Acetylcholinesterase as a biomarker of exposure to neurotoxic compounds in the clam Tapes philippinarum from the Lagoon of Venice

Measurement of acetylcholinesterase (AChE) is widely used as a biomarker of exposure to neurotoxic compounds in aquatic environments. In the present study, AChE activity was monitored in the clam Tapes philippinarum in the Lagoon of Venice, with the aim of evaluating the possible exposure of animals to neurotoxic compounds. Two sampling strategies were followed. In the first case, temporal and spatial variations of AChE activity were measured in clams collected seasonally (from October 2003 to June 2004) in 3 sites of the Lagoon: Campalto (site 1), near a sewage treatment plant, Marghera (site 2), a highly polluted area, and Poveglia (site 3), a reference site. The condition index (CI) of clams was also calculated. In the second case, AChE activity was measured in clams collected at the end of seasonal samplings in 12 sites located throughout the Lagoon: areas licensed for clam farming (sites 5 to 11), nearby canals characterised by waste waters from agricultural areas (sites 4, 12, 13, 14), and a reference site outside the Lagoon (site 15). Results revealed a significant reduction in AChE activity in clams from Marghera in October, January and April, indicating their probable exposure to neurotoxic compounds originating in the industrial zone of Porto Marghera. A seasonal trend in AChE activity was also observed, enzyme activity being higher in January and lower in June. Clams from Campalto generally had higher CI values than those from Marghera and Poveglia. However, no particular correlation between AChE activity and CI was found. Lastly, marked differences in AChE activity were recorded when comparing enzyme activity of clams from various sites in the Lagoon of Venice with those of clams collected at site 15. Significant reductions in AChE activity were observed in animals collected in both nearby canals and licensed areas, indicating the homogeneous spatial distribution of potentially neurotoxic compounds throughout the Lagoon. It is probable that sediment mobilisation caused by clam harvesting and the passage of shipping plays an important role in resuspension of persistent contaminants having neurotoxic activity.     

Molecular and biochemical biomarkers responses in the mussel Mytilus edulis collected from Southern Brazil coast

Marine ecosystems are typically subjected to a variety of stressors containing complex xenobiotics mixtures. This study aims to evaluate the responses of molecular and biochemical biomarkers in the mussel Mytilus edulis providing data for environmental monitoring programs development in Southern Brazil. Mussels were collected at a polluted site, near Patos Lagoon outfall, and at a control site. Gills, muscle and mantle samples were used for biomarker determinations. Mussels collected at the polluted site significantly increased superoxide dismutase (SOD) and glutathione S-transferase (GST) activities and decreased catalase (CAT) activity. Moreover, an increase in sod1, gstπ and hsp70 mRNA expression was observed. Overall, biochemical and molecular biomarkers responses were observed, but these responses varied depending on the analyzed tissue. These results indicate possible contaminants effects on organisms and the need for effective environmental monitoring programs in this ecosystem.     

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.     

Oxidative stress in the mussel Mytella guyanensis from polluted mangroves on Santa Catarina Island,Brazil

Digestive glands of the mangrove mussel Mytella guyanensis, collected at one non-polluted site (site 1) and two polluted sites (sites 2 and 3), were analysed for different antioxidant defenses, lipid peroxidation and DNA damage. Thiobarbituric acid-reactive substance (TBARS) and 8-oxo-7,8-dihydro-2'-deoxyguanosine levels were enhanced at the polluted sites. With the exception of superoxide dismutase, the activities of catalase and glutathione peroxidase were also higher at the polluted sites. Greater increases were observed in glutathione reductase, glutathione S-transferase and etoxyresorufine-O-deethylase activities at the polluted sites. Conversely, reduced glutathione content was higher at the control site. Trace metal contents in mussels collected at polluted sites were increased compared to the control site, and there were strong positive correlations between TBARS and Cu and Pb contents. M. guyanensis is routinely exposed to an oxidative stress condition at both polluted sites, and considering xenobiotic bioaccumulation in bivalve molluscs, the mangrove mussel represents an excellent bioindicator for environmental monitoring studies.     

