Responses of B-esterase enzymes in oysters (Crassostrea gigas) transplanted to pesticide contaminated bays form the Ebro Delta (NE,Spain)

Marine bivalves such as oysters are widely used as bioindicators to monitor marine coastal pollution. This study aimed to use B-esterase activity responses in oysters (Crassostrea gigas) cultured in Ebro Delta bays to monitor environmental effects of pesticides. The B esterases investigated were acetylcholinesterase, propionylcholinesterase, and carboxylesterase and their activities were measured in adductor muscle and gills from oysters transplanted in Ebro Delta bays where the are traditionally grown. Enzyme activities were related with physico-chemical parameters and pesticide levels measured in water. Cholinesterase activities measured in gills were unaffected across sites and periods. Conversely, carboxylesterase activities in oyster gills varied across periods and sites and were negatively correlated with residue levels of organophoshporous and carbamate pesticides in water. Therefore, inhibition of carboxylesterase activities can be considered a good indicator of exposure to anti-cholinergic pesticides in oysters.     

Sublethal impact of short term exposure to the organophosphate pesticide azamethiphos in the marine mollusc Mytilus edulis

Concern has been raised that the increased use of pesticides in intensive aquaculture practices may cause adverse sublethal effects to non-target aquatic species. Azamethiphos is an organophosphate (OP) pesticide used to combat sea lice infestations in farmed salmonids. Here, the sublethal impact on the blue mussel, Mytilus edulis, of short term exposure to azamethiphos was determined. The testing regime included biomarkers of exposure (acetylcholinesterase activity), cytotoxicity (neutral red retention), immune function (phagocytic index) and physiological condition (feeding rate). The distribution and sensitivity of M. edulis acetylcholinesterase to inhibition by azamethiphos was first determined, yielding IC(50) values of 0.736 and 1.30 mg l(-1) for gill and haemolymph, respectively. Exposure of mussels to 0.1 mg l(-1) azamethiphos for periods of up to 24h caused a significant reduction in acetylcholinesterase activity in both the haemolymph (P<0.0002) and the gill (P<0.002), alteration in cell viability (P<0.02) and decrease in phagocytic index (P<0.03). The feeding rate remained unaffected. The results support the hypothesis that, in addition to its neurotoxic effects, azamethiphos can modulate haemocyte function and immune defence in M. edulis at environmentally relevant concentrations after only a few hours.     

Toxic contaminants and their biological effects in coastal waters of Xiamen,China. II. Biomarkers and embryo malformation rates as indicators of pollution stress in fish

Baseline information is presented on embryo malformation rate and biomarkers in fish as indicators of sub-lethal stress caused by pollution in coastal waters of Xiamen, PR China. Fish and eggs were sampled from several areas in Xiamen coastal waters (Xiamen Harbour, Maluan and Tongan Bays and East Channel), where varying levels of pollutant input have been documented. Comparative sampling was done at a "cleaner" reference site at Dongshan Island. Embryonic malformation rates, which indicate general water quality, varied with location and species of fish, and exceeded background levels for unpolluted waters (assumed approximately 5%) by up to eightfold at some sites. Generally, sites around Xiamen Harbour show signs of poor water quality having highest mean levels of embryo deformity (20-30%) and these decreased towards open waters (Tongan Bay, Eastern Channel) where abnormalities approached background levels. An indication that toxic contaminants may be having a localised effect in the region, particularly in the harbour was reinforced by the biomarker assays. However, activities of the biomarkers ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase in fish livers indicate no clear pattern, and there is no evidence that fish from the four sampling areas have been more or less exposed to PAHs and other compounds that induce these biomarkers. Antioxidant biomarkers (glutathione peroxidase, catalase, superoxide dismutase, and reduced glutathione) suggest that exposure to xenobiotics appears to be lowest in Dongshan and Maluan and highest in the harbour and Tongan. Inhibition of acetylcholinesterase in fish muscle indicated possible effects by organophosphate and carbamate pesticides in Xiamen waters and these effects may be greatest in the area of the harbour.     

