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The pressure for more energy demand is a strong driver for the construction of offshore oil and gas platforms. This also applies to the Adriatic Sea where, since 1960, several gas fields have been commissioned. The continuous expansion of the gas industry may potentially have detrimental effects on marine communities. This may become particularly clear in such a peculiar semi-enclosed basin as the Adriatic Sea, which contains ecosystems that are subject to intensive anthropic pressures, such as unsustainable fishing and inputs of environmental pollutants. To date, still scant information is available on the environmental effects of gas platforms. Thus, to comply with the national Italian legislation and to support the development of protocols for the monitoring of offshore activities, multidisciplinary studies have been conducted and performed by CNR-ISMAR in cooperation with some private and public research institutes. A multidisciplinary approach based on chemical, biological and ecological investigations was performed to evaluate both the water column and the sediment quality near a gas platform. The overall results indicate moderate effects of platform-related environmental stress.

Despite an increasing number of surveys describing adverse effects of contaminated sediments on marine organisms, few studies have addressed protists. In this study, the free-crawling marine ciliate Euplotes crassus was evaluated as the test organism for the screening of sediment toxicity using sediments from both coastal and estuarine sites of the Venice Lagoon (Marghera harbour [MH], Valle Millecampi [MV], Murano island [MI] and Lido inlet [LI]). Two endpoints of high ecological value, mortality (Mry) and replication rate (RpR), were assessed in combination with the two sublethal biomarkers of stress, endocytotic rate (Ecy) and lysosomal membrane stability (NRRT). The results showed a significant inhibition of RpR, Ecy and NRRT paralleled by a small and insignificantly increased Mry of the exposed specimens. Our results thus demonstrate that only a combination of mortality and sublethal biomarkers was able to characterise an exposure-related stress syndrome. The suite of biomarkers described here was also able to detect and resolve a pollution-induced stress syndrome at an early stage of pollution. The contamination level of the sediments was assessed using chemical analysis, by estimating bioavailability and by computing a toxic pressure coefficient (TPC) to account for potential additive effects of different pollutants. The observed biological responses were consistent with the contamination levels in sediments, suggesting a high potential for using Protozoa in bioassays to assess environmental risk in coastal marine systems.

As a consequence of the growing global demand of energy supplies, intense oil and gas exploration and exploitation programs have been carried out worldwide especially within the marine environments. The release of oil-derived compounds in the sea from anthropogenic sources both as effluents or accidental spill is perceived as a major environmental concern. An approach based on a combination of biomarkers and the distribution of some classes of environmentally relevant pollutants was used to investigate the occurrence of a stress syndrome in mussels (Mytilus galloprovincialis) collected at three gas platforms placed in two distinct oceanographic districts within the Adriatic Sea. Biological responses were integrated by a ranking algorithm which demonstrated both a range of biological effects reflecting exposure gradients and a temporal related trend in the investigated responses. The overall results demonstrate a moderate to absent pollution from studied gas platforms with low but remarkable biological disturbance in sentinel organisms.

The aim of this study was to evaluate the susceptibility to pollutant mediated oxidative stress of the Mediterranean mussel Mytilus galloprovincialis in the Venice lagoon (Italy). In June 2003, mussels from a farm were transplanted to eight sites in the lagoon for five weeks. Oxidative stress responses were measured by: (i) total oxyradical scavenging capacity (TOSC) assay, for an overall evaluation of the oxidative stress response capability; (ii) catalase (CAT), as a key enzyme involved in the antioxidant defence system; (iii) malondialdehyde (MDA), as an indicator of lipid peroxidation, to evaluate an oxidative damage; (iv) metallothioneins (MTs), as they play a role in the antioxidant defence. The TOSC analysis revealed a reduced capability to eliminate: (i) peroxyl radical in mussels transplanted at Palude della Rosa, Valle Millecampi and Chioggia; (ii) hydroxyl radical at Campalto and Valle Millecampi; (iii) peroxynitrite at Valle Millecampi. Inhibition in CAT activity, observed in all the monitored sites, confirms the presence of an oxidative pressure in transplanted mussels. In addition, Pearson correlation analysis was performed in order to observe possible links between the various parameters. The PCA was a powerful tool to discriminate impacted sites, suggesting that the mussels transplanted throughout the Venice lagoon were subjected to different levels of oxidative pressure. Furthermore, it provided an easy and useful tool to summarize the obtained results.

