Loss of diversity of ammonia-oxidizing bacteria correlates with increasing salinity in an estuary system

Ammonia-oxidizing bacteria (AOB) play an important role in nitrogen cycling in estuaries, but little is known about AOB diversity, distribution and activity in relation to the chemical and physical changes encountered in estuary systems. Although estuarine salinity gradients are well recognized to influence microbial community structure, few studies have examined the influence of varying salinity on the diversity and stability of AOB populations. To investigate these relationships, we collected sediment samples from low-, mid- and high-salinity sites in Plum Island Sound estuary, MA, during spring and late summer over 3 years. Ammonia-oxidizing bacteria distribution and diversity were assessed by terminal restriction fragment length polymorphism (TRFLP) analysis of the ammonia monooxygenase (amoA) gene, and fragments were identified by screening amoA clone libraries constructed from each site. Most striking was the stability and low diversity of the AOB community at the high-salinity site, showing little variability over 3 years. Ammonia-oxidizing bacteria at the high-salinity site were not closely related to any cultured AOB, but were most similar to Nitrosospira spp. Ammonia-oxidizing bacteria at the mid- and low-salinity sites were distributed among Nitrosospira-like sequences and sequences related to Nitrosomonas ureae/oligotropha and Nitrosomonas sp. Nm143. Our study suggests that salinity is a strong environmental control on AOB diversity and distribution in this estuary.     

Metal-ion-mediated oxidative stress in the gill homogenate of rainbow trout (Oncorhynchus mykiss)

Human activities play a major role in toxic and carcinogenic metal pollution of the environment. This study was undertaken to evaluate the effects of copper and mercury at the 400-to 1000-μM concentration range on some biochemical markers of oxidative stress, such as lipid peroxidation (LPO), glutathione-S-transferase (GST) activity, and reduced glutathione (GSH) content in the rainbow trout gill homogenates with or without supplementation of manganese, selenium, and bovine serum albumin (BSA). The integrity of DNA was also measured to assess metal ion toxicity. The results showed that the LPO and specific activity of GST were elevated. This indicated that cell-protecting antioxidant mechanisms were overtaxed and could not prevent membrane peroxidation. Following the addition of metals, the GSH content was also significantly reduced in a concentration-dependent manner. Mercury was found to be more effective than copper. The application of antioxidants proved beneficial in inhibiting LPO, reducing GST activity, and elevating the GSH levels in the gill samples. Manganese was more effective than selenium and BSA. Surprisingly, when BSA (1.0%) was added to the gill homogenates treated with a 1000-μM concentration of metal ions, instead of alleviating malondialdehyde (MDA) generation, a drastic elevation in the MDA levels, alleviation in GST activity, and a further decrease in glutathione (GSH) levels were observed, which were most likely the result of pro-oxidant activity of BSA. The results also indicated that mercury and copper functioned as genotoxic pollutants, which altered the DNA integrity by inducing the single and double-stranded DNA breaks in the gill cell nuclei. Collectively, toxicity of metal ions is related to the depletion of GSH content and inhibition of antioxidant enzyme GST, resulting in the propagation of LPO and DNA damage.     

The growth of grey mullet in a rural and urbanized Irish estuary

The condition of grey mullet from the urbanized Coliigan estuary was higher than that of mullet from the rural Barnawee estuary and those of several other samples described in the literature (P<0·05). It is suggested that their better condition was due to dietary supplementation from creamery and tannery effluent in the urbanized estuary. As a result of their distinctive condiiion, indirect evidence is provided for homing in this species, at this site.     

