粒径和包裹物对纳米银在海洋微藻中的毒性影响

纳米银由于其独特的抗菌特性广泛应用于工商业领域.通过研究3种不同粒径和包裹物的纳米银对典型海洋微藻中肋骨条藻(Skeletonema costatum)的毒性效应,进一步丰富纳米银对藻类毒性效应及其作用机制的认识.结果表明3种纳米银对硅藻细胞生长都有不同程度的生长抑制效应,其毒性大小依次为10 nm-OA10 nm-PVP20 nm-PVP.在细胞毒性方面,相同粒径的聚乙烯吡咯烷酮(PVP)包裹的纳米银比油胺包裹(OA)的纳米银毒性更小;而相同包裹物的10 nm和20 nm两种尺寸的纳米银的毒性效应主要取决于其在水环境中的粒径和浓度,在浓度小于500μg·L~(-1)时,大粒径的纳米银毒性更大;在浓度≥500μg·L~(-1)时,小粒径的纳米银毒性更大.在光合毒性方面,50μg·L~(-1)的10 nm-PVP显著上调3Hfcp A的表达(P0.05)和显著下调D1的表达(P0.05),10 nm-OA在浓度为500μg·L~(-1)显著上调3Hfcp A的表达(P0.05),而20nm-PVP处理组没有显著性变化,说明光合作用相关基因的表达对小粒径PVP包裹的纳米银更敏感.本研究结果表明,纳米银对海洋微藻的毒性是由粒径大小、包裹物类型、暴露介质以及暴露浓度等多方面因素共同决定的.纳米银的粒径越小毒性越大,粒径对纳米银的毒性效应影响很大;而包裹物通过影响纳米银在水介质中的粒径大小间接影响纳米银的毒性效应.     

两种粒径纳米银对Nitrosomonas europaea的毒性效应

为明晰不同粒径纳米银（AgNPs）对欧洲亚硝化毛杆菌（Nitrosomonas europaea）的毒性效应,采用室内培养方式,探究10nm和50nm的AgNPs对N.europaea生长、氮转化能力、细胞结构、活性氧生成和功能基因表达的影响.结果表明,AgNPs暴露抑制N.europaea生长,随着暴露时间的延长,细菌生长抑制率增加,在4h达到最大值;培养基中NH₄⁺向NO₂^-转化速率减缓,N.europaea的铵态氮转化能力降低;扫描电镜（SEM）图像显示AgNPs造成部分细菌表面塌陷且有孔洞,细胞膜受损严重;透射电镜（TEM）图像显示AgNPs造成细菌内部核物质消融,细胞质膜界限模糊;流式细胞仪（FCM）检测发现AgNPs增加细胞内活性氧的生成;qRT-PCR技术对AgNPs暴露后N.europaea功能基因amoA、hao、merA表达进行测定,发现AgNPs抑制N.europaea功能基因的转录表达.综上所述,AgNPs通过与细胞膜相互作用和产生氧化应激损伤N.europaea,抑制amoA和merA的表达,进而影响铵态氮转化过程,且小粒径AgNPs的毒性强于大粒径.     

纳米银对小球藻光合作用和呼吸作用的影响

使用硼氢化钠还原硝酸银,聚乙烯醇(PVA)作为分散剂,制备出粒径为(7 ± 3)nm的纳米银,分别使用计数法和溶解氧法,研究了纳米银对小球藻(Chlorella vulgaris)生长、光合作用和呼吸作用的影响,并调查了对叶绿素a的抑制状况.结果显示,黑暗条件下加入3mg/L的纳米银,基本抑制了小球藻的呼吸作用,当暴露于4mg/L的纳米银时,小球藻生长的抑制率为93%;而光照条件下加入10mg/L纳米银时,才能抑制其光合作用,此时对小球藻生长的抑制率达到90%.光照时,叶绿素a在10mg/L纳米银的作用下,抑制率达到77%.研究表明了纳米银对小球藻呼吸作用有很强的抑制作用,对光合作用的影响可能通过抑制叶绿素a的合成或破坏叶绿体的结构来完成;光照能够明显减弱纳米银的毒性.     

