莫凌，吳江平，張云，邢巧，林彰文，羅孝俊，麥碧嫻

1. 海南省環(huán)境科學研究院，海口 570100 2. 中國科學院廣州地球化學研究所 有機地球化學國家重點實驗室，廣州 510640 3. 華南師范大學 生命科學學院，廣州 510631

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電子垃圾拆解地翠鳥對多氯聯(lián)苯的累積及風險評估

莫凌1，吳江平2,，張云3，邢巧1，林彰文1，羅孝俊2，麥碧嫻2

1. 海南省環(huán)境科學研究院，海口 570100 2. 中國科學院廣州地球化學研究所 有機地球化學國家重點實驗室，廣州 510640 3. 華南師范大學 生命科學學院，廣州 510631

粗獷的電子垃圾拆解活動已造成當?shù)匾吧锒嗦嚷?lián)苯(PCBs)嚴重污染，但PCBs在野生鳥類中的生物累積特征及潛在的毒害作用研究較少。本研究采集了廣東省某電子垃圾拆解地翠鳥(Alcedo atthis)及其食物(各種小型魚類)樣品，研究翠鳥對PCBs的累積特征、生物放大效應及毒性風險。翠鳥肌肉中PCBs中值含量為220 μg·g-1脂重，比其他報道值高1～3個數(shù)量級。計算的生物放大因子(BMF)顯示，大部分PCB單體的BMF值都大于1，表明翠鳥對PCBs具有生物放大效應。計算的共面PCBs毒性當量(TEQs)范圍為39～23 600 pg·g-1濕重，已經(jīng)達到或超過了影響某些鳥類生殖或發(fā)育障礙的報道值。上述結果表明，電子垃圾拆卸活動已經(jīng)造成了當?shù)卮澍BPCBs嚴重污染，PCBs污染物對電子垃圾拆解地翠鳥及其他野生生物的毒性效應尚需進一步研究。

多氯聯(lián)苯；鳥類；生物積累；生物放大；電子垃圾

Received 14 November 2015 accepted 22 December 2015

多氯聯(lián)苯(polychlorinated biphenyls，PCBs)是一類人工合成的多氯芳香烴類化合物。由于PCBs具有物理化學性質穩(wěn)定、耐熱性、電絕緣性等特點，常用作熱載體、絕緣材料、溶劑等廣泛添加于電子電器等產(chǎn)品中[1]。這些產(chǎn)品在使用過程中或廢棄后PCBs很容易進入到環(huán)境。研究證實，PCBs在環(huán)境中具有持久性、可生物累積性和較高的生物毒性[2-3]。2001年5月，PCBs被列入《斯德哥爾摩公約》持久性有機污染物(POPs)首批受控名單，在全球禁用[4]。

隨著PCBs的禁用，全球PCBs主要生產(chǎn)和使用地區(qū)(主要為北美和歐洲)環(huán)境中PCBs含量顯著下降[5]。然而，作為全球廢棄電子電器產(chǎn)品(電子垃圾)主要集中地亞洲和非洲一些國家環(huán)境中PCBs含量卻有上升的趨勢[5-6]。PCBs在電子垃圾集中地的環(huán)境行為及其生態(tài)風險評估已引起了廣泛關注。我們的初步研究發(fā)現(xiàn)，廣東省某電子垃圾拆解地野生魚類體內的PCBs含量很高[7]，這些污染物是否傳遞到高營養(yǎng)級生物(如食魚鳥)及其對這些生物可能的毒害作用尚不清楚。本研究在廣東省某電子垃圾拆解地同時采集了翠鳥(Alcedo atthis)及其食物(3種小型魚類)樣品，通過測定這些樣品中PCBs的含量，研究了翠鳥對PCBs的生物放大效應，并評估了這些污染物對翠鳥潛在的毒性效應。

1 材料與方法 (Materials and methods)