肿瘤相关抗原基因HCA520重组蛋白的表达纯化及其肝癌患者血清中相应抗体的分析

HCA5 2 0是用SEREX(serologicalidentificationofrecombinantcDNAexpressingcloning)方法 ,即用肝癌病人血清从肝癌组织cDNA表达文库中筛选得到的肝癌相关性抗原编码基因 .利用RT PCR技术检测了HCA5 2 0mRNA在各种组织中的分布情况 ,构建了GST融合表达载体并且用亲和层析的方法纯化表达的融合蛋白 .最后用Western印迹检测了重组蛋白的免疫反应性 ,用斑点印迹检测了肝癌病人血清中HCA5 2 0的天然抗体的存在情况 .结果表明 ,HCA5 2 0在各种组织中呈丰度差异分布 ,构建好的pGEX 4T 3重组载体经IPTG诱导后高效表达GST HCA5 2 0融合蛋白 ,其分子量约 4 9kD .经GST Agarose亲和层析 ,重组蛋白得到高度纯化 .Western印迹证实 ,纯化蛋白为目的重组蛋白 ,重组蛋白具有与天然蛋白相同或相似的免疫反应性 .斑点印迹分析表明 ,2 0份肝癌病人血清中有 1份HCA5 2 0抗体阳性 ,而 4份正常人均为阴性 .HCA5 2 0的足量提供 ,可用以研究其在致癌中的作用 ,并可以免疫动物制备抗体 .它作为抗原 ,分析其抗体在不同肿瘤病人中的表达情况 ,评估其在临床肿瘤诊断中的作用     

Tissue-specific oxidative stress responses in fish exposed to 2,4-D and azinphosmethyl

Species- and tissue-specific defenses against the possibility of oxidative stress and lipid peroxidation were compared in adult fish, Oreochromis niloticus and Cyprinus carpio, exposed to 2,4-dichlorophenoxyacetic acid (2,4-D), azinphosmethyl and their combination for 96 h. Superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase activities were monitored in kidney, brain and gill. In all exposure groups there was a marked increase in SOD activity in gill tissues in both fish species, while it was at the control level in other tissues. The highest elevation of SOD activity by combined treatment was observed in C. carpio. Individual and combined treatments caused an elevation in catalase and GPx activities in kidney of C. carpio. Catalase activity was unaffected in brain of O. niloticus, while GPx activity was decreased after all treatments. Glutathione S-transferase (GST) activity was higher than the control levels in kidney of both fish exposed to pesticides. No significant changes were observed in malondialdehyde level in kidney and brain of C. carpio. Our results indicate that the toxicities of azinphosmethyl and 2,4-D may be related to oxidative stress. Also, the results show that SOD activity in gill and GST activity in kidney may be used as biomarkers for pollution monitoring and indicate that the activities of certain biomarkers in C. carpio are more sensitive to pesticides than those in O. niloticus.     

Phenotypic plasticity and population differentiation in seeds and seedlings of the grass Anthoxanthum odoratum

Summary Seeds of Anthoxanthum odoratum were transplanted reciprocally between a xeric and a mesic field population that were genetically differentiated in adult traits. In one experiment seeds were reciprocally buried in bags in the soil, in a second experiment seeds were reciprocally sown in small plots. For most traits, site effects were much larger than seed-source effects. Germination, emergence, mortality of buried seed and recruitment were significantly higher at the xeric site than at the mesic site, irrespective of population of origin. Seed dormancy, was significantly higher for seed originating from the mesic than from the xeric population. Seedling recruits originating from the xeric population tended to be larger at both sites. Fecundity of seedling recruits depended on the environment; fecundities of plants growing in the xeric site had more than double the fecundity of plants growing in the mesic site. Phenotypic plasticity rather than population differences determined variation in performance in the seed and seedling stages.     

Stability of Bacterial Composition and Activity in Different Salinity Waters in the Dynamic Patos Lagoon Estuary: Evidence from a Lagrangian-Like Approach

We employed a Lagrangian-like sampling design to evaluate bacterial community composition (BCC—using temporal temperature gel gradient electrophoresis), community-level physiological profiles (CLPP—using the EcoPlate? assay), and influencing factors in different salinity waters in the highly dynamic Patos Lagoon estuary (southern Brazil) and adjacent coastal zone. Samples were collected monthly by following limnetic–oligohaline (0–1), mesohaline (14–16), and polyhaline (28–31) waters for 1 year. The BCC was specific for each salinity range, whereas the CLPPs were similar for mesohaline and polyhaline waters, and both were different from the limnetic–oligohaline samples. The limnetic–oligohaline waters displayed an oxidation capacity for almost all organic substrates tested, whereas the mesohaline and polyhaline waters presented lower numbers of oxidized substrates, suggesting that potential activities of bacteria increased from the polyhaline to oligohaline waters. However, the polyhaline samples showed a higher utilization of some simple carbohydrates, amino acids, and polymers, indicating a shortage of inorganic nutrients (especially nitrogen) and organic substrates in coastal saltwater. The hypothesis of bacterial nitrogen limitation was corroborated by the higher Nuse index (an EcoPlate?-based nitrogen limitation indicator) in the polyhaline waters and the importance of NO₂ ^?, NO₃ ^?, low-molecular-weight substances, and the low-molecular-weight:high-molecular-weight substances ratio, indicated by the canonical correspondence analyses (CCAs). Our results demonstrate the important stability of microbial community composition and potential metabolic activity in the different water salinity ranges, which are independent of the region and time of the year of sample collection in the estuary. This is a quite unexpected result for a dynamic environment such as the Patos Lagoon estuary.     