纳米银对聚磷菌吸磷和释磷的影响及毒性效应

利用硼氢化钠还原硝酸银,并使用聚乙烯醇（PVA）作为分散剂,制备出分散良好、粒径为（14±3）nm的纳米银颗粒,考察了其对聚磷菌（Microlunatus phosphovorus）好氧吸磷和厌氧释磷的影响,以及产生的毒性效应.结果表明,在好氧状态下,7mg/L的纳米银能够完全抑制聚磷菌的生长（P <0.01）,达到10mg/L时才能完全抑制聚磷菌的吸磷能力（P=0.01）;在厌氧状态下,大于20mg/L的纳米银才使聚磷菌释磷能力受到部分抑制（P <0.05）.活性氧簇（ROS）和扫描电子显微镜（SEM）的检测结果表明,纳米银使细菌体内ROS水平降低,部分细菌菌体表面塌陷,这说明,纳米银不但可以毒害聚磷菌菌体表面,还可以降低菌内ROS水平.     

纳米银和银离子对土壤中硝化微生物及其氨氧化速率的影响

为研究纳米银对土壤硝化微生物及其氮转化的影响,采用土壤培养方式,对不同剂量纳米银(10、50、100 mg·kg~(-1))和银离子(1、5、10 mg·kg~(-1))暴露下黄棕壤和水稻土硝化细菌数量、土壤酶活性、amoA基因丰度、NH_4~+-N与NO_3~--N含量变化以及土壤潜在氨氧化速率进行研究.结果表明,纳米银和银离子暴露后,2种土壤亚硝酸细菌和硝酸细菌数量显著减少;土壤酶活性受到抑制,对脲酶的影响大于过氧化氢酶;土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)的amoA基因丰度均降低,对AOB基因丰度的影响大于AOA.在2种土壤中外源添加(NH_4)_2SO_4时,随着纳米银和银离子暴露剂量增加,土壤NH_4~+-N含量累积,NO_3~--N含量减少,氨氧化速率降低,铵态氮向硝态氮的转化受阻.综上所述,纳米银和银离子对土壤硝化微生物产生毒害作用并影响铵态氮转化,且影响程度与土壤理化性质有关.     

不同粒径纳米TiO2对大型溞的毒性效应及腐殖酸的调控作用

为评估纳米TiO₂在环境水体中的暴露风险,选用大型溞作为模式生物,研究了不同粒径纳米TiO₂（20、40、60和100 nm）对大型溞毒性效应的影响,并探究了腐殖酸对不同粒径纳米TiO₂毒性效应的调控作用.结果表明,粒径是影响纳米TiO₂颗粒毒性效应的重要因素,以大型溞半数致死时间（LT₅₀）为指标,不同粒径纳米TiO₂对大型溞的毒性作用强弱顺序依次为：20 nm颗粒 > 40 nm颗粒 > 60 nm颗粒 > 100 nm颗粒（p<0.05）.腐殖酸的存在可以显著降低纳米TiO₂颗粒对大型溞的毒性作用,腐殖酸对小尺寸纳米TiO₂颗粒的毒性抑制作用更为明显（p<0.05）.大型溞体内ROS水平与抗氧化系统相关酶活分析表明,纳米TiO₂导致大型溞体内活性氧自由基（ROS）浓度升高是其产生毒性作用的重要原因,腐殖酸的存在可以显著降低大型溞体内由于纳米TiO₂暴露而引起的ROS浓度上升（p<0.05）,进而减轻纳米TiO₂对大型溞的毒性作用.此外,腐殖酸可以减小不同粒径纳米TiO₂之间的毒性差异.本研究结果可为纳米TiO₂在环境水体中的暴露风险评估提供参考依据.     

甲胺磷、乙草胺和铜单一与复合污染对黑土环境安全的胁迫研究

以黑土为环境介质,通过急性毒性试验法研究了东北黑土区普遍存在的2种农用化学品甲胺磷和乙草胺与重金属Cu对赤子爱胜蚓(Eiseniafoetida)的单一与复合毒性效应.单一毒性试验结果表明,3者对蚯蚓均有毒性,顺序为甲胺磷>乙草胺>Cu.复合毒性实验结果表明,2种有机农药与重金属Cu复合毒性效应十分复杂,与不同的浓度组合及染毒历时有关,一般随时间的延长,毒性加剧.2种有机农药通过不同途径毒害蚯蚓,复合毒性效应表现为协同作用.可见,3者对土壤生态系统环境安全性和土壤健康质量存在潜在危害,同时这几种污染物的共存进一步加大了潜在危害性,且复合毒性效应与各组浓度组合及污染暴露时间密切相关.     

