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The oligotrophy of the southern Adriatic Sea is characterized by seasonal stratification which enables nutrient supply to the euphotic layer. A set of interdisciplinary methods was used to elucidate the diversity and co-dependency of bacterio- and phytoplankton of the water column during the stratification period of July 2021. A total of 95 taxa were determined by microscopy: 58 diatoms, 27 dinoflagellates, 6 coccolithophores, and 4 other autotrophs, which included Chlorophyceae, Chrysophyceae, and Cryptophytes. Nanophytoplankton abundances were higher in comparison to microphytoplankton. The prokaryotic plankton community as revealed by HTS was dominated by Proteobacteria (41–73%), Bacteroidota (9.5–27%), and cyanobacteria (1–10%), while the eukaryotic plankton community was composed of parasitic Syndiniales (45–80%), Ochrophyta (2–18%), Ciliophora (2–21%), Chlorophytes (2–4%), Haptophytes (1–4%), Bacillariophyta (1–13%), Pelagophyta (0.5–12%) and Chrysophyta (0.5–3%). Flow cytometry analysis has recorded Prochlorococcus and photosynthetic picoeukaryotes as more abundant in deep chlorophyll maximum (DCM), and Synechococcus and heterotrophic bacteria as most abundant in surface and thermocline layers. Surface, thermocline, and DCM layers were distinct considering community diversity, temperature, and nutrient correlations, while extreme nutrient values at the beginning of the investigating period indicated a possible nutrient flux. Nutrient and temperature were recognized as the main environmental drivers of phytoplankton and bacterioplankton community abundance.

Dissolved Organic Matter (DOM), the largest pools of reduced carbon on Earth, represents the major source of energy for micro-heterotrophs, starting the microbial loop. In the open ocean, most of the autochthonous DOM is labile and is therefore respired within days, but a small fraction, called recalcitrant, persists for years to millennia. There are large oligotrophic areas in the oceans, such as the North Atlantic subtropical gyre and the Mediterranean Sea, where the decoupling between production and consumption of DOM leads to its accumulation with implications for the microbial loop.The main goal of this study is to investigate the putative functioning of the microbial loop all across the Mediterranean Sea following the main path of the Atlantic water entering through the Gibraltar Strait. Dissolved Organic Carbon (DOC) concentration and fluorescence of chromophoric DOM were measured together with heterotrophic prokaryotes, nanoflagellates and virus abundance by flow cytometry. Samples were collected during the MSM72 oceanographic cruise carried out in March-April 2018. Results show that in the western Mediterranean Sea the heterotrophic prokaryotes are relatively abundant and DOM is efficiently removed, suggesting an active transfer of carbon to the higher trophic levels. In contrast, in the eastern Mediterranean Sea DOM accumulates, prokaryote abundance is low and low nucleic acid prokaryotes dominates, suggesting a malfunctioning of the microbial loop leading to a less efficient carbon transfer to the food web. Following the core of the Atlantic Water flowing eastward, as the nutricline deepens, an ecological succession in the phytoplankton communities (from picoeukaryotes to cyanobacteria) is observed, together with a depletion of the fluorescent components in the DOM pool, reflecting changes in the DOM quality.Different hypotheses can be formulated to explain these observations: the microbial communities could be limited by nutrient availability, by an enhanced top-down pressure by grazers or viruses or by the quality of DOM that could be recalcitrant due to abiotic processes such as photobleaching or to the progressive use of its labile fraction by micro-heterotrophs.With the upcoming increase in water column stratification, a change in the quality of DOM due to the combined effect of abiotic (e.g., photobleaching) and biotic (e.g., change in the phytoplankton community) is expected with consequences for the functioning of the microbial loop. The questions about the malfunctioning of the microbial loop and its implications in a global change scenario have puzzled the scientists for years and the Mediterranean Sea is a well-suited natural laboratory to answer them.

