銅-磺胺嘧啶復(fù)合脅迫對蔬菜種子發(fā)芽的急性毒性效應(yīng)

金彩霞, 司曉薇, 毛蕾, 樊靜

河南師范大學(xué)環(huán)境學(xué)院 黃淮水環(huán)境與污染防治省部共建教育部重點實驗室 河南省環(huán)境污染控制重點實驗室, 新鄉(xiāng) 453007

銅-磺胺嘧啶復(fù)合脅迫對蔬菜種子發(fā)芽的急性毒性效應(yīng)

金彩霞*, 司曉薇, 毛蕾, 樊靜

河南師范大學(xué)環(huán)境學(xué)院 黃淮水環(huán)境與污染防治省部共建教育部重點實驗室 河南省環(huán)境污染控制重點實驗室, 新鄉(xiāng) 453007

研究了黃潮土中常用獸藥磺胺嘧啶(SD)與重金屬銅(Cu)單一及復(fù)合污染對小白菜和西紅柿種子發(fā)芽(發(fā)芽率、根伸長、芽伸長)的影響，分析了土壤中藥物濃度與作物生長抑制的劑量-效應(yīng)關(guān)系及復(fù)合污染的毒性效應(yīng)。結(jié)果表明，無論是在Cu或SD單一污染物作用下，根伸長和芽伸長抑制率與污染物濃度顯著相關(guān)(P<0.01)，發(fā)芽抑制率與污染物濃度不相關(guān)(P >0.05)；污染物對根伸長及芽伸長的抑制高于對種子發(fā)芽的抑制；SD對2種作物的毒性效應(yīng)明顯強于Cu，SD對小白菜和西紅柿根伸長的ID50(抑制率為50%時污染物濃度)分別為1.7和33.6 mg·kg-1，而Cu為273.6和457.7 mg·kg-1。Cu-SD復(fù)合污染，對西紅柿的根和芽伸長的拮抗作用顯著(P<0.01)；但對小白菜來說，在低濃度Cu(100 mg·kg-1)作用下，二者的聯(lián)合拮抗作用不顯著(P>0.05)，隨Cu的增加，拮抗作用顯著(P<0.01)。

磺胺嘧啶；Cu；小白菜；西紅柿；發(fā)芽抑制率；復(fù)合污染；生態(tài)毒性

隨著集約化、規(guī)?；B(yǎng)殖業(yè)的快速發(fā)展，大量獸藥被用來防治各種畜禽疾病。研究表明，大約有60%～90%獸藥隨動物糞便排出體外，含有大量獸藥的養(yǎng)殖場糞尿作為有機肥施入農(nóng)田后可造成農(nóng)田土壤污染[1]。這些獸藥在土壤中累積，進而影響其中的植物、動物和微生物的正常生命活動，最終將影響人類的身體健康[2]。因此，有關(guān)獸藥對植物的毒性效應(yīng)已受到研究者的關(guān)注[3]。

磺胺類藥物(SAs)是一類以對氨基苯磺酰胺為基本化學(xué)結(jié)構(gòu)的合成抗菌藥物，是我國生產(chǎn)和使用量最大的獸藥之一。新鮮豬糞中磺胺藥物可高達20～40 mg·kg-1，每畝農(nóng)田每年將有高達幾百克的磺胺藥物施入[4]。邰義萍等[5]通過對廣東省畜牧糞便中磺胺類抗生素含量的分布研究發(fā)現(xiàn)，豬糞中4種磺胺類藥物總含量在1.9259～13.3995 mg·kg-1之間，平均為4.4039 mg·kg-1，其中以磺胺甲基咪定和磺胺甲噁唑為主；牛糞中磺胺類化合物檢出率在90%以上，平均含量為3.7877 mg·kg-1，以磺胺甲噁唑和磺胺甲基嘧啶為主。劉峰等[6]調(diào)查發(fā)現(xiàn)，福建省廈門市和莆田市的3種磺胺類抗生素污染程度為磺胺二甲嘧啶>磺胺甲噁唑>磺胺嘧啶，所有樣品中均檢出磺胺類抗生素，畜牧糞便中平均值15.1 μg·kg-1。Cu是植物生長發(fā)育所必需的營養(yǎng)元素，但若超量亦會造成污染。其可以通過工業(yè)排放、化肥施用、污水灌溉、污泥農(nóng)用等多種途徑進入農(nóng)田生態(tài)系統(tǒng)，影響作物的生長、發(fā)育并通過食物鏈富集到動物體和人體，從而帶來危害[7]。因此，農(nóng)田生態(tài)系統(tǒng)中磺胺類藥物-銅的復(fù)合污染較普遍。而目前針對此類復(fù)合污染開展的生態(tài)毒理研究較少，有必要加強相關(guān)方面的研究。

本研究以黃潮土作為供試土壤，研究了Cu-SD單一及復(fù)合污染對西紅柿和小白菜種子根伸長、芽伸長和發(fā)芽率的影響。通過作物種子生長受抑制程度，評價Cu-SD單一及復(fù)合污染的生態(tài)毒性及可能對農(nóng)業(yè)生產(chǎn)帶來的環(huán)境風(fēng)險。

1 材料與方法(Materials and methods)

1.1 實驗材料

1.1.1 藥品

磺胺嘧啶(SD)原藥，純度93%，購自北京博亞華牧業(yè)科技有限公司；CuSO4·5H2O 分析純，廣州化學(xué)試劑廠生產(chǎn)。

1.1.2 供試種子和土壤

小白菜(Raphanus sativus，品種為上海青)，發(fā)芽率為98%；西紅柿(Cyphomandra betacea)合作908，發(fā)芽率為95%，均購自新鄉(xiāng)市種子公司。供試土壤采自河南師范大學(xué)校內(nèi)生物實驗田0～20 cm表土，其主要理化性質(zhì)見表1。