利用斑马鱼胚胎研究污水处理工艺中DOM与DBPs的关系

以模式生物斑马鱼胚胎为试验材料,通过暴露试验研究了污水处理工艺不同阶段出水中溶解性有机物(DOM)与消毒副产物(DBPs)毒性效应关系,揭示污水处理工艺中DBPs前体物DOM的生成变化规律以及对再生水生态安全性的影响。结果表明,污水处理各阶段DOM和DBPs水样对斑马鱼胚胎均有不同程度的致死效应,改良A2/O工艺段水样暴露的斑马鱼胚胎死亡率较大,DOM暴露的斑马鱼胚胎致死毒性效应低于DBPs,且DBPs对斑马鱼胚胎的半致死毒性效应较大;污水处理过程中增加的腐殖酸类物质主要来自于好氧过程微生物的生化合成;污水处理各阶段水样中腐殖酸类物质含量水平与斑马鱼胚胎的致死毒性效应存在正相关关系,因此污水生物处理会增大水中消毒前体物(DOM)特别是腐殖酸类物质的含量,从而增加了DBPs的产生量和再生水的生物毒性及环境风险。     

聚苯乙烯微塑料对糖尿病小鼠肾脏的影响

以糖尿病小鼠为模式生物,探究了100nm和5μm的聚苯乙烯微塑料(PSMPs)(200μg/L)通过饮水暴露28d后对小鼠肾脏的毒性.结果表明,糖尿病小鼠对PSMPs的暴露更加敏感,100nm和5μm PSMPs暴露导致糖尿病小鼠肾脏出现明显的炎性细胞浸润和淤血等病理损伤,且100nm PSMPs对肾脏造成的病理损伤更为严重.此外100nm PSMPs暴露显著加剧了糖尿病小鼠肾脏炎症因子(TNF-α和IL-6)、超氧化物歧化酶(SOD)和血清肌酐表达水平,并导致肾脏代谢紊乱.与5μm PSMPs相比,100nm PSMPs暴露呈现出的小粒径毒性效应可能与纳米级别PSMPs更易透过肠道屏障在肾脏中累积有关.此外,肾脏代谢通路紊乱引起的糖酵解能力受损和能量代谢水平降低也增加了PSMPs对糖尿病小鼠肾脏的负荷.     

基于毒理基因组学的PBDE-209和镉联合毒性效应研究

多污染物引发的联合毒性是环境污染物风险评价中的重要问题之一,然而此类研究相对不足.多溴联苯醚和重金属镉可在多种环境介质中共存,虽对其各自单独暴露所产生的毒性已有相对明确的认识,但对二者联合毒性效应及其分子机制的探索仍十分有限.生物信息学的发展为污染物的健康效应研究提供了一种新的思路.为此,本研究基于比较毒理基因组学数据库(CTD)分析了十溴联苯醚和镉在单独或共同暴露下所影响的基因和相关疾病,推测两种污染物对胎儿红系造血过程产生的潜在影响,并进一步整合外源红系细胞基因表达数据库中的基因芯片确证二者共同影响的基因集合.同时,通过对这些基因集合进行生物学过程和信号通路富集分析,筛选出潜在的靶基因JAK3和AKT2,并在无显著细胞毒性的暴露条件下,通过体外细胞实验验证了十溴联苯醚和镉对这两个靶基因表达的干扰.本研究结果可为预防多溴联苯醚和重金属的联合暴露风险提供一定的科学依据,同时,CTD数据库和外源特定疾病发生发展相关基因表达数据库联合使用,并结合传统毒理学研究手段的评价模式,可为污染物的共同暴露与特定疾病的相关性及分子机理的探索在研究思路和技术手段上提供参考.     

Water chemistry controlled aggregation and photo-transformation of silver nanoparticles in environmental waters

The inevitable release of engineered silver nanoparticles (AgNPs) into aquatic environments has drawn great concerns about its environmental toxicity and safety. Although aggregation and transformation play crucial roles in the transport and toxicity of AgNPs, how the water chemistry of environmental waters influences the aggregation and transformation of engineered AgNPs is still not well understood. In this study, the aggregation of polyvinylpyrrolidone (PVP) coated AgNPs was investigated in eight typical environmental water samples (with different ionic strengths, hardness, and dissolved organic matter (DOM) concentrations) by using UV–visible spectroscopy and dynamic light scattering. Raman spectroscopy was applied to probe the interaction of DOM with the surface of AgNPs. Further, the photo-transformation and morphology changes of AgNPs in environmental waters were studied by UV–visible spectroscopy, inductively coupled plasma mass spectrometry, and transmission electron microscopy. The results suggested that both electrolytes (especially Ca^2 + and Mg^2 +) and DOM in the surface waters are key parameters for AgNP aggregation, and sunlight could accelerate the morphology change, aggregation, and further sedimentation of AgNPs. This water chemistry controlled aggregation and photo-transformation should have significant environmental impacts on the transport and toxicity of AgNPs in the aquatic environments.     