The sea urchin Paracentrotus lividus (P. lividus) was exposed to either virgin or biofilm-covered polystyrene microbeads (micro-PS, 45 μm) in order to test the effect of microbial colonization on the uptake, biodistribution, and immune response. The biofilm was dominated by bacteria, as detected by scanning electron microscopy and 16S rRNA sequencing. A higher internalization rate of colonized micro-PS inside sea urchins compared to virgin ones was detected, suggesting a role of the plastisphere in the interaction. Colonized and virgin micro-PS showed the same biodistribution pattern by accumulating mainly in the digestive system with higher levels and faster egestion rates for the colonized. However, a significant increase of catalase and total antioxidant activity was observed only in the digestive system of colonized micro-PS-exposed individuals. Colonized micro-PS also induced a significant decrease in the number of coelomocytes with a significant increase in vibratile cells, compared to control and virgin micro-PS-exposed animals. Moreover, a general time-dependent increase in the red/white amoebocytes ratio and reactive oxygen species and a decrease in nitrogen ones were observed upon exposure to both colonized and virgin micro-PS. Overall, micro-PS colonization clearly affected the uptake and toxicological responses of the Mediterranean sea urchin P. lividus in comparison to virgin micro-PS.

Ocean life—from viruses to whales—is built from “biomolecules.” Biomolecules such as DNA infuse each drop of ocean water, grain of sediment, and breath of ocean air. The Ocean Biomolecular Observing Network (OBON) is developing a global collaboration that will allow science and society to understand ocean life like never before. The program will transform how we sense, harvest, protect, and manage ocean life using molecular techniques, as it faces multiple stresses including pollution, habitat loss, and climate change. It will also help communities detect biological hazards such as harmful algal blooms and pathogens, and be a key component of next-generation ocean observing systems. OBON will encourage continuous standardization and intercalibration of methods and data interoperability to help enhance and future-proof capabilities. OBON's objectives are: 1) to build a coastal-to-open ocean multi-omics biodiversity observing system; 2) to develop and transfer capacity between partners; 3) to enhance marine ecosystem digitization and modelling and 4) to coordinate action on pressing scientific, management, and policy questions.

The sea urchin

Plankton are a pivotal component of the diversity and functioning of coastal marine ecosystems. A long time-series of observations is the best tool to trace their patterns and variability over multiple scales, ultimately providing a sound foundation for assessing, modelling and predicting the effects of anthropogenic and natural environmental changes on pelagic communities. At the same time, a long time-series constitutes a formidable asset for different kinds of research on specific questions that emerge from the observations, whereby the results of these complementary studies provide precious interpretative tools that augment the informative value of the data collected. In this paper, we review more than 140 studies that have been developed around a Mediterranean plankton time series gathered in the Gulf of Naples at the station LTER-MC since 1984. These studies have addressed different topics concerning marine plankton, which have included: i) seasonal patterns and trends; ii) taxonomic diversity, with a focus on key or harmful algal species and the discovery of many new taxa; iii) molecular diversity of selected species, groups of species or the whole planktonic community; iv) life cycles of several phyto- and zooplankton species; and v) interactions among species through trophic relationships, parasites and viruses. Overall, the products of this research demonstrate the great value of time series besides the record of fluctuations and trends, and highlight their primary role in the development of the scientific knowledge of plankton much beyond the local scale.

Estuarine processes play a key role in determining the amount and quality of land-derived dissolved organic matter (DOM) reaching the oceans. Microbial-mediated reactions can affect the concentration, quality, and bioavailability of DOM within an estuary. In this study, we investigated biological DOM removal in a small estuary and its variability in two contrasting seasons (spring and autumn) characterized by natural differences in the concentration and quality of the riverine DOM. Two incubation experiments were carried out using natural DOM and heterotrophic prokaryotes community collected at the estuary in March and September. Dissolved organic carbon (DOC) concentration, DOM fluorescence, and the heterotrophic prokaryotes abundance (HPA) showed marked differences between the two seasons. These parameters were followed through time for up to two months. Despite the marked differences in the initial conditions, the DOC removal rates were surprisingly similar in the two periods (16 µM DOC month−1 in March and 18 µM DOC month−1 in September), with the biggest removal in the first 48 h. The trend of fluorescent DOM (FDOM) during the incubation showed marked differences between the two periods. In March, the net removal of all the FDOM components was observed consistently with the decrease in DOC, whereas, in September, the net production of humic-like substances was observed.