The use of cholinesterase as potential biomarker: In vitro characterization in the polychaete Capitella teleta

The ecological relevance of polychaetes coupled with their easy culture and maintenance in the laboratory, has led them to become increasingly used in marine ecotoxicological studies, raising the need to validate frequently applied monitoring tools at various biological levels. The present study was aimed to characterize the cholinesterases (ChE) activity in the polychaete Capitella teleta, using three substrates (acetylthiocholine iodide, propionylthiocholine iodide, and S-butyrylthiocholine iodide) and four known inhibitors (eserine hemisulfate, BW284c51, iso-OMPA and chlorpyrifos-oxon). Results showed that most of the measured cholinesterase activity was acetylcholinesterase (AChE). Inhibition of enzyme kinetic experiments denoted that sensitivity of C. teleta’s ChE to the organophosphorous metabolite chlorpyrifos-oxon (IC₅₀ = 60.72 nM) was analogous to some fish species. This study highlights the relevance of ChE characterization before its use as a biomarker in ecotoxicology and biomonitoring studies.     

Integrated use of biomarkers (acetylcholinesterase and antioxidant enzymes activities) in Mytilus galloprovincialis and Mullus barbatus in an Italian coastal marine area

The use of biomarkers to evaluate the biological effects of chemical pollutants in marine organisms represents a recent tool in the monitoring field responding to the need to detect and assess the effects of chemical contaminants on the biota. The aim of the present work was the field application of the integrated use of acetylcholinesterase (AChE) and antioxidant enzymes (catalase--CAT, glutathione peroxidase--GSH-Px), for detecting the possible exposure/effect induced by chemical pollutants in native marine organisms from a coastal marine area, represented by Salento Peninsula (Italy), that shows a coastline of high environmental value, but under constant urban pressure, including agriculture activities, widely diffused in the whole hinterland. Eight sampling stations were chosen: four not urbanized areas considered "uncontaminated" controls and four clearly exposed to anthropogenic impact. The bioindicator species studied were a sessile invertebrate, Mytilus galloprovincialis, and a benthic teleost fish, Mullus barbatus.AChE activity in M. galloprovincialis revealed significant differences among places; the minimum values observed (3.9+/-1.8 nmolmin(-1)mg(-1)) was about 50% reduced with respect to the maximum found (11.4+/-0.9 nmolmin(-1)mg(-1)). The reduction in AChE activity observed in two control stations could be explained by the leaching of pesticides into the sea from the agricultural lands. Moreover, the inhibition of AChE activity by heavy metals besides pesticides, can also explain the reduction of the enzymatic activity observed in an industrialized and harbour area. In M. galloprovincialis AChE activity showed a significant inverse correlation with catalase activity but not with glutathione peroxidase that did not significantly change in animals sampled from the eight stations. Also in M. barbatus AChE activity showed significant differences among places; it was inversely correlated with liver GSH-Px activity, but not with catalase activity, that did not show any significantly variation in animals sampled in the different stations.In conclusion, the integrated use of AChE and antioxidant enzymes (catalase or glutathione peroxidase) in M. galloprovincialis and M. barbatus, two species living in different compartment of marine coastal ecosystem, can find a useful application within the framework of marine coastal environment monitoring programs for detecting the possible exposure/effect induced by chemical pollutants, including pesticides, on living marine organisms.     

Contaminant concentrations, biochemical and hematological biomarkers in blood of West Indian manatees Trichechus manatus from Brazil

The West Indian manatee Trichechus manatus is threatened with extinction in Brazil, and this study focused on nondestructive blood samples analyzed for metals, polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs), as well as biochemical and hematological biomarkers. Studied manatees were kept at Projeto Peixe-Boi headquarters in Pernambuco State, and at two natural areas in estuaries where they are released to the wild. Manatees kept at the natural estuary in Paraiba State have blood concentrations of Al, Pb, Cd, Sn that are 11, 7, 8 and 23 times greater, respectively, than the concentrations found in blood of animals from the same species in Florida, USA. An inhibition of butyrylcholinesterase in manatees kept at the two reintroduction sites in Alagoas and Paraiba States indicated possible exposure of the animals to cholinesterase inhibitor insecticides. PCBs and OCPs were not detected. Results from this study will help delineate conservation efforts in the region.     

