紫贻贝长期暴露于亚致死剂量的林丹和阿特拉津下的组织学反应

为开发快速、敏感的生物标记物以监测海洋贝类生物中是否存在农药,我们检测了双壳动物紫贻贝长期暴露在亚致死剂量的丙体六氯环乙烷(林丹,γ-HCH)和2-氯4-乙胺基-6异丙胺基-1,3,5-三氮苯(阿特拉津)下的组织学变化。紫贻贝容易积累环境中的杀虫剂,因此,本研究旨在阐明农药的生物积累与组织病理学效应之间的关系。利用GC/MSD分析法定期对贻贝和水样中的林丹与阿特拉津含量进行测定。将贻贝在实验室中培养21天,以使其代谢适应于带有水质控制的封闭式不间断流动系统。随后,30只贝暴露于亚致死剂量的林丹(0.9 mg/L)或阿特拉津(3.583 mg/L)溶液中56天。实验期间,控制重要的参数,比如温度和盐度分别控制在18℃和34‰。在处理28天和56天后取样,检测组织学损坏及吸收的农药量。暴露的紫贻贝每克干重分别能聚集约304.8-372.0μg/g林丹和83.3-137.4μg/g阿特拉津。组织学改变高度集中在鳃的上皮和外套膜组织;上皮与相邻的组织形成分离状态。组织病理学结果显示,抗性机制的激活使紫贻贝能在亚致死压力下存活。组织病理效应范围从浸润反应到以血淋巴细胞出现间质细胞反应为特征。因此,在农药聚集部位的组织学和超微结构的改变是敏感的,并与农药的生物积累具有正相关关系,说明这些改变可能作为农药暴露的生物标记物。     

重金属镉与铜在棕尾别麻蝇体内的积累和排泄研究

为了研究镉(Cadmium,Cd)和铜(Copper,Cu)在棕尾别麻蝇Boettcherisca peregrina (Diptera:Sarcophagidae)生长发育过程中积累和排泄的动态变化,在室内给棕尾别麻蝇初产幼虫连续饲喂含不同浓度Cd2'或Cu2+的人工饲料,每隔24 h测定幼虫对Cd2+或Cu2+积累和排泄及变态过程中的变化.结果表明:棕尾别麻蝇幼虫取食含有Cd2+的饲料后,大部分Cd2+积累在体内,少部分通过排泄系统排出.Cd2+在幼虫体内的积累量随着取食时间的延长而增加,幼虫取食含200 μg/g Cd2+饲料后,体内Cd2+积累量高于取食含100 μg/g、400 μg/g和800 μg/g Cd2+饲料后体内Cd2+积累量,幼虫对Cd2+排泄量亦有相同的规律.Cd2+在幼虫、蛹和刚羽化成虫中的含量依次降低.棕尾别麻蝇幼虫取食含Cu2+的饲料后,其体内Cu2+积累量小于排泄量,而且体内积累量和排泄量随着取食时间的延长而增加,Cu2+积累量和排泄量与取食饲料中的Cu2+浓度均不成正比.在变态过程中,Cu2+也不断被排泄,体内Cu2+含量随之下降.     

阿特拉津及其降解菌的使用对土壤微生物群落的影响

比较了阿特拉津及降解菌株BTAH1的使用对土壤微生物的影响.结果表明,在实验周期内阿特拉津对土壤微生物的代谢作用有较明显的刺激作用,与空白土壤(未施用阿特拉津和降解菌)相比,对照土壤(施用50mg·kg^-1土阿特拉津)呼吸强度显著增加,且土壤中的阿特拉津浓度对土壤NH₄⁺-N和NO₃^--N浓度的影响显著.降解菌BTAH1可在1周内降解土壤中98%以上的阿特拉津,从而使土壤呼吸强度有所下降,土壤中NH₄⁺-N和NO₃^--N的浓度基本与空白土壤持平,对微生物量C和微生物量N影响不显著;放线菌和真菌数量也基本与空白持平,细菌数量较高.对土壤细菌的16SrDNA文库的ARDRA分析发现,阿特拉津及其降解菌的使用对土壤细菌群落结构有一定程度的影响,阿特拉津的使用会降低细菌群落的多样性,而降解菌的使用会恢复土壤细菌的多样性.     