39 pages, 6 figures, 2 tables, This paper presents major gaps and challenges for implementing the UN Decade of Ocean Science for Sustainable Development (2021-2030) in the Mediterranean region. The authors make recommendations on the scientific knowledge needs and co-design actions identified during two consultations, part of the Decade preparatory-phase, framing them in the Mediterranean Sea’s unique environmental and socio-economic perspectives. According to the ‘Mediterranean State of the Environment and Development Report 2020’ by the United Nations Environment Programme Mediterranean Action Plan and despite notable progress, the Mediterranean region is not on track to achieve and fully implement the Sustainable Development Goals of Agenda 2030. Key factors are the cumulative effect of multiple human-induced pressures that threaten the ecosystem resources and services in the global change scenario. The basin, identified as a climate change vulnerability hotspot, is exposed to pollution and rising impacts of climate change. This affects mainly the coastal zones, at increasing risk of extreme events and their negative effects of unsustainable management of key economic assets. Transitioning to a sustainable blue economy is the key for the marine environment’s health and the nourishment of future generations. This challenging context, offering the opportunity of enhancing the knowledge to define science-based measures as well as narrowing the gaps between the Northen and Southern shores, calls for a joint (re)action. The paper reviews the state of the art of Mediterranean Sea science knowledge, sets of trends, capacity development needs, specific challenges, and recommendations for each Decade’s societal outcome. In the conclusions, the proposal for a Mediterranean regional programme in the framework of the Ocean Decade is addressed. The core objective relies on integrating and improving the existing ocean-knowledge, Ocean Literacy, and ocean observing capacities building on international cooperation to reach the “Mediterranean Sea that we want”, With the institutional support of the ‘Severo OchoaCentre of Excellence’ accreditation (CEX2019-000928-S)

This study investigated the biogeography, the presence and diversity of potentially harmful taxa harbored, and potential interactions between and within bacterial and eukaryotic domains of life on plastic debris in the Mediterranean. Using a combination of high-throughput DNA sequencing (HTS), Causal Network Analysis, and Scanning Electron Microscopy (SEM), we show regional differences and gradients in the Mediterranean microbial communities associated with marine litter, positive causal effects between microbes including between and within domains of life, and how these might impact the marine ecosystems surrounding them. Adjacent seas within the Mediterranean region showed a gradient in the microbial communities on plastic with non-overlapping endpoints (Adriatic and Ligurian Seas). The largest predicted inter-domain effects included positive effects of a novel red-algal Plastisphere member on its potential microbiome community. Freshwater and marine samples housed a diversity of fungi including some related to disease-causing microbes. Algal species related to those responsible for Harmful Blooms (HABs) were also observed on plastic pieces including members of genera not previously reported on Plastic Marine Debris (PMD).

Phytoplankton is the primary source of Dissolved Organic Matter (DOM) to the oceans. DOM is mainly released by extracellular exudation and used by heterotrophic prokaryotes to synthesise biomass and recycle inorganic nutrients. DOM released by phytoplankton is mainly composed by carbohydrates, proteins and lipids that are thought to be labile and by humic substances that are thought to be recalcitrant and thus resistant to bacterial degradation. There are a lot of uncertainties regarding the biological lability of exudates and the role of DOM released by phytoplankton in the marine carbon cycle. In this study, cultures of the diatom P. tricornutum were produced under axenic conditions and Dissolved Organic Carbon (DOC) concentration, Excitation-Emission matrices (EEMs) and cell density were measured with time in order to follow the release of DOM during the different growth phases. Exudates were then inoculated with a marine microbial community for 24 days, DOC removal and FDOM transformation were followed with time in the exudates and in the permeate (< 3k Da; Low Molecular Weight, LMW) and retentate (> 3k Da; High Molecular Weight, HMW) fractions. Heterotrophic prokaryotes abundance was also followed during the incubations. Our results show that ~75% of the total DOC pool was LMW. After 24 days, 28% of the initial DOC pool was removed. Fluorescence indicate high lability of protein-like molecules and degradation of bigger proteins into smaller peptides before their removal. The production of humic-like and flavin-like substances was also observed.