Copper accumulation by bacteria and transfer to scallop larvae

A bacterial copper-resistant strain was isolated from a hatchery-conditioned adult of the scallop Argopecten purpuratus and was identified as Vibrio sp. according to its physiological characteristics. The lowest concentration of Cu2+ required for its complete inhibition in VNSS medium was 50 microg ml(-1). The Vibrio strain was found to accumulate copper, exhibiting cellular and loosely bound copper levels of 201.14 and 493.21 microg g(-1) dry weight, respectively, after 24 h of incubation in VNSS medium supplemented with 15 microg ml(-1) of Cu2+, with cellular concentration factors of 10.17 and 14.35 after 12 and 24 h of exposure. When a scallop larvae culture was exposed to a concentration of 6.46 x 10(2) CFU ml(-1) of Cu-enriched Vibrio, they accumulated 20.42 +/- 1.12 and 30.96 +/- 1.85 microg Cu2+ g(-1) dry weight after 12 and 24 h, respectively. This study suggests that bacterial copper accumulation could be very active in marine environments increasing the occurrence of copper transfer to marine food chains.     

A geochemical record of environmental changes in sediments from Sishili Bay,northern Yellow Sea,China: Anthropogenic influence on organic matter sources and composition over the last 100 years

Total organic carbon (TOC), total nitrogen (TN), δ¹³C and δ¹⁵N were measured in sediment cores at three sites in Sishili Bay, China, to track the impacts of anthropogenic activities on the coastal environment over the last 100 years. The increased TOC and TN in the upper section of sediment cores indicated a eutrophic process since 1975. In comparison, the TOC and TN in the sediment core near to a scallop aquaculture area displayed a much slower increase, indicating the contribution of scallop aquaculture in mitigating eutrophication. Combined information from δ¹³C, δ¹⁵N and TOC:TN indicated an increased terrestrial signal, although organic matter sources in Sishili Bay featured a mixture of terrestrial and marine sources, with phytoplankton being dominant. Increased fertilizer use since 1970s contributed to the eutrophic process in Sishili Bay since 1975, and increased sewage discharge from 1990s has added to this process.     

Action Sites of Cypermethrin,a Synthetic Pyrethroid in the Snail Lymnaea acuminata

24 h of in vivo exposure to 40 and 80 = LC50 of the synthetic pyrethroid cypermethrin significantly reduced the acetylcholinesterase, cytochrome oxidase and lactic dehydrogenase activity in the nervous tissue of L. acuminata. Succinic dehydrogenase activity increased significantly after the same treatment. Fifteen minutes in vitro exposure to different doses of cypermethrin also significantly inhibited cytochrome oxidase and lactic dehydrogenase activity. However, acetylcholinesterase and succinic dehydrogenase activities remain unaltered. It is concluded that the cypermethrin on the one hand causes repititive discharges at the nerve terminal and on the other hand blocks the electron transport chain.     

Ecotoxicological effects of a semi-submerged municipal dump (Castle harbour, Bermuda) on the Calico scallop Argopecten gibbus

A biomarker study was undertaken using the Calico scallop Argopecten gibbus to assess the ecotoxicological effects of a semi-submerged municipal dump on the adjacent patch reef lagoon ecosystem (Castle harbour, Bermuda). Caged scallops were deployed in situ for 2 months at various distances from the dump (50 m, 900 m and 2.7 km) and at a reference site (14 km). A suite of biomarkers comprising metallothionein (MT), lipid peroxidation (LPO), vitellin-like proteins (Vn), glutathione S-transferase (GST), DNA strand breaks and condition factor (CF) were investigated in various tissues of the scallop (gill, digestive gland and gonad). Levels of heavy metals were also measured in the whole scallop soft tissue. While there was some variation in response between tissues, in general the results indicated that the dump was negatively impacting scallops deployed in the adjacent marine environment: elevated levels of MT, DNA strand breaks, Vn and GST and reduced condition factor were found for scallops deployed nearest to the dump and at the site 1.5 km from this point source of contamination (Tuckers town) in Castle harbour, with respect to the reference site, North Rock (although this exhibited some degree of metal contamination). The gills from scallops deployed at the dump site were the most responsive tissue, with the highest expression of MT, LPO and DNA damage. This study indicates the potential of the Calico scallop as a convenient bioindicator species in the marine tropical benthos.     

