NADPH: Phenanthrenequinone oxidoreductase in channel catfish (Ictalurus punctatus)

The toxicity of certain polycyclic aromatic hydrocarbons has been related to the phenomenon of oxidative stress. Recent studies in our laboratory have demonstrated that 9,10-phenanthrenequinone (PQ) is an efficient redox cycling agent in cytosols of several organs of the channel catfish (Ictalurus punctatus). Stomach cytosol demonstrated a high level of dicoumarol-sensitive PQ reduction which was at least tenfold greater than that seen in kidney, liver or gill. In the presence of stomach cytosol, PQ stimulated production of Superoxide union in-vitro at rates several fold higher than did menadione, a model redox active quinone. The active component from stomach cytosol has been purified, and is termed NADPH: phenanthrenequinone oxidoreductase, which the authors believe represents a novel quinone-reducing enzyme in this species.     

腐殖酸抑制纳米银对菲律宾蛤仔鳃的氧化胁迫和遗传毒性

为了解DOM与AgNPs的相互作用及其对AgNPs毒性的影响,本文以菲律宾蛤仔为受试生物、以水环境广泛存在腐殖酸(Humic acid,HA)为代表性有机质。通过贝类体内抗氧化系统的生物标志物(过氧化氢酶(Catalase,CAT)、超氧化物歧化酶(Superoxide dismutase,SOD)、金属硫蛋白(Metallothionein,MT)和丙二醛(Malondialdehyde,MDA))的响应,研究了HA对AgNPs生物效应的影响。结果表明,HA显著增加了AgNPs的稳定性,抑制了AgNPs的聚集。AgNPs(20μg·L^-1)暴露3 d后,蛤仔鳃组织中Ag的蓄积显著上升,并随着时间的延长不断升高。AgNPs暴露第3、7天,不同浓度的HA条件下,鳃组织中Ag的蓄积无显著差异,但显著地低于无HA存在的AgNPs暴露组。CAT活性和MDA含量仅在无HA存在的AgNPs暴露组出现显著升高,其余各组之间无显著差异。表征DNA损伤程度的Olive尾矩(OTM)和尾长(TL)在无HA存在的AgNPs暴露组中随时间的延长不断上升。HA显著降低了AgNPs的对鳃组织细胞的遗传毒性。结果表明,HA通过增加AgNPs在介质中的稳定性而显著地抑制了鳃组织对Ag的蓄积,从而减弱了AgNPs对鳃组织的氧化胁迫和遗传毒性。结果证明了有机质含量等环境理化条件对纳米毒性存在显著影响。     

Effects of oxidative stress on protein thiols and disulphides in Mytilus edulis revealed by proteomics: actin and protein disulphide isomerase are redox targets

Many proteins contain cysteines which are sensitive to oxidation. This is sometimes reversible through interaction with glutathione, glutaredoxin or thioredoxin systems making these proteins potential sensors of oxidative stress. In this study we analysed whether there was an increase in mixed disulphide bond (-S-S-) formation in the blue mussel Mytilus edulis in response to menadione. This was achieved by initially blocking reduced thiols with N-ethylmaleimide, -S-S- were then reduced with dithiothreitol (DTT) and labelled with 5-iodoacetamidofluorescein (5-IAF). Free -SHs were also labelled directly with 5-IAF. Separations were performed on 1D or 2D SDS PAGE and images analysed. There was an increase in -S-S- in response to menadione and detection of changes in oxidised proteins was easier than that of changes in the amount of reduced proteins. Protein disulphide isomerase (PDI) was labelled both as -SH and -S-S-, underlining its involvement in the redox status of the animal. A glutathione transferase (GST P1-1) forms an inter-chain disulphide bridge in response to menadione.     

Determinants of species susceptibility to oxidative stress: A comparison of channel catfish and brown bullhead

Xenobiotic-mediated productions of reactive oxygen species, via enzymemediated redox cycling, have been implicated in a variety of toxicological phenomena including lipid peroxidation, enzyme inactivation and oxidative DNA damage leading to cancer. A comparison was undertaken of two benthic freshwater fish species that appear to differ markedly in their susceptibility to chemical carcinogenesis—the channel catfish (Ictalurus punctatus) and the more cancer-prone brown bullhead (Ameriurus nebulosus)-in terms of basic biochemical characteristics related to oxidative stress. This has included analysis of microsomal redox cycling of model xenobiotics (e.g. menadione) as well as antioxidant and other detoxifying enzymes in hepatic tissue of the two species. In addition, endpoints of oxidative stress, such as altered glutathione status and oxidative DNA damage, were examined. These studies have revealed numerous qualitative and quantitative differences between the two species both in terms of basal enzyme activities and in species response to model prooxidants. For example, bullhead appear to possess a greater capacity for microsomal redox cycling of xenobiotics, but have glutathione-dependent defense systems less able to withstand oxidative challenge. These and other interspecific differences have allowed for an improved understanding of the basic mechanisms which may underly species susceptibility to oxidative stress and critical manifestations such as cancer.     