In this study, a spatial and temporal survey at three sites located in the “canals” of the Venice historic centre (Italy) and at a reference site was undertaken to evaluate stress effects on mussels sampled in the Venice urban area, where raw sewage is discharged without treatment directly into the water. A battery of biomarkers (metallothionein, micronuclei, condition index and survival in air) was used to evaluate the stress condition of the animals. At the same time the alkali-labile phosphate assay (ALP) was performed in mussel' hemolymph with the aim to find an estrogenic effect biomarker in this mussel species. Biomarker results showed an impairment of the general health condition in the mussels coming from the urban area, in agreement with the chemical analysis. Significantly higher level of the ALP was found in male mussels sampled in April in the urban area, in comparison with the ones from the reference site ( P 0.001 ). Finally, the PCA proved an easy and useful tool to summarize the obtained results, also able to classify the data to indicate a pollution gradient in the Venice urban area.

The Venice Lagoon has been the focus of many environmental studies due to its heavy contaminant load derived from a variety of sources, including industrial activity, oil tanker traffic, and waste runoff from the mainland. In recent years, there has been increasing concern about water quality in the urban areas of Venice related to the discharge of untreated sewage directly into canals, adding to the pollutant load already existing in these areas. One way of gauging the impact of these chemicals is monitoring the local fauna. In the search for good indicators of water quality in the Venice urban area, two physiological indices for mussels-survival in air and condition index-have been evaluated. In 2002, a seasonal study was undertaken; mussels (Mytilus galloprovincialis) were collected in three sites located in the canals of the city and a clean reference site. At the same time, two transplantation experiments were performed, deploying farmed mussels for 1,2, 4, and 12 weeks in the historic centre of Venice and also at a reference site. Data from survival in air test and the condition index of native and transplanted mussels are reported. The results suggest an impact of the complex mixture of pollutants on mussel health status. Both native mussels and those transplanted to the urban area showed reduced survivability in air and decreased condition index values, indicating a less healthy status in animals from the canals of the city. Data are discussed in relation to pollutant bioaccumulation.

A multiple biomarker approach was adopted in a seasonal study carried out in the Lagoon of Venice, with the double aim of evaluating the natural and anthropic stresses influencing the biological responses of mussel ( Mytilus galloprovincialis ) and of assessing the effects due to spatial rather than temporal variations. Biochemical (aldehyde dehydrogenase and catalase activities), cellular (neutral red retention time) and physiological (survival in air and condition index) biomarkers were determined in mussels collected in four differently impacted lagoon areas. Multivariate analysis showed that samples were distributed mainly according to temperature and four seasonal groups were identified. The combination of spatial and temporal information enabled us to distinguish physiological variations due to natural causes from those due to anthropic stress, and to identify the sampling period when several biomarkers are less influenced by both natural and endogenous factors.

The aim of this study was to investigate the cellular and biochemical response of mussels, Mytilus galloprovincialis, transplanted from a relatively pristine site to a polluted one and vice versa in the Lagoon of Venice (northeast of Italy) and to apply auto-metallography, a rapid and sensitive histochemical technique, to determine the bio-available fraction of heavy metals accumulated in the body tissues of organisms. Animal digestive glands have been used for morphological analyses (lysosomal volume, surface and numerical density, mean epithelial thickness, mean diverticular and luminar radius), autometallographical black silver deposits quantification and biochemical assays (superoxide dismutase and catalase activity). Furthermore, heavy metal content was determined by atomic absorption spectrophotometry (AAS) using standard procedures. The overall results indicate a direct influence of the environment in the thinning of the digestive cells and in the increasing number of lysosomes in mussels from the more polluted site. These data are in agreement with the metal content in digestive cell lysosomes as determined by autometallography, whereas AAS measurements show less significant differences.