Effects of the Black River (Cape,South Africa) on the distribution and survival of marine psammofauna

The physicochemistry of the Black River and adjacent marine beach, in terms of tidal, daily and seasonal fluctuations of temperature and salinity, presents a highly stressed environment exacerbated by canalization of the river mouth. Riverine pollution further stresses this environment, producing low densities and diversities of marine infauna adjacent to the river. No fauna were found in the river mouth where typical estuarine conditions exist only during high tide. Tolerance of the dominant marine species, Cerebratulus fuscus, to temperature, salinity and river water combinations showed that mortality was more rapid after exposure to river water dilutions than after exposure to distilled water dilutions. Computer-generated models suggested that this species should survive conditions closer to the river than were observed. This confirmed additional stress due to pollution carried in the river from urban and industrial drainage. Lower river water temperatures during winter appear to ameliorate pollution stress, enhancing survivability of the infauna.     

Preliminary results of biomarker responses in zooplankton of brackish environments

Zooplankton is an essential component of the food chains of marine and brackish environments. Ecotoxicological risk to zooplankton, evaluated by the biomarker approach, can be used as an early warning signal of risk to the health of marine ecosystems. The aim of this project is to evaluate the application of certain biomarkers in zooplankton. In the present paper, laboratory and field studies were performed with the copepods Acartia margalefi, Acartia latisetosa and the mysid Siriella clausi. Acetylcholinesterase activity (AChE) was determined in homogenates of whole organisms. Mean AChE activity was 10.05 micromol min g(-1) for A. margalefi, 3.30 micromol min g(-1) for A. latisetosa and 79.70 micromol min g(-1) for S. clausi. A. margalefi and S. clausi also showed a linear increase in enzyme activity with increasing concentrations of samples. Preliminary laboratory work was also carried out with specimens of A. latisetosa, which were exposed to an organophosphorus insecticide (parathion). Moderate inhibition of AChE (19%) with respect to controls was observed. AChE activities were two orders of magnitude higher in the zooplankton than in hemolymph samples of the decapod Carcinus aestuarii (Fossi et al., 1996) indicating that these species have a high metabolic rate, which makes them suitable for biomarker studies.     

Delta13C and delta15N shifts in benthic invertebrates exposed to sewage from McMurdo Station, Antarctica

In an effort to identify biomonitors for contamination of Antarctic marine benthos by sewage, this study determines whether the US Antarctic Program’s McMurdo Station produces a benthic sewage footprint and whether resident megafauna are assimilating sewage-derived material. We identified strong C and N isotopic gradients in benthic sediment as a function of downstream distance from McMurdo Station’s point-source sewage addition. Sediment C and N isotope ratios approached marine background levels at the sampling end-point 612 m downcurrent. Based on isotope abundances in their tissues, at least some sewage C and N were assimilated by the sedentary, suspension feeding soft coral Alcyonium antarcticum, ascidian Cnemidocarpa verrucosa and bivalve Laternula elliptica. However, as inferred by tissue-sediment differences in downstream isotope trends, such assimilation was not in proportion to sewage exposure and input, therefore implying non-generalist feeding behavior by these species. In contrast, the motile, generalist feeding sea urchin Sterechinus neumayeri, sea star Odontaster validus and ribbon worm Parborlasia corrugatus showed isotopic evidence of sewage C and N assimilation roughly in proportion to sewage input. We recommend these generalist feeders for further use as biomonitors at this site now that sewage treatment has been implemented. As these species are circumpolar in distribution, they may also prove useful elsewhere in the Antarctic.     

Integrated use of biomarkers (superoxide dismutase, catalase and lipid peroxidation) in mussels Mytilus galloprovincialis for assessing heavy metals' pollution in coastal areas from the Saronikos Gulf of Greece