Molecular toxicity and defense mechanisms induced by silver nanoparticles in Drosophila melanogaster

The widely use of silver nanoparticles (AgNPs) as antimicrobial agents gives rise to potential environmental risks. AgNPs exposure have been reported to cause toxicity in animals. Nevertheless, the known mechanisms of AgNPs toxicity are still limited. In this study, we systematically investigated the toxicity of AgNPs exposure using Drosophila melanogaster. We show here that AgNPs significantly decreased Drosophila fecundity, the third-instar larvae weight and rates of pupation and eclosion in a dose-dependent manner. AgNPs reduced fat body cell viability in MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays. AgNPs caused DNA damage in hemocytes and S2 cells. Interestingly, the mRNA levels of the entire metallothionein gene family were increased under AgNPs exposure as determined by RNA-seq analysis and validated by qRT-PCR, indicating that Drosophila responded to the metal toxicity of AgNPs by producing metallothioneins for detoxification. These findings provide a better understanding of the mechanisms of AgNPs toxicity and may provide clues to effect on other organisms, including humans.     

具有不同粒径和相同表面结构纳米银颗粒的制备及表征

具有不同粒径相同表面结构AgNPs（nano-silver,纳米银）的可控合成是开展AgNPs毒性研究和风险评估的基础,也是材料制备领域的难点之一.采取化学还原的方法,使用AgNO₃（硝酸银）作为反应前体,使用TSC（trisodium citrate,柠檬酸三钠）和NaBH₄（sodium borohydride,硼氢化钠）作为稳定剂及还原剂,通过优化剂量比和反应条件等合成参数,一步式原位反应生成不同粒径的AgNPs.利用TEM（透射电子显微镜）、UV-Vis（紫外可见分光光谱）、ICP-MS（电感耦合等离子体质谱）、FT-IR（傅里叶转换红外光谱）和DLS（动态光散射）等技术综合表征了合成纳米颗粒的形貌和结构性质.结果表明:①TEM结果显示,3种AgNPs均为球形且粒径分别为12、25和33 nm.②UV-Vis表征结果显示,所得产物在391~408 nm之间有较强吸收,说明合成产物为AgNPs.③利用ICP-MS测试样品中未反应的ρ（Ag+）,得出该制备方法具有高产率（>99%）.④DLS结果证实了合成的AgNPs在水溶液中带负电荷且具有较窄的粒度分布.⑤FT-IR结果显示,所制备的AgNPs表面结构一致,具有碳碳双键、酯基、羧基和羟基等官能团,在材料制备的过程中,溶液的初始配比、反应时间及环境条件都会对反应结果产生很大影响.研究显示,通过化学还原方法制备的AgNPs具有方法简便、重现性好、产率高和单分散性的特点,所制备的系列AgNPs颗粒表面结构一致,具有良好的化学稳定性.      

十溴二苯醚及其降解产物对浮游生物的毒性

十溴二苯醚(BDE-209)是我国环境中主要的多溴二苯醚(PBDEs)同系物.为研究BDE-209及其降解产物对水环境的影响,以初级消费者浮游动物大型蚤(Daphnia magna)和初级生产者浮游植物水华微囊藻(Microcystis flos-aquae)为染毒对象,研究BDE-209及其降解不同阶段产物对浮游生物的毒性.结果表明,大型蚤方面,繁殖毒性大于生长毒性,48 h半数致死浓度(48h-LC50)大小为:还原降解中间产物(0.80 mg·L~(-1),高毒)BDE-209(8.74 mg·L~(-1),中毒)还原降解终产物(15.27 mg·L~(-1),低毒),还原-氧化降解终产物的死亡率与溶剂空白一致,表明其基本无毒.水华微囊藻方面,染毒物质的毒性大小顺序与大型蚤一致,1 mg·L~(-1)的BDE-209、还原中间产物、还原终产物及还原-氧化终产物对水华微囊藻的抑制率分别为15.7%、93.7%、6.6%和1.3%.BDE-209降解过程中易生成毒性较大的中间产物,彻底还原脱溴可降低其毒性,后续辅以氧化降解,可消除其环境毒性.     