A survey to evaluate the impact of organic contaminants on the mussel Mytilus galloprovincialis in the Venice Lagoon, Italy was carried out in May 1993. M. galloprovincialis were sampled from putative moderately contaminated (Alberoni, Lio Grande, Crevan), urban (Salute) and industrial (CVE) sites in the Venice Lagoon, and from a clean reference site (Plataforma) in the adjacent Adriatic Sea. Measurements comprised (i) whole-tissue body burdens of aliphatic hydrocarbons, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and other organochlorines (DDTs, hexachlorocyclohexanes and hexachlorobenzene); and (ii) digestive gland microsomal cytochrome P450 (CYP)-dependent monooxygenase system (i.e. total CYP and cytochrome P4501A (CYP1A)-immunopositive protein levels, benzo(a)pyrene hydroxylase (BPH) activity) as a specific biomarker of impact by organic contaminants. Chemical analysis identified a contaminant gradient with Plataforma as the cleanest and CVE followed by Salute as the most contaminated extremes. No elevation of total CYP content or CYP1A-immunopositive protein level was seen at any of the lagoon sites compared with Plataforma. In contrast, BPH activity and BPH turnover (i.e. BPH activity per amount total CYP) were respectively 1- and 2.5-fold higher at CVE than Plataforma (Plt; 0.05), and indicated to be higher (up to 1-fold) at all the other lagoon sites compared with Plataforma. Correlation was seen between BPH activity and tissue levels of total aliphatic hydrocarbons (r = 0.94-0.98), but not between the former and total PAHs or PCBs. The results are consistent with other studies in the area and indicate greatest biological impact of contaminants was at CVE followed by the other lagoon sites, with a possible genotoxic role for the elevated BPH activity in the formation of bulky DNA-adducts.

In this study, the Active BioMonitoring (ABM) approach has been investigated using species of indigenous (from a pristine site) and transplanted (to a polluted area) hard clam Mercenaria mercenaria as an indicator organism of water/sediment pollution in Tampa Bay, Florida, a highly urbanized and industrialized subtropical estuary. A range of possible induced stress indices was selected and used on the basis of potential pollutant toxicity mechanisms including specific enzymatic responses (MFO as BPH and NADPH cytochrome c reductase, antioxidant enzymes as SOD and caltalase, PDH) and general cellular damage (lysosomal latency, histopathological lesions). The organic sediment contamination compared to the animal body burden revealed bioaccumulation activity in transplanted clams. The marked increases of BPH and catalase activities, together with the significant reduction of lysosomal membrane stability in transplanted animals, suggested their potential for application as biomarkers in subtropical coastal pollution monitoring.

Western blot analysis of microsomes and partially purified cytochrome P450 (CYP) from digestive gland of Mytilus edulis was carried out using polyclonal antibodies to hepatic Perca fluviatilis CYP1A, Oncorhynchus mykiss CYP3A and rat CYP2B, CYP2E and CYP4A isoforms. Multiple CYP bands were detected in partially purified CYP compared to single bands for microsomes for anti-CYP1A, anti-CYP2B, anti-CYP2E and anti-CYP3A. In contrast, anti-CYP4A showed two distinct bands for both. The apparent molecular weights in kD (mean +/- range or S.D.; n = 2-4) for partially purified CYP were 42.5 +/- 0.5 and 48.1 +/- 0.3 (2 bands, anti-CYP1A); 67.4 +/- 0.7, 52.8 +/- 0.6, 44.5 +/- 2.5 (3 bands, anti-CYP3A); 52.8 +/- 0.7, 48.1 +/- 1.1 and 43.9 +/- 1.1 (3 bands, anti-CYP2B); 52.7 +/- 0.8 and 47.2 +/- 0.2 (2 bands, anti-CYP2E); 50.9 +/- 0.3 and 44.1 +/- 0.2 kD (2 bands, anti-CYP4A). Digestive gland microsomes of Mytilus galloprovincialis from a polluted compared to a clean field site showed higher levels of bands recognised by anti-CYP1A, anti-CYP2E and anti-CYP4A, but not anti-CYP2B and anti-CYP3A (P0.05), indicative of independent regulation of different CYP forms. Overall, the apparent molecular weight and field studies indicate at least five different digestive gland CYP forms.

The purpose of this study was to identify possible relationships between biochemical—Catalase, Superoxide dismutase (SOD) and Propionaldehyde dehydrogenase (PDH)—and histochemical—Neutral Red Retention time (NRR)—parameters of mussels ( Mytilus galloprovincialis ) and chemical (Hg, Pb, Cd, Zn, Cr, DDTs and PCBs) contaminants from the most polluted area of the Venice Lagoon which lies around Porto Marghera (industrial pollution) and the city of Venice (domestic pollution). Sampling of mussels was undertaken at the beginning of March 1995 and repeated in March 1996, just before spawning, when tissue concentrations of chemical pollutants reach their annual maxima. The results of the organic contaminant analysis showed generally good agreement between sites for the two sampling periods, with values broadly 10% lower during 1996 as compared to 1995. Metal contaminant concentrations in tissues revealed greater variability than organic compounds. Distribution pattern of biological parameters were more complex and variable. The apparent lack of significant positive correlations between antioxidant enzyme activities and pollutant body burdens as well as the low values of NRR are consistent with the present debate on the use of these biological indices as markers of contaminant-mediated stress for molluscs.

