密度分离法提取土壤中微塑料的优化

为探究一种简单、经济和可靠的微塑料分离提取手段,设立了11种(F1~F11)具有不同体积比的饱和NaCl和饱和NaI溶液的混合溶液,将混合液作为浮选液来提取土壤中的4种类型(聚乙烯,PE;聚苯乙烯,PS;聚氯乙烯,PVC;聚对苯二甲酸乙二醇酯,PET)微塑料,同时对浮选过程进行了改进.结果表明:当单独使用NaCl溶液浮选时,微塑料的总提取率为55.83%,但随着NaI所占体积比的不断增加,微塑料的总提取率基本呈上升趋势,在NaCl:NaI=1:1时总提取率超过了90%,在只有NaI的条件下总提取率高达96.67%;在F1~F11的任意条件下,PE和PS表现为较高的提取率,均超过了86.67%;PVC和PET在单独使用NaCl溶液浮选时提取率极低,在NaCl:NaI=1:1条件下提取率分别为93.33%和90%,接近单独使用NaI溶液浮选时的提取率;F1~F11任意条件下,低密度微塑料PE和PS的提取率之和均高于高密度微塑料PVC和PET的提取率之和,但在F6~F11条件下提取率差值不大.根据提取结果并结合经济成本等多方面因素考虑,建议使用NaCl:NaI=1:1的混合溶液来分离土壤中的微塑料.     

滇池近岸水体微塑料污染与富营养化的相关性

内陆淡水湖泊微塑料污染受到广泛关注,但与水体富营养化的相关性尚不明晰.在滇池沿岸布设24个采样点,研究了近岸表层水体微塑料丰度、聚合物成分、粒径、颜色及形态,同步测定富营养化相关水质指标并计算富营养化状态指数.结果表明,滇池近岸水体微塑料丰度在800~6000 n·m^-3之间,平均为2867 n·m^-3,检出的聚合物类型主要是聚对苯二甲酸乙二醇酯（polyethylene terephthalate,PET）、聚醚型聚氨酯（polyetherurethane,PEU）、聚丙烯（polypropylene,PP）、聚乙烯（polyethylene,PE）和聚醋酸乙烯酯（polyvinyl acetate,PVAc）.微塑料粒径以0.2~0.5 mm为主,纤维状微塑料是最主要的检出形态,其次为碎片和薄膜.24个监测点中,处于重度、中度、轻度富营养化和中营养水平的样点数分别占8.33%、58.33%、29.17%和4.17%,主要污染物为总氮（TN）.相关性分析发现滇池近岸水体微塑料丰度与TN质量浓度呈极显著正相关（P<0.01）,与叶绿素a（Chl-a）呈负相关,但未达到显著性水平（P>0.05）,靠近昆明市主城区的北岸水体中微塑料丰度和TN质量浓度均显著高于其他三岸,主要来源于污水处理厂尾水排放.     

大辽河沉积物中微塑料的污染特征

以大辽河为研究对象,选取8个典型区域的沉积物,采用密度浮选法,结合体视显微镜及μ-FTIR,进行大辽河沉积物中微塑料的组成及分布特征研究.结果表明,大辽河沉积物微塑料平均丰度为（66.67±79.93）个/kg;其中位于鞍山海城市的DL5丰度最高,为（193.33±172.43）个/kg,辽阳县柳壕镇的DL6丰度最低,为（20.00±34.64）个/kg;微塑料颜色以白色、黑色、黄色为主（83.75%）,形状以纤维、薄膜、碎片为主（91.25%）,粒径以（1000~2000）μm为主（33.75%）.通过红外光谱分析可得出,大辽河沉积物中薄膜类微塑料的主要成分是聚酰胺（PA）、聚丙烯（PP）和聚乙烯（PE）,碎片类的主要成分是聚乙烯（PE）和聚丙烯（PP）,泡沫类和颗粒类的主要成分是聚苯乙烯（PS）,纤维类的主要成分是人造丝（RY）及聚对苯二甲酸乙二醇酯（PET）.总体上,该研究区域沉积物中微塑料的丰度处于中等偏低水平.本研究可为我国淡水河流沉积物中微塑料的污染分布研究提供基础科学数据.     

辽河流域土壤中微(中)塑料的丰度、特征及潜在来源

以辽河流域为研究对象,基于密度浮选原理,结合体视显微镜及显微红外光谱（μ-FTIR）,对辽河流域土壤中微（中）塑料的形貌、丰度及分布特征进行了系统地研究,并合理推测了影响微（中）塑料分布的主要因素及潜在来源.结果表明,土壤微（中）塑料平均丰度为（145.83±211.46） n·kg^-1.微（中）塑料以碎片（46.00%）、<1000 μm （39.57%）、聚丙烯（PP）（41.71%）和白色（46.86%）为主.其中,聚乙烯（PE）主要形状是薄膜和碎片（96.91%）,PP主要形状是碎片（85.62%）,聚苯乙烯（PS）主要形状是泡沫,人造丝（Rayon）、聚酯纤维（PES）和聚对苯二甲酸乙二醇酯（PET）微（中）塑料以纤维状为主（>85%）.工业活动（快递公司、塑料厂、服装厂）、农业活动（地膜覆盖、污水灌溉、污泥堆肥）、人口密度和污水处理厂等都可能会导致土壤中微（中）塑料丰度的较高水平.污水排放及其灌溉、塑料制品（农药瓶、化肥包装袋、塑料农膜、编织袋等）和用于装饰、包装、运输的泡沫材料等是辽河流域土壤中微（中）塑料的潜在来源.     

