GLOSSary: the GLobal Ocean 16S subunit web accessible resource.

BACKGROUND: Environmental metagenomics is a challenging approach that is exponentially spreading in the scientific community to investigate taxonomic diversity and possible functions of the biological components. The massive amount of sequence data produced, often endowed with rich environmental metadata, needs suitable computational tools to fully explore the embedded information. Bioinformatics plays a key role in providing methodologies to manage, process and mine molecular data, integrated with environmental metagenomics collections. One such relevant example is represented by the Tara Ocean Project. RESULTS: We considered the Tara 16S miTAGs released by the consortium, representing raw sequences from a shotgun metagenomics approach with similarities to 16S rRNA genes. We generated assembled 16S rDNA sequences, which were classified according to their lengths, the possible presence of chimeric reads, the putative taxonomic affiliation. The dataset was included in GLOSSary (the GLobal Ocean 16S Subunit web accessible resource), a bioinformatics platform to organize environmental metagenomics data. The aims of this work were: i) to present alternative computational approaches to manage challenging metagenomics data; ii) to set up user friendly web-based platforms to allow the integration of environmental metagenomics sequences and of the associated metadata; iii) to implement an appropriate bioinformatics platform supporting the analysis of 16S rDNA sequences exploiting reference datasets, such as the SILVA database. We organized the data in a next-generation NoSQL "schema-less" database, allowing flexible organization of large amounts of data and supporting native geospatial queries. A web interface was developed to permit an interactive exploration and a visual geographical localization of the data, either raw miTAG reads or 16S contigs, from our processing pipeline. Information on unassembled sequences is also available. The taxonomic affiliations of contigs and miTAGs, and the spatial distribution of the sampling sites and their associated sequence libraries, as they are contained in the Tara metadata, can be explored by a query interface, which allows both textual and visual investigations. In addition, all the sequence data were made available for a dedicated BLAST-based web application alongside the SILVA collection. CONCLUSIONS: GLOSSary provides an expandable bioinformatics environment, able to support the scientific community in current and forthcoming environmental metagenomics analyses.

Extracellular DNA can preserve the genetic signatures of present and past viral infection events in deep hypersaline anoxic basins.

Deep hypersaline anoxic basins (DHABs) of the Mediterranean Sea are among the most extreme ecosystems on Earth and host abundant, active and diversified prokaryotic assemblages. However, factors influencing biodiversity and ecosystem functioning are still largely unknown. We investigated, for the first time, the impact of viruses on the prokaryotic assemblages and dynamics of extracellular DNA pool in the sediments of La Medee, the largest DHAB found on Earth. We also compared, in La Medee and L'Atalante sediments, the diversity of prokaryotic 16S rDNA sequences contained in the extracellular DNA released by virus-induced prokaryotic mortality. We found that DHAB sediments are hot-spots of viral infections, which largely contribute to the release of high amounts of extracellular DNA. DNase activities in DHAB sediments were much higher than other extracellular enzymatic activities, suggesting that extracellular DNA released from killed prokaryotes can be the most suitable trophic resource for benthic prokaryotes. Preserved extracellular DNA pools, which contained novel and diversified gene sequences, were very similar between the DHABs but dissimilar from the respective microbial DNA pools. We conclude that the strong viral impact in DHAB sediments influences the genetic composition of extracellular DNA, which can preserve the signatures of present and past infections.

Pollutant Pb burden in Mediterranean Centroscymnus coelolepis deep-sea sharks.

We report lead (Pb) analyses in juvenile (n = 37; mean length = 24.7 ±â€¯2.3 cm) and adult (n = 16; mean length = 52.3 ±â€¯9.3 cm) Centroscymnus coelolepis Mediterranean deep-sea sharks that are compared to Pb content in bathy-demersal, pelagic and shallow coastal sharks. Median Pb concentrations of C. coelolepis muscle (0.009-0.056 wet ppm) and liver (0.023-0.061 wet ppm) are among the lowest encountered in shark records. Stable Pb isotope imprints in adult C. coelolepis muscles highlight that most of Pb in C. coelolepis is from human origin. Lead isotopes reveal the persistence of gasoline Pb emitted in the 1970s in low-turnover adult shark's muscle while associated liver imprints are in equilibrium with recent pollutant Pb signatures suggesting an efficient pollutant Pb turnover metabolism. The comparison of Pb distribution between adult and juvenile cohorts suggests the role of dietary exposure and possible maternal offloading of Pb during gestation, likely associated to vitellogenesis in this aplacental viviparous deep-sea shark.

