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文章信息
- 侯淼淼, 王春伶, 徐椿森, 邱宁, 夏治俊, 苏良霞, 王剑伟
- HOU Miaomiao, WANG Chunling, XU Chunsen, QIU Ning, XIA Zhijun, SU Liangxia, WANG Jianwei
- 聚苯乙烯微塑料暴露对稀有鲫仔鱼生长的影响
- Effects of Polystyrene Microplastics Exposure on the Growth of Gobiocypris rarus Larvae
- 四川动物, 2020, 39(2): 140-147
- Sichuan Journal of Zoology, 2020, 39(2): 140-147
- 10.11984/j.issn.1000-7083.20190390
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文章历史
- 收稿日期: 2019-11-15
- 接受日期: 2019-12-31
2. 中国科学院水生生物研究所, 武汉 430072;
3. 中国科学院大学, 北京 100049
2. Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China;
3. University of Chinese Academy of Sciences, Beijing 100049, China
塑料因具有防水、耐用、质轻、绝缘、抗腐蚀等诸多优点,在人类生产生活中得到广泛使用。据统计,全球每年的塑料使用量不少于3×108 t,但塑料制品在自然环境下难以降解,大量使用导致生态环境中的废弃塑料或塑料制品越来越多。截止2014年,全球仅海洋表面漂浮的塑料垃圾超过2.5×105 t(Eriksen et al., 2014)。环境中的塑料经过物理、化学或生物作用可分解为更小的塑料颗粒或碎片,加上直接排入环境中的微小塑料颗粒或碎片,造成了环境中严重的微塑料污染(Thompson et al., 2004)。微塑料通常指尺寸在100 nm~5 mm的小塑料碎片,这一定义被多数人认同和采用(Law & Thompson,2014;丁剑楠等,2017;Toussaint et al., 2019)。调查结果表明,从南极到北极、从内陆到海洋、从水体到沉积物,皆能发现不同程度的微塑料污染(Ivar do Sul et al., 2014;Wagner & Lambert,2017;徐向荣等,2018),甚至在人体中也检测到了微塑料的存在(van Cauwenberghe & Janssen,2014;Smith-Llera,2019)。
作为一类新型污染物,微塑料已受到国内外学者和公众的高度关注,关于微塑料污染的研究已成为目前国际上的研究热点之一(Koelmans et al., 2016;Toussaint et al., 2019)。近年来,有关微塑料来源、类型、分布以及对水生生物毒理学效应等方面的研究报道日益增多(Andrady,2011;丁剑楠等,2017;陈启晴等,2018)。已有研究表明,微塑料能够赋存于水生生物体内,并通过物理性损伤、载体效应(如增塑剂释放、富集其他污染物等)、生物积累与食物链传递等途径危害水生生物健康(Oliveira et al., 2013;Wright et al., 2013;Batel et al., 2016),在个体水平(如摄食率、繁殖率、存活率、生长速率等)、组织水平(如炎症反应、氧化损伤、脂肪空泡等)、细胞水平(如肝细胞坏死等)、基因水平(如内分泌干扰基因表达变化等)表现出毒性效应(von Moos et al., 2012;Hämer et al., 2014;Rochman et al., 2014;周倩等,2015)。但总体来看,关于微塑料本身对水生生物毒性效应的研究较少,鱼类仅见于斑马鱼 Danio rerio、青鳉 Oryzias latipes、挪威舌齿鲈 Dicentrarchus labrax等少数物种(Rochman et al., 2013, 2014;Mazurais et al., 2015;Lu et al., 2016;Lei et al., 2018),因此,亟需积累更多的鱼类毒理学数据,进一步解析微塑料毒性作用机制,以期更准确评价环境中微塑料的生态风险。
稀有
受试生物为稀有
暴露试验参照美国环境保护局的黑头软口鲦 Pimephales promelas 仔鱼7 d生长存活试验(USEPA,2002),试验容器为500 mL圆形玻璃结晶皿,每个试验容器放试验液400 mL、初孵仔鱼30尾,每个浓度设置3个平行,为暴露组,并设置空白对照组。结晶皿置于控温的水浴中,实验期间每天监测水温。暴露期间每天10: 00和16: 00投喂新孵丰年虫 Artemia nauplii 无节幼体,每天更换90%以上的暴露溶液,换液前使用HACH水质分析仪(HQ30d,美国)随机抽测3个结晶皿中的溶解氧。7 d后统计仔鱼的存活率并将存活个体全部用100 mg·L-1中性MS-222麻醉后在解剖镜下测量全长,从每个浓度组随机选取3尾仔鱼放置于荧光显微镜下拍摄带荧光信号的照片,并使用ImageJ对荧光强度进行半定量分析。之后从每个结晶皿中随机挑选10尾鱼再进行7 d净化试验。净化试验中各浓度组更新溶液为与空白对照组相同的稀释水,其他控制条件与7 d亚慢性毒性试验期间相同。净化7 d后从每个浓度组随机挑选3尾仔鱼用MS-222麻醉后置于荧光显微镜下观察荧光微塑料的分布情况,拍摄照片并分析荧光强度。