海水中多环芳烃的暴露浓度对鲈鱼体内脂质过氧化程度的影响

本文以鲈鱼为实验材料，以鲈鱼体内的丙二醛含量为指标研究了在水样中16种多环芳烃组分和外加芘为实验试剂的条件下，鲈鱼体内不同器官、组织的脂质过氧化程度的变化．实验结果表明：（1）在鲈鱼鳃、全血中的丙二醛浓度较高，而肝脏的丙二醛浓度较低。（2）水样中多环芳烃及外加芘的暴露能导致鲈鱼肝的丙二醛浓度升高，这表明多环芳烃以及芘的暴露能引起鲈鱼肝内脂质过氧化程度的提高。（3）水体中多环芳烃及外加芘浓度的变化对鲈鱼肝的丙二醛浓度的影响具有可逆性，且鲈鱼肝的丙二醛浓度与水环境的多环芳烃（包括芘）暴露浓度呈良好的正相关关系。这说明：鲈鱼肝的丙二醛浓度可作为指示水环境的多环芳烃污染程度的指标。由于生物体内的丙二醛浓度受多种因素的影响，在使用丙二醛浓度指标来指示海洋环境的多环芳烃污染时。最好与其他指标配合使用，以更准确地指示海洋环境多环芳烃污染。     

Oxygen radical formation in anoxic transgression and anoxia-reoxygenation: Foe or phantom? Experiments with a hypoxia tolerant bivalve

Intertidal blue mussels, Mytilus edulis, experience hypoxia reoxygenation during tidal emersion and resubmersion cycles, and this is often suggested to represent a major stress for the animals, especially for their respiratory tissues, the gills. We exposed mussels to experimental short and prolonged anoxia and subsequent reoxygenation and analyzed the respiratory response in excised gill tissue and the effects of treatment on reactive oxygen species (mainly ROS: superoxide anion, O₂^·− and hydrogen peroxide, H₂O₂), formation using live imaging techniques and confocal microscopy. Our aim was to understand if this “natural stress” would indeed produce oxidative damage and whether antioxidant defenses are induced under anoxia, to prevent oxidative damage during reoxygenation. Exposure to declining pO₂ in the respiration chamber caused an increase of gill metabolic rate between 21 and 10 kPa, a pO₂ range in which whole animal respiration is reported to be oxyregulating. Exposure of the animals to severe anoxia caused an onset of anaerobiosis (succinate accumulation) and shifted high and low critical p_c values (p_c1: onset of oxyregulation in gills, p_c2: switch from oxyregulation to oxyconformity) to higher pO₂. Concentrations of both ROS decreased strongly during anoxic exposure of the mussels and increased upon reoxygenation. This ROS burst induced lipid peroxidation in the mantle, but neither were protein carbonyl levels increased (oxidative damage in the protein fraction), nor did the tissue glutathione concentration change in the gills. Further, analysis of apoptosis markers indicated no induction of cell death in the gills. To our knowledge, this is the first paper that directly measures ROS formation during anoxia reoxygenation in mussels. We conclude that hypoxia tolerant intertidal mussels do not suffer major oxidative stress in gill and mantle tissues under these experimental conditions.     

Effect of seasonality on oxidative stress responses and metal accumulation in soft tissues of Aulacomya atra,a mussel from the South Atlantic Patagonian coast

This study investigated the effects of pollution and its interaction with temperature on the oxidative status of the ribbed mussel Aulacomya atra in the southern Atlantic Patagonian coast. Animals were collected from four sites with different degree and type of human activity impact, during the summer and winter of 2011. Seawater chromium, copper, manganese, nickel and zinc concentrations were measured, as well as metal accumulation, lipid peroxidation, protein oxidation, reduced glutathione levels, and enzymatic activities of superoxide dismutase and glutathione-S-transferase in gills and digestive glands.Metal bioaccumulation and oxidative stress responses in both tissues were generally higher in mussels from harbor areas. Water temperature had a remarkable effect on gill SOD activity and protein oxidation during winter in mussels from all locations.Methodologically, we conclude that measuring both metal bioaccumulation and oxidative stress responses allowed for a more accurate assessment of the biological effects of metal present in seawater.     

