中国四种土壤跳虫对重金属汞的响应

刘传栋，苗秀莲，贾少波，张广征，单丽英

聊城大学生命科学学院 生态学与生物多样性山东省重点实验室，聊城 252059

中国四种土壤跳虫对重金属汞的响应

刘传栋，苗秀莲，贾少波*，张广征，单丽英

聊城大学生命科学学院 生态学与生物多样性山东省重点实验室，聊城 252059

汞；跳虫；半致死浓度；慢性毒性实验；低剂量刺激效应

近期由环境保护部和国土资源部发布的《全国土壤污染状况调查公报》(2014年4月17日)表明，中国目前国土污染形势严峻，其中重金属对耕地的污染涉及粮食安全，尤其需要关注。我国污水灌溉中重金属Hg含量的高低与相对应的灌区土壤中重金属的累积量多少基本一致，目前受重金属污染的耕地已经有1/6左右，如何进行耕地重金属污染的监测、综合防治和生态修复，成为迫切需要考虑和解决的问题[1-4]。土壤动物作为土壤生态系统中的重要组成部分和土壤健康状况的体现者，选择其成员作为模式动物是探索土壤生态系统毒理学的重要途径。

1 材料和方法(Materials and methods)

1.1 仪器与试剂

仪器：RTOP多段编程人工气候箱(杭州托普仪器有限公司)，SZX12体视显微镜(Olympus公司)，BX51正置显微镜(Olympus公司)，BSA224S电子天平(Sartorius公司)。试剂：氯化汞(HgCl2)等化学试剂为国产分析纯试剂。

1.2 实验材料

整个实验及先期培养均在上口径120 mm、下口径88 mm、高65 mm的专用培养瓶(透明聚丙烯容器)中进行，培养瓶底部覆盖一层0.5 cm厚的琼脂培养基(1.5%)，顶部有透明聚丙烯盖封口。

1.3 实验方法

1.3.1 LC50急性毒性试验(存活试验)

暴露期间喂食含毒酵母糊，其中Hg2+含量对应6个实验浓度组分别为0.0、0.4、2.0、10、50、100 mg·kg-1干重。初始每隔12 h观察一次，24 h后每隔1天观察一次跟踪观察到第7天，并做好记录。死亡的判定标准是触之不动。

1.3.2 28 d慢性毒性试验

根据预试验结果，设置6个浓度0.05、0.5、1.0、2.0、4.0、8.0 mg·L-1的染毒组和一个对照组(0 mg·L-1)进行28 d慢性毒性试验。每组设3个重复。培养基染毒过程及暴露期间的实验方法与LC50急性毒性试验相同。同LC50急性毒性试验，暴露期间喂食含毒酵母糊，其Hg2+干重含量分别对应染毒组浓度。每周开口通气并补充少量蒸馏水。幼虫计数方法采用显微拍照定格后人工识别计数。

1.4 数据处理

数据用平均值±标准差(mean±SD)表示。采用SPSS Statistics 19.0统计分析软件；在95%的置信水平进行单因素方差分析(One-Way ANOVA)，Duncan法对平均值进行多重比较，p<0.05为差异显著；采用Bliss法(机率单位加权回归法)[14-15]计算汞(Hg2+)对跳虫的半致死浓度(LC50)。

2 结 果(Results)

2.1 LC50急性毒性实验

2.1.1 Hg急性毒性期间的跳虫行为观察

2.1.2 Hg对4种跳虫的24 h-LC50和72 h-LC50

2.1.3 急性实验中四种跳虫7 d内在不同Hg2+浓度下的存活情况

表1 Hg对四种跳虫24 h, 72 h的半致死浓度Table 1 24 h, 72 h-LC50 values of Hg to four species of Collembolans

表2 不同浓度Hg对曲毛裸长()7 d存活影响的F值和p值Table 2 F values and p values of different concentrations of Hg on survival of S. curviseta for 7 d

图1 不同浓度Hg对曲毛裸长()7 d存活的影响注：图柱上字母的不同表示差异达到 0.05 的显著水平。Fig. 1 Effect of different concentrations of Hg on survival of S. curviseta for 7 dNote: Different letters on columns mean significant difference at p<0.05.