Effects on hatching in Littorina littorea after an oil spill

One year after an accidental spill of approx. 2000 t of Iranian crude oil on the west coast of Norway, specimens of the gastropod Littorina littorea were collected from the oil polluted shore and allowed to spawn in the laboratory. Snails were also collected from an unpolluted reference site. The oil contamination seemed to have no detectable effects on fertilization. However, the hatching success of the veliger larvae was significally less in the ‘oil polluted’ population. Some highly abnormal eggs were also observed in this egg population.     

Comparison of the effects of pollution on the marine bivalve Donax trunculus in the vicinity of polluted sites with specimens from a clean reference site (Mediterranean Sea)

The physiological and biochemical characteristics of tissue samples of the marine mussel, Donax trunculus, from an oil-polluted site (Qiryat Yam) and from a site adjacent to an industrial factory (Frutarom) producing polyvinyl chloride (PVC) were compared with samples from a clean reference site (Akko). All sites were located along the sandy shores of the Israeli Mediterranean Sea. The mussels from the oil-polluted site showed increased activity of the system of active transport of organic anions (SATOA) in the gills and the renocardial organ and also of the multixenobiotic resistance transporter (MXR) in the gills. In contrast, samples collected near the PVC factory showed a decrease in SATOA activity and no increase in the activity of MXR in the gills. Specimens from the reference site demonstrated a redox state of equilibrium between energy production and utilization, while in Donax from both the oil polluted and the PVC-polluted sites, the mitochondrial redox state reflected intensive consumption of energy. No significant changes were found in the activity of reduced glutathione s-transferase (GST) in the cytosolic fraction of the digestive gland of Donax collected from any of the three sites. The data demonstrate a differential increase in the anti-chemical defense systems and an intensification of energy metabolism in the mussels exposed to pollution.     

P/Ca in coral skeleton as a geochemical proxy for seawater phosphorus variation in Daya Bay, northern South China Sea

The P/Ca ratio in coral skeletons is considered to be a direct proxy for the nutrient P in seawater. We examined the reliability of this proxy by analyzing P/Ca in a Porites coral collected from a eutrophic area in the northern South China Sea. P concentrations were significantly higher compared to previously reported values from pristine and open seas, corresponding to the elevated nutrients from the study site. We compared coral P/Ca against recent in-situ records of seawater P concentrations. Our results show that P/Ca was primarily a function of TP sw rather than PO₄sw, and that the signal of skeletal P included not only phosphate, but also organic phosphorus. Besides the form of skeletal P, sub-sampling and analytical procedures and the distinctive nutrient regime were the most reasonable explanations for our results. We suggest that total P in coral skeletons may be an efficient proxy for seawater P variations and associated phytoplankton dynamics in eutrophic environments.     

Coral death from sewage and phosphate pollution at Aqaba,Red Sea

Localized pollution of coral reef areas is occurring at Aqaba, Red Sea, as a result of sewage discharge, and as a result of spillage of phosphate dust during loading of phosphate mineral onto ships. The rate of death of colonies of the coral Stylophora pistillata was found to be 4–5 times as great in the polluted area as in a control area. Coral damage in the control area is generally caused by grazing or by extreme low tide, but the cause of coral death in the polluted area was not readily apparent. The growth of algae, both on the damaged corals, and on glass slides placed out in the reef, was greatly stimulated in the polluted area, but it appeared that such algal growth was not the direct cause of coral death. Corals in the polluted area may be under stress because of reduced light intensity, inhibition of calcification by excess phosphate, and increased sediment load. It was found that in the polluted area there was a greater weight of sediment settling on the glass slides for a given weight of algae. But in addition, since algal growth was faster in the polluted area, the sediment load was increased by the sediment trapping capacity of the enhanced algal growth. Thus it is suggested that increased algal growth stimulated by increased nutrient concentrations may be important in greatly increasing the sediment load experienced by corals.     

