog, 2011, 6(3): 247−254.(in Chinese)
[38]	赵晓祥, 陈琪, 庄惠生. 壬基酚对赤子爱胜蚓的生态毒理学研究 [J]. 生态环境, 2006, 15(6):1185−1187. ZHAO X X, CHEN Q, ZHUANG H S. Ecotoxicology research of nonylphenol to Eisenia foetida [J]. Ecology and Environment, 2006, 15(6): 1185−1187.(in Chinese)
[39]	沈生荣, 杨贤强, 赵保路, 等. 茶多酚体外助氧化作用的自由基机理 [J]. 茶叶科学, 1992, 12(2):145−150. SHEN S R, YANG X Q, ZHAO B L, et al. Radical mechanism of prooxidant effect on tea polyphenols(TP)in vitro [J]. Journal of Tea Science, 1992, 12(2): 145−150.(in Chinese)
[40]	袁根良, 蒋丽, 殷光玲. 茶多酚急性毒性试验研究 [J]. 食品安全质量检测学报, 2015, 6(9):3730−3733. YUAN G L, JIANG L, YIN G L. Experimental study on acute toxicity of tea polyphenols [J]. Journal of Food Safety and Quality, 2015, 6(9): 3730−3733.(in Chinese)