1.2 實驗方法

1.2.1 預(yù)備試驗

土壤采自河南師范大學(xué)生物園，屬于未受任何化學(xué)品污染的潔凈土壤，其理化性質(zhì)如下：pH 6.31；陽離子交換量12.26 cmol·kg-1；TP 0.04 mg·kg-1；TK 0.18 mg·kg-1；有機質(zhì)1.65%；粘粒13.4%；粉粒24.2%。土壤風(fēng)干研磨，過2 mm篩(10目)，備用。稱取50 g 備用土壤于90 mm直徑的玻璃培養(yǎng)皿中，加入不同劑量的SD和Cu，并用去離子水調(diào)節(jié)土壤含水量至田間最大持水量的60%，將其置于恒溫培養(yǎng)箱中25 ℃下平衡48 h后，用鑷子將植物種子均勻播種于土壤中(種子放置時，保持種子胚根末端和生長方向呈直線)，蓋好玻璃培養(yǎng)皿，置于恒溫培養(yǎng)箱25 ℃暗處培養(yǎng)。當(dāng)對照種子發(fā)芽率>90%，根長度為20 mm時，實驗結(jié)束。確定種子根和芽伸長抑制濃度，使其抑制率(ID)在10%～60%之間，開始正式試驗。

表1 供試土壤的主要理化性質(zhì)Table 1 Main physicochemical properties of the soil samples

注: 1)陽離子交換量; 2)總磷; 3)總鉀; 4)凱氏氮。

Note: 1) Cation exchange capacity, 2) Total phosphorus, 3) Total potassium, 4) Kjeldahl nitrogen.

抑制率(ID)可通過式(1)計算：

ID = (1-B/A)×100%

(1)

式中，A為對照根或芽的伸長，mm；B為處理組中根或芽的伸長，mm。

1.2.2 正式試驗

1.2.2.1 單一污染實驗

根據(jù)預(yù)備實驗結(jié)果，在種子發(fā)芽和根伸長IC達到抑制率10%～60%范圍內(nèi)，設(shè)置6個不同處理濃度(見表2)，每個處理20粒種子，設(shè)3次重復(fù)。實驗結(jié)束時，測定各處理的根長(根長的測定以胚軸與根之間的過點開始)和芽長，計算各指標(biāo)的平均值及標(biāo)準偏差，并繪制劑量-抑制率曲線，進行回歸分析，計算ID50。

1.2.2.2 復(fù)合污染實驗

為了與單一污染實驗相對應(yīng)，小白菜和西紅柿的SD濃度同樣設(shè)定為0.5、0.8、1.0、1.5、2.0、2.5和2.0、5.0、10.0、15.0、20.0、25.0 mg·kg-1。Cu的濃度設(shè)置為0、100.0、300.0 mg·kg-1，每個處理20粒種子，3次重復(fù)。

1.3 數(shù)據(jù)分析

采用SPSS17.0和Origin7.5數(shù)據(jù)處理軟件(美國Microcal Software公司)對數(shù)據(jù)進行方差分析及回歸分析，計算出ID50。對劑量-效應(yīng)曲線進行擬合并作圖。實驗數(shù)據(jù)結(jié)果采用平均值±標(biāo)準差(X±SD)表示，差異顯著性水平為P<0.05。

2 結(jié)果與討論(Results and discussion)

2.1 SD對西紅柿和小白菜根伸長和芽伸長的影響

從圖1和表3可見，土壤中SD的濃度與西紅柿和小白菜的根/芽伸長抑制率之間呈線性相關(guān)。從表3可以看出，無論是西紅柿或小白菜，SD對根伸長的抑制都大于對芽伸長的抑制。從ID50可以得出，小白菜根對SD相對于西紅柿根更敏感，當(dāng)土壤中的SD濃度為1.77 mg·kg-1時，小白菜根伸長的抑制率即達到50%，而土壤中的SD濃度為33.64 mg·kg-1，西紅柿根的抑制率才達到50%。由相關(guān)分析可知，西紅柿和小白菜根伸長抑制率與SD濃度的線性相關(guān)性(R2=0.9678，R2=0.9401)要好于芽伸長 (R2=0.9108，R2=0.7921)。此外，方差分析表明，土壤中SD的濃度與西紅柿根/芽伸長和小白菜的根伸長之間具有顯著相關(guān)性(P<0.01)，而與小白菜芽伸長之間不具有相關(guān)性(P>0.05)。

磺胺類藥物對西紅柿和小白菜根/芽伸長產(chǎn)生較強的毒害效應(yīng)。其毒性主要是植物體內(nèi)該污染物與葉酸相互競爭的結(jié)果，葉酸與嘌呤合成有關(guān)，而嘌呤是細胞分裂素和脫落酸的前體，因此,植物吸收獸藥而降低了對葉酸的吸收，從而影響了其正常的生理功能[8-9]。在SD的脅迫下，作物可能因吸收藥物，體內(nèi)缺乏細胞生長所需的葉酸，故根和芽伸長受到較強的抑制，但其具體作用機制有待于進一步研究。

2.2 Cu對西紅柿和小白菜根伸長和芽伸長的影響

由圖2及表4可知，土壤中Cu濃度與西紅柿和小白菜的根/芽伸長抑制率之間具有顯著線性相關(guān)性(P<0.01)。當(dāng)土壤中Cu濃度為100 mg·kg-1時, 西紅柿和小白菜的根伸長抑制率分別為-3.56%和-1.24%，此時Cu促進了作物的生長。而在高濃度Cu脅迫下，作物受害癥狀明顯，根伸長受到顯著抑制，Cu濃度為250 mg·kg-1時，小白菜根伸長抑制率為48.3%，當(dāng)Cu濃度達到500 mg·kg-1時，西紅柿根伸長抑制率為53.0%。由ID50可知，與SD對西紅柿和小白菜的影響類似，Cu對根伸長的抑制大于對芽伸長的抑制，且小白菜根對土壤中Cu濃度要比西紅柿根更敏感。與SD對作物種子發(fā)芽影響相同，在試驗濃度范圍內(nèi)，土壤中Cu的濃度對西紅柿和小白菜種子發(fā)芽抑制率無顯著影響(P>0.05)。

表2 單一因素作用下兩種污染物的實驗濃度Table 2 The concentration of two drugs in the single exposure (mg·kg-1)

圖1 SD污染對西紅柿和小白菜根/芽伸長的影響

表3 作物根/芽伸長與土壤中SD濃度的關(guān)系Table 3 Relationship between inhibitory rate of shoot/root elongation and concentration of SD