1.1 樣品采集與前處理

翠鳥是一種食魚性鳥類，主要是以其棲息地附近水體中各種小型魚類為食[8]。翠鳥(A. atthis)樣品(n = 22)于2010年5月到2010年7月在廣東省某電子垃圾拆解地池塘邊用網(wǎng)捕法采集，鳥類采集經(jīng)廣東省林業(yè)局批準。同時在池塘中采集了中國斗魚(Macropodus opercularis，n = 9)、食蚊魚(Gambusia affinis，n = 11)和馬口魚(Opsariichthys bidens，n = 9) 3種小型魚類。鳥類樣品采集后用N2進行安樂死。所有樣品低溫運輸至實驗室后，-20 ℃冷凍保存待分析。

翠鳥樣品解剖后，取肌肉組織。每種魚類樣品(整魚樣)隨機混合成3個混合樣。取約4 g翠鳥肌肉組織和8 g魚類樣品，冷凍干燥并混勻后，加入回收率指示物CB 30、CB 65和CB 204后，用正己烷/丙酮混合溶劑(1/1，V/V)索氏抽提48 h。抽提液一部分用于測定脂肪含量(重量法)。余下抽提液濃縮至1 mL左右，經(jīng)凝膠滲透色譜柱(GPC)去除脂肪和其他干擾物。洗脫液濃縮至1～2 mL后，過復合硅膠柱(酸性硅膠:中性硅膠=8:8)凈化，最后濃縮定容至200 μL，加入內標(CB 24、CB 82和CB 198)。樣品的抽提及凈化具體方法參見文獻[7]。

1.2 儀器測定

75種PCBs單體用安捷倫氣相色譜質譜聯(lián)用儀(Agilent 6890 GC-5975B Series MS)測定。采用EI源、選擇性離子掃描模式(SIM)，使用色譜柱DB-5MS (60 m × 0.25 mm i.d.，0.25 μm film thickness；J&W Scientific, Folsom，CA)進行分離。選擇無分流進樣，進樣量為1 μL。載氣為高純氦氣，柱流速為1.50 mL·min-1。升溫程序：起始溫度120 ℃，6 ℃·min-1升溫至180 ℃，1 ℃·min-1升溫至240 ℃，然后6 ℃·min-1升溫至290 ℃并保留17 min。進樣口、離子源溫度和界面溫度分別為290 ℃、250 ℃和290 ℃。目標化合物采用內標法(6點校正曲線)定量。

1.3 質量保證與質量控制(QA/QC)

QA/QC體系主要包括回收率指示物添加、程序空白、加標空白、基質加標、樣品重復樣測定等。程序空白樣中有少量PCB 118和PCB 138檢出，實際樣品進行了相應扣除。樣品中PCB 30、PCB 65和PCB 204的回收率分別為83% ± 16%、94% ±17%和94% ± 15%，定量結果未經(jīng)回收率校正?？瞻准訕撕突|加標中PCBs單體(20種PCBs)的回收率范圍分別為75%～107%和75%～104%。樣品平行樣中所有目標化合物的相對標準偏差均小于15%。PCBs的最低檢測限(LOQs)按方法空白中各單體的含量平均值加3倍標準偏差計算。對于空白中沒有檢測出的目標化合物，按5倍信噪比(S/N)計算。樣品中PCBs的檢測限為0.01～0.20 μg·g-1脂重。

1.4 生物放大因子(BMF)計算

普通翠鳥體內中各PCB單體的生物放大因子(BMF)按照下式計算：

BMF=C翠鳥/C食物

式中C翠鳥為化合物在翠鳥中的濃度，單位為μg·g-1脂重，C食物為翠鳥食物(3種小魚)中對應化合物的濃度，單位為μg·g-1脂重。

1.5 毒性當量(TEQ)計算

利用聯(lián)合國衛(wèi)生組織提出的鳥類二噁英類化合物毒性當量因子(TEFs)[7]，計算了幾種主要的共面PCB單體(包括PCB 77、PCB 81、PCB 105、PCB 114、PCB 118、PCB 123、PCB 126、PCB 156、PCB 167和PCB 169)的毒性當量(TEQ)。其計算公式如下：