Antimutagenic and antioxidant activities of quebracho phenolics (Schinopsis balansae) recovered from tannery wastewaters

Quebracho extracts are used in tannery due to their high concentration of phenolics. The Mexican tannery industry uses around 450 kg/m(3) of which, 150 kg/m(3) remains in wastewaters and are discharged in drain pipe systems or rivers. The quebracho phenolics recovered from tannery wastewater (QPTW) was characterized by HPLC. The antimutagenic and antioxidant activities as well as the microbiological quality were evaluated. Total phenolic content of QPTW was 621mg catechin equivalent/g sample. Gallic and protocatechuic acids were the major components characterized by HPLC. QPTW showed an inhibition range on aflatoxin B(1) mutagenicity from 16 to 60% and was dose-dependent. Antioxidant activity (defined as beta-carotene bleaching) of QPTW (64.4%) at a dose of 12.3mg/mL was similar to that of BHT (68.7%) at a dose of 0.33 mg/mL, but lower than Trolox (90.8% at a dose of 2.5mg/mL); meanwhile antiradical activity (measured as reduction of DPPH) (60.8%) was higher than that of BHT (50.8%) and Trolox (34.2%). Quebracho residues were demonstrated to be an outstanding source of phenolic acids and for research and industrial uses.     

The occurrence, distribution and toxicity of cyanobacteria in the estuary of Lagoa dos Patos, Rio Grande do Sul

Several blooms of Microcystis aeruginosa have been observed in the Patos Lagoon estuary during the last fifteen years without a proper investigation of their ecological importance or possible toxicity. The present study has identified and quantified the presence of cyanobacteria in the Patos Lagoon estuary, particularly of M. aeruginosa. During this survey, identification and quantification of the main phytoplankton groups were done in relation to geographical distribution in the estuary. The presence of M. aeruginosa colonies in the estuarine region confirmed their superficial distribution throughout the estuarine waters during twelve months with a maximum of 1, 3.10(6) cells. L-1 in December, 1994 and a minimum of 1, 5.10(5) cells. L-1 in August, 1995 and also confirmed that M. aeruginosa originated from waters in the north of the estuary. The period of the highest cell and colonies densities was coincident with high chlorophyll-a levels in surface waters. Toxicity of M. aeruginosa bloom material was determined by bioassay and concentrations of hepatotoxins microcystins were identified by HPLC-DAD. M. aeruginosa blooms were considered highly toxic, presenting a 24 h-LD50 lower than 100 mg.Kg-1 b.w. and a toxin content higher than 1 microgram.mg-1 d.w. Several microcystin variants were found in the extracts with microcystin-LR predominating.     

环境污染对小清河口大型底栖动物多样性的影响

利用2008年7月小清河口和黄河口各8个站位的底栖动物调查资料,运用多种多样性指数对两河口底栖动物群落多样性特征进行了比较,并运用Spearman相关分析分析了14个环境因素和底栖动物群落多样性特征的相关关系。结果显示,小清河口共有大型底栖动物4门6纲17目35科37属38种,平均分类差异指数为85.7,分类差异变异指数为585.6,黄河口共有大型底栖动物6门9纲24目42科45属48种,平均分类差异指数为89.3,分类差异变异指数为435.8。小清河口的多样性指数(H’)仅约为黄河口的1/3。Spearman相关分析表明,H’与铵盐、硝酸盐、铜和汞含量呈负相关性,说明底栖动物群落多样性随着富营养化程度的提高而相应降低,营养物质氮的大量输入和重金属的污染对小清河口大型底栖动物群落产生了一定负面影响。ABC曲线法分析显示,小清河口底栖动物的丰度优势度曲线与生物量优势度曲线相交,表明小清河口底栖动物群落处于中度干扰状态;BOPA指数显示,小清河口8个站位中有4个站位受到不同程度的环境污染,生活污水和工业废水的排放可能对小清河口底栖动物生态特征带来巨大影响,加强工业和生活污水排放管理,强化清洁生产,加强工业污染源的治理力度,对恢复小清河流域水质、改善生态环境具有重要意义。     