Cu是植物生長發(fā)育所必需的營養(yǎng)元素，它參與植物光合作用，并且是超氧化物歧化酶、多酚氧化酶、細胞色素氧化酶和抗壞血酸氧化酶等組成成分[10]，當(dāng) Cu 過量時，又會對植物產(chǎn)生毒害從而使植物根伸長受阻、根莖生物量減少等[11], 其原因可能是過量的 Cu 使細胞膜的通透性下降，導(dǎo)致Cu滲入細胞膜而產(chǎn)生毒害作用，進而影響細胞器的結(jié)構(gòu)和功能。此外，有研究表明，Cu 能夠破壞植物根尖細胞有絲分裂過程中紡錘體的形成，從而使根尖細胞無法正常分裂，最終阻礙根的伸長[12-13]。

從2種作物的ID50值可知，無論是在 SD或Cu的脅迫下，污染物對小白菜的生態(tài)毒性效應(yīng)要強于西紅柿。此外，SD 對作物的生態(tài)毒性遠大于 Cu。Cu 對小白菜根伸長的ID50值是SD 的154 倍；Cu對于西紅柿根伸長的ID50值是SD的13.6倍。因此，相對于傳統(tǒng)重金屬污染，獸藥抗生素作為新型的環(huán)境污染物，對生態(tài)環(huán)境可能造成的危害應(yīng)引起重視。

圖2 Cu污染對西紅柿和小白菜根/芽伸長的影響

表4 作物根/芽伸長與土壤中Cu濃度的關(guān)系Table 4 Relationship between inhibitory rate of shoot/root elongation and concentration of Cu

2.3 Cu-SD復(fù)合污染對西紅柿和小白菜種子根伸長和芽伸長的影響

由圖3和表5可知，Cu-SD復(fù)合污染時，土壤中SD含量與西紅柿根伸長和芽伸長抑制率之間具有顯著相關(guān)性(P <0.01)，但當(dāng)Cu濃度為300 mg·kg-1時，SD含量與西紅柿根伸長抑制率之間相關(guān)性不顯著(P >0.05)，而與西紅柿芽伸長抑制率之間卻顯著相關(guān)(P <0.01)。土壤中SD含量與西紅柿根伸長和芽伸長抑制率之間相關(guān)關(guān)系，隨著Cu濃度的增加而變化， SD與西紅柿根伸長抑制率間的相關(guān)性隨Cu濃度增加而減小，而與芽伸長抑制率之間的相關(guān)性增大。在Cu-SD復(fù)合作用下，Cu與SD對西紅柿根/芽伸長有明顯的拮抗作用。Cu的加入降低了SD對西紅柿根伸長和芽伸長的抑制作用，隨著Cu加入量的增加，拮抗作用增強。當(dāng)土壤中SD濃度為20 mg·kg-1時，在SD單一作用下，西紅柿根/芽伸長抑制率分別為33.6%和21.4%；當(dāng)加入100 mg·kg-1的Cu時，西紅柿根/芽伸長抑制率分別減少到18.8%和14.3%,；當(dāng)其與300 mg·kg-1的Cu復(fù)合時，西紅柿根/芽伸長抑制率分別減少到13.3%和13.9%。由回歸方程可知，在Cu濃度分別為0、100、300 mg·kg-1西紅柿的根伸長抑制率達到50%時，土壤中SD濃度隨Cu濃度的增加而增加，其ID50分別為33.6、75.5和160.3 mg·kg-1。

由圖4和表5可知，SD和Cu復(fù)合污染與小白菜根/芽伸長抑制率相關(guān)性達到顯著水平(P <0.05)，當(dāng)Cu濃度為300 mg·kg-1時，小白菜芽伸長抑制率和土壤中SD濃度具有極顯著相關(guān)性(P <0.01)。由R2可知，在SD單一作用下，SD與小白菜的根伸長相關(guān)性較好，但與芽伸長抑制率相關(guān)性較差，隨Cu濃度的增加，SD與小白菜根伸長抑制率之間的相關(guān)性變差。土壤中Cu濃度為100 mg·kg-1或300 mg·kg-1時，對于根伸長抑制率來說，Cu與SD表現(xiàn)為拮抗作用。二者復(fù)合污染對小白菜芽伸長抑制率的影響比較復(fù)雜，當(dāng)Cu的濃度為100 mg·kg-1時，這兩種污染物之間為協(xié)同作用；當(dāng)Cu濃度為300 mg·kg-1時則為拮抗作用。由圖4可知，與單一污染相比，在相同的SD濃度下，加入Cu后，小白菜根伸長抑制率降低，但是小白菜芽伸長抑制率卻明顯升高，這說明Cu的加入降低了SD對根的毒性卻增加了對芽的毒性。此外，在Cu濃度分別為0、100、 300 mg·kg-1時，對根的ID50(SD)分別為1.8、14.7和4.7 mg·kg-1。由此得出，土壤中添加Cu后，SD對小白菜的毒性有降低的趨勢。

圖3 Cu-SD復(fù)合污染對西紅柿根伸長和芽伸長的毒性效應(yīng)

表5 在相同濃度Cu作用下西紅柿和小白菜根/芽伸長抑制率與土壤中SD的關(guān)系Table 5 Relationships between inhibitory rate of root/shoot elongation of tomato and Chinese cabbage and concentration of SD at the same concentration of Cu

圖4 Cu-SD復(fù)合污染對小白菜根伸長和芽伸長的毒性效應(yīng)

