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Marine and coastal quality assessment, based on test batteries involving a wide array of endpoints, organisms and test matrices, needs for setting up toxicity indices that integrate multiple toxicological measures for decision-making processes and that classify the continuous toxicity response into discrete categories according to the European Water Framework Directive. Two toxicity indices were developed for the lagoon environment such as the Venice Lagoon. Stepwise procedure included: the construction of a database that identified test-matrix pairs (indicators); the selection of a minimum number of ecotoxicological indicators, called toxicological core metrics (CMs-tox) on the basis of specific criteria; the development of toxicity scores for each CM-tox; the integration of the CMs-tox into two indices, the Toxicity Effect Index (TEI), based on the transformation of Toxic Unit (TU) data that were integrated as logarithmic sum, and the Weighted Average Toxicity Index (WATI), starting from toxicity classes integrated as weighted mean. Results from the indices are compared; advantages and drawbacks of both approaches are discussed., (Copyright © 2010. Published by Elsevier Ltd.)

Porewater plays an important role in sediment toxicity assessment using bioassays, but the most reliable extracting method and the potential contribution of confounding factors to the real toxicity need to be studied. The applicability of bioassays with the early life stages of Paracentrotus lividus, Mytilus galloprovincialis and Crassostrea gigas on porewaters extracted by centrifugation from the Venice Lagoon (Italy) is evaluated and demonstrated: toxicity tests can discriminate the toxicity of porewaters from sites with different kinds and levels of pollution and, using toxicity scores, data are classified in five toxicity classes. Sulphides do not represent a confounding factor in porewater toxicity; in contrast ammonia exhibited some concentrations above the toxicity threshold for sea urchin embryos.

Bioassays are routinely employed for sediment quality assessment. In order to be able to effectively use Bioassays responses in regulatory and management frameworks, toxicity scores, which rank toxicity data in defined classes that are continuous and difficult to interpret, should be reliable and suitable tools to support decisions about the presence or absence of toxicity in tested samples and on how toxic a sample is. A statistical approach is needed to define thresholds for toxicity scores. The Minimum Significance Difference (MSD) criterion allowed the evaluation of toxicity thresholds for each test-matrix and organism pair, based on large sets of experimental data. The MSD values were normalized with respect to the control, ranked in ascending order, and the 90th percentile was identified; the Toxicity Threshold (TT) was calculated by subtracting the 90th percentile from 100 and the Toxicity Limit (TL) was estimated as the percentage of control response multiplied by TT. Taking into account sample responses normalized with respect to control (S), when S > TL, the sample is considered nontoxic; when S

This work reports some considerations on the possible contribution of sulfide and ammonia to the toxicity of elutriate samples of sediments from the Venice lagoon, tested with a battery of bioassays using early life stages of the sea urchin Paracentrotus lividus and the oyster Crassostrea gigas. A comparison of ammonia or sulfide concentration in the test matrix, matrix toxicity, and the sensitivity limit of bioassays for ammonia or sulfide were used in evaluating toxicity data. Results highlighted that sperm cell and embryo toxicity of elutriates were not affected by sulfides. Neither was any direct relationship shown between elutriate toxicity and ammonia concentration. Most elutriates had ammonia concentrations below the sensitivity limit of acute test methods, while the more sensitive subchronic toxicity tests were affected by ammonia interference in some samples.

A simple and rapid in vitro toxicological assay, utilizing submitochondrial particles (SMP), has been used to evaluate the toxic effects of fifteen herbicides belonging to the phenol and phenoxyalkanoic acid chemical classes. The SMP assay allows the quantitative evaluation of the toxicity of compounds with different mechanisms of action: uncouplers, inhibitors of the enzyme complexes involved in reverse electron transfer and in oxidative phosphorylation and chemicals that alter the membrane structure. The two groups of herbicides showed different levels of toxicity. For phenol derivatives, EC50 values ranged from 0.16 microM (ioxynil) to 6.7 microM (2,4-dinitrophenol), whereas for phenoxy herbicides EC50 values ranged from 21 microM (2,4,5-trichlorophenoxyacetic acid, 2,4,5-T) to 110 microM (4-chloro-2-methylphenoxyacetic acid, MCPA). On the average, the toxicity of phenolic compounds is greater than that of phenoxyalkanoic acids by two orders of magnitude. Quantitative structure-activity relationships (QSAR) were developed between EC50 values and various molecular descriptors. The results suggest the existence of different mechanisms of action for the two classes of compounds. The findings obtained for phenolic herbicides are consistent with a protonophoric uncoupling mechanism, whereas for phenoxy herbicides a non-specific mode of action at membrane level can be hypothesized.