DNA adducts as indicators of genotoxic exposure in indigenous and transplanted mussels, Mytilus edulis L. from Icelandic coastal sites

Indigenous mussels, Mytilus edulis, were collected at sites with supposed different amounts of pollution; Reykjavík harbour, Keflavík harbour, Grafarvogur and Hvalfj?rdur (reference), along the south-western coast of Iceland in March 2000. Mussels from Hvalfj?rdur and Reykjavík harbour were also collected in August the same year. Additionally, mussels were transplanted from the reference site to Reykjavík harbour for 6 weeks during both winter and summer for comparison. DNA adducts were analysed by 32P-post-labelling in gills and digestive gland. Highest adduct levels were found in gill tissue from indigenous mussels collected in Reykjavík harbour. Adduct levels in both tissues from mussels collected at the reference site were below or very close to the detection limit during winter, but seemed to increase a little during summer. Mussels from sites with supposed intermediate pollution had intermediate levels of DNA adducts in gills but did not differ from Reykjavík harbour in digestive gland. No increase in adduct levels was observed in mussels transplanted from the reference site to Reykjavík harbour, except for a slight increase in digestive gland during winter. This study shows that 32P-post-labelling analysis of DNA adducts is sensitive enough to be used on indigenous mussels from relatively pristine areas and that adduct levels are increased in harbours/urban sites. However, transplantation of mussels from a clean site to the harbour for 6 weeks did not result in increased adduct levels in gills, the tissue with the highest adduct levels. The results also indicate that seasonal variation in adduct levels may occur.     

Genotoxic and mutagenic activity of environmental air samples from different rural, urban and industrial sites in Flanders, Belgium

The present study reports mutagenic and genotoxic activities associated with ambient air collected at 15 sites characteristic for urban, industrial or rural conditions in Flanders. Airborne particulates (PM10) and semi-volatile compounds were collected on quartz filters (QF) and polyurethane foam (PUF) cartridges using a high-volume sampling device. The mutagenic and genotoxic potency of the organic extracts--Soxhlet extraction with acetone--was determined by use of the Salmonella mutagenicity standard plate-incorporation assay and the Vitotox assay, respectively. Concentrations of 16 polycyclic aromatic hydrocarbons (PAHs) in the extracts were determined by reversed-phase high-performance liquid chromatography (HPLC). Ambient air samples contained significant PAH levels and mutagenic activities at all 15 sites: direct mutagenicity of up to 47 revertants per cubic meter was found in the QF extracts and more limited activity of up to 11 rev m(-3) in the PUF extracts. Metabolic activation of PUF extracts resulted in an important increase in mutagenic activity, up to 30 rev m(-3), but no such increase was observed for QF extracts. The highest values were observed outside large cities at industrial sites and at a rural site contaminated by pollution from a chemical plant at a distance of 4 km. Also at the background location near the North Sea a significant mutagenic activity was measured in the QF extracts (+S9: 9 rev m(-3); -S9: 7 rev m(-3)). Apparently, there is in Flanders a significant background exposure level to airborne mutagenicity, even in areas with limited or no nearby pollution sources. Based on the concentrations of 10 mutagenic PAHs and supposing additivity of their specific mutagenicities, only a few percent (mean 3%) of the observed indirect mutagenic activity could be explained. This implies that most mutagenic activity originated from other substances that were not identified or measured in our chemical analysis. This underscores the importance of bio-monitoring measurements.     

Development and characterization of novel cell lines from Etroplus suratensis and their applications in virology, toxicology and gene expression