Genotoxicity of copper oxide and silver nanoparticles in the mussel Mytilus galloprovincialis

Though there is some information on cytotoxicity of copper nanoparticles and silver nanoparticles on human cell lines, there is no information on their genotoxic and cytotoxic behaviour in bivalve molluscs. The aim of this study was to investigate the genotoxic impact of copper oxide and silver nanoparticles using mussels Mytilus galloprovincialis. Mussels were exposed to 10 μg L⁻¹ of CuO nanoparticles and Cu²⁺ and Ag nanoparticles and Ag⁺ for 15 days to assess genotoxic effects in hemocytes using the comet assay. The results obtained indicated that copper and silver forms (nanoparticles and ionic) induced DNA damage in hemolymph cells and a time-response effect was evident when compared to unexposed mussels. Ionic forms presented higher genotoxicity than nanoparticles, suggesting different mechanisms of action that may be mediated through oxidative stress. DNA strand breaks proved to be a useful biomarker of exposure to genotoxic effects of CuO and Ag nanoparticles in marine molluscs.     

不同粒径TiO_2颗粒对海洋微藻的毒性效应

本实验以新月菱形藻为受试生物,研究了低浓度不同粒径TiO2颗粒(21nm、60nm和400nm)对海洋微藻生长、抗氧化酶活性(超氧化物歧化酶SOD、过氧化氢酶CAT和过氧化物酶POD)、脂质过氧化产物(MDA)含量的影响,并测定了相应的活性氧自由基(ROS)的含量,初步探讨了TiO2颗粒对海洋微藻的作用机制。结果表明,1mg/L TiO2颗粒对新月菱形藻生长的抑制作用随着粒径的减小而逐渐增强,第48h、72h、96h呈现出显著的纳米效应。TiO2颗粒可以诱导藻细胞内ROS的含量增加,对藻细胞产生氧化胁迫,新月菱形藻的抗氧化酶活性发生应激响应,以清除过量的ROS,但剩余的ROS对藻细胞产生氧化损伤,导致MDA含量升高,并且纳米级TiO2颗粒对新月菱形藻的氧化损伤大于微米级颗粒。在不同粒径TiO2颗粒的胁迫下,藻细胞SOD和CAT活性的响应也存在差异。本研究将为开展人工纳米材料对海洋生态系统影响的潜在风险评估提供科学依据。     

斑节对虾GPx3a基因的分子克隆及表达分析

谷胱甘肽过氧化物酶(Glutathione peroxidase,GPx)是生物体内重要的抗氧化酶,能防止过氧化氢对生物体的氧化应激。该研究利用RACE技术获得了斑节对虾(Penaeus monodon)GPx3a(PmGPx3a)的全长c DNA序列,进行了相关生物信息学分析。作者利用荧光定量PCR方法研究了PmGPx3a在斑节对虾不同组织的表达情况。探究了PmGPx3a在不同胁迫条件下(盐度、重金属和细菌)的表达情况。结果表明PmGPx3a c DNA全长1135 bp,其中开放阅读框(ORF)长651 bp,预测编码216个氨基酸。PmGPx3a推导的氨基酸序列与其他动物的GPx3a氨基酸序列具有高度一致性。实时定量PCR结果显示,在高低盐胁迫下,PmGPx3a在肝胰腺中相对表达量都为上升的(p0.05)。在铜、锌、铬胁迫中,鳃中的PmGPx3a的相对表达量总体呈现下降趋势,在肝胰腺中呈现上升趋势。在哈维弧菌(Vibrio harveyi)刺激下,PmGPx3a在血淋巴中的相对表达量总体呈现上升趋势,24 h后表达量最大,显著高于对照组2.2倍(P0.05)。以上研究结果表明,PmGPx3a基因参与了斑节对虾对环境胁迫和氧化应激的适应性反应。     

Effects of nanoparticles in Mytilus edulis gills and hepatopancreas - a new threat to marine life?