Liu等[14]研究了農(nóng)藥氯氰菊酯(CPM)與Cu復(fù)合污染對白菜種子發(fā)芽的影響，結(jié)果表明：土壤中CPM-Cu復(fù)合對白菜根伸長的影響要小于單一Cu的作用，這與本實驗研究結(jié)果相同。本實驗中SD和Cu復(fù)合污染對西紅柿根和芽伸長的抑制作用和小白菜根伸長的抑制作用整體上要小于SD單一作用，這可能與污染物和作物之間的作用機制有關(guān)。目前關(guān)于復(fù)合污染的作用機理主要有競爭結(jié)合位點、影響生物酶的活性、干擾生物正常的生理過程、改變生物細胞結(jié)構(gòu)和功能、螯合(或絡(luò)合)和沉淀作用以及干擾生物大分子的結(jié)構(gòu)和功能等幾種學(xué)說[15-17]。張娟等[19]研究表明，土壤環(huán)境中有機質(zhì)含量是影響磺胺嘧啶吸附行為的重要因素，土壤中有機物與重金屬的復(fù)合污染也可能會降低重金屬毒性[18]，在SD的脅迫下，土壤因吸附、解吸SD，某些土壤表面特性發(fā)生改變，可能吸附更多的Cu而降低其生物有效性，減少了作物根對SD和Cu的吸收，從而降低了二者復(fù)合污染對作物的毒害作用。Inaba及Takenaka[20]研究表明，人工合成螯合劑和有機酸能夠降低Cu的生物毒性。兩種不同的化合物交互作用時，其自身的基團會發(fā)生作用從而改變其化學(xué)結(jié)構(gòu)[21]，可能導(dǎo)致拮抗作用?；前奉愃幬锟膳cCu生成鹽沉淀[22-23]，SD中含有對氨基苯磺酰胺等，這些基團可能與Cu形成穩(wěn)定的SD- Cu復(fù)合體，從而使SD的毒性減低甚至失去活性。而SD和Cu復(fù)合污染對小白菜芽伸長的抑制作用卻大于單一SD作用機制有待于進一步的研究。總之，有機化學(xué)物質(zhì)與重金屬間的聯(lián)合作用機制很復(fù)雜，其毒性一般由藥物的交互作用和物質(zhì)間的濃度組合以及污染物本身的化學(xué)性質(zhì)來決定，在一定條件下，污染物暴露的濃度組合關(guān)系甚至起決定作用[24]。

綜上所述，可知：

在SD或Cu單一污染條件下，土壤中污染物濃度與小白菜和西紅柿根伸長和芽伸長抑制率顯著相關(guān)(P <0.01)，污染物對2種作物的根伸長抑制率大于芽伸長抑制率和種子發(fā)芽率。

從藥物對作物生長抑制的強弱及ID50值來看，SD對2種作物的生態(tài)毒性效應(yīng)遠強于重金屬Cu；在本實驗中，小白菜相對于西紅柿更敏感些。

Cu-SD復(fù)合污染對西紅柿生長的聯(lián)合作用主要體現(xiàn)為拮抗作用，隨著SD濃度的增加，二者對西紅柿的聯(lián)合拮抗效應(yīng)增加，而對小白菜根伸長的聯(lián)合作用體現(xiàn)為拮抗作用，對芽伸長主要體現(xiàn)為協(xié)同作用。

[1] Meyer M T, Bumgarner J E, Varns J L, et al. Use of radioimmunoassay as a screen for antibiotics in confined animal feeding operations and confirmation by liquid chromatography/mass spectrometry [J]. Science of the Total Environment, 2000, 248(2-3): 181-187

[2] Wu T, Li S F, Dai G, et al. Research process of veterinary drug residues of fluoroquinolones in animal food [J]. China Animal Health, 2012(3): 19-22

[3] Boxall A B A, Johnson P, Smith E J, et al. Uptake of veterinary medicines from soils into crops [J]. Journal of Agricultural and Food Chemistry, 2006, 54(6): 2288-2297

[4] Burkhardt M, Stoob K, Stamm C, et al. Veterinary antibiotics in animal slurries - A new environmental issue in grassland research [J]. Grassland Science in Europe, 2004, 9: 322-324

[5] 邰義萍, 羅曉棟, 莫測輝, 等. 廣東省畜牧糞便中喹諾酮類和磺胺類抗生素的含量與分布特征研究[J]. 環(huán)境科學(xué), 2011, 32(4): 1188-1193

Tai Y P, Luo X D, Mo C H, et al. Occurrence of quinolone and sulfonamide antibiotics in swine and cattle manures from large-scale feeding operation of Guanggong Province [J]. Environmental Science, 2011, 32(4): 1188-1193(in Chinese)

[6] 劉鋒, 廖德潤, 李可, 等. 畜禽養(yǎng)殖基地磺胺類喹諾酮類和大環(huán)內(nèi)酯類抗生素污染特征[J]. 農(nóng)業(yè)環(huán)境科學(xué)學(xué)報, 2013, 32(4): 847-853

Liu F, Miu D R, Li K, et al. Pollution characteris of the sulfonamindes, quinolones and macrolides in the samples collected from livestock and poultry feedlots [J]. Journal of Agro-Environment Science, 2013, 32(4): 847-853 (in Chinese)

[7] 王小驪, 張永志, 王剛軍, 等. 蔬菜中有害重金屬元素污染研究進展[J]. 浙江農(nóng)業(yè)學(xué)報, 2004, 16(5): 259-262

Wang X L, Zhang Y Z, Wang G J, et al. Advance of study on harmful heavy metal pollution on vegetables [J]. Acta Agriculture Zhejiangensis, 2004, 16(5): 259- 262 (in Chinese)

[8] 金彩霞, 劉軍軍, 陳秋穎, 等. 獸藥污染土壤對小麥和白菜根伸長抑制的毒性效應(yīng)[J]. 農(nóng)業(yè)環(huán)境科學(xué)學(xué)報, 2009, 28(7): 1358-1362

Jin C X, Liu J J, Chen Q Y, et al. Toxicological effects of veterinary drugs in soil on the inhibition of root elongation of wheat and Chinese cabbages [J]. Journal of Agro-Environment Science, 2009, 28(7): 1358-1362 (in Chinese)

[9] Jin C X, Chen Q Y, Sun R L, et al. Ecotoxic effects of sulfadiazine sodium, sulfamonomethoxine sodium and enrofloxacin on wheat, Chinese cabbage and tomato [J]. Ecotoxicology, 2009, 18: 878-885

[10] 薛艷, 沈振國, 周東美. 蔬菜對土壤重金屬吸收的差異與機理[J]. 土壤, 2005, 37(1): 32-36

Xue Y, Shen Z G, Zhou D M. Difference in heavy mental uptake between various vegetables and its mechanism [J]. Soil, 2005, 37(1): 32-36 (in Chinese)

[11] 鄭愛珍, 劉傳平, 沈振國. 銅對白菜、青菜生長的影響[J]. 北方園藝, 2005(2): 42-43

Zheng A Z, Liu C P, Shen Z G. Influences of copper on cabbage, and Chinese cabbage grown [J]. Northern Horticulture, 2005(2): 42-43 (in Chinese)

[12] 錢曉薇. 硫酸銅對蠶豆根尖細胞有絲分裂的影響[J]. 細胞生物學(xué)雜志, 2004, 26(1): 171-175

Qian X W. The effect of copper sulphate on mitosis of Vicia faba root tip cells [J]. Chinese Journal of Cell Biology, 2004, 26(1): 171-175 (in Chinese)