基于g-C3 N4研究环境中甲氧苄啶的光降解行为及其毒性

为研究残留抗生素在不同环境介质中的可见光降解行为特征,借助非金属和生物相容的石墨相氮化碳（g-C₃N₄）构建恒温恒湿和快速模拟可见光降解的实验环境,考察甲氧苄啶（TMP）进入不同环境（水、大气颗粒物和土壤）中的光降解过程与机制,并深入探讨TMP降解前后毒性变化规律.结果表明,用氙灯模拟太阳光和g-C₃N₄共存下,水、大气颗粒物和土壤中TMP经光照3 h后降解率分别为89.2%、35.8%和16.9%,同时发现参与水体TMP光降解的主要活性物质是·OH,而在大气颗粒物和土壤介质中光降解过程主要受·O₂^-控制.与大气颗粒物、土壤相比,水中TMP光解后可生成较为稳定的羰基化中间产物（m/z,305）,水生毒性试验结果表明TMP光解产物对斜生栅藻表现出较原药更强的毒性,可显著抑制藻细胞生长率、叶绿素a和b的含量,并引起细胞氧化应激.     

沿江化工园区污水处理厂出水对体外培养的大鼠睾丸细胞生殖毒性研究

应用基于原代培养的大鼠睾丸细胞体外毒性测试方法,通过支持细胞、生精细胞及间质细胞这3种细胞的形态、活力、胞外乳酸脱氢酶（LDH）活力、细胞早期凋亡坏死率和睾酮分泌量等指标对沿江化工园区污水处理厂出水的生殖毒性进行了研究. 结果表明,新开发的沿江化工园区A和省级沿江化工园区B污水处理厂出水有机提取物浓缩后均能造成不同程度的大鼠睾丸细胞生殖毒性,园区A提取物能明显影响支持细胞（p<0.01）、生精细胞（p<0.05）和间质细胞（p<0.05）的细胞活力,影响支持细胞和生精细胞胞外LDH活力（p<0.01）,且其毒性明显高于园区B. 暴露导致支持细胞处于凋亡早期,而生精细胞比较敏感,已到了凋亡晚期甚至有小团细胞坏死,虽然间质细胞的形态没有显著变化但园区A有机提取物已明显抑制了间质细胞睾酮的分泌（p<0.01）. 研究表明,通过基于原代培养的大鼠睾丸细胞体外毒性测试方法能够对污水处理厂出水的生殖毒性做出综合评价,该方法可以作为现行国家污水综合排放化学监测的有效补充.     

Sunlight-driven reduction of silver ion to silver nanoparticle by organic matter mitigates the acute toxicity of silver to Daphnia magna

Due to the unique antibacterial activities, silver nanoparticles (AgNPs) have been extensively used in commercial products. Anthropogenic activities have released considerable AgNPs as well as highly toxic silver ion (Ag⁺) into the aquatic environment. Our recent study revealed that ubiquitous natural organic matter (NOM) could reduce Ag⁺ to AgNP under natural sunlight. However, the toxic effect of this process is not well understood. In this work, we prepared mixture solution of Ag⁺ and AgNPs with varied Ag⁺% through the sunlight-driven reduction of Ag⁺ by NOM and investigated the acute toxicity of the solutions on Daphnia magna. Formation of AgNPs was demonstrated and characterized by comprehensive techniques and the fraction of unconverted Ag⁺ was determined by ultrafiltration-inductively coupled plasma mass spectrometry determination. The formation of AgNPs enhanced significantly with the increasing of solution pH and cumulative photosynthetically active radiation of sunlight. The toxicity of the resulting solution was further investigated by using freshwater crustacean D. magna as a model and an 8 hr-median lethal concentration (LC₅₀) demonstrated that the reduction of Ag⁺ by NOM to AgNPs significantly mitigated the acute toxicity of silver. These results highlight the importance of sunlight and NOM in the fate, transformation and toxicity of Ag⁺ and AgNPs, and further indicate that the acute toxicity of AgNPs should be mainly ascribed to the dissolved Ag⁺ from AgNPs.     