Bacteria are the most important microorganisms in the world oceans, accounting for up to 75% of the total biomass. They are responsible for fundamental biogeochemical processes and therefore often used as ecological indicators. In this study, bacteria were quantified by flow cytometry and their diversity assessed by High Throughput Sequencing (HTS) in the southern Adriatic Sea. The most abundant bacterial groups were also quantified by qPCR. The samples were collected from the surface to the seabed over a total of 16 different depths at four stations during the late winter BIOTA (BIO-Tracing Adriatic water masses) cruise conducted in March 2016. The investigated area showed unusual water mass properties and was characterized by a shallow mixed layer, which differed from the usual winter convection conditions, typical of middle-altitude ecosystems and important for the seasonal picoplankton dynamics of this area. Heterotrophic bacteria were separated into HNA (relative High Nucleic Acid content) and LNA (Low Nucleic Acid content) subpopulations with abundances up to 1.8 × 105 and 8.8 × 105 cells mL−1, respectively. HNA dominated at offshore stations reaching their maximum at depths below the euphotic zone. The bacterial community was dominated by Alphaproteobacteria, accounting for >40% of the total sequence reads and were mainly represented by the SAR11 clade (90.84%), followed by Marinimicrobia (18% of the total sequence reads), mainly represented by clade SAR406 (8.44%). Distinctive bacterial groups were found in the euphotic layer (Bacteroidetes and Actinobacteria) and aphotic layer samples (Deltaproteobacteria, Marinimicrobia, Chloroflexi, Acidobacteria and Planctomycetes). Results of the qPCR analyses further confirmed HTS results with highest abundances obtained for Alphaproteobacteria, followed by Gammaproteobacteria and Bacteroidetes. The adopted multiple approach, combining different molecular tools, critically supported by optics and flow cytometry, reveal changes in the bacterial assemblages during the unusual thermohaline conditions observed in 2016 in the southern Adriatic Sea.

Diatoms produce and release polyunsaturated aldehydes (PUAs) during senescence in culture and at the end of blooms in nature and these compounds play different ecological roles, as infochemicals, allelochemicals and pheromones In order to elucidate the toxic effects of PUAs, we isolated six bacterial strains from the Mediterranean Sea during a diatom bloom and tested their tolerance to PUA in terms of growth and cell membrane properties. Based upon 16S rRNA sequencing, these bacteria were assigned to the genera Pseudomonas , Sufflavibacter, Halomonas , Vibrio , Idiomarina , and Labrenzia . Growth of these strains was reduced by 50% (EC 50 ) at PUA concentrations ranging from 600 to 1700 μM of 2E,4E/Z-heptadienal (HEPTA), 400–800 μM of 2E, 4E/Z-octadienal (OCTA), and 70–400 μM of 2E, 4E/Z-decadienal (DECA). Two of these strains, Vibrio sp. and Halomonas , sp. were also investigated for membrane fatty acid composition in terms of adaptive modifications of their degree of saturation (ratio between saturated and unsaturated fatty acids) by GC-FID. A direct correlation between hydrophobicity and PUA toxicity was observed, and these bacteria were also found to react to PUAs by increasing the degree of saturation of their membranes fatty acids. Tested PUAs were 4-fold more toxic than the well-investigated n -alkanols, most probably due to their additional chemical aldehyde toxicity to disrupting proteins by the formation of Schiff's bases, and therefore, they act as very toxic and effective poison, probably accumulating in cytoplasmic membranes because of their high hydrophobicity.