Environmental DNA metabarcoding for benthic monitoring: A review of sediment sampling and DNA extraction methods.

Environmental DNA (eDNA) metabarcoding (parallel sequencing of DNA/RNA for identification of whole communities within a targeted group) is revolutionizing the field of aquatic biomonitoring. To date, most metabarcoding studies aiming to assess the ecological status of aquatic ecosystems have focused on water eDNA and macroinvertebrate bulk samples. However, the eDNA metabarcoding has also been applied to soft sediment samples, mainly for assessing microbial or meiofaunal biota. Compared to classical methodologies based on manual sorting and morphological identification of benthic taxa, eDNA metabarcoding offers potentially important advantages for assessing the environmental quality of sediments. The methods and protocols utilized for sediment eDNA metabarcoding can vary considerably among studies, and standardization efforts are needed to improve their robustness, comparability and use within regulatory frameworks. Here, we review the available information on eDNA metabarcoding applied to sediment samples, with a focus on sampling, preservation, and DNA extraction steps. We discuss challenges specific to sediment eDNA analysis, including the variety of different sources and states of eDNA and its persistence in the sediment. This paper aims to identify good-practice strategies and facilitate method harmonization for routine use of sediment eDNA in future benthic monitoring.

Preservation, origin and genetic imprint of extracellular DNA in permanently anoxic deep-sea sediments.

Molecular approaches that target the total DNA pool recovered from permanently anoxic marine ecosystems have revealed an extraordinary diversity of prokaryotes and unicellular eukaryotes. However, the presence of gene sequences contained within the extracellular DNA pool is still largely neglected. We have investigated the preservation, origin and genetic imprint of extracellular DNA recovered from permanently anoxic deep-sea sediments of the Black Sea. Despite high DNase activities, huge amounts of total extracellular DNA were found in both the surface and subsurface sediment layers, suggesting reduced availability of the extracellular DNA pool to nuclease degradation. The reduced degradation of the total extracellular DNA was confirmed by its low decay rate and the high accumulation in the deeper sediment layers. The copy numbers of 16S and 18S rDNA contained within the extracellular DNA pool in both the surface and subsurface sediment layers was very high, indicating that permanently anoxic sediments of the deep Black Sea are hot spots of preserved extracellular gene sequences. The extracellular DNA recovered from these sediment layers also contained highly diversified 18S rDNA sequences. These were not only representative of the major protistan lineages, but also of new very divergent lineages, branching as independent clades at the base of the tree. Our findings indicate that the extracellular DNA pool is a major archive of present/past eukaryotic gene sequences, and they highlight the importance of integrating molecular cell-oriented approaches with molecular analyses of the extracellular DNA pool, for a better assessment of microbial diversity and temporal changes in marine benthic ecosystems.

Anthropogenic noise and biological sounds in a heavily industrialized coastal area (Gulf of Naples, Mediterranean Sea).

Underwater noise is one of the most widespread threats to the world oceans. Its negative impact on fauna is nowadays well established, but baseline data to be used in management and monitoring programs are still largely lacking. In particular, the acoustic assessment of human-impacted marine coastal areas provides complementary information on the health status of marine ecosystems. The objective of our study was to provide a baseline of underwater noise levels and biological sounds at two sites within the Gulf of Naples (Italy), one of which is located in Bagnoli-Coroglio, a Site of National Interest (SIN) for its high contamination levels. Within the SIN, sounds were recorded both before and during sediment coring activities (vibrocorer sampling), in order to investigate the potential acoustic impact due to such operations. Acoustic recordings were analyzed following the European Marine Strategy Framework Directive indications as defined in the frame of the Descriptor 11. Results reported here show that the investigated area is characterized by a high anthropogenic noise pressure. Ambient noise levels were principally driven by shipping noise and biological sounds of invertebrates (e.g., snapping shrimps). Sounds referable to other biological activity were difficult to detect because heavily masked by shipping noise. Coring activity determined a substantial introduction of additional noise at a local spatial scale. This study expands underwater noise baseline data to be further implemented in future monitoring programs of coastal areas affected by anthropogenic impacts. In addition, it proposes new cues for using underwater acoustic monitoring tools to complement traditional methodologies for evaluating health status of ecosystems and for investigating recovery rates after restoration/reclamation programs.