[13] Arduini I, Godbold D L, Onnis A. Cadmium and copperchange root growth and morphology of Pinus pinea and Pinus pinaster seedlings [J]. Physiologia Plantarum, 1994, 92(4): 675-680

[14] Liu T F, Wang T, Sun C, et al. Single and joint toxicity of cypermethrin and copper on Chinese cabbage (Pakchoi) seeds [J]. Journal of Hazardous Materials, 2009, 163: 344-348

[15] Stewart A R, Malley D F. Effect of metal mixture (Cu, Zn, Pb and Ni) on cadmium partitioning in littoral sediments and its accumulation by the freshwater macrophytc Eriocaùlon septangulàre [J]. Environmental Toxicology and Chemistry, 1999, 18(3): 436-447

[16] Sharma S S, Schat H, Vooijs R, et al. Combination toxicology of copper, zinc, and cadmium in binary mixtures: Concentration dependent antagonistic, nonadditive, and synergistic effects on root growth in Silene vulgaris [J]. Environmental Toxicology and Chemistry, 1999, 18(2): 348-355

[17] 王米道, 程鳳俠, 司友斌. 銅與草甘膦復(fù)合污染對小麥種子發(fā)芽與根伸長的抑制作用[J]. 生態(tài)毒理學(xué)報, 2009, 4(4): 591-596

Wang M D, Cheng F X, Si Y B. The inhibition of the combined pollution of copper and glyphosate to the seed germination and root elongation of wheat [J]. Asian Journal of Ecotoxicology, 2009, 4(4): 591-596

[18] Chao L, Zhou Q X, Chen S, et al. Single and joint stress of acetochlor and Pb on three agricultural crops in Northeast China [J]. Journal of Environmental Science, 2007, 19: 719-724

[19] 張娟, 郭文錄, 孫成. 磺胺嘧啶在黑土及其不同粒徑組分中的吸附行為研究[J]. 農(nóng)業(yè)環(huán)境科學(xué)學(xué)報, 2011, 30(2): 301-306

Zhang J, Guo W L, Sun C. Asorption behavior of sulfadiazine on black soil and its different particle sizes [J]. Journal of Agro-Environment Science, 2011, 30(2): 301-306

[20] Inaba S, Takenaka C. Effects of dissolved organic matter on toxicity and bioavailability of copper for lettuce sprouts [J]. Environment International, 2005, 31: 603-608

[21] Chauhan L K S, Gupta S K. Combined cytogenetic and ultrastructural effects of substituted urea herbicides and synthetic pyrethroid insecticide on the root meristem cells of Allium cepa [J]. Pesticide Biochemistry and Physiology, 2005, 82: 27-35

[22] 霍燕蘭. 藥物分析技術(shù)[M]. 北京: 化學(xué)工業(yè)出版社, 2005

Huo Y L. Drug Analysis Technology [M]. Beijing: Chemical Industry Press, 2005

[23] 金彩霞, 劉軍軍, 鮑林林, 等. 磺胺間甲氧嘧啶-鎘復(fù)合污染對作物種子發(fā)芽的影響[J]. 中國環(huán)境科學(xué), 2010, 30(6): 839-844

Jin C X, Liu J J, Bao L L, et al. Joint toxicity of sulfamonomethoxine and Cd on seed germination and root elongation of crops in soil [J]. China Environmental Science, 2010, 30(6): 839-844

[24] 周啟星, 程云, 張倩茹, 等. 復(fù)合污染生態(tài)毒理效應(yīng)的定量分析關(guān)系[J]. 中國科學(xué)(C輯), 2003, 33(6): 566-573

Zhou Q X, Cheng Y, Zhang Q R, et al. Quantitative analysis of the relationship between ecotoxicological effects of complex pollution [J]. Science China, 2003, 33(6): 566-573

◆

欄目簡介

“國際優(yōu)秀期刊論文精選”是《生態(tài)毒理學(xué)報》2015年10月推出的一個全新欄目。近年來，生態(tài)毒理學(xué)在國內(nèi)發(fā)展迅速，國際權(quán)威期刊上刊載了很多中國學(xué)者新穎獨特、系統(tǒng)深入的研究成果，這些成果或可啟迪科研新思路，或可服務(wù)政府管理，或可助力企業(yè)創(chuàng)新，但因語言原因，在國內(nèi)的傳播受到一定限制。同時，越來越多的華人科學(xué)家已成為各國科研領(lǐng)域的中堅力量，身在海外的他們希望自己的研究成果在祖國得到更廣泛的傳播和應(yīng)用。另一方面，國內(nèi)學(xué)者以中文發(fā)表的優(yōu)秀研究成果也需要在更大的國際學(xué)術(shù)舞臺上推廣傳播。因此，《生態(tài)毒理學(xué)報》將陸續(xù)與國際權(quán)威期刊開展合作，向?qū)W報的忠實讀者推介以英文發(fā)表的優(yōu)秀科研成果，并將作者以中文發(fā)表的優(yōu)秀科研成果推廣到國際平臺上參與學(xué)術(shù)交流。

目前，《生態(tài)毒理學(xué)報》已與Environmental Toxicology and Chemistry (ET&C)達成協(xié)議，互相推薦介紹對方發(fā)表的優(yōu)秀論文，并有幸邀請到南京大學(xué)的張效偉教授(個人簡介)擔(dān)綱“國際優(yōu)秀期刊論文精選”欄目的特邀主編。

ET&C期刊上與之對應(yīng)的欄目名稱為“Global Spotlight”，ET&C的主編G. Allen Burton, Jr教授親自參與這個欄目的策劃和編輯。

張效偉教授和Burton教授從雙方合作策劃開始直到欄目的最終出版都付出了寶貴的時間和精力，在此向兩位教授致以最誠摯的謝意！

Environmental Toxicology and Chemistry (ET&C)是一本在毒性學(xué)和環(huán)境科學(xué)領(lǐng)域都廣受贊譽的權(quán)威期刊，由The Society of Environmental Toxicology and Chemistry (SETAC)主辦，創(chuàng)刊于1982年，主要報道環(huán)境毒理學(xué)、環(huán)境化學(xué)及生態(tài)風(fēng)險評價方面的研究成果。該刊物影響因子為3.225，在毒理學(xué)領(lǐng)域88本SCI期刊中排名23位，在環(huán)境科學(xué)領(lǐng)域223本SCI期刊中排名45位。