参考他人关于微塑料环境浓度调查和微塑料对水生生物的毒性效应的相关研究(Zhao et al., 2014;Lu et al., 2016;涂烨楠,2018)设置浓度组,7 d亚慢性毒性试验中暴露浓度分别为0.055 μg·L-1、0.55 μg·L-1、5.5 μg·L-1、55 μg·L-1、550 μg·L-1,采用血球计数板法实测微塑料浓度(丰度)。
1.3 数据统计与分析采用SPSS Statistics 20进行数据分析。运用单因素方差分析(One-Way ANOVA)和Dunnett's多重比较分析空白对照组与暴露组在存活率、全长等指标上的差异水平;采用独立样本t检验分析空白对照组与暴露组体内荧光强度的差异性;所有结果表示为x±SD,当P<0.05时认为差异有统计学意义。使用ImageJ对荧光强度进行半定量分析,使用Origin 8.5及PhotoShop CS6作图。
2 结果 2.1 PS微塑料对稀有暴露期间水温控制在24~26 ℃(24.9 ℃±0.45 ℃)、溶解氧>4 mg·L-1(7.41 mg·L-1±0.37 mg·L-1)、空白对照组累计死亡率 < 20%,符合7 d亚慢性毒性试验质量控制要求。3种粒径PS微塑料对稀有
PS微塑料粒径/μm | 质量浓度 /(μg·L-1) |
丰度 /(items/L) |
存活率 /% |
7 d仔鱼 全长/mm |
0.1 | 0 | 0 | 87.78±5.09a | 5.78±0.34a |
0.055 | 1.00×108 | 87.78±1.92a | 5.52±0.47a | |
0.55 | 1.00×109 | 76.67±23.09a | 5.68±0.48a | |
5.5 | 1.00×1010 | 80.00±17.64a | 5.74±0.36a | |
55 | 1.00×1011 | 86.67±3.34a | 5.76±0.32a | |
550 | 1.00×1012 | 71.11±21.69a | 5.61±0.47a | |
1 | 0 | 0 | 95.56±1.93a | 5.89±0.39a |
0.055 | (1.13±0.29)×105 | 91.11±5.09a | 6.09±0.38a | |
0.55 | (1.05±0.24)×106 | 94.45±3.85a | 6.01±0.37a | |
5.5 | (1.06±0.23)×107 | 91.11±3.85a | 5.96±0.32a | |
55 | (1.02±0.10)×108 | 87.78±5.09a | 5.84±0.42a | |
550 | (1.07±0.04)×109 | 92.22±7.70a | 6.01±0.41a | |
10 | 0 | 0 | 93.33±3.34a | 5.90±0.55a |
0.055 | (1.37±0.77)×102 | 93.33±0.00a | 5.96±0.66a | |
0.55 | (1.12±0.20)×103 | 83.33±8.82a | 5.74±0.63a | |
5.5 | (0.98±0.23)×104 | 94.44±1.93a | 5.98±0.71a | |
55 | (1.00±0.20)×105 | 83.33±6.67a | 5.90±0.55a | |
550 | (1.05±0.31)×106 | 90.00±6.67a | 5.73±0.55a | |
注:同一列中具有不同字母表示差异有统计学意义(P<0.05);0.1 μm的微塑料颗粒在显微镜下难以计数,此处为根据质量浓度换算的标称丰度 Notes:Different letters in the same column indicate there is a significant difference (P < 0.05); as it is difficult to count 0.1 μm microplastic particles under the microscope,the nominal abundance was calculated by mass concentration |
暴露7 d后,除空白对照组外,各暴露组仔鱼体内均发现荧光微塑料赋存(图版Ⅰ:A、C、E)。微塑料主要存在于暴露组仔鱼的鳃部和消化道,在身体其他部位或组织中未见。荧光半定量分析表明,荧光强度与暴露浓度正相关(图 1),其回归方程为:0.1 μm PS微塑料:y=8 819.9e0.820 5lgx(R2=0.970 3,P < 0.05);1 μm PS微塑料:y=3 448.9e1.024 2lgx(R2=0.900 1,P < 0.05);10 μm PS微塑料:y=7 373.7e0.639 6lgx(R2=0.825 3,P < 0.05)。
净化7 d后,各组仔鱼体内PS微塑料均显著减少(图版Ⅰ:B、D、F),在0.055 μg·L-1、0.55 μg·L-1浓度暴露后,仔鱼对3种粒径微塑料的7 d清除率达到100%,其他3个较高浓度暴露后的清除率也在60%以上,但微塑料粒径与暴露后清除效果关系并不明确(图 1)。
3 讨论 3.1 PS微塑料在稀有稀有
已报道的研究证明微塑料能够在个体可观察水平对水生生物产生毒性效应。从文献报道的情况看,已经证实在浮游动物(Lee et al., 2013;Hämer et al., 2014)、贻贝(von Moos et al., 2012)和斑马鱼(Lei et al., 2018)等的生长、存活等个体水平上产生毒性效应。本文在个体水平上未检测到微塑料的毒性效应,其原因可能是:(1)PS微塑料对稀有
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