大理河流域微塑料空间分布及其来源

微塑料（直径<5 mm的塑料颗粒）作为一种新型环境污染物,对土壤及水生生态系统构成了严重威胁.本研究基于黄河中游大理河流域采集的21个沉积泥沙和12个农田土壤样品,分析了大理河流域不同介质中微塑料赋存特征和组成差异,揭示了不同介质中微塑料来源及 沉积泥沙中微塑料丰度的影响因素.结果表明,大理河流域沉积泥沙和农田土壤中都受到微塑料的严重污染,其丰度分别为440~1190个·kg^-1和1180~2930个·kg^-1,其中,小粒径微塑料（粒径<500 μm）为主要组成,分别占泥沙和土壤中微塑料颗粒总数的50.12%和72.5%.纤维是泥沙和土壤中微塑料最常见的形状,分别为66.95%和48.3%;薄膜和小球微塑料在农田土壤中较多,比沉积泥沙中高6.51%和13.83%.通过拉曼 光谱仪测试,沉积泥沙中微塑料聚合物组成主要为聚对苯二甲酸乙二醇酯（PET）和聚丙烯（PP）,农田土壤中主要微塑料聚合物组成为聚乙烯（PE）.农用地膜是农田土壤中主要微塑料来源,生活污水和固体废物是泥沙中微塑料的主要来源.此外,沉积泥沙中微塑料丰度的主要影响 因素为速效磷含量和径流流速.本研究结果可为深入了解黄河中游流域微塑料污染状况提供参考.     

海州湾潮间带沙蚕对沉积物微塑料的指示作用

2018年7月于海州湾潮滩设置3个断面共9个站位,通过对海州湾潮滩沉积物中以及沙蚕体内微塑料的丰度和形态特征的研究,探讨了沙蚕体内微塑料的来源,以及沙蚕对潮滩沉积物微塑料的指示作用.结果表明,潮滩沉积物中微塑料的平均丰度为（0.49±0.17） n·g^-1,处于国内近岸环境研究的较高水平.所有检测到的微塑料中,最为丰富的形态和颜色类型分别为纤维和黑灰色,材质以聚乙烯（polyethylene,PE）、聚酯纤维（polyester,PET）和聚苯乙烯（polystyrene,PS）为主.沙蚕中微塑料检出率为77.78%~86.67%,平均丰度为（6.68±2.21） n·ind^-1,其丰度与个体质量显著正相关（r=0.42,P=0.002）,个体质量1.5 g以上的沙蚕中微塑料丰度显著高于<0.5 g和0.5~1 g两个组别（F₃=141.029,P=0.000）,微塑料形态以黑色或蓝色小纤维为主,主要材质同样为聚乙烯和聚酯纤维.通过对0~3 mm范围内的微塑料各项特征分析发现,沉积物与沙蚕中的微塑料丰度强相关（r=0.79,P=0.01）,其主要形态组成（r=0.90,P=0.035）和材质组成（r=0.73,P=0.024）同样显著相关,表明沙蚕会摄取沉积环境中的微塑料,存在与沉积物之间的微塑料交换,沙蚕作为沉积物中微塑料污染指示生物物种是可行的.     

厦门市筼筜污水处理厂中微塑料的特征研究

滨海城市污水处理厂作为微塑料进入海洋的途径之一,对其微塑料污染物特征展开研究,有助于认识和了解人类活动对海洋环境的影响,但目前国内对该问题的研究还相对较少。本研究以厦门筼筜污水处理厂为例,设计改进采样装置和实验方法,对其进出水以及污泥中的微塑料污染物特征进行研究。结果表明,该污水处理厂进水中微塑料浓度为1.703个/L,初级处理出水为1.090个/L,二级处理出水为0.324个/L,去除率分别为35.99%、80.97%。初级处理污泥微塑料浓度为2.14×103个/kg,二级处理污泥为6.62×103个/kg。该污水处理厂污水中微塑料形态以纤维（43.89%）为主,颗粒（32.85%）次之;颜色以白色（34.95%）为主,透明（22.74%）次之;主要成分以PP（21.17%）为主,PS（18.39%）次之。污泥中微塑料形态以碎片（68.9%）为主,颗粒（34.78%）次之;颜色以黄色（39.13%）为主,黑色（17.39%）和白色（13.04%）次之;主要成分以PP（34.78%）为主,PE（26.09%）次之。每日经过该污水厂排入厦门西海域的微塑料数量约为9.72×104个,数量巨大不容忽视。本研究结果表明污水处理厂在减少城市微塑料污染物进入海洋方面发挥了重要作用,但仍具有较大的提升空间。     