Chemical contamination can promote turnover diversity of benthic prokaryotic assemblages: The case study of the Bagnoli-Coroglio bay (southern Tyrrhenian Sea).

Chemical contamination of marine ecosystems represents a major concern for the detrimental consequences at different levels of biological organization. However, the impact of chronic contamination on the diversity and assemblage composition of benthic prokaryotes is still largely unknown, and this limits our understanding of the potential implications on ecosystem functioning. The Bagnoli-Coroglio bay (Gulf of Naples, Tyrrhenian Sea) is a typical example of coastal area heavily contaminated by metals and hydrocarbons, released for decades by industrial activities, which ceased at the beginning of nineties. In the present study we analyzed the abundance, diversity and assemblage composition of benthic prokaryotic assemblages at increasing distance from the historical source of contamination in relation to the heavy hydrocarbons (C > 12), polycyclic aromatic hydrocarbons (PAHs) and heavy metal concentrations in the sediments. Prokaryotic abundance in the sediments differed among sites, and was mostly driven by environmental factors rather than by contamination levels. Conversely, the richness of prokaryotic taxa was relatively high in all samples, was driven by contamination levels and decreased significantly with increasing contamination (15-38%). Moreover, our results indicate large variations in the composition of the benthic prokaryotic assemblages among sites, mostly explained by the different levels and types of chemical contaminants found in the sediments. Overall, our findings suggest that chemical contaminants, even after decades from the end of their release, can profoundly influence the richness and turnover diversity of the benthic prokaryotic assemblages, in turn promoting a high diversification of the benthic bacterial and archaeal assemblages by selecting those lineages more adapted to specific mixtures of different contaminants. Our results open new perspectives for understanding of the long-term effects of chemical contamination on the benthic prokaryotic assemblages and the ecological processes they mediate.

Impact of historical contamination on meiofaunal assemblages: The case study of the Bagnoli-Coroglio Bay (southern Tyrrhenian Sea).

The effects of contaminants on marine organisms have been documented since decades, but the long-term responses and recovery rates of benthic communities to mixtures of contaminants, several years after the cessation of industrial activities, need to be further investigated. Bagnoli-Coroglio Bay (Gulf of Naples, Tyrrhenian Sea) is a typical example of historically contaminated coastal area due to industrial activities stopped at the beginning of nineties. In the present study we carried out a fine spatial scale analysis of the distribution of meiofaunal (and nematodes) assemblages along five bathymetric transects located at increasing distance from the historical source of contamination in relation with the polycyclic aromatic hydrocarbon and heavy metal concentrations present in the sediment. Meiofaunal abundance and biomass changed widely along transects but independent from the distance from the source of contamination. Even when the contamination levels were expected to induce significant detrimental biological consequences, meiofaunal abundance and biomass were similar to those reported in unpolluted benthic coastal areas worldwide. Conversely, biodiversity in terms of meiofaunal taxa richness was generally low (range: 5-8 taxa in 12 of the overall 15 stations investigated). This was explained by the lack of sensitive groups such as ostracods, gastrotrichs and tardigrades commonly encountered in benthic coastal ecosystems, thus reflecting an overall poor/moderate environmental quality of the investigated area. Nematode (structural and functional) diversity was also low, particularly at stations characterized by higher contamination levels. At the same time, nematode species composition did not change significantly among stations suggesting a widespread effect of contaminants able to reduce the variability (i.e., turnover diversity) within the assemblages of the whole study area. Overall, our results indicate that even decades after the cessation of contaminant emissions, benthic biodiversity was affected in terms of both meiofaunal taxa and nematode species. These findings strongly reinforce the call for reducing sources of chronic pollution in marine ecosystems and provide new insights for a better understanding of the ecological recovery of historically contaminated marine environments.