20 pages, 1 figure, 6 tables, The induction of a cytochrome P450 with immunochemical similarities to CYP1A, and accompanying changes in microsomal NADPH-dependent benzo[a]pyrene (BaP) metabolism, were examined in digestive gland of the common mussel (Mytilus galloprovincialis L.) with exposure to 20 ppb water-borne polychlorobiphenyl (PCB) mixture (Arochlor 1254) for 4 or 10 days, or 4 days after a single injection into the mantle cavity of the mixed-type inducer PCB congener 2,2′,3,4,4′,5′-hexachlorobiphenyl (CB-138; 2.5 μg g−1 wet weight). Whole animal tissue levels of PCB following water-column exposure or injection were similar to those for mussel species from polluted field sites, viz. 0.8 to 1.9 μg g−1 wet weight. Levels of microsomal CYP1A-immunopositive protein increased 59% (CB-138) and 72% (Arochlor 1254; 10 days exposure) as determined by Western blot analysis using polyclonal antibodies to hepatic CYP1A of perch (Perca fluviatilis). No changes were seen in levels of digestive gland CYP1A-like mRNA 4 days after injection of CB-138 as determined by Northern analysis using cDNA to hepatic CYP1A1 of rainbow trout (Oncorhynchus mykiss). The increases in levels of CYP1A-immunopositive protein were accompanied by a shift in microsomal NADPH-dependent BaP metabolism towards phenol and diol and away from dione production, the former increasing from 32 to 85% of total free metabolites. The marked decrease in dione production (which is the major BaP metabolite formed in control microsomes) resulted in no increase in total microsomal BaP metabolism with exposure to PCBs. The Type I ligand α-naphthoflavone markedly inhibited microsomal phenol but had no affect on dione production, whereas the Type II ligand clotrimazole markedly inhibited dione, but had much less effect on phenol production. The overall results are interpreted in terms of the existence of an inducible CYP1A-like enzyme catalysing predominantly 2-electron monooxygenation leading to epoxide (and hence phenol and diol) formation, and a constitutive non-inducible cytochrome P450 catalysing predominantly 1-electron oxidation leading to dione formation. Both Arochlor 1254 or CB-138 produced cellular damage in the digestive gland in the form of decreased epithelial digestive cell height and decreased lysosomal membrane stability

Mussels (Mytilus galloprovincialis) were sampled from six sites in the Venice Lagoon in May 1993 for measurement of their physiological responses (clearance rate, absorption efficiency and respiration rate) under standard laboratory conditions. Scope for growth (SFG) calculated for a ‘standardised ration level’ (0.4 mg particulate organic matter l−1) reflected the underlying pollution induced stress caused by the toxicants accumulated in the body tissues. Mussels from Chioggia had the highest SFG (16 J g−1 h−1) with slightly lower rates at Lio Grande, Crevan and Alberoni (9–10 J g−1 h−1), whereas mussels from the inner part of the lagoon (Salute and Canale Vittorio Emanuele) had significantly reduced SFG values (2 and −4 J g−1 h−1) which indicated severe pollution induced stress. There were significant negative correlations between SFG and the tissue concentrations of petroleum hydrocarbons, PCBs, DDT and HCH, but there were no significant correlations between SFG and the tissue concentrations of metals (Cd, Co, Cr, Cu, Fe, Hg, Mn, Fe, Ni, Pb and Zn). SFG was also calculated on the basis of measured POM levels prevailing at each site and this demonstrated that the inhibitory effects of toxicants on the growth of mussels living in the more polluted inner sites may be partially offset by the positive effects of eutrophication and the higher food/ration levels at these sites. There were also differences in the physiological responses and SFG of mussels from the two main cultivation sites within the Venice Lagoon (i.e. Alberoni < Chioggia) and these reflected the higher contaminant levels and lower food levels at Alberoni.