阿特拉津和毒死蜱对东北小鲵蝌蚪氧化应激的影响

为了研究阿特拉津和毒死蜱单一及联合暴露对东北小鲵(Hynobius leechii)蝌蚪抗氧化酶活性的影响,实验选择不同浓度、不同时间点阿特拉津和毒死蜱单一及联合暴露对东北小鲵蝌蚪超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)及丙二醛(MDA)的影响。结果表明,随着暴露时间的增加及各处理组浓度的升高,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)活性明显降低,丙二醛(MDA)水平明显上升,从而说明阿特拉津和毒死蜱单一及联合暴露会对东北小鲵蝌蚪的抗氧化酶系活性产生影响,进而产生毒理效应。当暴露21 d后,通过恢复实验,发现恢复末期与暴露末期相比,低浓度组和中浓度组上述3种酶活性变化不显著,而高浓度组上述3酶活性均有明显改善。     

基因工程菌对阿特拉津的生物转化及其影响因素

研究考察了基因工程菌转化阿特拉津的共代谢碳源、转化动力学和影响因素。结果表明,作为共代谢碳源,葡萄糖优于乙酸盐,碳源浓度对转化影响不大,对工程菌生长影响显著。阿特拉津比转化速率与工程菌初始密度无关,与阿特拉津初始浓度有关,用Monod方程拟合转化动力学,求得方程参数为V_(max)=0．168/h,Ks= 30．49mg/L。降低温度会显著降低阿特拉津比转化速率;偏碱性的条件下,阿特拉津转化率较高,酸性条件严重抑制阿特拉津转化;盐度在一定范围内不影响转化活性;Co~(2 )、Fe~(2 )、Fe~(3 )和Zn~(2 )促进阿特拉津转化,Mn~(2 )、Ni~(2 )和Cu~(2 )抑制阿特拉津转化。菌体细胞对阿特拉津的吸附和转化作用呈正相关关系。     

农田生态系统中除草剂阿特拉津的环境行为及其模型研究进展

阿特拉津是世界范围内广泛使用的除草剂,在曾经使用阿特拉津的国家的地下水、河流和湖泊中已经检测到阿特拉津的残留,长期暴露在该有机污染物的环境中,势必对动物的生殖与繁衍及人体的健康产生不良影响。介绍了阿特拉津国内外的研究情况,着重评述了阿特拉津的吸附机制与影响因素、化学降解、生物降解、生态毒理学评价以及模型对其环境行为的描述,目的旨在帮助理解田间阿特拉津迁移机制及其如何污染地下水。农田生态系统中阿特拉津的迁移规律决定其环境行为,它是进行环境和健康风险评价的基础,目前应用较多的迁移模型是经典的对流-扩散方程,该模型已广泛运用于水-土系统中化学物质的迁移,且室内模拟的实验结果能较好解释田间现象,因此,开展阿特拉津在稳定流场饱和/非饱和室内土柱中的混合置换实验及批量平衡吸附实验,可获得反映阿特拉津迁移的穿透曲线特性及阿特拉津吸附性能的分配系数,结合数学模型拟合阿特拉津在土壤中非平衡迁移曲线,用拟合参数预测不同深度土壤中阿特拉津浓度变化和累积淋溶量动态规律,应该是今后研究工作的重点。     

阿特拉津对雄性鲫鱼血清雌二醇含量的影响

在实验室条件下研究了除草剂阿特拉津对鲫鱼(Carassius auratus)的急性毒性以及在不同暴露浓度和不同作用时间下低剂量阿特拉津对雄性鲫鱼血清雌二醇含量的影响.结果表明:阿特拉津对鲫鱼的24、48和96 h的LC50分别为229.33、146.17和105.94 mg·L-1,安全浓度为10.59 mg·L-1;长期(24 d)暴露时,低浓度(0.1～5.0 mg·L-1)阿特拉津对雄性鲫鱼血清雌二醇的合成具有诱导作用,且1.0 mg·L-1的诱导作用最强,与对照组相比血清雌二醇含量增加了84.1%;高浓度(10.0 mg·L-1)阿特拉津对雄性鲫鱼血清雌二醇的合成具有抑制作用;在5.0 mg·L-1浓度下,各个处理时间的血清雌二醇含量均高于对照组,其中第14天时雌二醇含量最高,比对照组增加50%(P＜0.01).血清雌二醇含量的变化直接影响雄性鲫鱼的生殖功能和性征变化,因富集阿特拉津而导致血清雌二醇浓度发生显著变化的雄性鲫鱼可能会出现一定的生殖缺陷.     