Assessing the efficiency and eco-sustainability of bioremediation strategies for the reclamation of highly contaminated marine sediments.

Coastal sediments subjected to high anthropogenic impacts can accumulate large amounts of polycyclic aromatic hydrocarbons (PAHs) and metals, demanding effective and eco-sustainable remediation solutions. In this study, we carried out bioremediation experiments on marine sediments highly contaminated with PAHs and metals. In particular, we investigated the effects of biostimulation (by the addition of inorganic nutrients), bioaugmentation (by the addition of fungi belonging to Aspergillus sp.) and microbial fuel cell-based strategies on PAH degradation and on changes in metal partitioning. Results reported here indicate that all biotreatments determined a significant decrease of PAH concentrations (at least 60%) in a relatively short time interval (few weeks) and that biostimulation was the most effective approach (>90%). Biostimulation determined a faster degradation rate of high than low molecular weight PAHs, indicating a preferential biodegradation of specific PAH congeners. At the same time, the biotreatments changed the partitioning of metals, including their solubilization, suggesting the need of parallel environmental risk assessment. Our findings also suggest that ex situ biotreatments can have a lower carbon footprint than current management options of contaminated sediments (i.e., landfill disposal and/or disposal in confined aquatic facilities), but integration with other strategies for metal removal (e.g. through bioleaching) from sediments is needed for their safe re-use. Overall, results presented here provide new insights into the development of effective and eco-sustainable bioremediation strategies for the reclamation of highly contaminated marine sediments.

Multiple human pressures in coastal habitats: variation of meiofaunal assemblages associated with sewage discharge in a post-industrial area.

Marine ecosystems are globally threatened by human activities, but some areas, such as those affected by abandoned industrial plants, show an overlap of acute and chronic impacts, which determine a considerable deterioration of their health status. Here we report the results of a research conducted on coastal sewers that discharge their loads in the highly contaminated area of Bagnoli-Coroglio (Tyrrhenian Sea, Western Mediterranean). The sampling area is characterized by heavy industrial activities (a steel plant using coal, iron and limestone) started in 1905 and ceased in 1990, which left widespread heavy metals and hydrocarbon contamination. After taking into account the potential influence of sediment grain size ranges through their inclusion as covariates in the analysis, we tested the potential impact of sewage discharge on the total abundance and multivariate structure of meiofaunal assemblages, as well as on the abundance of single taxa. The organic matter was analysed in terms of total phytopigment and biopolymeric carbon concentrations. Nematoda, Copepoda (including their nauplii), and Tardigrada were the most abundant meiofaunal taxa at all sites, but nematodes did not show a consistent pattern relative to the sewage outfalls. However, the sewer located in the historically most contaminated area showed a minimal abundance of all taxa, including nematodes, while copepods were relatively less abundant at the two southernmost sewers. Comparing the north vs. south site of the sewers, higher meiofaunal abundances were observed in the southward part, likely as a result of the local circulation. The results of this study indicate the general adaptation of meiofauna to multiple stressors (sewage discharge, superimposed to chronic industrial contamination) and its likely modulation by other local processes. They also provide relevant baseline information for future restoration interventions that would take into account the spatial variation of target organisms as needed.

Damage and degradation rates of extracellular DNA in marine sediments: implications for the preservation of gene sequences.

The extracellular DNA pool in marine sediments is the largest reservoir of DNA of the world oceans and it potentially represents an archive of genetic information and gene sequences involved in natural transformation processes. However, no information is at present available for the gene sequences contained in the extracellular DNA and for the factors that influence their preservation. In the present study, we investigated the depurination and degradation rates of extracellular DNA in a variety of marine sediment samples characterized by different ages (up to 10,000 years) and environmental conditions according to the presence, abundance and diversity of prokaryotic gene sequences. We provide evidence that depurination of extracellular DNA in these sediments depends upon the different environmental factors that act synergistically and proceeds at much slower rates than those theoretically predicted or estimated for terrestrial ecosystems. These findings suggest that depurination in marine sediments is not the main process that limits extracellular DNA survival. Conversely, DNase activities were high suggesting a more relevant role of biologically driven processes. Amplifiable prokaryotic 16S rDNA sequences were present in most benthic systems analysed, independent of depurination and degradation rates and of the ages of the sediment samples. Additional molecular analyses revealed that the extracellular DNA pool is characterized by relatively low-copy numbers of prokaryotic 16S rDNA sequences that are highly diversified. Overall, our results suggest that the extracellular DNA pool in marine sediments represents a repository of genetic information, which can be used for improving our understanding of the biodiversity, functioning and evolution of ecosystems over different timescales.