Biomarkers of exposure and effect in the Brazilian flounder Paralichthys orbignyanus (Teleostei: Paralichthyidae) from the Patos Lagoon estuary (Southern Brazil)

Biomarkers of exposure (liver metallothionein-like proteins content and catalase and glutathione S-transferase activities) and effect (liver lipoperoxidation and blood cell DNA damage) of contaminants were analyzed in the Brazilian flounder Paralichthys orbignyanus from the Patos Lagoon estuary (Southern Brazil). Flounders were collected for a year in two sites: "Coroa do Boi" (polluted site) and "Saco do Justino" (non-polluted site). Results indicated that micronucleated cells frequency was the best biomarker to distinguish flounders from the two sites. Taken together, data from DNA damage analyses (micronucleus test and comet assay) indicated that flounders from the non-polluted site efficiently repaired the DNA breaks, contrary to those from the polluted site, which probably had their DNA repair system inhibited or exhausted. Furthermore, data from enzyme activities (catalase and GST) and lipid peroxidation indicated that flounders from the polluted site were under oxidative stress in summer and autumn.     

Does use of tropical beaches by tourists and island residents result in damage to fringing coral reefs? A case study in Moorea French Polynesia

Although coral reefs worldwide are subject to increasing global threats, humans also impact coral reefs directly through localized activities such as snorkeling, kayaking and fishing. We investigated five sites on the northern shore of Moorea, French Polynesia, and quantified the number of visitors on the beach and in shallow water. In field surveys, we measured total coral cover and colony sizes of two common genera, Porites and Acropora, a massive and branching morphology, respectively. One site, which hosted over an order of magnitude more people than the other four, had significantly less total coral cover and supported very little branching Acropora. In addition, size frequency distributions of both the branching and massive genera were skewed toward smaller colony sizes at the high use site. Our results demonstrated that the use of tropical beaches may result in less coral cover, with branching colonies rare and small.     

Oxidative stress, genotoxicity and histopathology biomarker responses in mullet (Mugil cephalus) and sea bass (Dicentrarchus labrax) liver from Bizerte Lagoon (Tunisia)

The aim of the study was to evaluate the impact of environmental contaminants on oxidative stress, genotoxic and histopathologic biomarkers in liver of mullet (Mugil cephalus) and sea bass (Dicentrarchus labrax) collected from a polluted coastal lagoon (Bizerte Lagoon) in comparison to a reference site (the Mediterranean Sea). Antioxidant enzyme activities were lower in fish from the polluted site compared with fish from the reference site, suggesting deficiency of the antioxidant system to compensate for oxidative stress. DNA damage was higher in both fish species from the contaminated site indicating genotoxic effects. The liver histopathological analysis revealed alterations in fish from Bizerte Lagoon. Hepatocytes from both fish species featured extensive lipid-type vacuolation and membrane disruption. Results suggest that the selected biomarkers in both fish species are useful for the assessment of pollution impacts in coastal environments influenced by multiple pollution sources.     

Lysosomal membrane stability and metallothioneins in digestive gland of Mussels (Mytilus galloprovincialis Lam.) as biomarkers in a field study

The lysosomal membrane destabilization and the metallothionein content in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.), collected along the east coast of the North Adriatic (Istrian and Kvarnerine coast, Croatia), were examined over a period of four years (1996–1999). The lysosomal membrane stability, as a biomarker of general stress, showed that the membrane labilization period in mussels from polluted, urban- and industrial-related areas was significantly decreased (p<0.05) when compared to mussels from control, clean sea water sites. In the harbour of Rijeka, the most contaminated site, the lysosomal membrane stability was reduced by more than 70% compared to the control. This method also proved to be a useful biomarker for detection of additional stress caused by short-term hypoxia that occurred once during this study inside the polluted and periodically quite eutrophic Pula Harbour. The concentration of metallothioneins in the mussel digestive gland, as a specific biomarker of exposure to heavy metals, did not reveal significant differences (p<0.05) between sites covered by this study.     

Understanding ship-grounding impacts on a coral reef: potential effects of anti-foulant paint contamination on coral recruitment