阿特拉津降解菌株AD111的分离鉴定及其降解特性

【背景】玉豆轮作过程中,玉米田中长残留除草剂阿特拉津易对下茬大豆作物产生不良影响。【目的】从黑龙江省安达市的农田土筛选一株能适应该土壤环境生长的阿特拉津降解菌并研究其降解特性。【方法】利用富集培养法,分离、筛选一株阿特拉津高效降解菌并结合外观形态、生理生化及16SrRNA基因序列测定对其进行鉴定,通过单一变量法设置不同的碳源、pH、温度和阿特拉津浓度,研究降解菌株最佳发酵及降解条件。【结果】得到一株在BSM-G中能够以阿特拉津为唯一氮源生长的高效阿特拉津降解菌AD111,鉴定为马德普拉塔无色小杆菌(Achromobacter marplatensis)。菌株AD111降解阿特拉津的最适温度为35℃,最适pH为8.0,最佳碳源为蔗糖,24 h内对浓度为50 mg/L的阿特拉津降解率达到99.7%,对300 mg/L的阿特拉津降解率达到81.9%。【结论】降解菌AD111具有较好的环境适应及阿特拉津降解能力,为解决黑龙江偏碱土壤中阿特拉津残留提供了良好的候选菌株。     

阿特拉津降解菌株的分离、鉴定和工业废水生物处理试验

用液体无机盐培养基富集培养法和无机盐平板直接分离法, 从生产阿特拉津的农药厂的废水和污泥混合物中分离到13个能以阿特拉津为唯一氮源生长的细菌菌株。通过16S rRNA基因序列分析, 11个菌株被鉴定为Arthrobacter spp., 2个菌株被鉴定为Pseudomonas spp.。对阿特拉津降解活力最高的Arthrobacter sp. AD30和Pseudomonas sp. AD39的降解基因组成和降解特性进行了详细研究。降解基因的PCR扩增表明, AD30和AD39都含有trzN-atzBC基因, 能将有毒的阿特拉津降解成无毒的氰尿酸。降解实验表明, 向阿特拉津浓度为200 mg/L的无机盐培养基中分别接种等量的AD30、AD39和这两个菌株的混合菌液, 30°C振荡培养48 h以后, 阿特拉津去除率分别为92.5%、97.9%和99.6%, 表明混合菌的降解效果好于单菌。用AD30和AD39的混合菌液接种阿特拉津浓度为176 mg/L的工业废水, 30°C振荡培养72 h以后, 99.1%的阿特拉津被去除, 表明混合菌株在阿特拉津工业废水的生物处理中有很好的应用潜力。     

工业废水中阿特拉津降解细菌的遗传和生理多样性

【目的】通过遗传学和生理学实验,揭示分离自工业废水的阿特拉津降解细菌具有遗传和生理多样性,为阐明阿特拉津生物降解的分子机理和阿特拉津降解细菌在污染环境生物修复中的应用提供新见解。【方法】用普通PCR方法检测菌株的阿特拉津降解基因,分析其降解基因组成;用基因组重复序列PCR技术(rep-PCR)分析降解菌株的基因组类型;用Western blot方法检测菌株阿特拉津降解途径的第一个酶三嗪水解酶(TrzN);用不同氮源(阿特拉津、莠灭净、扑草净、西玛津、氰草净、阿特拉通和氰尿酸)和碳源(蔗糖、葡萄糖、麦芽糖、乳糖、柠檬酸钠、乙酸钠和琥珀酸钠)培养降解菌株,通过检测培养液的OD600值,证明菌株能够利用的氮源和碳源种类。【结果】对分离自工业废水的27个阿特拉津降解菌株所进行的阿特拉津降解基因PCR检测表明,其降解基因组成分别为trzN-atzBC、trzN-atzABC和atzADEF;通过rep-PCR实验将27个阿特拉津降解菌株分为7个群;Western blot结果表明,27个菌株中有24个含有三嗪水解酶TrzN;氮源利用实验表明,2个菌株能够利用所有7种氮源生长,其余25个菌株只能利用其中的2-6种;碳源利用实验表明,10个菌株能够利用所有7种碳源生长,其余17个菌株只能利用其中的3-6种。【结论】分离自某工业废水的27株阿特拉津降解功能菌存在相当广泛的遗传和生理学上的多样性,trzN-atzABC降解基因组成为首次发现。     