TEQ= ∑(PCBi×TEFi)

式中PCBi和TEFi分別為某種共面PCB單體的濃度(pg·g-1濕重)和其TEF。

2 結果與討論(Results and discussion)

2.1 PCBs的含量與組成

翠鳥及3種小型魚類體內7種指示性PCBs及75種PCB單體總含量(ΣPCBs)見表1。翠鳥肌肉中ΣPCBs的濃度范圍為4.0～3 300 μg·g-1脂重(中值濃度為220 μg·g-1脂重)。目前，僅有一篇文獻報道了華南某自然保護區(qū)(中值1 800 ng·g-1脂重)和農(nóng)村區(qū)域(中值410 ng·g-1脂重)普通翠鳥中PCBs的含量[7]。但已有不少研究報道了其他食魚性鳥類肌肉中PCBs的含量。Luo等[9]研究的電子垃圾區(qū)池鷺肌肉中的PCBs的含量達到了120 000 ng·g-1脂重。比利時的牛背鷺肌肉中的濃度達到90 000 ng·g-1脂重[10]。Tanabe等[11]報道了采集于南印度濕地和沿海區(qū)域的白胸翡翠(翠鳥)肌肉中PCBs殘留濃度為400 ng·g-1脂重。Kunisne等[12]報道的日本北海道黑尾鷗肌肉濃度為2 700～11 000 ng·g-1脂重。采自日本Lake Biwa湖區(qū)和羅馬尼亞Danube Delta區(qū)域的普通鸕鶿肌肉中PCBs的濃度分別為2 900～7 700 000和700 ng·g-1脂重[12-14]。Braune等[15]研究了加拿大西部秋沙鴨和普通潛鳥胸部肌肉中PCBs的殘留情況，這2種鳥類中PCBs的殘留濃度為1.1～1 000 ng·g-1脂重。Elliott等[16]測定了英國、哥倫比亞和加拿大鵲鴨和棕脅秋沙鴨肌肉中PCBs含量，其濃度分別為1 600 ng·g-1脂重和78 ng·g-1脂重。相比于前人報道的食魚性鳥類肌肉中PCBs的殘留濃度，本研究翠鳥PCBs含量與日本Lake Biwa湖區(qū)的鸕鶿和中國電子垃圾拆解地池鷺相當(范圍為4.0～3 300 μg·g-1脂重)，但高于其他報道值1～3個數(shù)量級。翠鳥肌肉中PCBs含量也高于采集于同一電子垃圾拆卸區(qū)的其他7種水生鳥類(1 800～18 000 ng·g-1脂重)和1種陸生鳥類(7 300 ng·g-1脂重)以及華南某保護區(qū)4種陸生鳥類(45～1 770 ng·g-1脂重)肌肉中PCBs含量[9， 17-19]。雖然監(jiān)測的PCBs單體總數(shù)量的不同，會影響本文翠鳥中PCBs的殘留濃度與其他研究鳥類中濃度的對比，但是這些結果也初步表明，電子垃圾拆解地翠鳥已經(jīng)受到PCBs嚴重污染。此外，諸多因素如食性和攝食量、遷移性和生態(tài)位(營養(yǎng)級)等會影響不同鳥類對PCBs的累積。

表1 電子垃圾拆卸區(qū)翠鳥及3種小型魚類中PCBs含量(單位：μg·g-1脂重)

注：9 (3)表示樣品的個數(shù)，括號里面的數(shù)字表示混合后樣品的數(shù)量；3.5 (2.2～5.9)表示中值和范圍；∑PCBs表示75個PCBs單體的總濃度。

Note: 9 (3) is the number of individual samples collected; figures in brackets indicate analyses number of pooled samples when individual were pooled; 3.5 (2.2～5.9) means median and range; ∑PCBs is the sum concentrations of the 75 PCB congeners examined.

