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The OECD TG 215 method (2000) (C.14 method of EC Regulation 440/2008) was developed on the rainbow trout ( Oncorynchus mykiss ) to assess chronic toxicity (28 d) of chemicals on fish juveniles. It contemplates to use other well documented species identifying suitable conditions to evaluate their growth. OECD proposes the European sea bass ( Dicentrarchus labrax , L. 1758) as Mediterranean species among vertebrates recommended in the OECD guidelines for the toxicity testing of chemicals. In this context, our study is aimed to proposing the adaptation of the growth test (OECD TG 215, 2000) to D. labrax . For this purpose toxicity tests were performed with sodium dodecyl sulfate, a reference toxicant commonly used in fish toxicity assays. The main aspects of the testing procedure were reviewed: fish size (weight), environmental conditions, dilution water type, experimental design, loading rate and stocking density, feeding (food type and ration), test validity criteria. The experience gained from growth tests with the sea bass allows to promote its inclusion among the species to be used for the C.14 method.

Marine bioinvasions and other rapid biodiversity changes require today integrating existing monitoring tools with other complementary detection strategies to provide a more efficient management. Here we explored the efficacy of fishermen observations and traditional port surveys to effectively track the occurrence of both indigenous and non-indigenous megafauna in the Adriatic Sea. This consisted mainly of mobile taxa such as fishes, crustaceans and molluscs. Port surveys using traps and nets within 10 major Adriatic harbours, were compared with the information obtained from 153 interviews with local fishermen. Information gathered by traps and nets varied significantly and generally resulted of limited efficacy in exotic species detection. Interviews allowed tracking the occurrence of new species through time and space, providing complementary knowledge at the low cost. This combined approach improves our capability of being informed on the arrival of species of different origin, providing a more rational, improved basis for environmental management and decision making.

The United Nations recognised the transfer of invasive species across natural barriers as one of the greatest pressures to the world's oceans and seas. The BWM Convention sets the global standards on ballast water management (BWM) requirements, while recognising that regional and local specifics have to be considered for its effective implementation. In the Adriatic Sea cross-border activities were conducted to provide for essential information/data and tools to support a regionally coordinated implementation of the BWM Convention. This special issue contains 18 publications that include results and recommendations from studying the ballast water and management issues through sampling of ballast water on vessels, risk assessment for exemptions and BWM, biological and chemical port baseline surveys and monitoring conducted in ports along the Adriatic Sea coast, oceanographic conditions, ballast water sediment issues and their management in ports, and the implementation options of the BWM Convention through the Adriatic States' environmental law and institutions cooperation. Essential data and tools to support a regional approach in the implementation of the BWM Convention were provided, and are therefore available to the administrations of the Adriatic countries to enable protection of the Adriatic Sea environment, human health property and resources from negative impacts of ballast water being discharged in the area. Data, approaches and tools provided here may be helpful in any other region to support an effective BWM Convention implementation.

Port baseline surveys (PBS) provide species inventories in and around ports, with a focus on non-indigenous species that may have been introduced by vessels, primarily via ballast water. PBS are an essential tool to support effective management strategies for non-indigenous as well as native harmful aquatic organisms and pathogens (HAOP). This paper describes the methodology of PBS that were conducted in 12 Adriatic ports. The PBS employed existing protocols that were adapted to meet the characteristics of the Adriatic sites. Their results are reported in several papers included in this special issue, each of which is devoted to a specific community. An overview of existing surveys protocols – which provide valuable support to decision-making and to design effective monitoring of non-indigenous species – is also supplied.

Oxidant treatment of ballast water (BW) is commonly used in BW systems in order to minimize the transport of alien species. The release of disinfection by-products (DBPs) associated to the treatment of BW and cross-contamination of butyltin (BT) compounds through BW discharge is a topic of environmental concern. A chemical port baseline survey has been conducted in seven ports of the Adriatic Sea. Analysis have been performed on transplanted mussels, surface sediment, seawater, BW. Results showed an evidence of BT contamination, particularly in sediments, probably related to their illegal usage or to intensive shipping activities. Therefore, BW may act as a vector and contribute to re-buildup of BT contamination in the coastal regions. A baseline set of data concerning DBPs is provided, showing the preferential distribution of these compounds in the marine environment that will be useful for future considerations on monitoring and assessment of chemical contamination associated with BW.