Extracellular DNA as a genetic recorder of microbial diversity in benthic deep-sea ecosystems.

Extracellular DNA in deep-sea sediments represents a major repository of genes, which previously belonged to living organisms. However, the extent to which these extracellular genes influence current estimates of prokaryotic biodiversity is unknown. We investigated the abundance and diversity of 16S rDNA sequences contained within extracellular DNA from continental margins of different biogeographic regions. We also compared the taxonomic composition of microbial assemblages through the analysis of extracellular DNA and DNA associated with living cells. 16S rDNA contained in the extracellular DNA pool contributed up to 50% of the total 16S rDNA copy number determined in the sediments. Ca. 4% of extracellular Operational Taxonomic Units (OTUs) were shared among the different biogeographic regions revealing the presence of a core of preserved OTUs. A higher fraction of OTUs was exclusive of each region potentially due to its geographic and thermohaline characteristics. Ca. one third of the OTUs identified in the extracellular DNA were absent from living prokaryotic assemblages, possibly representing the signatures of past assemblages. Our findings expand the knowledge of the contribution of extracellular microbial sequences to current estimates of prokaryotic diversity obtained through the analyses of "environmental DNA", and open new perspectives for understanding microbial successions in benthic ecosystems.

Biosurfactant-induced remediation of contaminated marine sediments: Current knowledge and future perspectives.

The contamination of marine sediments is widespread in coastal regions of the world and represents a major concern for the potential detrimental consequences on ecosystems' health and provision of goods and services for human wellbeing. Thus, there is an urgent need to find sustainable and eco-compatible solutions for the remediation of contaminated sediments. Bioremediation is a low cost and environmental-friendly strategy with a high potential for the remediation of contaminated marine sediments. Here we review the potential application of biosurfactants produced by microbial taxa for the remediation of contaminated marine sediments and we discuss future research needs to develop efficient and eco-sustainable biosurfactant-based strategies for the recovery of contaminated marine sediments, in view of large-scale applications.

Assessing viral taxonomic composition in benthic marine ecosystems: reliability and efficiency of different bioinformatic tools for viral metagenomic analyses.

In benthic deep-sea ecosystems, which represent the largest biome on Earth, viruses have a recognised key ecological role, but their diversity is still largely unknown. Identifying the taxonomic composition of viruses is crucial for understanding virus-host interactions, their role in food web functioning and evolutionary processes. Here, we compared the performance of various bioinformatic tools (BLAST, MG-RAST, NBC, VMGAP, MetaVir, VIROME) for analysing the viral taxonomic composition in simulated viromes and viral metagenomes from different benthic deep-sea ecosystems. The analyses of simulated viromes indicate that all the BLAST tools, followed by MetaVir and VMGAP, are more reliable in the affiliation of viral sequences and strains. When analysing the environmental viromes, tBLASTx, MetaVir, VMGAP and VIROME showed a similar efficiency of sequence annotation; however, MetaVir and tBLASTx identified a higher number of viral strains. These latter tools also identified a wider range of viral families than the others, providing a wider view of viral taxonomic diversity in benthic deep-sea ecosystems. Our findings highlight strengths and weaknesses of available bioinformatic tools for investigating the taxonomic diversity of viruses in benthic ecosystems in order to improve our comprehension of viral diversity in the oceans and its relationships with host diversity and ecosystem functioning.

Exo-enzymatic activities and dissolved organic pools in relation with mucilage development in the Northern Adriatic Sea.