The 184 m cargo ship Bunga Teratai Satu collided with Sudbury Reef, part of the Great Barrier Reef and remained grounded for 12 days. The ship was re-floated only 3 days prior to the November 2000 mass coral spawning. No cargo or fuel was lost but the impact resulted in significant contamination of the reef with anti-foulant paint containing tributyltin (TBT), copper (Cu) and zinc (Zn). Larvae of the reef-building scleractinian coral Acropora microphthalma were exposed to various concentrations of sediment collected from the grounding site in replicated laboratory experiments. Two experiments were performed, both of which used varying ratios of contaminated and control site sediment in seawater as treatments. In the first experiment, the influence of contaminated sediment on larval competency was examined using metamorphosis bioassays. In the second, the effect of contaminated sediment upon larval recruitment on pre-conditioned terracotta tiles was assessed. In both experiments, sediment containing 8.0 mg kg(-1) TBT, 72 mg kg(-1) Cu and 92 mg kg(-1) Zn significantly inhibited larval settlement and metamorphosis. At this level of contamination larvae survived but contracted to a spherical shape and swimming and searching behaviour ceased. At higher contamination levels, 100% mortality was recorded. These results indicate that the contamination of sediment by anti-fouling paint at Sudbury Reef has the potential to significantly reduce coral recruitment in the immediate vicinity of the site and that this contamination may threaten the recovery of the resident coral community unless the paint is removed.     

Assessment of heavy metal pollution in Xuzhou urban topsoils by magnetic susceptibility measurements

The concentrations of four selected heavy metals (Pb, Zn, Ni and Cr) were measured on 167 topsoil samples collected from the city of Xuzhou, China via inductively coupled plasma mass spectroscopy (ICP-MS). It was found that Pb and Zn were principally derived from anthropogenic inputs whereas Cr and Ni distributions were mainly controlled by parent materials. The spatial distribution patterns of Pb, and Zn were similar to that from low field magnetic susceptibility (χlf), suggesting interaction among them. Two threshold values for magnetic susceptibility and frequency-dependent susceptibility percentage (χfd) were applied to discriminate between polluted and unpolluted samples according to their magnetic susceptibility, resulting in 109 samples populating the “polluted” subset. The Pb and Zn concentrations of the “polluted” subset were statistically significantly higher than those measured in the “unpolluted” one. The heavy metal concentrations were also investigated varying the magnetic susceptibility thresholds to change the “polluted” subset.     

Lysosomal Membrane Stability and Metallothioneins in Digestive Gland of Mussels (Mytilus galloprovincialis Lam.) as Biomarkers in a Field Study

The lysosomal membrane destabilization and the metallothionein content in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.), collected along the east coast of the North Adriatic (Istrian and Kvarnerine coast, Croatia), were examined over a period of four years (1996–1999). The lysosomal membrane stability, as a biomarker of general stress, showed that the membrane labilization period in mussels from polluted, urban- and industrial-related areas was significantly decreased (p<0.05) when compared to mussels from control, clean sea water sites. In the harbour of Rijeka, the most contaminated site, the lysosomal membrane stability was reduced by more than 70% compared to the control. This method also proved to be a useful biomarker for detection of additional stress caused by short-term hypoxia that occurred once during this study inside the polluted and periodically quite eutrophic Pula Harbour. The concentration of metallothioneins in the mussel digestive gland, as a specific biomarker of exposure to heavy metals, did not reveal significant differences (p<0.05) between sites covered by this study.     

The bacterial community composition of an active oil field in the northwestern Gulf of Mexico

The bacterial composition of the water column around two oil production platforms and a control site was examined. Samples were collected during three seasons of a 12-month sampling period three water depths were sampled at all stations occupied.No major differences were discovered in taxonomic or physiological makeup of bacterial populations of the oil field and control site. The genus Pseudomonas predominated at the oil field stations and the control. Bacterial numbers were lower for oil field stations than the control and generally decreased with depth. Microbial biomass estimates were consistently higher at the control site. Oil degrading and sulphur oxidizing bacteria were more numerous within the oil field, and their numbers decreased with distance from the platforms. Buccaneer crude oil did not adversely affect growth or attachment ability of oil field isolates.     

Effects of particulate matter released by a fish farm (Eilat,Red Sea) on survival and growth of Stylophora pistillata coral nubbins

This field study aims to determine whether increased levels of organically enriched particulate matter released by net pen fish farms (Eilat, Red Sea) would affect the growth of nubbins taken from the branching coral Stylophora pistillata. We followed the survival and growth of 1322 nubbins pruned from five colonies that were transplanted at a depth of 6 m in the vicinity of the fish cages and in a reference site, in front of the Interuniversity Institute (IUI). Nubbins were attached on U-shaped PVC plates in three orientations (up, vertical and down positions). After 50 days, survival was high in both localities and no difference was recorded between the spatial orientations. At the fish farm, however, burial of the nubbin's lateral growths and partial coverage of nubbins by settled particulate matter resulted in significant reduction of the lateral growth rates of nubbins settled in the up position as compared to the reference site. On the other hand, faster growth rates were recorded in the vertical set of nubbins at the fish farm when compared with the IUI site. These results strengthen the conclusion that physical effects, rather than nutrient enrichment, may constitute the main cause of stress for minute coral fragments (resembling coral recruits) growing near the vicinity of a fish farm.     