Physiological, anatomic and behavioural changes in the respiratory system of Salmo gairdneri Rich. On acute and chronic exposure to chlorothalonil

1. Various changes in anatomical, physiological and behavioural aspects of the respiratory system of Salmo gairdneri on exposure to chlorothalonil (TCIN) were examined, along with the relationship between LC50 response and oxygen concentration. 2. The TCIN LC50 was significantly lower at 5 mg/l oxygen concentration than at 8 mg/l. 3. Haematocrit was depressed on exposure to TCIN, whereas exposure to pentachlorophenol and lindane raised the haematocrit. Muscle lactate was not elevated in acutely exposed fish. 4. Ventilatory frequency was raised above 30 micrograms/l TCIN. 5. Gills of fish exposed to 2.0 micrograms/l TCIN for 24 days had reduced lamellar diffusive capacity, primarily caused by an increase in blood barrier thickness.     

The effect of chlorpyrifos on protein content and acid DNase activity in the mussel,Mytilus galloprovincialis

We evaluated the effects of short-term exposure to an organophosphate pesticide chlorpyrifos on the digestive gland and gills of the mussel Mytilus galloprovincialis. We studied metabolic activity by quantifying protein content and physiological function responses using acid DNase activity. The increase in protein content was observed in both the target tissues of mussels exposed to 0.03 μg/L chlorpyrifos when compared with control mussels. The pattern of acid DNase activity in digestive gland and gills indicated a tissue-specific response, although the lowest concentration of chlorpyrifos caused changes in acid DNase activity in both tissues. In the digestive gland, the increase of acid DNase activity was observed in mussel exposed to 0.03 μg/L chlorpyrifos, followed by decrease up to 100 μg/L chlorpyrifos. Enzyme activity in the gills showed a dose response effect. The results support the use of acid DNase activity in the digestive gland as a sensitive response to an environmentally relevant range of pesticide concentrations. It may also indicate an effect on mussel physiological status.     

Longitudinal Exposure to Selected Pesticides in Drinking Water

The presence and temporal fluctuations of concentrations of insecticides and herbicides in natural waters has been well documented. Little, however, is known about exposure to pesticides through drinking water for the general population. Concentrations often pesticides, including 4,4′DDE and atrazine, were measured up to six times at equally spaced intervals over a 1-year period in drinking water of 80 randomly selected residences in Maryland. Atrazine was detected in 228 (57.9%) of the drinking water samples with a mean of 0.15?µg/L, with standard deviation 0.12?µg/L, median 0.17?µg/L, and range <0.037 to 0.46?µg/L. 4,4′DDE was found in 22 (5.6%) water samples; no other target analytes were detected. Concentrations of atrazine in drinking water were found to vary over a 12-month period with the greatest concentrations in the late summer and fall and the lowest in the early spring. Atrazine concentrations in drinking water were influenced more by differences in levels among residences than by time of year. Seven-day average exposures and exposures per unit body weight to atrazine in drinking water exhibited a similar temporal pattern. Among individuals, drinking water consumption rate was a more important determinant of atrazine exposure (µg/d) and exposure per unit body weight (µg/kg/d) than atrazine concentration in drinking water or body weight.     

Evaluation of the lethal and sub-lethal toxicity and potential endocrine disrupting effect of nonylphenol on the zebra mussel (Dreissena polymorpha)

Nonylphenol (NP) is commonly found in surface waters nearby municipal wastewater treatment plants and was shown to have endocrine disrupting effects in aquatic organisms. The purpose of this study was to investigate the toxicity and potential endocrine disrupting effects of NP on the freshwater zebra mussel (Dreissena polymorpha). Toxicity assessment yielded LC(50) values of 3.68, 2.19 and 1.62 mg L(-1) after 15, 35 and 50 days of exposure, respectively. LC(10) values of 1.6, 1.11 and 0.68 mg L(-1) were respectively obtained for similar exposure periods. At concentrations >5 mg L(-1), mortality effects were significant, as were those relating to attachment and siphon extension (indicating filtration), both general indicators of health. Endocrine disruption effects were investigated after a prolonged exposure (112 d) to 5 and 500 microg L(-1) NP by measuring Vitellin (Vn)-like protein levels using the alkali-labile phosphate (ALP) assay and gel electrophoresis (GE). An increase in ALP levels was observed in both male and female mussels, although only marginal owing to a significant decrease in the mussels' health indicated by its condition, during the experiment. These levels, however, increased proportionally with NP concentration. Using solid phase thin-layer chromatography, we confirmed increased levels of the steroid cholesterol and evidence of NP uptake. Cholesterol levels in gonad tissue proved to be a more responsive biomarker of exposure to NP than levels of ALP. Further implications relating to the occurrence of endocrine disruption in the zebra mussel are discussed.     