A laboratory study was carried out exposing mussels (Mytilus sp.) to linear alkylbenzene sulphonate (LAS) (6 mg litre−1), Cd (0.05 mg litre−1) and LAS plus Cd at the same concentrations. The aim was to assess the use of several histopathological and biochemical indices as potential biomarkers of the impact of these xenobiotics in the digestive gland of molluscs. Treated mussels actively accumulated Cd in the digestive gland compared with controls (p ⩽ 0.01), the highest levels occurring after 30 days of exposure in the group treated with Cd plus LAS. Among several histological alterations screened in digestive gland tissues the thickness of digestive tubules in Cd treated animals decreased more markedly (p ⩽ 0.01) than in LAS exposed mussels. As for biochemical parameters, the investigated antioxidant enzyme activities, superoxide dismutase (SOD), catalase, DT-diaphorase and glutathione peroxidase (GPX) did not show any significant induction due to these xenobiotics. However, a slight decrease of the antioxidant defences of the animals was detected after 30 days of exposure to contaminants.

This paper presents the results of investigations on the suitability of Euplotes crassus, an interstitial marine ciliate, to be used as model organism in ecotoxicology and thereafter to evaluate the toxicity of estuarine and coastal sediments upon laboratory exposure. Nowadays, anthropogenic activities have resulted in accumulation of metals and organic pollutants in the environment as well as in the food chain hence leading to serious ecological and human health problems. This may pose a risk to benthic and epibenthic organisms and it is crucial to discover toxicity tests that will identify adverse effects of sediment-associated chemicals on benthic organisms. Due to their nature as a eukaryotic cell/organism and their position in the food web, ciliated protozoa are suitable models for evaluating the effects of pollution on aquatic communities. Lethal and sublethal effects of exposure to inorganic and organic pollutants were tested on the cell mortality, replication rate, lysosomal membrane stability and endocytosis rate of E. crassus. Increasing nominal concentrations of individual and mixtures of mercury, copper, and benzo(a)pyrene were investigated in this study as they might be bioavailable in naturally occurring polluted sites. A significant decrease in the mean replication rate (p

Detecting the anthropogenic impacts of offshore gas platforms requires reliable tools, because the traditional evaluation based only on chemical analyses is neither appropriate nor sufficiently sensitive. Thus, a 3-year monitoring project was carried out to evaluate the impact of a platform based on a chemical-biological approach. Benthic communities are investigated as they are widely used to monitor the effects of marine impacts because the organisms are mostly sessile and integrate the effects of pollutants over time. Changes in benthic infauna, sediments and water quality, as well as biota bioaccumulation, caused by drilling and platform operations were evaluated experimentally. Furthermore, mussels (Mytilus galloprovincialis) were collected seasonally from the platform legs, both close to and far from the sacrificial anodes and at a control site. Responses of biomarkers of exposure and effect were related to the average levels of polycyclic and aliphatic hydrocarbons, organic matter content and heavy metals in bivalve tissues. Our data suggested that a slight perturbation can be detected only by integrating the results of chemical analyses on water and sediments and those obtained from benthic biological surveys.

Despite a large number of gas platforms existing in the Adriatic Sea, which is a semi-enclosed basin characterized by a slow turnover rate and increasing industrial as well as other anthropogenic activities, the effects of these structures on the aquatic ecosystem require further investigation. Since 1998, multidisciplinary studies have been performed by CNR-ISMAR to comply with legislation and to support the development of protocols for the monitoring of offshore activities in the Adriatic Sea. The present study was developed to implement a biomonitoring plan to assess the ecotoxicological effects of the extraction activities of an off-shore gas platform. Biomarkers were evaluated in mussels collected from the platform in relation to physiological stress, DNA damage, cellular damage, oxidative stress and exposure effects. Organic contaminants and trace element bioaccumulation were also assessed in the soft body of the mussels to correlate bioaccumulation of pollutants with biomarker responses. The results indicate an absence of platform-related environmental stress.

A toxicity identification evaluation phase-I (TIE-1) procedure was carried out on five pore water samples extracted from sediments of the Venice Lagoon previously investigated to assess both chemical contamination and toxic effects on the biota. Two different sequential TIE procedures were tested. A first sequence (TIE-1) provided for adding Na2S2O3, adding Na-EDTA, filtering, elution through a C18-SPE column and removing ammonia using the macroalgae Ulva rigida Agardh 1823, while a second procedure (TIE-2) was set up using U. rigida treatment for ammonia removal as first step, keeping unchanged the sequence of the other manipulations. Two different exposure time to the macroalgae were tested (3-h and 15-h). Sperm-cell toxicity test with the echinoid Paracentrotus lividus and embryotoxicity tests with the bivalves Mytilus galloprovincialis and Crassostrea gigas were performed on pore-water samples to assess the effect of the sequential treatments on the overall toxicity. The results confirmed that ammonia contribution to toxicity is strong in most of the samples and that metals, specially Cu, are of concern at least in three sites. The TIE-2 procedure provided more reliable results for the samples characterized by high ammonia contribution to the overall toxicity, whereas the results of TIE-1 and TIE-2 were equivalent for the samples where ammonia contribution was not prevailing. Chemical analyses and test results showed that a 3-h U. rigida exposure is suitable to remove ammonia toxicity minimizing potential metal up-take.