Impacts of the Nakhodka heavy-oil spill on an intertidal ecosystem: an approach to impact evaluation using geographical information system

A major heavy-oil spill from the Russian tanker Nakhodka occurred in the Sea of Japan on 2 January 1997. We investigated the impacts of this spill on a rocky intertidal ecosystem along the southern coast of the Sea of Japan. We selected Imago-Ura Cove as our study site to observe temporal changes along the oiled shore, because minimal cleaning effort was made in this area. Field surveys were conducted every autumn and spring from 1997 to 2000. We measured coverage by macroalgae in 1 x 1-m(2) quadrats and counted the animals in 5 x 5-m(2) quadrats along the intertidal zone. Changes in the ecosystem caused by the oil spill were analyzed by applying a geographical information system (GIS) to the Sea of Japan for the first time. The GIS showed that following the accident there were heavily oiled areas in sheltered regions, but these decreased over the three years. It also showed that coverage by macroalgae and the number of animals increased, although some species of algae with microscopic sporophyte generations, and some populations of perennial shellfish, remained stable or decreased during the study period. GIS was able to trace temporal changes in intertidal communities resulting from the impacts of heavy oil on flora and fauna at a spatial scale of 10-100 m. GIS is thus a practical tool for visualizing, analyzing, and monitoring changes in an ecosystem polluted by oil, taking into account topographic differences along the coastline.     

Antagonism between coral pathogen Vibrio coralliilyticus and other bacteria in the gastric cavity of scleractinian coral Galaxea fascicularis

Scleractinian corals host numerous microbial symbionts with different types of interactions. The gastric cavity of scleractinian coral, as a semiclosed subenvironment with distinct chemical characteristics(e.g., dissolved O2, pH, alkalinity, and nutrients), harbors a distinct microbial community and a diverse array of bacteria that can be pathogenic or beneficial. Galaxea fascicularis is one of the dominant massive scleractinian coral species on inshore fringing reefs in the northern South China Sea.Although the abundance of coral-associated bacteria has been investigated in G. fascicularis, less is known about the microorganisms in the gastric cavity. In this study, we specially isolated cultivable bacterial strains from the gastric cavity of G.fascicularis collected from Hainan Island using a noninvasive sampling approach. Among the 101 representative bacterial strains, one Vibrio coralliilyticus strain, SCSIO 43001, was found to be a temperature-dependent opportunistic pathogen of G.fascicularis. The antagonistic activity between the 100 strains and V. coralliilyticus SCSIO 43001 was tested using a modified Burkholder diffusion assay. Our results showed that V. coralliilyticus SCSIO 43001 inhibits the growth of Erythrobacter flavus and Sphingomonas yabuuchiae. Additionally, we found that three Pseudoalteromonas strains showed moderate to high antibacterial activity against V. coralliilyticus SCSIO 43001 and several other coral-associated Gram-negative bacterial strains.These results suggest that competition between the coral pathogen and other bacteria also occurs in the gastric cavity of coral, and Pseudoalteromonas strains in the gastric cavity of G. fascicularis may provide a protective role in the defense against coinhabiting coral pathogens at elevated temperature.     

Heavy metals in marine nematodes: Uptake,tissue distribution and loss of copper and zinc

The uptake and loss of copper and zinc was studied in two species of marine nematode from a non-polluted site and in one species from a polluted site. Copper and zinc concentrations were measured during and after accumulation from seawater in six different tissues of the nematodes. Most of the metal taken up was found to be associated with the cuticle and with a group of tissues including the hypodermis. The importance of the different tissues in heavy metal metabolism by the nematodes is discussed together with the possible routes of uptake of the metals into the animals.