Summary Microalgae play a major role as primary producers in aquatic ecosystems. Cell signalling regulates their interactions with the environment and other organisms, yet this process in phytoplankton is poorly defined. Using the marine planktonic diatom Pseudo‐nitzschia multistriata, we investigated the cell response to cues released during sexual reproduction, an event that demands strong regulatory mechanisms and impacts on population dynamics.We sequenced the genome of P. multistriata and performed phylogenomic and transcriptomic analyses, which allowed the definition of gene gains and losses, horizontal gene transfers, conservation and evolutionary rate of sex‐related genes. We also identified a small number of conserved noncoding elements.Sexual reproduction impacted on cell cycle progression and induced an asymmetric response of the opposite mating types. G protein‐coupled receptors and cyclic guanosine monophosphate (cGMP) are implicated in the response to sexual cues, which overall entails a modulation of cell cycle, meiosis‐related and nutrient transporter genes, suggesting a fine control of nutrient uptake even under nutrient‐replete conditions.The controllable life cycle and the genome sequence of P. multistriata allow the reconstruction of changes occurring in diatoms in a key phase of their life cycle, providing hints on the evolution and putative function of their genes and empowering studies on sexual reproduction.

The Gulf of Naples is a dynamical area with intense exchanges between offshore oligotrophic and coastal eutrophic waters with frequent freshwater inputs. The Sarno River, one of the most polluted rivers in Europe, strongly contributes to the pollution of the area, discharging high amounts of heavy metals and organic wastes from heavily cultivated and industrial areas. This paper reports on the diversity and community structure of the marine residential Bacteria and Archaea of the Gulf of Naples in an area close to the river Sarno plume and investigates their small-scale taxonomic diversity and expression patterns as a proxy of potential metabolic activity using metagenomics and metatranscriptomics. Bacteria and Archaea were mainly represented by marine clades, with only minor contributors from freshwater ones. The community was dominated by Alpha- and Gammaproteobacteria, of which Rhodospirillales, Pelagibacteriales, and Oceanospirilalles were most represented. However, Alteromonadales and Rhodobacterales were the most active, despite their relative lower abundance, suggesting that they are important for overall ecosystem functioning and nutrient cycling. Nitrification and a reversed form of dissimilatory sulfate reduction were the major metabolic processes found in the metatrascriptomes and were mainly associated to Nitrosopumilales and Pelagibacter, respectively. No clear indication of transcripts related to stress induced by heavy metals or organic pollutants was found. In general, despite the high loads of pollutants discharged continuously by the Sarno River, the microbial community did not show marks of stress-induced changes neither structural nor functional, thus suggesting that this river has little or no effect on the planktonic bacterial community of the Gulf of Naples.

Regions of freshwater influence are affected by a constant interplay of different environmental factors that rule their seasonal and interannual highly variable dynamics. Riverine freshwater discharges alter the buoyancy of the local water masses, generating dynamic fronts and increasing water column stratification and nutrient availability. In such a heterogeneous context the paradigms for biological-physical interactions driving biogeochemical dynamics in the open seas do not always apply. We present here data from 12 surveys spanning 12 years (from 2004 to 2016) in the Po River prodelta area, which is highly representative of the above-described complexity, as it conveys the second largest freshwater discharge to the Mediterranean Sea. In particular, we focused on the microbially-mediated organic matter processing by estimating the numerical abundances of non-pigmented (‘heterotrophic’) and photoautotrophic (Synechococcus) prokaryotes, heterotrophic production and the activity of the exoenzymes beta-glucosidase, alkaline phosphatase and leucine aminopeptidase. The abundance of Synechococcus and heterotrophic prokaryotes as well as microbial activities were positively correlated to temperature and chlorophyll a concentration. Concomitantly, each variable was negatively correlated to salinity. In high-temperature, high salinity and low chlorophyll a samples, the heterotrophic production was limited and deviated from the typical linearity of the Arrhenius' law, suggesting resource limitation rather than temperature control. A multivariate analysis performed to analyze the pattern of organic matter degradation/utilization highlighted 11 distinct clusters of functional diversity. Temperature, salinity and chlorophyll a concentration data alone were able to explain most of the functional cluster separation as evidenced by a regression tree analysis. For freshwater-influenced samples, salinity was the only variable affecting metabolic patterns, whereas in typically marine waters, at salinity ≥37, metabolic profiles were determined by specific combinations of all 3 parameters.