圖1 電子垃圾拆卸區(qū)翠鳥和其食物中PCBs的同系物分布模式Fig. 1 Congener profiles of PCBs in the kingfishers and their prey fish species collected from an e-waste recycling site

圖2 翠鳥/魚類的3種捕食關系中PCBs的BMFs值Fig. 2 Biomagnification factors (BMFs) for PCB congeners derived from the three predator/prey pairs

翠鳥及其食物(3種小魚)中PCBs同系物組成模式如圖1所示。翠鳥肌肉中CB 118是最主要的同系物，占ΣPCBs的16% ± 1.5%。CB 28/31、CB 153、CB 101和CB 138也是主要的同系物，共占ΣPCBs的47% ± 1.0%。翠鳥的食物與翠鳥有著相似的PBDEs同系物分布模式，其中CB 118、CB 153、CB 101和CB 138是最主要的同系物。本研究普通翠鳥肌肉中的PCB同系物組成特征與之前報道的華南某自然保護區(qū)和農(nóng)村區(qū)域普通翠鳥中PCBs的同系物相同[7]，都以CB 118、CB 28/31和CB 153為最主要同系物。本研究翠鳥體內PCBs同系物組成與電子垃圾拆卸區(qū)其他水生鳥類PCBs同系物組成也基本相同，都是以5～6氯等低氯代的PCBs單體為主[9]，但與陸生鳥類PCB的同系物組成特征(主要是以5～8氯為主PCBs單體為主)具有較大的差異[17， 20]。水生鳥類和陸生鳥類生活環(huán)境和食性的不同，可能是導致這種差異的主要原因。相對于高氯代的PCB單體，低氯代的PCBs具有較高的水溶性，更容易在水生生物體內富集，造成翠鳥體內較高含量的低氯代PCB單體。

表2 電子垃圾拆卸區(qū)普通翠鳥中共面PCBs的毒性當量值(單位：pg·g-1濕重)

表3 文獻報道的PCBs的TEQs對鳥類的毒性參考值

2.2 PCBs的生物放大

為了調查PCBs單體在普通翠鳥食物鏈中可能的生物放大效應，我們計算了這些化合物的生物放大因子(BMFs)(圖2)。大部分的PCB單體的BMF值都大于1(ΣPCBs的平均BMF值為1.1～2.5)，表明翠鳥對這些PCB單體產(chǎn)生了生物放大效應。通過對比PCB單體在3種捕食關系中的BMFs發(fā)現(xiàn)，翠鳥/中國斗魚和翠鳥/食蚊魚之間BMF值都大于1，而PCB少數(shù)單體(CB 49、CB87/115、CB110、CB156、CB190/170)在翠鳥/馬口魚之間BMF值小于1，通過查詢文獻，普通翠鳥其食物中99%都是以2.3 cm左右小型淡水魚類(最大也能達到12.5 cm)[8]，但是本次樣品中馬口魚的平均長度為7.0[21]，較其他2種魚類長。因此，馬口魚可能不是翠鳥的主要食物之一，不同魚類在翠鳥食物中的比例可能是造成這種差異的原因。本研究計算的PCBs的BMF值與之前報道的華南某自然保護區(qū)普通翠鳥對這些污染物的BMF值基本相似(ΣPCBs的平均BMF值為1.1～1.4)[7]。Drouillard等[22]采用腸道累積放大的方法預測了環(huán)鴿(Streptopelia risoria)對PCBs的生物放大效果，預測的BMF范圍為18.5～33.8，高于我們當前的研究值。野外實驗條件和實驗室預測條件的差別以及物種間對于PCBs不同的累積特性，可能是導致不同研究BMF值不盡相同的主要原因。