The results of five experiments with the marine fish Zosterisessor ophiocephalus (Pall.) exposed to relatively low concentrations of PCBs (Aroclor 1254) in water and fed with uncontaminated and contaminated mussel tissues are reported. The data obtained indicate that PCBs accumulated in fish liver are responsible for induction of hepatic benzo[a]pyrene hydroxylase (BPH) activity as well as for the higher cytochrome P-450 content in PCB treated fish. It also appears that, when MFO activity is related to PCB liver content, the relationship seems to be relatively independent of exposure and/or administration modalities.

Total copper (Cu(T)), copper ion activity (pCu) and the copper complexation capacity (CuCC) were determined in samples of seawater collected in July 2003 from the Venice Lagoon. Cu(T) and CuCC showed considerable spatial variability: Cu(T) ranged from 1.8 to 70.0nM, whereas the CuCC varied from 195 to 573nM. pCu values varied from 11.6 to 12.6 and are consistent with those previously reported in estuarine and coastal areas (10.9-14.1). The range of Cu(T) values compares well with those reported in the past in the lagoon and in the adjacent Adriatic Sea. The highest concentrations of Cu(T) were found in samples collected near the industrial area of Porto Marghera, whereas the lowest were measured near the Chioggia and Malamocco inlets, where an intense tidally-driven renewal of seawater takes place. Although CuCC showed a high degree of spatial variability, the values recorded in the Venice Lagoon are comparable to those reported in other estuarine systems. In addition, CuCC was positively correlated with dissolved organic carbon (DOC), suggesting that organic ligands responsible for Cu complexation are part of the bulk organic matter pool in the lagoon. The CuCC:Cu(T) molar ratio was, on average 55:1, indicating that a large excess of complexation capacity exists in the Venice Lagoon. The high levels of CuCC and the narrow range of pCu indicates the importance of the role played by organic ligands in controlling the free ion Cu concentrations in the lagoon, and as a consequence, regulating its availability and/or toxicity.

Oxidative stress related investigations to monitor the impact of the pollutant discharges into the Venice lagoon (Italy) originating from anthropogenic activities (raw sewage water, agricultural and industrial effluents, oil tanker traffic), on marine organisms have classically been carried out by analyzing specific, single antioxidants (i.e. catalase, superoxide dismutase). In this paper, two studies are reported where the total oxyradical scavenging capacity assay (TOSC) was selected and measured toward peroxyl, and hydroxyl free radicals, and peroxynitrite in mussels (Mytilus galloprovincialis) transplanted into the Venice city and throughout the lagoon to measure the biological effects of anthropogenic activities. In the first experiment, mussels from a clean site (farm) were transplanted to the urban area of Venice for 0, 1, 2 and 4 weeks; cytosolic TOSC toward peroxyl and hydroxyl free radicals, and peroxynitrite revealed that the transplantation process caused a stress (handling stress, anoxia, oxidative burst) resulting in a reduction of TOSC in both control and urban sites, therefore, preventing clear interpretation of the data after one week. At week 2, a significant TOSC reduction (P < 0.05) toward peroxyl and hydroxyl radicals in the urban site revealed that mussels experienced oxidative pressure exerted by pollutants. Most TOSC values returned to initial levels at week 4; however, TOSC induction was noticed in the control group toward peroxyl and hydroxyl radicals while in the exposed group it was not indicating an inhibition of the oxidative metabolism. In the second experiment, mussels were deployed at seven different sites throughout the lagoon. After five weeks of exposure, significant TOSC reduction was measured (P < 0.05) toward peroxyl for Palude della Rosa, Chioggia and Valle Millecampi, toward hydroxyl radicals for Valle Millecampi and Campalto and toward peroxynitrite for Valle Millecampi. Although these data indicate a depletion of the low molecular weight scavengers, additional biomarkers are needed to draw a conclusion on the health of the mussels. TOSC was proved to be an interesting health index parameter to measure pollution impact in a transplantation study provided that the mussels are exposed for two weeks and a control is run in parallel.