(ET&C)的“Global Spotlight”欄目第一期中精選了《生態(tài)毒理學(xué)報》“環(huán)境基準研究進展”專輯(2015年第1期)中的3篇文章，分別是：

1)張遠,丁森,趙茜等撰寫的”基于野外數(shù)據(jù)建立大型底棲動物電導(dǎo)率水質(zhì)基準的可行性探討”(生態(tài)毒理學(xué)報, 2015, 10(1): 204-214)

2)王穎,馮承蓮,吳豐昌等撰寫的“物種敏感度分布的非參數(shù)核密度估計模型”(生態(tài)毒理學(xué)報, 2015, 10(1): 215-224)

3)金小偉, 王子健，王業(yè)耀等撰寫的“淡水水生態(tài)基準方法學(xué)研究：繁殖/生殖毒性類化合物水生態(tài)基準探討”(生態(tài)毒理學(xué)報, 2015, 10(1): 31-39)

第一期

1. 腐殖酸可降低納米銀的水生生物毒性

Humic substances alleviate the aquatic toxicity of polyvinylpyrrolidone-coated silver nanoparticles to organisms of different trophic levels

內(nèi)容提要：本文作者主要研究了腐殖酸對聚乙烯吡咯烷酮包覆的納米銀顆粒(polyvinylpyrrolidone-coated AgNPs)毒性的影響，受試生物涵蓋了水生系統(tǒng)不同的營養(yǎng)級別，包括藻類(Raphidocelis subcapitata)、水蚤類(Chydorus sphaericus)以及淡水魚類(Danio rerio)。結(jié)果顯示，腐殖酸可降低AgNPs對本研究中所有水生生物的毒性，并具有明顯的劑量效應(yīng)關(guān)系。原因為：1)腐殖酸使AgNPs表面帶有更多負電荷，這阻礙了AgNPs與藻細胞的接觸，使毒性降低；2)腐殖酸抑制了AgNPs中Ag+的溶出，而本研究顯示自由Ag+的毒性高于團聚的納米銀顆粒。

詳情請見：http://onlinelibrary.wiley.com/doi/10.1002/etc.2936/full

Abstract: The present study investigated how humic substances (HS) modify the aquatic toxicity of silver nanoparticles (AgNPs) as these particles agglomerate in water and interact with HS. An alga species (Raphidocelis subcapitata), a cladoceran species (Chydorus sphaericus), and a freshwater fish larva (Danio rerio), representing organisms of different trophic levels, were exposed to colloids of the polyvinylpyrrolidone-coated AgNPs in the presence and absence of HS. Results show that the presence of HS alleviated the aquatic toxicity of the AgNP colloids to all the organisms in a dose-dependent manner. The particle size distribution of the AgNPs' colloidal particles shifted to lower values due to the presence of HS, implying that the decrease in the toxicity of the AgNP colloids cannot be explained by the variation of agglomeration size. The surface charge of the AgNPs was found to be more negative in the presence of high concentrations of HS, suggesting an electrostatic barrier by which HS might limit interactions between particles and algae cells; indeed, this effect reduced the algae toxicity. Observations on silver ions (Ag+) release show that HS inhibit AgNP dissolution, depending on the concentrations of HS. When toxic effects were expressed as a function of each Ag-species, toxicity of the free Ag+was found to be much higher than that of the agglomerated particles.

View more in http://onlinelibrary.wiley.com/doi/10.1002/etc.2936/full

精選自：Zhuang Wang, Joris T.K. Quik, Lan Song, Evert-Jan Van Den Brandhof, MarjaWouterse and Willie J.G.M. Peijnenburg. Humic substances alleviate the aquatic toxicity of polyvinylpyrrolidone-coated silver nanoparticles to organisms of different trophic levels. Environmental Toxicology and Chemistry: Volume 34, Issue 6, pages 1239-1245, June 2015. DOI: 10.1002/etc.2936

2. “生物有效性測量”與“全沉積物毒性識別評估(TIE)”聯(lián)用有效識別出沉積物中主要致毒污染物

Identifying the causes of sediment-associated toxicity in urban waterways in South China: Incorporating bioavailabillity-based measurements into whole-sediment toxicity identification evaluation

內(nèi)容提要：沉積物中污染物種類繁多，準確判斷其中產(chǎn)生生物毒性的主要來源是個難點，本文作者先采用TIE法初步判斷出主要致毒污染物為有機物和重金屬(毒性描述階段(相I))，傳統(tǒng)的毒性單位分析結(jié)果顯示Cr、Cu、Ni、Pb和Zn主要致毒重金屬，而氯氰菊酯、氯氟氰菊酯、溴氰菊酯和氟蟲腈為主要致毒有機物中(毒性鑒定階段(相II))。采用4步分級提取法和Tenax提取法分析了重金屬和有機物的生物有效性。生物有效性毒性單位分析更加準確地鎖定了毒性主要貢獻重金屬為Zn、Ni和Pb，毒性主要貢獻有機物為氯氰菊酯、氯氟氰菊酯和氟蟲腈。沉積物的稀釋降低了重金屬的毒性并使其毒性貢獻鑒定變得復(fù)雜，生物有效性測量可以有效地提高TIE結(jié)果的準確性。

詳情請見：http://onlinelibrary.wiley.com/doi/10.1002/etc.2970/full

Abstract: Sediments in urban waterways of Guangzhou, China, were contaminated by a variety of chemicals and showed prevalent toxicity to benthic organisms (see Figure 1). A combination of whole-sediment toxicity identification evaluation (TIE) and bioavailability-based extraction was used to identify the causes of sediment toxicity. Of the 6 sediment samples collected, 4 caused 100% mortality to Chironomus dilutes in 10-d bioassays, and the potential toxicants were assessed using TIE in these sediments after dilution. The results of phase I characterization showed that organic contaminants were the principal contributors to the mortality of the midges in 2 sediments and that metals and organics jointly caused the mortality in the other 2 sediments. Ammonia played no role in the mortality for any samples. Conventional toxic unit analysis in phase II testing identified Cr, Cu, Ni, Pb, and Zn as the toxic metals, with cypermethrin, lambda-cyhalothrin, deltamethrin, and fipronils being the toxic organics. To improve the accuracy of identifying the toxicants, 4-step sequential extraction and Tenax extraction were conducted to analyze the bioavailability of the metals and organics, respectively. Bioavailable toxic unit analysis narrowed the list of toxic contributors, and the putative toxicants included 3 metals (Zn, Ni, and Pb) and 3 pesticides (cypermethrin, lambda-cyhalothrin, and fipronils). Metals contributed to the mortality in all sediments, but sediment dilution reduced the toxicity and confounded the characterization of toxicity contribution from metals in 2 sediments in phase I. Incorporating bioavailability-based measurements into whole-sediment TIE improved the accuracy of identifying the causative toxicants in urban waterways where multiple stressors occurred and contributed to sediment toxicity jointly.