Every day new extraordinary properties of nanoparticles (a billionth of a meter) are discovered and worldwide millions are invested into nanotechnology and nanomaterials. Risks to marine organisms are still not fully understood and biomarkers to detect health effects are not implemented, yet. We used the filter feeding blue mussel as a model to analyse uptake and effects of nanoparticles from glass wool, a new absorbent material suggested for use in floating oil spill barriers. In both, gills and hepatopancreas we analysed uptake of nanomaterials by transmission electronmicroscopy (TEM) in unstained ultrathin sections over a period of up to 16 days. Lysosomal stability and lipofuscin content as general indicators of cellular pathology and oxidative stress were also measured. As portals of uptake, diffusion and endocytosis were identified resulting in nanoparticle accumulation in endocytotic vesicles, lysosomes, mitochondria and nuclei. Dramatic decrease of lysosomal membrane stability occurred after 12h of exposure. Lysosomal damage was followed by excessive lipofuscin accumulation indicative of severe oxidative stress. Increased phagocytosis by granulocytes, autophagy and finally apoptosis of epithelial cells of gills and primary and secondary digestive tubules epithelial cells indicated progressive cell death. These pathological responses are regarded as general indices of toxic cell injury and oxidative stress. By the combinational use of biomakers with the ultrastructural localisation of nanoparticle deposition, final evidence of cause-effect relationships is delivered.     

军曹鱼（Rachycentron canadum）幼鱼对环境低氧胁迫氧化应激与能量利用指标的响应

本文探究环境低氧对军曹鱼(Rachycentron canadum)氧化应激和能量利用指标的影响，为军曹鱼的健康养殖提供参考依据。通过设置低氧胁迫–恢复实验，将军曹鱼幼鱼(平均体质量(220.67±20.73)g)在低氧((2.64±0.25)mg/L)胁迫3 h及复氧((6.34±0.15)mg/L)8 h、24 h和48 h后，测定其肝脏和肌肉组织的氧化应激与能量利用指标。结果显示，低氧胁迫后，肝脏中丙二醛（Malondialdehyde，MDA)、过氧化氢酶(Catalase，CAT)和谷胱甘肽还原酶(Glutathione Reductase，GR)活力均显著低于对照组(p<0.05)，乳酸脱氢酶(Lactate Dehydrogenase，LDH)活性显著高于对照组(p <0.05)；肌肉中MDA和脂质过氧化物(Lipid Peroxidase，LPO)活性均显著低于对照组(p<0.05)，超氧化物歧化酶(Superoxide Dismutase，SOD)和LDH活性均显著高于对照组(p<0.05)；肌糖原和肝糖原含量极显著低于对照组(p<0.01)。复氧过程中，肝脏和肌肉中MDA、LPO、SOD、CAT、谷胱甘肽过氧化物酶(Glutathione Peroxidase，GPx)和GR含量均出现不同程度的升高；肝糖原在复氧24 h后显著高于对照组(p<0.05)，复氧48 h后显著低于对照组(p<0.05)；肌糖原在复氧8 h、24 h和48 h后均显著低于对照组(p<0.05)。研究表明，低氧胁迫能够对军曹鱼幼鱼机体造成一定的氧化损伤，肝脏和肌肉组织的酶活力和能量供应发生变化；低氧胁迫后的再复氧环境，对机体造成更为强烈的氧化损伤，可通过自身生理调节逐渐恢复到正常水平。     

不同长牡蛎群体对海水酸化的生理响应差异分析

海水酸化暴露可对海洋生物产生多层面的影响。本研究以潮间带野生与潮下带养殖长牡蛎(不同生境背景)的不同组织(鳃、外套膜及消化腺)为研究对象,分析在室内调控p CO2模拟海水酸化暴露条件下,其基础代谢活动、能量代谢以及氧化应激相关指标的变化情况。结果显示:海水酸化暴露后,两种长牡蛎(Crassostrea gigas)的基础代谢过程均受到了一定抑制作用且受影响程度差异明显。潮间带野生与潮下带养殖长牡蛎的关键生理过程(能量代谢及氧化应激)对海水酸化暴露存在不同的响应变化,表明两种长牡蛎应对海水酸化的调节机制可能存在差异。依据PLS-DA分析结果显示,在所有生理指标中,对样本的差异贡献较高(VIP值1)的指标为:SDH、AST、ATPase、ATP含量、糖原含量、CAT、GST及SOD,表明海水酸化暴露后,在两种长牡蛎的3种组织中上述指标的响应变化程度更大。综合评价分析多个生理指标的整体变化揭示:在海水酸化暴露条件下,潮间带野生长牡蛎比潮下带养殖长牡蛎对海水酸化的生理响应更为剧烈;相比于鳃及消化腺组织,长牡蛎外套膜组织可能受影响更大。     

Levels of cellular glutathione and metallothionein affect the toxicity of oxidative stressors in an established carp cell line.