图2 不同浓度Hg对小原等节()7 d存活的影响Fig. 2 Effect of different concentrations of Hg on survival of P. minuta for 7 d

暴露时间TimeofexposureF值Fvaluesp值pvalues12h168.750.0001d2064.500.0003d686.330.0005d893.10.0007d1014.600.000

表4 不同浓度Hg对四刺泡角()7 d存活影响的F值和p值Table 4 F values and p values of different concentrations of Hg on survival of C. duplicispinosa for 7 d

图3 不同浓度Hg对四刺泡角()7 d存活的影响Fig. 3 Effect of different concentrations of Hg on survival of C. duplicispinosa for 7 d

图4 不同浓度Hg对茉莉花长角()7 d存活的影响Fig. 4 Effect of different concentrations of Hg on survival of Entomobrya sp. for 7 d

暴露时间TimeofexposureF值Fvaluesp值pvalues12h16.240.0001d31.380.0003d60.590.0005d91.090.0007d58.320.000

2.2 28 d慢性毒性试验

图5 不同浓度Hg对茉莉花长角()28 d的影响Fig. 5 Effect of different concentrations of Hg on survival of Entomobrya sp.for 28 days

图6 不同浓度Hg对小原等节()28 d的影响Fig. 6 Effect of different concentrations of Hg on survival of P. minuta for 28 days

暴露时间TimeofexposureF值Fvaluesp值pvalues1d12.250.0007d6.980.00114d23.070.00021d13.100.00028d6.880.001

图7 不同汞浓度下28 d时茉莉花长角()和小原等节()的幼虫数量与成虫数量比注：折线图上的**表示差异达到 0.01 的极显著水平。Fig. 7 Ratios of live offspring to adults of Entomobrya sp. and P. minuta on Hg concentrations for 28 daysNote: ** means significant difference at p<0.01.

3 讨 论(Discussion)

3.1 不同Hg2+浓度对跳虫7 d内存活的影响

3.2 低浓度汞对跳虫存活和繁殖的影响

3.3 跳虫的24 h，72 h-LC50变化及其适应性意义

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ResponsesofFourSpeciesofChineseSoilCollembolatoHg

Liu Chuandong, Miao Xiulian, Jia Shaobo*, Zhang Guangzheng, Shan Liying

Shandong Key Laboratory of Ecology and Biodiversity, School of Life Sciences, Liaocheng University, Liaocheng 252059, China

15 May 2014accepted19 June 2014

To study the toxic effects of Hg on diverse species of Collembola, four Chinese dominant species of collembolans, Sinella curviseta, Ceratophysella duplicispinosa, Proisotoma minuta and Entomobrya sp. were exposed to six different concentrations of Hg for 24 h, 72 h LC50measurement. The LC50values for S. curviseta, C. duplicispinosa, P. minuta and Entomobrya sp. were calculated according to the Bliss method to be 92.42, 36.85, 34.93, and 47.56 mg·L-1, and 95% confidence limits were 68.57～168.6, 29.61～43.53, 25.57～44.94 and 34.33～65.11 mg·L-1, respectively, while the corresponding 72 h-LC50values were 21.92, 20.17, 18.44, and 15.99 mg L-1, and 95% confidence limits were 16.18～29.82, 6.64～29.81, 13.87～26.45 and 10.03～21.37 mg·L-1. At the same time, the Proisotoma minuta and Entomobrya sp. were exposed to Hg of six different lower concentrations for 28 d survival and reproduction experiment to determine the numbers of adults and offspring. The result shows that Entomobrya sp. and S. curviseta have higher tolerance for Hg, followed with P. minuta, while C. duplicispinosa ranks last. In addition, P. minuta display some hormesis at about 0.05 mg·L-1of Hg2+.

Hg; collembola; LC50; chronic toxicity test; hormesis

国家自然科学基金(No. 31272383) ；山东省自然科学基金(No. ZR2010CM018)

刘传栋(1976-)，男，硕士，研究方向为动物生态毒理学，E-mail: lcliucd@189.cn；

*通讯作者(Corresponding author)，E-mail: jiasbio@163.com

10.7524/AJE.1673-5897-20140515008

2014-05-15录用日期：2014-06-19

1673-5897(2014)5-964-08

： X171.5

： A

贾少波(1959—)，男，生态学博士，教授，主要研究方向为动物学及生态学。

刘传栋, 苗秀莲, 贾少波, 等. 中国四种土壤跳虫对重金属汞的响应[J]. 生态毒理学报，2014, 9(5): 964-971

Liu C D, Miao X L, Jia S B, et al. Responses of four species of Chinese soil collembola to Hg [J]. Asian Journal of Ecotoxicology, 2014, 9(5): 964-971 (in Chinese)