A field study was carried out in the Lagoon of Venice (north-east Italy) with the aim of evaluating the potential use of lysosomal destabilisation as a biomarker of anthropogenic stress in the autochthonous mussel Mytilus galloprovincialis. Two different approaches were adopted in biomonitoring six sites in the Lagoon, evaluating indigenous populations of mussels and organisms transplanted from a reference site and checked at several points in time. Lysosomal membrane stability was investigated by means of two tests: neutral red retention assay (NRRA) for evaluating haemocyte lysosomes and lysosomal latency test (LLT) for digestive cell lysosomes. Results indicate that the lysosomal response measured in haemocytes according to NRRA is a more valuable biomarker of anthropogenic stress in the framework both of passive and active biomonitoring in marine coastal environments.

A number of biochemical markers and a physiological index were measured in mussels, Mytilus galloprovincialis, transplanted or native to five different contaminated sites in the lagoon of Venice. Mussels from Pellestrina, a reference site in the adjacent Adriatic Sea, were transplanted for 6 weeks to areas of the lagoon where indigenous mussels were also collected. As biochemical indices, superoxide dismutase (SOD), catalase, aldehyde dehydrogenase (ADH) and NADPH cytochrome c reductase (NADPHcred) were measured in mussel digestive gland; survival in air as a physiological index was also determined. Biomarker responses varied among sites and between indigenous and transplanted animals. Significant induction of catalase and SOD was shown in animals transplanted to the urban sites of Salute and Chioggia, respectively. In indigenous mussels, induction of SOD and NADPHcred was seen in animals from the polluted site of Treporti and the heavily contaminated industrial area of Marghera. The overall biochemical data indicate significantly higher activity for ADH in transplanted animals in comparison with indigenous ones which, in contrast, present an increase in SOD. As regard survival in air, control mussels did not seem to be healthier in comparison either with transplanted or indigenous ones, suggesting that pollution has no effect on this parameter.

In this study the clam, Tapes philippinarum, a filter-feeding bivalve living in soft bottoms, was used to investigate the water/sediment pollution in Venice Lagoon (Venice, Italy), a heavily urbanised and industrialised area. To this end, clams collected from indigenous populations in a clean farming area (Val Doga) were transplanted into a relatively low contaminated site (Palude della Rosa) and in a heavily polluted area (Porto Marghera), for 5 weeks. A range of possible induced stress indices were measured, on the basis of potential toxicity mechanisms, including specific enzymatic activities [Bap hydroxylase and NADPH cytochrome c reductase, superoxide dismutase, catalase, glutathione peroxidase and propionaldehyde dehydrogenase (PDH)] and general cellular responses (lysosomal latency). Moreover, chemical analyses (polychlorinated biphenyls, polycyclic aromatic hydrocarbon, DDTs) were carried out on sediments and animal tissues at the beginning and end of the transplantation experiments. The chemical results show an active bio-accumulation activity from sediment in clams translocated to the most polluted area. Correspondingly, biological data indicate marked effects on PDH activity and latency in T. philippinarum from the same site. In contrast, no similar pattern has been observed in the other considered indices, possibly due to antagonistic effects of the complex contaminant mixture present in the environment.

The response of Crassostrea virginica to a complex mixture of toxic contaminants was studied at four sites in Laguna de Terminos, Mexico. Contaminants assessed were heavy metals, organochlorine compounds, and hydrocarbons. Biomarkers (cholinesterase activity, neutral red retention, and metallothionein), histopathology and prevalence of Perkinsus marinus were used to evaluate the effects of contaminants. The resultant contaminant body burden was moderately as high as a whole, and biomarker levels were also moderate. Oedema in the gills and systemic haemocytosis were the only histopathological lesions observed. P. marinus was never evidenced in the oyster tissues examined. Statistical results from principal components analysis show that metallothioneins are positively correlated with copper and chromium, neutral red and oedema with chlordanes, pentachloroanisol, drins, polychlorobiphenyls and total pesticides, total hydrocarbons and polycyclic aromatic hydrocarbons, whereas cholinesterase activity is negatively correlated to Cu and Cr. The biomarkers used were sensitive indicators responding to moderate levels of pollutants.

(1986). Hydrobiology and Fish biology. Bollettino di zoologia: Vol. 53, No. sup001, pp. 93-100.