View more in http://onlinelibrary.wiley.com/doi/10.1002/etc.2970/full

精選自：Xiaoyi Yi, Huizhen Li, Ping Ma and Jing You. Identifying the causes of sediment-associated toxicity in urban waterways in South China: Incorporating bioavailabillity-based measurements into whole-sediment toxicity identification evaluation. Environmental Toxicology and Chemistry: Volume 34, Issue 8, pages 1744-1750, August 2015. DOI: 10.1002/etc.2970

圖1 “生物有效性測量”與“全沉積物毒性識別評估(TIE)”聯(lián)用有效識別出沉積物中主要致毒污染物

3. 蛋白組學(xué)分析揭示出阿特拉津?qū)Ψ侵拮傅纳扯拘栽从谄湔T導(dǎo)生殖細胞凋亡并干擾緊密連接蛋白和代謝相關(guān)信號通路

Proteomics analysis ofXenopuslaevisgonad tissue following chronic exposure to atrazine

內(nèi)容提要：阿特拉津可對雄性脊椎動物生殖系統(tǒng)造成不良影響，但其中分子機制尚不明了。本文作者將非洲爪蟾(Xenopus laevis)暴露于100 ppb的阿特拉津120 d，利用同位素標(biāo)記相對和絕對定量(iTRAQ)技術(shù)檢測了非洲爪蟾睪丸和卵巢中蛋白圖譜的變化，結(jié)果顯示100 ppb阿特拉津可影響爪蟾發(fā)育，延遲和阻礙雄性細精管的形成。組學(xué)分析顯示，睪丸中143種以及卵巢中121種蛋白均表達異常，這些蛋白與細胞凋亡、細胞間緊密連接以及代謝途徑相關(guān)。

詳情請見http://onlinelibrary.wiley.com/doi/10.1002/etc.2980/full

Abstract: Atrazine is the most commonly detected pesticide contaminant in ground and surface water. Previous studies have shown that atrazine is an endocrine disruptor owing to its adverse effects on the male reproductive system in several vertebrates, but very few molecular mechanisms for these effects have been revealed. In the present study, Xenopus laevis were exposed to 100 ppb of atrazine for 120 d, and then the isobaric tags for relative and absolute quantitation (iTRAQ) technique was used to detect global changes in protein profiles of the testes and ovaries. The results showed that 100 ppb of atrazine exposure adversely affected the growth of X. laevis and did not induce hermaphroditism but delayed or prevented the development of male seminiferous tubules. Proteomic analysis showed that atrazine altered expression of 143 and 121 proteins in the testes and ovaries, respectively, and most of them are involved in cellular and metabolic processes and biological regulation based on their biological processes. In addition, apoptosis, tight junctions, and metabolic pathways were significantly altered in the atrazine-treated gonads. Based on the above results, it is postulated that the reproductive toxicity of atrazine may be the result of disruption of tight junctions and metabolic signaling pathways and/or induction of apoptosis in germ cells.

View more in http://onlinelibrary.wiley.com/doi/10.1002/etc.2980/full

精選自：Xiuping Chen, Jiamei Wang, Haojun Zhu, Jiatong Ding and YufaPeng.Proteomics analysis of Xenopus laevis gonad tissue following chronic exposure to atrazine. Environmental Toxicology and Chemistry: Volume 34, Issue 8, pages 1744-1750, August 2015. DOI: 10.1002/etc.2980

4. 基于非本地物種毒性數(shù)據(jù)推導(dǎo)出的水質(zhì)基準可為本地物種提供適當(dāng)保護嗎？

Do water quality criteria based on nonnative species provide appropriate protection for native species?

內(nèi)容提要：基于非本地物種的毒性數(shù)據(jù)推導(dǎo)出的水質(zhì)基準一直存在爭議，質(zhì)疑者提出應(yīng)有針對不同生物區(qū)系的水質(zhì)基準。而這個問題在中國尤為突出。本文作者搜集了38種污染物對本地物種和非本地物種的毒性數(shù)據(jù)，比較了這些生物的HC5和物種敏感度分布，結(jié)果顯示，基于非本地物種推導(dǎo)的水質(zhì)基準有74%的確定性可以有效保護中國的水生態(tài)系統(tǒng)。如果采用評價因子(AF=10)來校正基準，那么可以獲得90%的確定性。

詳情請見：http://onlinelibrary.wiley.com/doi/10.1002/etc.2985/full

Abstract: The potential use of toxicity data for nonnative species to derive water quality criteria is controversial because it is sometimes questioned whether criteria based on species from one geographical region provide appropriate protection for species in a different region. However, this is an important concept for the development of Chinese water quality criteria or standards. Data were assembled on 38 chemicals for which values were available for both native and nonnative species. Sensitivities of these organisms were compared based on the 5% hazardous concentration values and the species sensitivity distribution from a literature review. Results of the present study's analysis showed that there is approximately 74% certainty that use of nonnative species to generate water quality criteria would be sufficiently protective of aquatic ecosystems in China. Without applying any assessment factor to the water quality criteria generated from nonnative species, the uncertainty would be 26% when the native Chinese species might be under protection. Applying an assessment factor of 10 would offer adequate protection to native Chinese species for approximately 90% of tested chemicals and thus reduce the uncertainty from 26% to 10%.