We tested the hypothesis that the appearance of mucilage in the Northern Adriatic Sea was related with the accumulation of dissolved organic compounds released by intensive enzymatic activities and not utilized as direct substrate for microbial growth. To do this enzymatic activities and dissolved organic and inorganic pools in periods characterized by the presence of mucilage and in the same seasons but in absence of mucilage were compared. Extracellular enzymatic activities (aminopeptidase, beta-glucosidase and alkaline phosphatase), nutrient pool concentrations (total dissolved nitrogen, dissolved organic nitrogen, total dissolved phosphorus, dissolved organic phosphorus) and the biochemical composition of particulate and dissolved organic matter (in terms of proteins and carbohydrates) were determined on a monthly basis over a period of 3 years. Aminopeptidase and alkaline phosphatase activities displayed higher values in springs preceding the appearance of mucilage than in spring when no mucilage was observed. Beta-Glucosidase activity showed significantly higher values in summer periods characterized by the massive production of mucilage than in summers without mucilage events. The months preceding mucilage events were also characterized by an increase of the alkaline phosphatase to aminopeptidase activity ratio and by a significant accumulation of dissolved proteins. These findings, together with the significant increase of the DON/DOP ratio, suggest that mucilage formation is favoured by the deficiency of organic P. The present study provides compelling evidences that mucilage formation is favoured by the unbalance between organic matter mobilization by enzymatic activities and the accumulation of labile dissolved organic-N compounds.

Nucleic acid (DNA, RNA) quantification and RNA/DNA ratio determination in marine sediments: comparison of spectrophotometric, fluorometric, and HighPerformance liquid chromatography methods and estimation of detrital DNA

In this study, we compared three methods for extraction and quantification of RNA and DNA from marine sediments: (i) a spectrophotometric method using the diphenylamine assay; (ii) a fluorometric method utilizing selective fluorochromes (thiazole orange for total nucleic acids and Hoechst 33258 for DNA); and (iii) a high-pressure liquid chromatography (HPLC) method which uses a specific column to separate RNA and DNA and UV absorption of the nucleic acids for quantification. Sediment samples were collected in the oligotrophic Cretan Sea (eastern Mediterranean, from 40 to 1,540 m in depth) and compared to the distribution and composition of the benthic microbial assemblages (i.e., bacteria and microprotozoa). DNA concentrations measured spectrophotometrically and by HPLC were not significantly different, while fluorometric yields were significantly lower. Such differences appear mainly due to fact that the stain-DNA complex is strongly dependent on the DNA composition and structure. RNA concentrations determined by the three methods displayed some differences; fluorometric and spectrophotometric methods obtain RNA concentration by difference and therefore may be biased by DNA estimates. By contrast, the HPLC method provides independent assessments of RNA and DNA concentrations. We tentatively estimated the contribution of the detrital DNA to the total DNA pools in two ways. The two calculations provided quite similar results indicating that the majority of the DNA pool in the deep-sea sediments was detrital. Microbial RNA generally accounted for almost the entire sedimentary RNA pools below 100-m depth. RNA concentrations were found to decrease along the Cretan shelf and slope. The RNA/DNA ratio calculated by using fluorometric DNA concentrations was significantly correlated with values of sediment community oxygen consumption only below 100-m depth (dominated by the microbial biomass). These data suggest that the RNA/DNA ratio, based on fluorometric estimates of DNA, can be used as an indicator of benthic metabolic activity, but only when metazoan contribution to the microbial DNA is negligible.

Small-Scale Distribution of Bacteria, Enzymatic Activities, and Organic Matter in Coastal Sediments.