Samples of Mytilus galloprovincialis collected in different sites of the Venice lagoon (Italy) were investigated for total arsenic concentrations by ICP-AES and for single arsenic species by HPLC-ICP-MS. For this purpose, an analytical procedure for the sensitive and efficient speciation of the arsenic species As(III), As(V), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenobetaine (AB), arsenocholine (AC), and four arsenosugars was optimised. The total arsenic and the single arsenic species were determined in both the hepatopancreas (digestive gland) and the remaining soft tissues in order to verify the different arsenic accumulation in the body parts of mussels. Arsenic compounds were extracted from the mussels with a methanol/water mixture; the extracts were evaporated to dryness, redissolved in water, and chromatographed in an anion-exchange column, a Hamilton PRP-X100. Only small quantities or traces of inorganic arsenic were detected in the mussels. The majority of arsenic compounds detected in the extracts were organic species, with a predominance of arsenobetaine and of an arsenosugar. In addition, a greater arsenic accumulation in the digestive glands of mussels was observed.

Some compounds of environmental interest were investigated using the toxicity test with early life stage of bivalve molluscs. Three pesticides (Carbofuran, Atrazine and Malathion), for which literature data are available only for some species of oysters, were tested with Mytilus galloprovincialis (Lmk). The toxicity of copper was investigated both with M. galloprovincialis and with Crassostrea gigas (Thunberg). Bioassays highlighted a good sensitivity towards the compounds, with EC50 values in the order of mg l(-1) for the pesticides and of microg l(-1) for copper. Comparison with other toxicity tests for aquatic environments showed that embryotoxicity test with bivalve is slightly more sensitive than embryotoxicity test with the sea urchin Paracentrotus lividus (Lmk), remarkably more sensitive than Microtox test but less sensitive than tests with the copepod Tigriopus brevicornis.

Sperm cell and embryo toxicity tests with the sea urchin Paracentrotus lividus were performed to assess the toxicity of sulfide, which is considered a confounding factor in toxicity tests. For improved information on the sensitivity of these methods to sulfide, experiments were performed in the same aerobic conditions used for testing environmental samples, with sulfide concentrations being monitored at the same time by cathodic stripping voltammetry. New toxicity data for sulfide expressed as median effective concentration (EC50) and no-observed-effect concentration (NOEC) are reported. The EC50 value for the embryo toxicity test (total sulfide at 0.43 mg/L) was three times lower than for the sperm cell test (total sulfide at 1.20 mg/L), and the NOEC values were similar (on the order of total sulfide at 10(-1) mg/L) for both tests. The decrease in sulfide concentration during the bioassay as a consequence of possible oxidation of sulfide by dissolved oxygen was determined by voltammetric analysis, indicating a half-life of about 50 min in the presence of gametes. To check the influence of sulfide concentrations on toxicity effects in real samples, toxicity (with the sperm cell toxicity test) and chemical analyses also were performed in pore-water samples collected with an in situ sampler in sediments of the Lagoon of Venice (Italy). A highly positive correlation between increased acute toxicity and increased sulfide concentration was found. Examination of data revealed that sulfide is a real confounding factor in toxicity testing in anoxic environmental samples containing concentrations above the sensitivity limit of the method.

Since the late 1980s the lagoon of Venice, a shallow Mediterranean coastal area, has experienced strong environmental changes. Macroalgae, which were the predominant primary producers of the lagoon, reduced markedly, but neither phytoplankton nor seagrasses replaced them. Temporal and spatial changes in macroalgal standing crop (SC) and phytoplankton concentration were investigated between 1987 and 1998. Maps of macroalgal SC show a marked declining trend. Biomass in fresh weight decreased from: 558 ktonnes in 1987, to 85 ktonnes in 1993 and to 8.7 ktonnes in 1998. As a whole, the biomass in 1998 was only 1.6% of the biomass recorded in 1987. Similarly the macroalgal net (NPP) and gross (GPP) primary production decreased from ca. 1502 and 9721 ktonnes year(-1) to ca. 44 and 229 ktonnes year(-1), respectively. In the early 1990s the clam Tapes philippinarum Adams & Reeve and seagrasses, especially Zostera marina Linnaeus, colonised the bottoms free of macroalgae, but the development of intense clam-fishing activities prevented both phytoplankton blooms and seagrass spreading. Maps of chlorophyll a drawn according to data collected in parallel to macroalgal standing crop show unchanged concentrations. Macroalgae changes are enhanced by comparing annual trends in four areas of the central lagoon during 1989-1992 and 1998-1999. In those areas phytoplankton also decreased significantly. Marked changes of some environmental variables strongly associated with the primary production were recorded both during the lagoon mapping and in the areas studied on a yearly basis.