A Pb-resistant bacterial strain (named hereinafter Pb15) has been isolated from highly polluted marine sediments at the Sarno River mouth, Italy, using an enrichment culture to which Pb(II) 0.48 mmol l(-1) were added. 16S rRNA gene sequencing (Sanger) allowed assignment of the isolate to the genus Bacillus, with Bacillus pumilus as the closest species. The isolate is resistant to Pb(II) with a minimum inhibitory concentration (MIC) of 4.8 mmol l(-1) and is also resistant to Cd(II) and Mn(II) with MIC of 2.22 mmol l(-1) and 18.20 mmol l(-1), respectively. Inductively coupled plasma atomic emission spectrometry (ICP-AES) showed that Pb inoculated in the growth medium is absorbed by the bacterial cells at removal efficiencies of 31.02% and 28.21% in the presence of 0.48 mmol l(-1) or 1.20 mmol l(-1) Pb(II), respectively. Strain Pb15 forms a brown and compact biofilm when grown in presence of Pb(II). Scanning Electron Microscopy (SEM) coupled with Energy Dispersive X-ray Spectroscopy (SEM-EDS) confirm that the biofilm contains Pb, suggesting an active biosorption of this metal by the bacterial cells, sequestering 14% of inoculated Pb as evidenced by microscopic analyses. Altogether, these observations support evidence that strain Pb15 has potentials for being used in bioremediation of its native polluted sediments, with engineering solutions to be found in order to eliminate the adsorbed Pb before replacement of sediments in situ. (C) 2016 Elsevier B. V. All rights reserved.

High-throughput sequencing of Prasinovirus DNA polymerase and host green algal (Mamiellophyceae) ribosomal RNA genes was used to analyse the diversity and distribution of these taxa over a ∼10 000 km latitudinal section of the Indian Ocean. New viral and host groups were identified among the different trophic conditions observed, and highlighted that although unknown prasinoviruses are diverse, the cosmopolitan algal genera Bathycoccus, Micromonas and Ostreococcus represent a large proportion of the host diversity. While Prasinovirus communities were correlated to both the geography and the environment, host communities were not, perhaps because the genetic marker used lacked sufficient resolution. Nevertheless, analysis of single environmental variables showed that eutrophic conditions strongly influence the distributions of both hosts and viruses. Moreover, these communities were not correlated, in their composition or specific richness. These observations could result from antagonistic dynamics, such as that illustrated in a prey–predator model, and/or because hosts might be under a complex set of selective pressures. Both of these reasons must be considered to interpret environmental surveys of viruses and hosts, because covariation does not always imply interaction.

While the effects of industrial contamination in coastal areas may persist for years in benthos communities, plankton should not show permanent impairments because of their high spatial dynamics, fast turnover times and pronounced seasonality. To test this hypothesis, in 2019 we conducted five surveys in the Bay of Pozzuoli (Gulf of Naples, Mediterranean Sea), in front of a dismissed steel factory and in the adjacent inshore coastal waters. High seasonal variability was observed for bacteria, phytoplankton and mesozooplankton, whereas plankton spatial gradients were relatively smooth during each survey. Plankton biomass and diversity did not reveal any effects of past industrial activities not even at the innermost stations of the Bay, which however showed some signals of present anthropogenic pressure. Hydrodynamic and morphological features likely play a prominent role in maintaining a relatively good status of the plankton of the Bay, which hints at the relevance of coastal circulation and meteorological dynamics to revitalize areas impacted by human activities.