Mussels are used as sentinel organisms and bioindicators to evaluate the toxic effects of chemical pollutants in marine organisms, especially heavy metals, representing an important tool for biomonitoring environmental pollution in coastal areas. Antioxidant defence enzymes play an important role in cellular antioxidant defence systems and protect from oxidative damage by reactive oxygen species (ROS). Indigenous mussels Mytilus galloprovincialis of the Saronikos Gulf of Greece were used for monitoring heavy metal pollution in three polluted sites in the area and in one unpolluted site. Seasonal variations of the activity of antioxidant defence enzymes, superoxide dismutase (SOD) and catalase (CAT), as well as lipid peroxidation (LP) were measured as biomarkers in a period of three years in relation to concentrations of trace metals in their gills and mantle and compared to mussels from an unpolluted sampling site. SOD activity increased at least 2 fold at the polluted sites when compared to the control site (the high activity was recorded in the spring time). CAT activity was increased 2-3 times at the polluted sites, with high activity in the winter and spring time, compared to the control site. LP concentration was twice higher at the polluted sites, following the same seasonal pattern. Trace metals contents in mussels collected at polluted sites were 3-4 fold higher compared to the control site and showed moderate variations along the months, with a winter maximum followed by a summer pre-spawning minimum matching the seasonal trends of temperature and salinity. Our results showed that metal pollution in the Elefsis Bay (the most polluted coastal area) causes relatively medium levels of oxidative stress in tissues of mussels due to cellular oxy-radical generation. This study, which is the first in the area, showed that seasonal variations of the activity of antioxidant defence enzymes and LP concentrations in mussels can be used as potential biomarkers of toxicity for long-term monitoring in marine coastal ecosystems.     

Biomarker responses in caged rockfish (Sebastes schlegeli) from Masan Bay and Haegeumgang, South Korea

The purpose of this study was to compare enzymatic biomarker activities in fish caged at two sites, Masan Bay (contaminated) and Haeguemgang (reference). In the present study, ethoxyresorufin O-deethylase (EROD), brain acetyl cholinesterase (bAChE), muscle acetyl cholinesterase (mAChE) and butyryl cholinesterase (mBChE) in caged rockfish (Sebastes schlegeli) were measured 0, 1, 3, 7, 14, 21 and 30 days after caging. The level of CYP1A mRNA and Protein expression was induced higher in Masan Bay at 1, 3, 7, 14 and 30 days after caging. EROD activity in the caged fish was significantly higher in Masan Bay than in Haeguemgang 3 and 7 days after caging, but not at 14 and 30 days after caging. bAChE activity was significantly inhibited at 7 and 14 days after caging in Masan Bay. However, mBChE activity was not significantly inhibited during the experiment. Taken together, the data suggest that the caged fish were exposed, at least transiently, to CYP1A inducers and ChE inhibitors, which is consistent with our previous observations.     

Seasonal succession of cyanobacterial protease inhibitors and Daphnia magna genotypes in a eutrophic Swedish lake

Lakes are well known for having a pattern of seasonal succession of phytoplankton and zooplankton. The succession of different taxa of phytoplankton results in a succession of zooplankton taxa, and within the genus Daphnia, into a succession of different genotypes (clones). One cause for this succession of Daphnia clones might be the production of digestive protease inhibitors by cyanobacteria, which usually bloom in summer. Here we report seasonal changes in the frequency and the abundance of Daphnia magna haplotypes in a eutrophic lake, which developed a chymotrypsin-inhibitor-producing cyanobacterial bloom in May. These seasonal changes were not related to changes of biotic and abiotic lake parameters. However, a very high content of chymotrypsin inhibitors was observed in May (but not in other months). This was assumed to have exerted a strong punctual selection pressure on the Daphnia population and on the direct targets of the protease inhibitors, i.e. the digestive chymotrypsins of Daphnia. Actually, D. magna from before and during the cyanobacterial bloom showed a different protease pattern on activity stained SDS-PAGE in comparison to clones from the month after the bloom. However, no difference in tolerance, measured as IC₅₀ values, to inhibition by natural lake seston from May was found between the clones from before and after the bloom. Thus, the hypothesis that a seasonal adaptation of D. magna subpopulations from either April/May or June might have occurred could not be proven. This suggests that the Daphnia population investigated here is locally adapted to cyanobacterial protease inhibitors.     