Four novel cell lines from tissues of eye, gill, kidney and brain of Etroplus suratensis were developed and characterized. The cell lines of eye, gill, kidney and brain were sub-cultured for 245, 185, 170 and 90 passages, respectively, since 2008. These cell lines showed predominantly epithelial-like cells. Effects of temperature and foetal bovine serum concentration on the growth of these cell lines were examined and optimum growth was found at the temperature of 28° C with 20% foetal bovine serum. All the four cell lines were successfully cryopreserved and revived at different passage levels. Cell-cycle analysis of these cell lines was carried out by fluorescence-activated cell sorting. Polymerase chain reaction (PCR) products obtained from the cells and tissues of E. suratensis with primers specific to the conserved region of 16S ribosomal RNA and cytochrome oxidase I genes of E. suratensis revealed the origin of cell lines from E. suratensis. Antibodies raised against the tissues and cells of eye, kidney and gill were highly cross reacted to their specific tissue and cells of E. suratensis. Chromosomal analysis revealed that E. suratensis cells have a normal diploid karyotype with 2n = 48. The cells of these cell lines were successfully transfected with pEGFP vector DNA. The eye (IEE), gill (IEG) and kidney (IEK) cell lines were found to be susceptible to nodavirus but resistant to infectious pancreatic necrosis virus (IPNV). The cells of gill, kidney and eye were applied to test the cytotoxicity of tannery effluents.     

Soil Respiration and Bacterial Structure and Function after 17 Years of a Reciprocal Soil Transplant Experiment

The effects of climate change on soil organic matter—its structure, microbial community, carbon storage, and respiration response—remain uncertain and widely debated. In addition, the effects of climate changes on ecosystem structure and function are often modulated or delayed, meaning that short-term experiments are not sufficient to characterize ecosystem responses. This study capitalized on a long-term reciprocal soil transplant experiment to examine the response of dryland soils to climate change. The two transplant sites were separated by 500 m of elevation on the same mountain slope in eastern Washington state, USA, and had similar plant species and soil types. We resampled the original 1994 soil transplants and controls, measuring CO₂ production, temperature response, enzyme activity, and bacterial community structure after 17 years. Over a laboratory incubation of 100 days, reciprocally transplanted soils respired roughly equal cumulative amounts of carbon as non-transplanted controls from the same site. Soils transplanted from the hot, dry, lower site to the cooler and wetter (difference of -5°C monthly maximum air temperature, +50 mm yr^-1 precipitation) upper site exhibited almost no respiratory response to temperature (Q₁₀ of 1.1), but soils originally from the upper, cooler site had generally higher respiration rates. The bacterial community structure of transplants did not differ significantly from that of untransplanted controls, however. Slight differences in local climate between the upper and lower Rattlesnake locations, simulated with environmental control chambers during the incubation, thus prompted significant differences in microbial activity, with no observed change to bacterial structure. These results support the idea that environmental shifts can influence soil C through metabolic changes, and suggest that microbial populations responsible for soil heterotrophic respiration may be constrained in surprising ways, even as shorter- and longer-term soil microbial dynamics may be significantly different under changing climate.     

Characterization of sorption sites and differential stress response of microalgae isolates against tannery effluents from ranipet industrial area—An application towards phycoremediation

Phycoremediation ability of microalgae namely Oscillatoria acuminate and Phormidium irrigum were validated against the heavy metals from tannery effluent of Ranipet industrial area. The microalgae species were cultured in media containing tannery effluent in two different volumes and the parameters like specific growth rate, protein content and antioxidant enzyme activities were estimated. FTIR spectroscopy was carried out to know the sorption sites interaction. The antioxidant enzymes namely superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) contents were increased in microalgae species indicating the free radical scavenging mechanism under heavy metal stress. SOD activity was 0.502 and 0.378 units/gram fresh weight, CAT activity was 1.36 and 0.256 units/gram fresh weight, GSH activity was 1.286 and 1.232 units/gram fresh weight respectively in the effluent treated microalgae species. Bio sorption efficiency for Oscillatoria acuminate and Phormidium irrigum was 90% and 80% respectively. FTIR analysis revealed the interaction of microalgae species with chemical groups present in the tannery effluent. From the results, the microalgae Oscillatoria acuminate possess high antioxidant activity and bio sorption efficiency when compared to Phormidium irrigum and hence considered useful in treating heavy metals contaminated effluents.     

胶州湾海水,海洋生物体中重金属含量的研究

研究了胶州湾海水、海洋生物体中重金属含量 .结果表明 ,沿岸河口地区海水中重金属污染主要受沿岸工业排放废水的影响 ;海洋生物体内重金属含量不仅受海水中的重金属含量大小的支配 ,温度对其也有一定的影响 .海洋生物体中重金属含量为底栖生物大于鱼类 ,以沉积物为主要饵料的杂食性底栖动物内重金属含量大于鱼类 ;海洋生物对于Cu比其它重金属具有较强的富集能力 .     