Marine picoplankton, unicellular organisms with cell sizes up to 3 μm in diameter, numerically dominate marine ecosystems, encompassing Archaea, Bacteria, Eukarya (protists and fungi) as well as viruses. Autotrophic and heterotrophic picoplankton abundance and community composition with a focus on picoeukaryotes (PEs) were investigated in the winter of 2016 at three stations along a coast-to-offshore transect in the southern Adriatic Sea. Abundances were estimated by flow cytometry, while community composition by Illumina High-Throughput Sequencing (HTS) of 16S and 18S rRNA genes. The photosynthetic picoplankton diversity was also investigated by High-Performance Liquid Chromatography (HPLC) of liposoluble pigments. Heterotrophic bacteria and cyanobacteria (Prochlorococcus and Synechococcus) accounted for up to 7 × 105; 2.3 × 104 and 2.5 × 104 cells mL−1, respectively, while photosynthetic picoeukaryotes peaked with 3 × 103 cells mL−1. Prokaryotes, as revealed by HTS were dominated by Alphaproteobacteria (mainly SAR11, 44.91% of total 16S sequence reads), followed by Gammaproteobacteria (Oceanospirillales and Pseudomonadales, 14.96%), Bacteroidetes (mainly Flavobacteriales, 13%), Cyanobacteria (Prochlorococcus and Synechococcus, 9.52%), Marinimicrobia (SAR406, 7.97%), Deltaproteobacteria (SAR324, 3.83%), Actinobacteria (2.24%) and Chloroflexi (SAR202, 1.90%). Photosynthetic pigment concentrations were very low (12.12 μgL−1 at the most) and taxonomic pigments could be attributed to Prochlorococcus, Synechococcus, Prymnesiophyceae, Bacillariophyceae, Chrysophyceae, and Prasinophyceae. HTS data revealed that PEs were dominated by heterotrophs, such as Syndiniophyceae, parasitic dinoflagellates (79.67% of total 18S sequence reads), Dinophyceae (8.7%) and the radiolarians Collodaria belonging to Sphaerozoidae (22.1%) and Spumellaria (5.0%). On the other hand, photoautotrophs, including Chlorophyta (Mamiellophyceae, Prasinophyceae, Trebouxiophyceae, and Ulvophyceae), Stramenopiles (Bacillariophyta, Chrysophyceae, Dictyochophyceae, Pelagophyceae), photoautotrophic Cryptophyta and some Haptophyta (Prymnesiophyceae), did not exceed 5% of total sequence reads. This study provides the first snapshot of the PEs diversity in oligotrophic euphotic waters of the southern Adriatic Sea, hence setting the stage for large-scale surveying and characterization of the eukaryotic diversity in the entire basin.

Bacterioplankton are fundamental components of marine ecosystems and influence the entire biosphere by contributing to the global biogeochemical cycles of key elements. Yet, there is a significant gap in knowledge about their diversity and specific activities, as well as environmental factors that shape their community composition and function. Here, the distribution and diversity of surface bacterioplankton along the coastline of the Gulf of Naples (GON; Italy) were investigated using flow cytometry coupled with high-throughput sequencing of the 16S rRNA gene. Heterotrophic bacteria numerically dominated the bacterioplankton and comprised mainly Alphaproteobacteria , Gammaproteobacteria , and Bacteroidetes . Distinct communities occupied river-influenced, coastal, and offshore sites, as indicated by Bray-Curtis dissimilarity, distance metric (UniFrac), linear discriminant analysis effect size (LEfSe), and multivariate analyses. The heterogeneity in diversity and community composition was mainly due to salinity and changes in environmental conditions across sites, as defined by nutrient and chlorophyll a concentrations. Bacterioplankton communities were composed of a few dominant taxa and a large proportion (92%) of rare taxa (here defined as operational taxonomic units [OTUs] accounting for IMPORTANCE The study of bacterioplankton in coastal zones is of critical importance, considering that these areas are highly productive and anthropogenically impacted. Their richness and evenness, as well as their potential activity, are very important to assess ecosystem health and functioning. Here, we investigated bacterial distribution, community composition, and potential metabolic activity in the GON, which is an ideal test site due to its heterogeneous environment characterized by a complex hydrodynamics and terrestrial inputs of varied quantities and quality. Our study demonstrates that bacterioplankton communities in this region are highly diverse and strongly regulated by a combination of different environmental factors leading to their heterogeneous distribution, with the rare taxa contributing to a major proportion of diversity and shifts in community composition and potentially holding a key role in ecosystem functioning.