2.3 TEQ值

目前還未見PCBs對于翠鳥的毒性風險評價數(shù)據(jù)。我們計算了幾種主要的共面PCB單體(包括CB 77、CB 81、CB 105、CB 114、CB 118、CB 123、CB 156和CB 167)的毒性當量(TEQ)，并通過對比TEQs對其他水生鳥類的毒性參考值(TRVs)或風險評估的閾值，評估PCBs對普通翠鳥潛在的毒害作用。計算的TEQs范圍為39～24 000 pg·g-1濕重，平均值為3 700 pg·g-1濕重(表2)。較多研究報道了TEQs對鳥類的TRVs(表3)，超過這些參考值將對鳥類產(chǎn)生胚胎死亡和發(fā)育障礙等方面的影響[23]。與這些結果比較，電子垃圾拆卸區(qū)翠鳥體內的TEQs含量超過了大部分水生鳥類的TRV值。這一結果預示著電子垃圾拆卸區(qū)翠鳥體內的PCBs可能對其生殖和發(fā)育方面帶來潛在的毒害作用。

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Bioaccumulation and Risk Assessment of Polychlorinated Biphenyls in the Common Kingfisher (Alcedoatthis) from an Electronic Waste Recycling Site in South China

Mo Ling1, Wu Jiangping2,*, Zhang Yun3, Xing Qiao1, Lin Zhangwen1, Luo Xiaojun2, Mai Bixian2

1. Hainan Research Academy of Environmental Sciences, Haikou 570100, China 2. State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640, China 3. School of Life Science, South China Normal University, Guangzhou 510631, China

The wildlife from electronic waste (e-waste) sites have been heavily polluted by polychlorinated biphenyls (PCBs), due to the primitive e-waste recycling activities. However, information on the bioaccumulation and the toxic effects of PCBs in wild avian species from e-waste sites is limited. In the present study, we investigated the levels and congener profiles of PCBs in the common kingfisher (Alcedo atthis) from an e-waste recycling site in Guangdong Province, South China. Additionally, PCBs in the diet items including three fish species collected from the same sampling site were also examined, to evaluate the potential biomagnification of PCBs in the common kingfisher. Finally, we assessed the potential toxic effects of PCBs to these birds by estimating the toxic equivalent quantity (TEQ) of the co-planar PCBs. Elevated PCB residues (median = 220 μg·g-1lipid weight for total PCBs) were detected in the kingfishers, which were one to three orders of magnitude higher than the values previously reported in the species from other sampling sites. The calculated predator/prey biomagnification factors (BMFs) were greater than unity for most of the PCB congeners examined, suggesting biomagnification of these chemicals in the common kingfisher. The TEQ concentrations estimated in the common kingfisher ranged from 39 to 23 600 pg·g-1wet weight, with some of these values reaching or exceeding the levels known to impair bird reproduction and survival. Our results revealed that the common kingfisher from the e-waste recycling site has been heavily contaminated by PCBs. The need for further examination is warranted to determine the potential adverse effects resulting from the PCBs exposure, in the common kingfishers and other wildlife that are habitants of e-waste sites.

polychlorinated biphenyls; bird; bioaccumulation; biomagnification; electronic waste

10.7524/AJE.1673-5897.20151114001

海南省自然科學基金(20154176)；國家自然科學基金(41230639，41173109)

莫凌(1984-)，男，博士，助理研究員，高級工程師，研究方向為持久性有毒污染物的環(huán)境行為及毒害作用，E-mail: morning.ml@163.com

*通訊作者(Corresponding author)， E-mail: jpwu@gig.ac.cn

2015-11-14 錄用日期：2015-12-22

1673-5897(2016)2-155-08

X171.5

A

簡介：吳江平(1976-)，男，博士，副研究員，主要研究方向為持久性有毒污染物的生物累積、沿食物鏈傳遞及其毒害作用，近5年在國內外環(huán)境領域主流刊物發(fā)表文章50余篇，被SCI期刊引用共1200余次。

莫凌, 吳江平, 張云, 等. 電子垃圾拆解地翠鳥對多氯聯(lián)苯的累積及風險評估[J]. 生態(tài)毒理學報，2016, 11(2): 155-162

Mo L, Wu J P, Zhang Y, et al. Bioaccumulation and risk assessment of polychlorinated biphenyls in the common kingfisher (Alcedo atthis) from an electronic waste recycling site in south China [J]. Asian Journal of Ecotoxicology, 2016, 11(2): 155-162 (in Chinese)