劉傳棟，苗秀蓮，賈少波，張廣征，單麗英

聊城大學生命科學學院 生態(tài)學與生物多樣性山東省重點實驗室，聊城 252059

中國四種土壤跳蟲對重金屬汞的響應

劉傳棟，苗秀蓮，賈少波*，張廣征，單麗英

聊城大學生命科學學院 生態(tài)學與生物多樣性山東省重點實驗室，聊城 252059

汞；跳蟲；半致死濃度；慢性毒性實驗；低劑量刺激效應

近期由環(huán)境保護部和國土資源部發(fā)布的《全國土壤污染狀況調(diào)查公報》(2014年4月17日)表明，中國目前國土污染形勢嚴峻，其中重金屬對耕地的污染涉及糧食安全，尤其需要關(guān)注。我國污水灌溉中重金屬Hg含量的高低與相對應的灌區(qū)土壤中重金屬的累積量多少基本一致，目前受重金屬污染的耕地已經(jīng)有1/6左右，如何進行耕地重金屬污染的監(jiān)測、綜合防治和生態(tài)修復，成為迫切需要考慮和解決的問題[1-4]。土壤動物作為土壤生態(tài)系統(tǒng)中的重要組成部分和土壤健康狀況的體現(xiàn)者，選擇其成員作為模式動物是探索土壤生態(tài)系統(tǒng)毒理學的重要途徑。

1 材料和方法(Materials and methods)

1.1 儀器與試劑

儀器：RTOP多段編程人工氣候箱(杭州托普儀器有限公司)，SZX12體視顯微鏡(Olympus公司)，BX51正置顯微鏡(Olympus公司)，BSA224S電子天平(Sartorius公司)。試劑：氯化汞(HgCl2)等化學試劑為國產(chǎn)分析純試劑。

1.2 實驗材料

整個實驗及先期培養(yǎng)均在上口徑120 mm、下口徑88 mm、高65 mm的專用培養(yǎng)瓶(透明聚丙烯容器)中進行，培養(yǎng)瓶底部覆蓋一層0.5 cm厚的瓊脂培養(yǎng)基(1.5%)，頂部有透明聚丙烯蓋封口。

1.3 實驗方法

1.3.1 LC50急性毒性試驗(存活試驗)

暴露期間喂食含毒酵母糊，其中Hg2+含量對應6個實驗濃度組分別為0.0、0.4、2.0、10、50、100 mg·kg-1干重。初始每隔12 h觀察一次，24 h后每隔1天觀察一次跟蹤觀察到第7天，并做好記錄。死亡的判定標準是觸之不動。

1.3.2 28 d慢性毒性試驗

根據(jù)預試驗結(jié)果，設(shè)置6個濃度0.05、0.5、1.0、2.0、4.0、8.0 mg·L-1的染毒組和一個對照組(0 mg·L-1)進行28 d慢性毒性試驗。每組設(shè)3個重復。培養(yǎng)基染毒過程及暴露期間的實驗方法與LC50急性毒性試驗相同。同LC50急性毒性試驗，暴露期間喂食含毒酵母糊，其Hg2+干重含量分別對應染毒組濃度。每周開口通氣并補充少量蒸餾水。幼蟲計數(shù)方法采用顯微拍照定格后人工識別計數(shù)。

1.4 數(shù)據(jù)處理

數(shù)據(jù)用平均值±標準差(mean±SD)表示。采用SPSS Statistics 19.0統(tǒng)計分析軟件；在95%的置信水平進行單因素方差分析(One-Way ANOVA)，Duncan法對平均值進行多重比較，p<0.05為差異顯著；采用Bliss法(機率單位加權(quán)回歸法)[14-15]計算汞(Hg2+)對跳蟲的半致死濃度(LC50)。

2 結(jié) 果(Results)

2.1 LC50急性毒性實驗

2.1.1 Hg急性毒性期間的跳蟲行為觀察

2.1.2 Hg對4種跳蟲的24 h-LC50和72 h-LC50

2.1.3 急性實驗中四種跳蟲7 d內(nèi)在不同Hg2+濃度下的存活情況

表1 Hg對四種跳蟲24 h, 72 h的半致死濃度Table 1 24 h, 72 h-LC50 values of Hg to four species of Collembolans

表2 不同濃度Hg對曲毛裸長()7 d存活影響的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

圖1 不同濃度Hg對曲毛裸長()7 d存活的影響注：圖柱上字母的不同表示差異達到 0.05 的顯著水平。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.

圖2 不同濃度Hg對小原等節(jié)()7 d存活的影響Fig. 2 Effect of different concentrations of Hg on survival of P. minuta for 7 d

暴露時間TimeofexposureF值Fvaluesp值pvalues12h168.750.0001d2064.500.0003d686.330.0005d893.10.0007d1014.600.000

表4 不同濃度Hg對四刺泡角()7 d存活影響的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

圖3 不同濃度Hg對四刺泡角()7 d存活的影響Fig. 3 Effect of different concentrations of Hg on survival of C. duplicispinosa for 7 d

圖4 不同濃度Hg對茉莉花長角()7 d存活的影響Fig. 4 Effect of different concentrations of Hg on survival of Entomobrya sp. for 7 d

暴露時間TimeofexposureF值Fvaluesp值pvalues12h16.240.0001d31.380.0003d60.590.0005d91.090.0007d58.320.000

2.2 28 d慢性毒性試驗

圖5 不同濃度Hg對茉莉花長角()28 d的影響Fig. 5 Effect of different concentrations of Hg on survival of Entomobrya sp.for 28 days

圖6 不同濃度Hg對小原等節(jié)()28 d的影響Fig. 6 Effect of different concentrations of Hg on survival of P. minuta for 28 days

暴露時間TimeofexposureF值Fvaluesp值pvalues1d12.250.0007d6.980.00114d23.070.00021d13.100.00028d6.880.001

圖7 不同汞濃度下28 d時茉莉花長角()和小原等節(jié)()的幼蟲數(shù)量與成蟲數(shù)量比注：折線圖上的**表示差異達到 0.01 的極顯著水平。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 討 論(Discussion)

3.1 不同Hg2+濃度對跳蟲7 d內(nèi)存活的影響

3.2 低濃度汞對跳蟲存活和繁殖的影響

3.3 跳蟲的24 h，72 h-LC50變化及其適應性意義

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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

國家自然科學基金(No. 31272383) ；山東省自然科學基金(No. ZR2010CM018)

劉傳棟(1976-)，男，碩士，研究方向為動物生態(tài)毒理學，E-mail: lcliucd@189.cn；

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

10.7524/AJE.1673-5897-20140515008

2014-05-15錄用日期：2014-06-19

1673-5897(2014)5-964-08

： X171.5

： A

賈少波(1959—)，男，生態(tài)學博士，教授，主要研究方向為動物學及生態(tài)學。

劉傳棟, 苗秀蓮, 賈少波, 等. 中國四種土壤跳蟲對重金屬汞的響應[J]. 生態(tài)毒理學報，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)