Sperm cell and embryo toxicity tests using the Mediterranean sea urchin Paracentrotus lividus Lmk were performed to assess the toxicity of As3+, Cd2+, Cr3+, Ni2+, Pb2+, Cu2+, Zn2+, and Hg2+. The aim of this study was to improve information about the comparative sensitivity of sea urchin bioassays to the heavy metals, which are an important cause of contamination in the ecosystem of the Lagoon of Venice. Considering the data in mM/L, the order of toxicity is Hg2+ > Cu2+ > Zn2+ > As3+ > or = Cr3+ > or = Cd2+ > or = Pb2+ > or = Ni2+ for the sperm cell test and Hg2+ > or = Pb2+ > Cu2+ > Zn2+ > Cd2+ > Ni2+ > As3+ > or = Cr3+ for the embryo test. New toxicity data for metals expressed as median effective concentration (EC50) and no-observed-effect concentration (NOEC) are reported for the Mediterranean species. Accurate observations of embryotoxic effects at increasing metal concentrations were done, detecting some different behaviors in metal toxicity. Toxicity data compared with water column and pore-water concentrations recorded in the Lagoon of Venice (Italy) demonstrate the potential ability of bioassays using sea urchin to detect important contaminants in this ecosystem.

Two areas of the Venice lagoon populated by seagrasses (three stations covered by Cymodocea nodosa (Ucria) Asherson, Zostera marina Linnaeus, Zostera noltii Hornemann) or seaweeds (two stations: one covered by Ulva rigida C. Agardh and another at present without seaweed biomass) were monitored by means of six surveys over a year in order to study macrofaunal composition and seasonal changes. The seagrass stations showed a mean species richness (28-30 S m(-2)), individual abundance (1854-4018 N m(-2)) and biomass (22.3-37.7 g m(-2) ash-free-dry-weight, AFDW) ca. 3-8 times higher than those populated by seaweeds (10-15 S m(-2), 494-1395 N m(-2) and 5.6-13.7 g m(-2) AFDW). Differences among seagrass or seaweed stations were much lower. The Ulva-dominated station showed a macrofauna completely different both from the other stations and the communities recorded ca. 30 years ago, before the prolific growth of Ulva. In this station, frequent biomass decompositions and anoxic crises created critical conditions for life favouring organisms with reduced life cycles, younger individuals and the epifaunal species instead of the infaunal ones. In particular, Ulva grazers and scrapers such as Gammarus aequicauda Stock and Gibbula adriatica Philippi were found to be by far the most abundant species, whereas the taxa characteristic of the associations found in the past, in the presence of seagrasses or seaweeds and typical of low eutrophicated environments, appear strongly reduced. Marked differences in the macrophyte dominance and in the bio-physico-chemical variables which characterise the main environmental conditions of the Venice lagoon support the different distribution and composition of macrofaunal communities. Seaweed stations appear mainly governed by the seasonal cycles of these un-rooted macrophytes which, by alternating periods of production and decomposition, are responsible for the drastic reduction of macrofauna biodiversity and biomass. Conversely, seagrass stations exhibit a better oxidisation of the environment and show conditions more favourable for macrofauna colonisation, especially in the presence of macrophytes which are characterised by very well developed below-ground systems such as Cymodocea nodosa.

A reliable sperm cell toxicity test procedure has been developed for the Mediterranean sea urchin Paracentrotus lividus. The sensitivity and discriminatory ability of the test were investigated with regard to surfactants and their biotransformation products. Aromatic and aliphatic surfactants of anionic (linear alkylbenzene sulfonates [LAS]) and nonionic (alcohol polyethoxylates [AE] and nonylphenol polyethoxylates [NPE]) types and their aerobic biodegradation products, i.e., sulfophenylcarboxylates (SPC), polyethylene glycols (PEG), carboxylated polyethylene glycols (PEGC), carboxylated AE (AEC), and nonylphenol (NP), were examined in order to elucidate the influence of their molecular structure on toxicity. Experimental results reveal that the sperm cell test showed good discriminatory ability among all tested compounds, median effective concentration (EC50) values differing by about four orders of magnitude. The toxicity of anionic surfactants depends on the length of the alkyl chain and that of nonionic surfactants is due to their length and branching. Much lower toxicity was shown by aerobic biodegradation products in comparison with that of their parent compounds, with the exception of NP. The obtained EC50s were comparable with available literature data and constitute new toxicity data regarding surfactants for sea urchins.