This paper describes the methodological approach used for the development of an Early Warning System (EWS) for Non Indigenous Species (NIS) and ballast water management and summarizes the results obtained. The specific goals of the EWS are firstly to warn vessels to prevent loading of ballast water when critical biological conditions occur in ports and surrounding areas i.e. mass development or blooms of Harmful Aquatic Organisms and Pathogens (HAOP). Secondly, to warn environmental and health authorities when NIS or pathogens are present in ports or surrounding areas to enable an early response and an implementation of remediation measures. The EWS is designed to be used for implementing various parallel obligations, by taking into consideration different legal scopes, associated information and decision-making needs. The EWS was elaborated, tested in the Adriatic Sea and illustrated by two case studies. Although the EWS was developed with an Adriatic Sea focus, it is presented in a format so that it may be used as a model when establishing similar systems in other locations. The role of the various actors is discussed and recommendations on further developments of the EWS are presented. It was concluded that the EWS is a suitable tool to reduce the spread of potentially harmful and ballast water mediated species.

MAGALETTI, E., BORRELLO, P., SPADA, E., BATALONI, S., DI GIROLAMO, I and GIANI, M., 2011. Surveillance of Potentially Toxic Benthic Microalgae Along the Italian Coast. In: Micallef, A. (ed.), MCRR3– 2010 Conference Proceedings, Journal of Coastal Research, Special Issue, No. 61, pp. 353–358. Grosseto, Tuscany, Italy, ISSN 0749–0208. Since 1989, the Italian Ministry for the Environment, Land and Sea (MATTM) is carrying out monitoring programs to assess the ecological quality of the marine coastal environment. In the period 2001–2007, phytoplankton communities were investigated every 15 days in 81 areas along the Italian coast and from 2008 also the benthic microalgae communities are being investigated during the Summer months. In 2006, the Italian Minister for the Environment requested ISPRA to carry out activities in collaboration with the Regional Environmental Agencies (ARPA) aimed at achieving a better understanding of the distribution and bloom dynamics of Ostreopsis species in Italian coasta...

During the last years many authors have char-acterized the produced formation waters (PFWs) with respect to chemical compounds and toxic-ity. Most of data are related to PFWs collected on offshore platform after treatment process. The available results showed that the particulate phase had an influence on PFW toxicity. As-suming the toxicity of PFWs treated on platform, the aim of this paper is to study the toxicity of these PFWs after a further filtration treatment carried out in laboratory. For this purpose PFWs were sampled from three natural gas platforms located in the Adriatic Sea (Italy) below treat-ment system. The eco-toxicological bioassays have been conducted on test-organisms be-longing to different trophic levels such as bac-teria, algae, crustaceans and fishes. The PFWs resulted toxic according to an overall assess-ment obtained through the bioassays. Further-more, it has been possible to identify the spe-cies that were more sensitive to the tested PFWs, namely Tigriopus fulvus, Dicentrarchus labrax and Vibrio fischeri. Besides, a chemical char-acterization was reported related to the con-taminants present in the PFWs to go with eco-toxicological assessment. Barium, zinc and manganese showed the most concentrations among the metals and the lower molecular weight components were common among the organic compounds. Some differences among PFWs were observed both for toxicity and chemical composition. The highest toxicity was recorded in PFWs (PFW1 and PFW2) containing the highest concentrations of some metals (Ba, Mn and Zn) and/or BTEX.