Effect of mosquito killer insecticides on freshwater mussels

The effect of four insecticides (Fyfanon, K-Othrin, Unitox 7 and Unitox 20) was investigated on the freshwater mussels Anodonta cygnea L., Anodonta anatina L. and Unio pictorum L. The studies were performed in hard water (total salinity of 310 mg/l). The LC50 values for 24, 48, 72, 96 hr and 7 days were determined with a static test at 22 degrees C. All the three mussel species proved to be extremely tolerant against the insecticides tested. The effect of sublethal concentrations of these insecticides on the periodic activity of the mussels was also analyzed, comparing the changes of the active and resting periods. In a concentration of 0.1 microliter/l, all the four insecticides affected the periodic activity of U. pictorum, whereas in A. anatina three of them (Fyfanon, K-Othrin and Unitox 7) and in A. cygnea two of them (K-Othrin and Unitox 7) evoked alterations. This concentration is lower by three to five orders than those of the LC50 values of 96 hr.     

An environmental 1H NMR metabolomic study of the exposure of the marine mussel Mytilus edulis to atrazine, lindane, hypoxia and starvation

Marine mussels are useful, robust model organisms that have been widely used as biomonitors. In the natural environment they can be subjected simultaneously to a mixture of environmental stresses (hypoxia, starvation) and to pollutants. In this study Mytilus edulis was used to investigate the effects of two pesticides, atrazine and lindane, which have different modes of action but produce similar changes in behaviour (depression of ventilation and feeding) in the mussels, and can cause starvation and mild hypoxia. Acetonitrile/²H₂O (60/40% v/v) extracts of foot muscle from animals subjected to hypoxia, or starvation, or low or high doses of pesticide were analysed using ¹H NMR spectroscopy to produce metabolic fingerprints associated with these treatments. Discriminant analysis based on metabolites that showed significant differences between treated and control animals gave a clear separation between all treatment groups. The fingerprints of atrazine treated animals were clearly separated from those of animals that were starved or subjected to hypoxia. The high and low doses of atrazine were also well separated. Lindane treatment was separated from control animals in a dose dependent way, and was associated with an increase in the concentration of alanine, and a decrease in all of the other identified metabolites (including osmolytes). This is consistent with a general depression of activity. This limited study demonstrates the potential of the metabolomic approach to provide a separation of the effects of poisoning from those of environmental stress. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Cost of selective feeding by the blue mussel (Mytilus trossulus) as measured by respiration and ammonia excretion rates

The metabolic cost as measured by respiration and ammonia excretion rates associated with a selective as compared to a non-selective feeding behaviour was determined for the blue mussel, Mytilus trossulus. Mussels were challenged with four environmentally relevant seston matrices of different quality and quantity, which were known to evoke either a sorting response (i.e., selective feeding) where organic-rich particles were selected over inorganic particles as compared to no sorting (i.e., non-selective) where either inorganic or organic particles were ingested by the bivalve. Seston matrices were prepared by mixing known quantities of silt and algae such that the following extent of feeding responses would occur; no pre-sorting of ingested material, (1) no algae+50 mg silt l⁻¹, (2) 150×10⁶ cells l⁻¹ of algae+no silt, and, where pre-sorting of ingested material occurred, (3) 20×10⁶ cells l⁻¹ of algae and 20 mg l⁻¹ of silt, and (4) 150×10⁶ cells l⁻¹ of algae and 50 mg l ¹ of silt. A control, which represented basal metabolism consisting of mussels exposed only to filtered seawater (0.45 μm), was included for a total of five treatments. Mussel respiration and ammonia excretion rates were independent of whether mussels were pre-sorting or not sorting ingested material. Of the four matrices, only rates determined for conditions of maximum seston quality and quantity where pre-sorting of the seston occurred were significantly different from control mussels (p<0.05, ANOVA). Estimates of net energy used for feeding, where net energy is total energy intake (food) minus energy expenditure (energy lost through respiration and excreta as measured by ammonia excretion rates), indicated that feeding, whether selective or non-selective required only 0.92% of net energy intake. Hence, mussels appear to be highly adapted to a dynamic food environment with negligible costs associated with the feeding process, even when significant pre-selection of organic-rich particles occurs.     