View more in http://onlinelibrary.wiley.com/doi/10.1002/etc.2985/full

精選自：Xiaowei Jin, Zijian Wang, Yeyao Wang, YibingLv, Kaifeng Rao, Wei Jin, John P. Giesy and Kenneth M. Y. Leung. Do water quality criteria based on nonnative species provide appropriate protection for native species? Environmental Toxicology and Chemistry: Volume 34, Issue 8, pages 1744-1750, August. DOI: 10.1002/etc.2985

5. 納米銅顆粒的種間毒性關(guān)系

Assessing toxicity of copper nanoparticles across five cladoceran species

內(nèi)容提要：納米顆粒的種間毒性關(guān)系尚無明確結(jié)論，因此很難判斷納米顆粒對未進行毒性測試的物種的風(fēng)險如何。本文作者將5種水蚤(Daphnia magna, Daphnia pulex, Daphnia galeata, Ceriodaphnia dubia, Chydorus sphaericus )暴露于4種不同粒徑的納米銅顆粒(CuNPs)和1種亞微米銅顆粒的懸濁液中，考察物種的形態(tài)特征與CuNPs急性毒性的關(guān)系。結(jié)果顯示，桿狀的CuNPs比球狀的CuNPs毒性更低。納米銅顆粒與溶出銅離子均對CuNPs的毒性有貢獻，其中，當(dāng)新生蚤的體長、體表面積和身體體積更小時，5種懸濁液中顆粒的毒性更大。5種蚤的身體體積與5種CuNPs的毒性顯著相關(guān)(radj2>0.51, p< 0.001)，78-nm CuNPs與蚤身體體積的相關(guān)性最好 (radj2=0.95, p< 0.001)。這個研究可以為納米顆粒對有相似外形特征的物種進行毒性種間外推提供線索。

詳情請見：http://onlinelibrary.wiley.com/doi/10.1002/etc.3000/full

Abstract: As a result of ever increasing applications, nanoparticles will eventually end up in the environment. However, currently no common principle has been established to help understand the toxicity of nanoparticles (NPs) across species. Therefore, it is difficult to estimate the potential risks of nanoparticles to untested species in the environment. The authors exposed 4 different sizes of copper nanoparticles (CuNPs) and 1 submicron-sized copper particle to 5 cladoceran species (Daphnia magna, Daphnia pulex, Daphnia galeata, Ceriodaphnia dubia, Chydorus sphaericus ) to investigate whether morphological attributes of species can help to assess the acute toxicity of CuNPs across species. The results showed that rod-shaped CuNPs caused much lower toxicity to all species than spherical CuNPs. Both the particles and ions contributed to the total toxicity of the CuNP suspensions. Moreover, the toxicity caused by particles in 5 different copper suspensions increases with decreasing body length, surface area, and body volume of neonates of 5 cladoceran species. Especially the correlations between body volume of the 5 cladoceran species tested and the corresponding toxicity caused by 5 different CuNPs were statistically significant, and in all cases radj2was higher than 0.51 (p< 0.001). The highest correlation was found between body volume and the toxicity of the 78-nm CuNPs (radj2=0.95, p< 0.001). To conclude, the correlations between attributes of cladoceran species and the toxicity of CuNPs reported in the present study evoke the possibility to assess and extrapolate the toxicity of nanoparticles across species with similar attributes.

View more in http://onlinelibrary.wiley.com/doi/10.1002/etc.3000/full

精選自：Lan Song, Martina G. Vijver, Geert R. de Snoo and Willie J.G.M. Peijnenburg. Assessing toxicity of copper nanoparticles across five cladoceran species. Environmental Toxicology and Chemistry: Volume 34, Issue 8, pages 1744-1750, August 2015. DOI: 10.1002/etc.3000

Acute Toxic Effect of Cu-Sulfadiazine Combined Stress on the Germination of Vegetable Seeds

Jin Caixia*, Si Xiaowei， Mao Lei, Fan Jing

Henan Key Laboratory of Environmental Pollution Control, Key Laboratory for Yellow River and Huaihe River Water Environmental and Pollution Control, Ministry of Education, School of Environment, Henan Normal University, Xinxiang 453007, China

Received 17 October 2014 accepted 15 November 2014

This study reported the single and joint effects of commonly used sulfadiazine (SD) veterinary drug and copper (Cu) on the seed germination (including germination rate, root and shoot elongation) of pakchoi and tomato in yellow fluvo-aquic soil, analyzed the dose-response relationship between the drug concentration in soil and inhibitory action on crop growth, as well as the toxic effect of combined pollution. The results indicated that the inhibitory rate of root and shoot elongation correlated significantly with pollutant concentration (P<0.01), the inhibitory rate of germination and pollutant concentration were uncorrelated (P>0.05), whether in the presence of individual Cu or SD; the inhibitory effect of pollutants on root and shoot elongation was much stronger than that of the seed germination; the toxic effect of SD was much stronger than that of Cu, the ID50(half inhibition concentration) values of SD on root elongation of pakchoi and tomato were 1.7 and 33.6 mg·kg-1, respectively, while Cu were 273.6 and 457.7 mg·kg-1. The antagonism effect of Cu-SD combined pollution on the root and shoot elongation of the tomato was remarkable (P<0.01); but for the pakchoi, the combined antagonism effect was not significant when the Cu concentration was lower than 100 mg·kg-1(P>0.05), the antagonism effect was gradually notable as the increasing of the Cu concentration (P<0.01).

sulfadiazine; copper; pakchoi; tomato; germination inhibitory rate; combined pollution; ecotoxicology

中國博士后科學(xué)基金(20110491001)；國家自然科學(xué)基金(21107023)；河南省科技創(chuàng)新杰出人才計劃(144200510004)

(Corresponding author)， E-mail: heartjin0509@aliyun.com

10.7524/AJE.1673-5897.20141017001

2014-10-17錄用日期：2014-11-15

1673-5897(2015)5-164-08

X171.5

A

金彩霞(1976-)，女，博士，副教授，研究方向為環(huán)境化學(xué)、生態(tài)毒理學(xué)。

*

金彩霞, 司曉薇, 毛蕾. 銅-磺胺嘧啶復(fù)合脅迫下對蔬菜種子發(fā)芽的急性毒性效應(yīng)[J]. 生態(tài)毒理學(xué)報, 2015, 10(5): 164-171

Jin C X, Si X W, Mao L. Acute toxic effect of Cu-sulfadiazine combined stress on the germination of vegetable seeds [J]. Asian Journal of Ecotoxicology, 2015, 10(5): 164-171 (in Chinese)