For a toxicity assessment of substances entering the marine environment, it is preferable to carry out ecotoxicological tests on a base-set of taxa utilizing target species belonging to different trophic levels. In this study a battery composed of Vibrio fischeri (bacteria), Dunaliella tertiolecta (algae), Tigriopus fulvus (crustacea), Paracentrotus lividus (echinodermata), and Dicentrarchus labrax (pisces) was used for acute toxicity testing of sodium dodecyl sulfate (SDS). SDS is an anionic surfactant widely employed in industry, agriculture, and domestic usage and therefore is found in abundance in the environment, particularly in the sea. The mean values of EC50 obtained were 2.6, 4.8, 7.4, 3.2, 7.3 mg L(-1), respectively, for V. fischeri, D. tertiolecta, T. fulvus, P. lividus, and D. labrax. The results indicate the high acute toxicity of SDS with respect to all the trophic levels represented by the target species. In addition, they highlight the usefulness of employing a base-set of taxa rather than a single species in toxicological tests, in order to obtain more reliable information for the evaluation of toxicity and potential hazards to the marine environment of selected substances.

A laboratory study was performed on the extracellular production of carbohydrates by the marine diatoms Cylindrotheca closterium , Thalassiosira pseudonana and Skeletonema costatum. The investigation was aimed at elucidating the role of P-starvation and growth status on abundance and chemical characteristics of the released non-attached polysaccharides. Inorganic phosphorus depletion determined an increase of total polysaccharides in all species examined compared to nutrient-replete (complete f/2) conditions. The highest abundance of polysaccharides per unit cell was found in T. pseudonana (28.4 μmol C 10 − 6 cells), followed by C. closterium (2.56 μmol C 10 − 6 cells) and S. costatum (1.18 μmol C 10 − 6 cells). Maximum production rates were found at the transition between exponential and stationary growth phase. Gas-chromatographic analysis of the dissolved fraction showed glucose to be the most abundant monomer in exponentially growing, P-replete cultures (81.6%, 90% and 32% as molar percentage of total aldoses in C. closterium , T. pseudonana and S. costatum , respectively). A strong reduction in glucose was found in C. closterium , but not in T. pseudonana and S. costatum , under P-depleted conditions. Species-specific variations in the amount and aldose signatures of the released polysaccharides according to nutrient status and growth conditions can provide useful insights on the production and persistence of these organic compounds in the water column.

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish water and sediment. The choice of biological indicators must be based on reliable scientific knowledge and, possibly, on the availability of standardized procedures. In this article, we present a standardized protocol that used the marine crustacean Artemia to evaluate the toxicity of chemicals and/or of marine environmental matrices. Scientists propose that the brine shrimp (Artemia) is a suitable candidate for the development of a standard bioassay for worldwide utilization. A number of papers have been published on the toxic effects of various chemicals and toxicants on brine shrimp (Artemia). The major advantage of this crustacean for toxicity studies is the overall availability of the dry cysts; these can be immediately used in testing and difficult cultivation is not demanded1,2. Cyst-based toxicity assays are cheap, continuously available, simple and reliable and are thus an important answer to routine needs of toxicity screening, for industrial monitoring requirements or for regulatory purposes3. The proposed method involves the mortality as an endpoint. The numbers of survivors were counted and percentage of deaths were calculated. Larvae were considered dead if they did not exhibit any internal or external movement during several seconds of observation4. This procedure was standardized testing a reference substance (Sodium Dodecyl Sulfate); some results are reported in this work. This article accompanies a video that describes the performance of procedural toxicity testing, showing all the steps related to the protocol.

The aim of this study was to assess medium-term toxicity of weathered oil on European seabass. A mesocosm system reproducing an oil spill at sea was applied. Fish were collected after 48 h, 7, 30 and 60 days. Cyp1a gene transcription, EROD and UDPGT activities, bile PAHs metabolites and micronuclei frequency were investigated. A progressive disappearance of low molecular weight n-alkanes and PAHs in the water of the mesocosm occurred during the experimentation. Fishes exposed to oil displayed a significant increase of cyp1a expression and EROD activity during the entire experiment as well as higher concentrations of PAHs metabolites in bile. Micronulei frequency resulted significantly higher during all experiment in oil exposed sea bass compared to controls. The results highlight the environmental risk associated with the release of oil products at sea and confirm the adopted parameters as useful tools for studying the impact of accidental oil spills on fish.