The comparative toxicity of a variety of oxidative stressors was studied in the epithelioma papulosum cyprini line from carp using the neutral red cytotoxicity assay. LC50's decreased in the order t-butylhydroperoxide > hydrogen peroxide > diquat > paraquat. The cytotoxicity of hydrogen peroxide was significantly reduced when the cells were grown in L-15 medium rather than MEM and this could be attributed to elevated cellular glutathione and metallothionein levels and higher activities of GSH-dependent detoxification systems. The protective effect of metallothionein in radical scavenging was demonstrated by decreased toxicity of the redox-cycling toxicants, diquat and menadione after metallothionein levels had been pre-induced by Cd-exposure. This study demonstrates the relationship between toxic effects of oxidative stressors and expression of detoxification systems in fish.     

Application of the biomarker responses in scallop (Chlamys farreri) to assess metals and PAHs pollution in Jiaozhou Bay,China

Scallops (Chlamys farreri) were collected at three sites in the Jiaozhou Bay, China in April and July 2007. The responses of biomarkers were evaluated for assessment of physiological status of scallops and pollution in the areas. Compared with site S1, the activity of aryl hydrocarbon hydroxylase was induced notably (P < 0.05) and glutathione S-transferase activity was inhibited significantly (P < 0.05) in S2 and S3. The levels of DNA alkaline unwinding, DNA protein crosslinks, protein carbonyl and lipid peroxidation were tested and indicated the oxidative stress situation of scallops. There was significant difference of biomarker levels between sampling seasons (P < 0.05), and between digestive gland and gill (P < 0.05). The results provided the reference data for multiple-pollution assessment in the marine environment and indicated that tissue type and seasons affect biomarkers, therefore these factors should be taken into consideration when biomarkers are used for environmental assessment purposes.     

急性低氧胁迫对卵形鲳鲹选育群体血液生化指标的影响

本文研究了水环境急性低氧胁迫对卵形鲳鲹Trachinotus ovatus选育群体血液生化指标的影响。实验设计了急性低氧试验组和对照组,每组3个平行。结果显示,急性低氧胁迫后选育群体的血清离子含量与对照组相比都有不同程度的升高,其中钠、氯、钙浓度与对照组相比差异显著(p0.05),铁浓度极显著高于对照组(p0.01);血清蛋白、尿酸、肌酐、血脂、血糖等指标的差异较大,其中尿酸、肌酐极显著高于对照组(p0.01),总蛋白、总胆固醇显著低于对照组(p0.05);血清中肌酸激酶含量极显著高于对照组(p0.01),肌酸激酶同工酶显著高于对照组(p0.05),γ-谷氨酰转肽酶含量显著低于对照组(p0.05);其他指标均有不同程度的升高或降低,但差异不显著。研究表明,急性低氧胁迫造成卵形鲳鲹鳃、肾、心脏和肝脏损伤,免疫力下降。     

Biomarker responses in Macoma nasuta (Bivalvia) exposed to sediments from northern San Francisco Bay

Our study investigates biomarker responses and survival of Macoma nasuta exposed to sediments collected from six locations in northern San Francisco Bay. Biomarkers analyzed were stress proteins (hsp70) in gill, mantle and digestive gland, lysosomal membrane damage and histopathologic lesions. Sediments and clam tissues were analyzed for a comprehensive suite of heavy metals and trace organic pollutants. Sediment grain size and organic carbon content were determined. Clams accumulated metals, polyaromatic hydrocarbons (PAHs) and organochlorine pesticides (aldrin and p,p(')-DDT and its metabolites p,p(')-DDD and p,p(')-DDE). Pearson and Spearman correlation analysis revealed that mortality, hsp70 in gill and histopathologic lesion scores in gonads, and lysosomal membrane damage were significantly correlated with tissue concentrations of DDT and/or its metabolites. Tissue concentrations of metals, in particular nickel, chromium, and copper, were associated with macrophage aggregates in digestive gland and germ cell necrosis. Cadmium was linked to mortality and lysosomal membrane damage.     