Environmental fate and behavior of silver nanoparticles in natural estuarine systems

Silver nanoparticles (AgNPs) are widely used in many consumer products, whereas their environmental behaviors in natural aquatic systems remain unknown, especially in natural brackish media. Therefore, it is urgent to investigate the environmental fate of AgNPs in natural brackish waters. Here, we investigated the stability of citrate-coated AgNPs in natural brackish water collected from 6 different sites with distinct salinities in the Xinglinwan Reservoir, located in Xiamen City, southeast China. The obtained results showed that AgNP colloids remained stable in low-salinity waters, which was mainly determined by the effects of dissolved organic matter (DOM) promoting the stability of the nanoparticles. However, the environmental fate of AgNPs in high-salinity waters was dominated by the salinity or ionic strength, especially the free ion concentrations of Cl^?, SO₄^2?, or S^2?, resulting in rapid sedimentation and dissolution. In addition, both DOM and salinity contributed to the environmental behavior of AgNPs in moderate-salinity waters, ultimately resulting in either colloidal stability or sedimentation. Overall, these results may reveal that AgNPs remain relatively stable for a long period in low-salinity natural waters, and that the stability might gradually decrease as AgNPs are transferred from freshwaters through brackish waters and eventually end up in seawater along the bay. Our findings also further indicate that the toxicity and potential risks of AgNPs may present more serious threats to the environment and organisms in natural freshwaters than in natural estuarine systems or seawater.     

玛瑙河多环境介质和铜锈环棱螺体内微塑料的赋存特征

微塑料污染对水生态系统及人类健康危害大,为探究微塑料在不同环境介质中的赋存特征,选择长江一级支流玛瑙河为研究区域,通过现场采样、显微镜观察和傅里叶红外光谱测定等,对玛瑙河表层水体、沉积物、河岸带土壤和底栖动物铜锈环棱螺中微塑料的丰度、粒径、形状、颜色和组成类型进行了分析.结果表明,玛瑙河表层水体的微塑料平均丰度为（5.9±0.26）n·L^-1;上层沉积物中微塑料丰度（以干重计）为（1.35±0.1）n·g^-1,下层沉积物中微塑料丰度（以干重计）为（0.93±0.12）n·g^-1;近河岸带土壤中微塑料丰度（以干重计）为（0.68±0.16）n·g^-1,远河岸带土壤中微塑料丰度（以干重计）为（0.69±0.14）n·g^-1;铜锈环棱螺体内微塑料丰度为（2.06±0.25）n·g^-1.分析发现,上层沉积物和下层沉积物中微塑料丰度呈正相关;铜锈环棱螺体内微塑料丰度分别与上、下层沉积物中微塑料丰度呈正相关;近、远河岸带土壤中微塑料丰度具有相关性.各环境介质和铜锈环棱螺体内微塑料粒径大多<0.1mm,主要形态为纤维状和碎片状,颜色以蓝色和黑色为主,成分主要是聚丙烯（PP）和聚乙烯（PE）.研究发现,河岸带土壤中微塑料主要来源于农用塑料薄膜的破碎和分解.通过多环境介质调查和铜锈环棱螺体内微塑料的分析,探明了大型底栖动物体内微塑料的累积效应,可为全面了解微塑料潜在生态风险提供依据.     

Improving eco-efficiency in the steel industry: The ArcelorMittal Gent case

In addition to CO₂ released by the combustion of fossil fuel and leading to climate change, large steelworks emit pollutants that have other environmental impacts. ArcelorMittal Gent, an integrated steelwork producing ca. 5 × 10⁶ tons of steel per year, not only decreased its specific energy consumption and CO₂-emissions, but also reduced the environmental impact of its other emissions. This is illustrated by means of the evolution of 6 partial eco-efficiency indicators for the impact categories acidification, photo-oxidant formation, human toxicity, freshwater aquatic ecotoxicity, eutrophication and water use. The partial eco-efficiency indicators are eco-intensities, defined as the environmental impact in the respective impact category, divided by the amount of liquid steel produced. In the period 1995 – 2005 these indicators decreased by 45, 4, 52, 9, 11 and 33% respectively, whereas the steel production increased by 17%. The net impact of discharges of wastewater is negligible for human toxicity and is negative (concentrations lower than in the canal water used) for freshwater aquatic toxicity and eutrophication. For acidification, human toxicity (only emissions to air) and water use, the decoupling between environmental impact and production was absolute; for photo-oxidant formation, freshwater aquatic ecotoxicity (only emissions to air) and eutrophication, it was relative.