Diethylene glycol (DEG) is commonly used to dehydrate natural gas in off-shore extraction plants and to prevent formation of gas hydrates. It may be released into the sea accidentally or in discharged produced waters (PWs). PWs samples from off-shore gas platforms in the Adriatic Sea (Italy) have been used in this study. The objectives of the study were: a) to evaluate the toxicity of DEG for marine organisms; b) to evaluate if a high DEG content in PWs may alter their toxicity; c) to verify whether the DEG threshold concentration established by the Italian legislation (3.5 g/l) for PWs discharged at sea is safe for marine environment. Ten different species (Vibrio fischeri, Phaeodactylum tricornutum, Dunaliella tertiolecta, Brachionus plicatilis, Artemia franciscana, Tigropus fulvus, Mytilus galloprovincialis, Crassostrea gigas, Tapes philippinarum and Dicentrarchus labrax) have been exposed to DEG; four of these species were also exposed to PWs in combination with DEG. The results showed that: a) DEG is not toxic at levels normally detected in Adriatic PWs; b) DEG in combination with PW showed mainly additive or synergistic effects; c) short-term bioassays showed that the DEG limit of 3.5 g/l could be acceptable.

Cyclins can be useful cell cycle markers for growth rate studies on harmful algal blooms. In this study, a gene fragment corresponding to cyclin box was cloned for the brown tide alga Aureococcus anophagefferens. This algal gene fragment, designated as Btcycl1, was most similar to cyclin B. Oligopeptides based on the deduced amino acid sequence were synthesized and used to raise an antiserum that reacted on Western blots with a protein of about 63 kDa, the same size as cyclin B in other organisms. The cyclin B–like protein recognized by this antiserum, and the messenger RNA amplified using the primers, were more abundant in exponential cultures and decreased markedly in stationary cultures. This protein also appeared to be cell cycle dependent. Immunofluorescence labeling showed that this antiserum specifically stained a protein in Aureococcus cells and had no cross-reaction with bacteria that were present in the algal culture. The Btcycl1 sequence and the antiserum will provide a useful tool for studies on regulation of in situ growth rate for this brown tide alga.

The effect of P- and N-depletion on the production and molecular-level composition of extracellular carbohydrates released by the diatom Cylindrotheca fusiformis was examined in axenic batch cultures. Inorganic phosphorus depletion caused a higher release of organic carbon per cell compared to nutrient-replete and nitrogen-limited conditions. Gas-chromatographic analysis of the monosaccharide composition of the dissolved fraction showed galactose to be the major component of the exopolymers (33 – 42%). In the P-limited treatment, an increase of galactose content and a decrease of glucose was found, while the composition of the remaining saccharides was almost unaffected. The results point out the relevance of phosphorus limitation not only in causing an increase in total exopolysaccharide production, but also in affecting the neutral aldose composition of the dissolved carbohydrates released by C. fusiformis, with heteropolysaccharides being more abundant than glucan. The overall sugar composition of the ...

The growth and the structure of a coastal bacterioplankton community were monitored in short-term bottle experiments in order to investigate the bacterial uptake of extracellular organic carbon released by the diatom Cylindrotheca closterium grown under P-balanced and P-depleted conditions. Bacterial specific growth rates and carbon demand were significantly lower in the exudates from P-depleted algae (24% and 30% reduction, respectively). The origin of the extracellular carbon appeared also to affect the taxonomic composition of the bacterioplankton assemblage, mainly reducing the development of ?-Proteobacteria. This pattern of bacterial carbon uptake could contribute to a longer persistence of the exudates released in P-depleted conditions affecting the dynamics of the carbon cycle in marine environments. © 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.