During 1988, wild mussels were collected bimonthly from two stations having different pollution load, the first located close to the city of Venice and the second offshore in the Gulf of Venice, north-east Italy. Composite samples of digestive glands were analysed for their aliphatic and aromatic hydrocarbon content and for microsomal BPH, NADH-ferricyanide reductase (NADH-FERRIRED) and NADH- and NADPH-cytochrome c reductase (NAD(P)H-CYTCRED) activities. Results show that significant differences exist between the two stations in digestive gland hydrocarbon content while the enzymatic activity appears to be at the same level; as a consequence no significant correlations have been found between biochemical parameters and hydrocarbon concentrations in digestive glands.

From July, 1975 to November, 1976 mussels from the Laguna Veneta, north-east Italy, and the adjacent Adriatic Sea were collected bimonthly at ten stations and analysed for 3,4-benzopyrene (BaP) and perylene (Pe) content. Mean concentrations were in the range 12·0–135·1 μg/kg dry weight for BaP and 1·5–16·9 μg/kg for Pe, but values as high as 327 μg/kg for BaP and 71 μg/kg for Pe have been measured. The distribution of BaP and Pe closely matches previous results on petroleum hydrocarbon contamination in mussels from this area and indicates that the major portion of BaP and Pe in these bivalves is due principally to human activities.

The seasonal pattern of condition index, carbohydrate, lipid, petroleum hydrocarbons and chlorinated hydrocarbons in mussels of the Laguna Veneta (North‐East Italy) was studied during 1978. Condition index, carbohydrate and lipid showed a marked annual cycle fairly common to most mussel populations. Tissue concentrations of lipophilic pollutants, though showing some correspondance with lipid content, did not correlate with physiological parameters.

This study is focused on evaluating biological effects on the target species Chamelea gallina as a result of repeated exploitation of clam beds by hydraulic dredging. A biomarker approach was adopted and two physiological indices were applied: adenylate energy charge (AEC) (biochemical level) and reburrowing behaviour (organism level). The aim was to investigate the biological response to various types of fishing, i.e., commercial systems using water at high pressures (HP), and experimental systems at lower pressures (LP) in the field, and to evaluate the response of clams to mechanical stress in the laboratory. Field results showed that HP-treated clams exhibited significantly lower levels of AEC compared with LP treatments. A similar trend was shown by reburrowing behaviour; HP-treated clams reburrowed less. Laboratory results were less clear: a very low level of AEC was measured in both control and treated clams. These poor conditions in foot muscle did not indicate worsening at organism level, as no dead or dying clams were recorded throughout the experiment. However, repeated mechanical stress reduced the percentage of reburrowing clams, which suggested that harvesting may affect reburrowing behaviour.

Macrobenthic assemblages in Calcasieu Lake estuary (Louisiana) were sampled at 11 sites from October 1983 through November 1985. The sites were numerically dominated by subsurface-deposit feeders, consisting mostly of polychaetes. Greater densities of macrofauna were collected at the northern (upper) stations of the lake than were collected in West Cove or the southern stations. Abundances of polychaetes, oligochaetes, and amphipods at the upper lake stations accounted for most of the differences among stations. The numerical dominance by detritivores (97% of the fauna) and lack of strong sediment or salinity gradients across the estuary, resulted in an absence of temporal pattern in trophic structure of the macrofauna.

Trophic groups were used to investigate broad patterns and predict species interactions of macrobenthic assemblages in Calcasieu Estuary, Louisiana. Macrobenthic assemblages of the estuary were numerically dominated by deposit-feeding species. Surface-deposit feeders were the most abundant macrobenthos of the upper estuary, subsurface-deposit feeders dominated the lower estuary, and a trophically well-mixed assemblage inhabited the middle estuary. There were periodic shifts in species dominance within each region, but the shifts were without temporal pattern and seldom led to changes in trophic structure of the region. The macrobenthic assemblages of the upper estuary were dominated by several early-colonizing species which switched feeding modes with changes in water flow, suggesting that disturbances in the upper estuary had a direct effect on macrobenthic communities. Disturbances also may have eliminated macrobenthic species before they could establish a well-mixed community. The presence of trophically mixed communities in the middle estuary probably was indicative of fewer disturbances there. The dominance by subsurface-deposit feeders in the lower estuary probably indicates that sedimentary food in Calcasieu Lake was seldom utilized at the sediment-water interface. Rather, most food became buried and was available only to subsurface-deposit feeders.