Effect of Food Concentration Inside Eelgrass Beds on the Energy Balance of the Invasive Mussel Musculista Senhousia

Samples of Musculista senhousia, 20 mm in length, were gathered from San Diego Bay and fed on three different food concentrations: 0.2, 0.4 and 0.8 mg AFDW L m 1, and their energy balance studied. Food concentration ranges were selected to reflect those observed in the field where the invasive mussel M. senhousia co-occurs with the angiosperm, Zostera marina. The lowest and intermediate concentrations correspond to concentrations described in and over large eelgrass patches, respectively. The upper concentration is equivalent to that occurring during the spring phytoplankton blooms. Ingestion rates varied between 81.6 µg AFDW h m 1 for the mussels fed on the lowest concentration, 191.2 µg AFDW h m 1 when the concentration rose to 0.4 mg AFDW L m 1, and 164.0 µg AFDW h m 1 for the highest concentration. Food concentration had a significant effect on absorption efficiency, which was approximately 70% at the intermediate and upper concentrations, but rose to over 80% at the lowest concentration. The food absorption rate varied with food concentration in a manner similar to that of the ingestion rate, i.e., with a maximum value of 142.34 µg AFDW h m 1 for the mussels fed on the intermediate concentration, this being higher than the values for both the highest concentration (114.72 µg AFDW h m 1) and the lowest (65.89 µg AFDW h m 1). Respiration rates were 54 µg O 2 h m 1 for the mussels fed on the lowest concentration and 74 µg O 2 h m 1 for those fed on the intermediate concentration. These results show a close relationship between respiration and ingestion rates, which can be fitted to the following equation: RR = e (m 0.42 + 0.10 IR) (r = 0.5703, R 2 = 32.53, p <0.01). Scope for growth (SFG) estimations were higher for those mussels fed at the intermediate and upper concentrations (1.95 J h m 1 and 1.50 J h m 1, respectively), between which no significant difference was observed (p > 0.05), whilst SFG for the lowest food concentration was only 0.69 J h m 1. The physiological rate with the greatest effect on growth was the absorption rate. Its relation to SFG is described by the equation SFG = 0.67 + 0.90AR (r = 0.977, R 2 = 95.56%, p <0.01). The discussion of these results is based on the growth rates that have been described for this mussel in its natural environment where habitat structure, such as the canopy provided by the eelgrass, can affect the success of the invasive mussel.     

Impact of Engineered Zinc Oxide Nanoparticles on the Individual Performance of Mytilus galloprovincialis

The increased use of engineered nanoparticles (ENPs) in consumer products raises the concern of environmental release and subsequent impacts in natural communities. We tested for physiological and demographic impacts of ZnO, a prevalent metal oxide ENP, on the mussel Mytilus galloprovincialis. We exposed mussels of two size classes, <4.5 and ≥4.5 cm shell length, to 0.1–2 mg l⁻¹ ZnO ENPs in seawater for 12 wk, and measured the effect on mussel respiration, accumulation of Zn, growth, and survival. After 12 wk of exposure to ZnO ENPs, respiration rates of mussels increased with ZnO concentration. Mussels had up to three fold more Zn in tissues than control groups after 12 wk of exposure, but patterns of Zn accumulation varied with mussel size and Zn concentrations. Small mussels accumulated Zn 10 times faster than large mussels at 0.5 mg l⁻¹, while large mussels accumulated Zn four times faster than small mussels at 2 mg l⁻¹. Mussels exposed to 2 mg l⁻¹ ZnO grew 40% less than mussels in our control group for both size classes. Survival significantly decreased only in groups exposed to the highest ZnO concentration (2 mg l⁻¹) and was lower for small mussels than large. Our results indicate that ZnO ENPs are toxic to mussels but at levels unlikely to be reached in natural marine waters.