花鲈低氧诱导因子基因(hifs)的序列分析及低氧诱导表达

低氧诱导因子基因(hifs)是由α和β构成的异源二聚体转录因子,在生物低氧应答中发挥着重要作用。本实验对花鲈4个hifs基因进行了序列分析及组织表达检测,并检测了各基因在低氧诱导后的mRNA表达变化。研究表明,通过全基因组比对、进化树分析和共线性分析,共鉴定出花鲈的4个hifs基因,分别为hif1α、hif2α、hif3α、hif1β。组织表达结果显示,hifs基因具有组织特异性表达特征。低氧((1.56±0.24)mg/L)诱导下,hifs各基因的响应时间和响应程度存在一定差异。肝组织中,低氧胁迫3和6 h后,hif1α、hif2α、hif3α和hif1β表达量均出现了显著升高,表明hifα和hif1β基因可能共同起低氧调控作用;而低氧胁迫12 h后,仅hif1α表达量显著高于对照组,表明hif1α基因在肝组织低氧调控12 h内持续起作用。鳃组织中,低氧胁迫3 h后,hif1α、hif2α、hif3α基因表达量出现了显著升高,而低氧胁迫6 h后,hif3α和hif1β基因表达量显著升高,说明在鳃组织前3h的低氧胁迫中仅hifα基因起调控作用,而6 h后hif3α和hif1β基因共同调控低氧胁迫。常氧恢复阶段肝、鳃组织中各基因表达量均与对照组无显著差异。本研究结果表明,4个hifs基因的组织表达具有特异性,同时对低氧的响应时间和程度不同,研究结果弥补了花鲈hifs基因及低氧应答研究的空白。     

纳米银对青岛近海浮游细菌活性的影响研究

纳米银广泛用于抗菌材料、医疗设备和其他抗菌产品。在纳米银产品的生产、运输、消费和处置过程中,纳米银将不可避免地释放到自然环境中,而水生生态系统是纳米银在自然界中重要的汇之一。本文以青岛近海为研究对象,采集现场海水进行实验室模拟培养,通过测定(48 h)不同浓度纳米银(0.5、5和50 mg/L)短期暴露下对表层海水中浮游细菌死亡率、蛋白质含量、碱性磷酸酶活性及氨肽酶活性的变化,研究纳米银对近岸海区浮游细菌生长和活性的影响。研究结果表明,在纳米银胁迫下细菌生长和活性受到显著抑制(P<0.05),高浓度组在培养4 h后死亡率达到最高值88.20%,随着暴露时间延长,纳米银胁迫组活细菌数量缓慢恢复、死亡率逐渐减小;纳米银对碱性磷酸酶活性和氨肽酶活性的抑制效应分别在培养4和8 h后达到最高,分别为8.58%和32.55%,在培养16 h后,酶活性开始缓慢上升,表现出对纳米银胁迫的适应性;纳米银组的蛋白质含量在培养8 h后达到最低,相比于对照组降低了29.80%,在培养后期同样有所升高,蛋白质含量接近于对照组。由此可见,纳米银对青岛近海浮游细菌的生长和活性都具有显著的抑制效应;不同浓度纳米银对细菌的抑制效应不同,纳米银浓度越高,抑制效应越明显(P<0.05),具有明显的浓度梯度效应;在培养后期,碱性磷酸酶活性和氨肽酶活性都有恢复,蛋白质含量上升,细菌死亡率下降,表明细菌对纳米银胁迫表现出一定的适应性。     

Biomarker responses in Macoma nasuta (Bivalvia) exposed to sediments from northern San Francisco Bay

Our study investigates biomarker responses and survival of Macoma nasuta exposed to sediments collected from six locations in northern San Francisco Bay. Biomarkers analyzed were stress proteins (hsp70) in gill, mantle and digestive gland, lysosomal membrane damage and histopathologic lesions. Sediments and clam tissues were analyzed for a comprehensive suite of heavy metals and trace organic pollutants. Sediment grain size and organic carbon content were determined. Clams accumulated metals, polyaromatic hydrocarbons (PAHs) and organochlorine pesticides (aldrin and p,p^′-DDT and its metabolites p,p^′-DDD and p,p^′-DDE). Pearson and Spearman correlation analysis revealed that mortality, hsp70 in gill and histopathologic lesion scores in gonads, and lysosomal membrane damage were significantly correlated with tissue concentrations of DDT and/or its metabolites. Tissue concentrations of metals, in particular nickel, chromium, and copper, were associated with macrophage aggregates in digestive gland and germ cell necrosis. Cadmium was linked to mortality and lysosomal membrane damage.