Inhibitory effects of auxins and related substances on the activity of an Arabidopsis glutathione S-transferase isozyme expressed in Escherichia coli

Glutathione S-transferases (GSTs; EC 2.5.1.18) are encoded by a gene family. Some GSTs have the capacity to bind to indole-3-acetic acid (IAA), whereas the gene expression of other GSTs is regulated by auxin. In order to assess a possible physiological significance of the auxin binding of GST, we investigated effects of auxins on the activity of GST expressed in Escherichia coli. cDNA cloning was carried out for the fifth gene ( GST5 ) of GST in Arabidopsis. Although the deduced amino acid sequence of GST5 was remotely related to that of the other Arabidopsis GSTs (less than 20% identical), the GST5 protein (GST5) expressed in E. coli showed GST activity. Apparent K_m values of GST5 are 0.86 and 1.29 m M for glutathione (GSH) and 1-chloro-2,4-dinitrobenzene, respectively. IAA, 2,4-dichlorophenoxyacetic acid (2,4-D), 1-naphthaleneacetic acid (1-NAA) and 2-NAA inhibited the enzyme activity competitively with respect to GSH. The apparent K_i of IAA is 1.56 m M . Salicylic acid inhibited GST activity in a noncompetitive manner. 2,4-D was the most inhibitory among the tested chemicals. GST5 bound to GSH-immobilized agarose gel was effectively eluted by IAA. These results indicate that IAA and the related substances bind to GST5 at the GSH-binding site, and exclude the possibility that the compounds could be substrates for GST5. Although the K_i value of IAA is too high for any physiological consequences, it might be assumed that GST activity is modulated in vivo by an auxin-related substance(s). The steady-state level of the GST5 mRNA was increased by wounding, heat shock, and spraying buffer on the plant, but was not influenced by auxin treatment.     

胶州湾海水、海洋生物体中重金属含量的研究

研究了胶州湾海水、海洋生物体中重金属含量.结果表明,沿岸河口地区海水中重金属污染主要受沿岸工业排放废水的影响;海洋生物体内重金属含量不仅受海水中的重金属含量大小的支配,温度对其也有一定的影响.海洋生物体中重金属含量为底栖生物大于鱼类,以沉积物为主要饵料的杂食性底栖动物内重金属含量大于鱼类;海洋生物对于Cu比其它重金属具有较强的富集能力.     

Localization of thymocyte stem cell precursors in the pharyngeal endoderm of early amphibian embryos.

The embryonic germ layer derivation of thymic lymphocytes in the leopard frog (Rana pipiens) was invesigated. The ectoderm and mesoderm—but not the endoderm—from the developing gill bud was reciprocally and orthotopically transplanted between diploid and triploid chromosomally marked 72-hr old embryos. The lymphocytes which subsequently developed in the thymus were of host ploidy. Thus, the head mesenchyme adjacent to the thymic anlagen does not appear to be the source of the stem cells from which thymocytes differentiate. Rather, the stem cells can he localized in the endoderm of the pharyngeal pouch which initially gives rise to the thymic epithelium. These findings suggest either the endodermal origin of thymocytes or the very early migration of stem cells into the thymus anlagen prior to any thymus histogenesis.     

Multiple biomarkers of pollution effects in caged mussels on the Greek coastline

A suite of biomarkers was measured in caged mussels at areas impacted by different anthropogenic activities along the Greek coastline to assess biological effects of environmental pollution. Mussels were caged at coastal sites in the vicinity of major cities, in areas influenced by major industries, agricultural practices and in islands away from known sources of pollution. Biomarkers indicative of neurotoxicity (acetylcholinesterase, AchE), oxidative stress (catalase, CAT), phase II biotransformation of xenobiotics (glutathione S-transferase, GST), metal exposure (metallothioneins, MTs) and protein synthesis (RNA:DNA ratio) were measured to assess effects of various types of pollutants. AchE activity proved to be the most responsive biomarker with decreased values at sites influenced by agricultural, urban and industrial activities. Decreased CAT and GST activities and increased MTs levels were recorded at a number of anthropogenic-impacted sites. RNA:DNA ratio showed a biphasic response as both high and low values were found at impacted sites. Principal component analysis clearly distinguished sites receiving pollution inputs from non-polluted sites. The combination of the selected biomarkers used in caged mussels resulted useful in the assessment of the effects of environmental pollution.