The small-scale distribution of several structural (bacterial abundance, phytopigment, total and soluble protein, and carbohydrate content) and functional ecological variables (enzymatic activities, frequency of dividing cells) was investigated in coastal sediments during a spring bloom. For bacterial abundance, enzymatic activity, and organic matter determinations, samples were collected at 5-m depth from a sediment surface delimited by a 42 x 42-cm frame, divided into 49 squares. In order to test the influence of the bottom microtopography on the investigated variables, the size of this frame was defined to cover the distance between two subsequent ripples. As indicated by Fisher's index, benthic bacteria, enzymatic activities, proteins, carbohydrates, and their soluble fractions exhibited an aggregate distribution. Sampling size (i.e., sample unit of 36 cm2) was appropriate for all variables, except for chlorophyll a and frequency of dividing cells that displayed a contagious distribution. To estimate the reliability of the current sampling strategy, we compared the mean values from three randomly selected sample units with the average value of the entire sediment surface (i.e., 49 samples). For all variables reported in this study, the use of three replicates was representative of the mean values of the sampled area with a confidence limit within +/-20%. Bacterial population sizes did not correlate with their potential food sources (e.g., phytopigments, proteins, and carbohydrates), or with enzymatic activities, suggesting the presence of possible time lags between organic inputs and microbial response. Chlorophyll a during the spring bloom displayed much higher concentrations than phaeopigments and correlated with carbohydrates. Chlorophyll a distribution was autocorrelated and displayed a large patch size (1,134 cm2). Phaeopigments and proteins accumulated in the central depression of the ripple-mark structure, apparently depending upon a passive accumulation due to the reduced current action. In contrast, microphytobenthic biomass and bacterial numbers were highest in the two opposite ripples, suggesting that different driving forces operate selectively on the living components.

Assessing the trophic state and eutrophication of coastal marine systems: a new approach based on the biochemical composition of sediment organic matter.

We used a biochemical approach based on the analysis of the quality and quantity of sedimentary organic matter for identifying new descriptors of the trophic state and environmental quality of coastal marine systems. A large-scale study, including 99 stations, belonging to 33 transects, was carried out along 250 km of the Apulian coasts (Mediterranean Sea) in March and September 2000. The investigated area covered a wide range of anthropogenic impacts (industrial ports, tourist harbours, areas affected by power plants and industrial wastes, mariculture areas). Other sites, including marine protected areas (i.e., without any apparent impact), were used as "control". Water column and benthic parameters provided different indications and classifications of the trophic state of coastal marine systems. We found that phytopigment content of the sediments changed in response to all different sources of anthropogenic impact and resulted in a useful descriptor of the trophic state and environmental quality. Highest sediment chlorophyll-a concentrations, indicating conditions of increasing eutrophication, were found in areas impacted by the discharge of heated waters from a power plant. In particular, the contribution of the autotrophic biomass to the biopolymeric carbon pool appeared to be a good descriptor of the decreasing environmental quality. Independently from the sampling period or the pollution source such contribution was significantly lower in transects subjected to anthropogenic impact than in control areas. Differences in trophic conditions were evident both in terms of quantity (i.e., total organic matter content) and quality (i.e., biochemical composition) of sediment organic matter. In particular, sediment protein concentration appeared to be a good descriptor of the trophic state of the benthic systems at different spatial scales. Multivariate (MDS) analysis allowed identifying areas characterised by hypertrophic, eutrophic and meso-oligotrophic conditions and to define relative threshold levels. A classification of the trophic state of coastal systems based on protein and carbohydrate concentrations is proposed.

Impact of aquaculture on benthic virus-prokaryote interactions in the Mediterranean Sea.

We investigated the effects of organic enrichment due to the biodeposition from fish farms on benthic prokaryotic and viral abundance and production, viral-induced prokaryotic mortality, enzymatic activities and bacterial diversity. We compared four areas across the Mediterranean Sea, from Cyprus to Spain, and two different habitats: sediments covered by the seagrass Posidonia oceanica and soft-bottom unvegetated sediments. In several cases, the sediments beneath the cages showed higher prokaryotic and viral abundance and production, and higher rates of organic matter decomposition. However, the differences between impact and control sediments were not consistent at all regions and habitats. Benthic bacterial diversity was always lower below the cages, where high viral-induced bacterial mortality rates were also observed. The δ- and γ-Proteobacteria dominated in both impacted and control sediments, but the relative importance of sulphate-reducing δ-Proteobacteria increased beneath the cages. We conclude that aquaculture can have a significant impact on benthic prokaryotes and viruses, by stimulating prokaryotic metabolism and viral infections, reducing bacterial diversity and altering assemblage composition. However, these impacts vary depending upon the sediment type and the habitat characteristics.