Biomarker responses in caged rockfish (Sebastes schlegeli) from Masan Bay and Haegeumgang,South Korea

The purpose of this study was to compare enzymatic biomarker activities in fish caged at two sites, Masan Bay (contaminated) and Haeguemgang (reference). In the present study, ethoxyresorufin O-deethylase (EROD), brain acetyl cholinesterase (bAChE), muscle acetyl cholinesterase (mAChE) and butyryl cholinesterase (mBChE) in caged rockfish (Sebastes schlegeli) were measured 0, 1, 3, 7, 14, 21 and 30 days after caging. The level of CYP1A mRNA and Protein expression was induced higher in Masan Bay at 1, 3, 7, 14 and 30 days after caging. EROD activity in the caged fish was significantly higher in Masan Bay than in Haeguemgang 3 and 7 days after caging, but not at 14 and 30 days after caging. bAChE activity was significantly inhibited at 7 and 14 days after caging in Masan Bay. However, mBChE activity was not significantly inhibited during the experiment. Taken together, the data suggest that the caged fish were exposed, at least transiently, to CYP1A inducers and ChE inhibitors, which is consistent with our previous observations.     

富营养化对贵州红枫湖水库汞形态和分布特征的影响

通过对贵州省红枫湖水库中各种汞形态的空间分布及季节变化的研究,探讨了富营养化对各种汞形态分布特征的影响.红枫湖湖水总汞浓度在2.5-13.9ng/L之间,平均值为6.9ng/L.红枫湖水库中汞浓度的空间分布和季节变化均严重受到人为汞污染源的干扰.水体中汞在颗粒态和溶解态之间的分配,主要受内源有机质以及氧化还原条件的影响.由于水华现象,春季后五采样点大量繁殖的藻类吸附了大量的汞,从而改变了汞在水库中的分配和迁移.藻类的生长对水体中溶解气态汞浓度分布也有显著的影响.在夏季,总甲基汞和溶解态甲基汞在水体中的垂直分布表明:在富营养化较严重的后五采样点,水体中升高的甲基汞主要是由于水体中汞的甲基化过程;而在富营养化特征不明显的大坝,水体中升高的甲基汞主要来自沉积物甲基汞的释放.红枫湖水体中各种汞形态的分布特征表明,富营养化对汞的迁移转化影响显著,尤其是汞的甲基化过程.水体富营养化为汞的甲基化提供了有利条件,给水生生态环境及人体健康带来了潜在的威胁.     

Investigation of the mud crab (Scylla serrata) as a potential bio-monitoring species for tropical coastal marine environments of Australia

Mud crabs, Scylla serrata, were sampled from four estuaries (the Normanby, Herbert, Burdekin and Fitzroy Rivers) along the coast of northern Queensland, Australia, representing a pollution gradient from low to high contamination based upon previous chemical monitoring. Four biomarkers; glutathione-S-transferase (GST) activity, cholinesterase (ChE) inhibition and the urinary metabolite concentrations of naphthalene (NPH) and benzo-a-pyrene (BaP) were evaluated in S. serrata hepatopancreas, haemolymph and urine. Site-specific evidence of elevated GST activity and BaP metabolite concentrations and significant ChE inhibition was detected. Biomarker responses from this field study provide evidence of contaminant exposure of S. serrata from river estuaries along the coast of northern Queensland and indicate that further investigation is warranted. Based on the current results, and with further work on characterising the dose-response and seasonal variation in this species, mud crabs have great potential as indicator species for water quality and ecosystem monitoring programs across tropical coastal regions of Australia.     

Biomarker responses in caged rockfish (Sebastes schlegeli) from Masan Bay and Haegeumgang, South Korea

The purpose of this study was to compare enzymatic biomarker activities in fish caged at two sites, Masan Bay (contaminated) and Haeguemgang (reference). In the present study, ethoxyresorufin O-deethylase (EROD), brain acetyl cholinesterase (bAChE), muscle acetyl cholinesterase (mAChE) and butyryl cholinesterase (mBChE) in caged rockfish (Sebastes schlegeli) were measured 0, 1, 3, 7, 14, 21 and 30 days after caging. The level of CYP1A mRNA and Protein expression was induced higher in Masan Bay at 1, 3, 7, 14 and 30 days after caging. EROD activity in the caged fish was significantly higher in Masan Bay than in Haeguemgang 3 and 7 days after caging, but not at 14 and 30 days after caging. bAChE activity was significantly inhibited at 7 and 14 days after caging in Masan Bay. However, mBChE activity was not significantly inhibited during the experiment. Taken together, the data suggest that the caged fish were exposed, at least transiently, to CYP1A inducers and ChE inhibitors, which is consistent with our previous observations.