聚乙烯微塑料对盐渍化土壤微生物群落的影响

地膜覆盖保墒已成为盐渍化土壤种植中重要的农艺措施,而盐渍化与微塑料双重胁迫对土壤微生物的影响越来越受到重视.为探究聚乙烯微塑料对盐渍化土壤微生物群落的影响,通过室内模拟盐渍化土壤环境中微塑料污染的方法,探究不同类型（氯盐类和硫酸盐类）和不同含量（弱、中、强）的盐渍化土壤赋存不同丰度聚乙烯（PE）微塑料（土样干重的1%和4%）条件下对土壤微生物群落的影响.结果表明,PE微塑料会降低盐渍化土壤微生物群落多样性和丰富度,且硫酸盐类盐渍土处理受到的影响更强烈.赋存PE微塑料后不同处理微生物组成基本一致,但其相对丰度会发生变化,硫酸盐类盐渍土处理中各菌群相对丰度的变化较氯盐类盐渍土处理更强;门水平上,变形菌门相对丰度与赋存PE微塑料丰度呈正相关,而拟杆菌门、放线菌门和酸杆菌门相对丰度与赋存PE微塑料丰度呈负相关;科水平上,黄杆菌科、食碱菌科、盐单胞菌科和鞘脂单胞菌科相对丰度随赋存PE微塑料丰度增大而增大.KEGG代谢通路预测显示,赋存PE微塑料会降低微生物新陈代谢和遗传信息等功能相对丰度,硫酸盐类盐渍土对新陈代谢功能的抑制作用强于氯盐类盐渍土,而对遗传信息功能的抑制效果弱于氯盐类盐渍土;新陈代谢功能二级通道中氨基酸代谢、碳水化合物代谢、能量代谢等功能受到抑制,推测新陈代谢功能的降低可能是由于上述二级代谢通路相对丰度降低引起的.试验结果可为微塑料和盐渍化双重污染条件下对土壤环境的影响研究提供理论依据.     

附着在不同微塑料表面的藻类结构与群落组成

选取水环境中常见的4种微塑料介质（聚丙烯（PP）、聚氯乙烯（PVC）、聚乙烯（PE）、聚对苯二甲酸乙二醇酯（PET））和一种自然介质（鹅卵石）进行野外生物膜的培养,以探究不同类型介质表面附着的生物膜的表观结构和藻类群落组成的差异特征.结果表明,鹅卵石与微塑料介质上的生物膜的形貌结构及藻类含量与组成特性均存在一定差异：在微塑料介质表面附着的藻类叶绿素a浓度普遍低于自然介质.实验进一步发现,不同的微塑料类型同样会对生物膜藻类含量以及功能特性存在一定影响：PET片上附着的藻类叶绿素a浓度最高（613.7μg/L）,PP片上最低（492.5μg/L）;然而,PP片上藻类的光合作用最大量子产量最高（0.443）.以上结论说明微塑料会改变附着藻类的生长情况和初级生产力,进而可能影响生物膜在水体中的碳循环过程,对水体净化及污染治理产生一定影响.     

微塑料对牟氏角毛藻的毒理效应

本文选取聚对苯二甲酸乙二酯（PET）和聚苯乙烯（PS）微粒作为目标污染物,探讨其不同浓度（50 mg/L、100 mg/L和200 mg/L）和不同粒径（100 μm、175 μm和250 μm）对受试生物牟氏角毛藻（Chaetoceros muelleri）的影响。结果表明,3种粒径和浓度的微塑料（PET 和 PS ）均对微藻细胞密度产生抑制作用,PET（100 μm,200 mg/L）和PS（100 μm,200 mg/L）对微藻产生的最大抑制率分别为 37.3% 和 32.5%。微塑料浓度越大,对藻细胞密度的抑制作用越强;而粒径变化对藻细胞密度的抑制作用未呈现明显规律。PET对牟氏角毛藻细胞干重的影响较小,与对照组相比无显著性差异（P>0.05）,PS则对牟氏角毛藻细胞干重无影响。微塑料对牟氏角毛藻叶绿素a（Chl a）的抑制作用与其类型和暴露时间有关。PET对Chl a起抑制作用,PS则起促进作用。     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Alterations of organ histopathology and metallolhionein mRNA expression in silver barb, Puntius gonionotus during subchronic cadmium exposure

Common silver barb,Puntius gonionotus,exposed to the nominal concentration of 0.06 mg/L Cd for 60 d,were assessed for histopathological alterations(gills,liver and kidney),metal accumulation,and metallothionein(MT)mRNA expression.Fish exhibited pathological symptoms such as hypertrophy and hyperplasia of primary and secondary gill lamellae,vacuolization in hepatocytes,and prominent tubular and glomerular damage in the kidney.In addition,kidney accumulated the highest content of cadmium,more than gills and liver.Expression of MT mRNA was increased in both liver and kidney of treated fish.Hepatic MT levels remained high after fish were removed to Cd-free water.In contrast,MT expression in kidney was peaked after 28 d of treatment and drastically dropped when fish were removed to Cd-free water.The high concentrations of Cd in hepatic tissues indicated an accumulation site or permanent damage on this tissue.