This paper presents a winter carbon budget for the northern Adriatic Sea, obtained through direct measurements during two multidisciplinary cruises and literature data. A box model approach was adopted to integrate estimates of stocks and fluxes of carbon species over the total area. The oligotrophy at the basin scale and the start of primary productivity well before the onset of spring stratification were observed. In winter, the system underwent a complete reset, as the mixing of water masses erased any signal of previous hypoxia or anoxia episodes. The northern Adriatic Sea was phosphorus depleted with respect to C and N availability. This fact confirms the importance of mixing with deep-sea water for P supply to biological processes on the whole. Despite the abundant prokaryotic biomass, the microbial food web was less efficient in organic C production than phytoplankton. In the upper layer, the carbon produced by primary production exceeded the fraction respired by planktonic community smaller than 200 µm. On the contrary, respiration processes prevailed in the water column below the pycnocline. The carbon budget also proved that the northern Adriatic Sea can be an effective sink for atmospheric CO2 throughout the entire winter season.

The effects of 4E/Z-octadienal (OCTA) on ScDSP-1 and ScDSP-2 gene expression and reactive oxygen species (ROS) production were investigated in the marine diatom Skeletonema tropicum (formerly costatum) using qRTPCR and flow cytometry. ScDSP-1 and ScDSP-2 genes have been previously shown to be involved in cell death in ageing cells and in response to photosynthetic stress. OCTA induced a differential, concentration-dependent DSP gene expression associated to ROS production, 821.6 and 97.7 folds higher for ScDSP-1 and ScDSP-2, respectively. Among the concentrations tested, only 8 μM OCTA, which caused a reduction of 50% in cell concentrations at 24 h, was able to elicit an expression pattern consistent with a signalling role. Interestingly, only intermediate levels of reactive oxygen species (ROS) (i.e., 1.5±0.1 increase) were observed to be elicited by such concentration. These results suggest that ROS are key components of the molecular cascade triggered by polyunsaturated aldehydes (PUA) and leading to cell death. This could have implications for bloom final stages at sea, where PUA may act as effectors of diatom population dynamics through ROS acting as modulators.

Polyunsaturated aldehydes (PUA) are supposed to play critical roles in chemically-mediated plankton interactions. Laboratory studies suggest that they act as mediators of chemical defense and chemical communication. PUA are oxylipins containing an alpha,beta,gamma,delta-unsaturated aldehyde structure element and are mainly found in diatoms. We present here a detailed surface mapping of PUA during a spring bloom of the diatom Skeletonema marinoi in the Adriatic Sea. We monitored dissolved PUA, as well as particulate PUA, which are produced by phytoplankton after cell disintegration. Our survey revealed a patchy distribution of PUA and shows that at most stations S. marinoi is the major contributor to the overall PUA. Our data also suggest that lysis of a diatom bloom can contribute significantly to the dissolved PUA concentrations and that other producers, which are smaller in cell size compared to diatoms, have to be taken into account as well if the total PUA content of marine samples is considered. The analyses of samples collected in deeper water suggests that diatom contribution to PUA decreases with depth, while smaller-sized unidentified organisms take place as dominant contributors to the PUA concentrations.

The planktonic diatom Pseudo-nitzschia multistriata is a potentially toxic species recorded during late summer–autumn in the Gulf of Naples (Tyrrhenian Sea, Italy). We analysed the genetic structure by amplifying the internal transcribed spacer (ITS-1–5.8S–ITS-2) region of the ribosomal DNA of 44 strains isolated along 2 years. Polymorphism in the ITS region was detected by direct sequencing and the PCR-products from selected strains were thus cloned to assess intra-strain ITS variability. Strains clustered into three main types: type A and B – differing by 0.6% sequence divergence – and type A/B, showing both A and B variants within the same genome. The three types showed no differences in the large subunit sequences (LSU) of the rDNA, ultrastructure of the valve, secondary structure of ITS-1 and ITS-2, ploidy level and they were sexually compatible. Based on the results of these multiple approaches, we can state that the three ITS-types belong to the same reproductive unit (or “species” sensu Mayr [(1942). Systematics and the Origin of Species. Columbia University Press, New York]). We suggest that ITS polymorphism in P. multistriata may be related to the contemporary occurrence of different but still inbreeding populations which either diverged recently or originated in different geographic areas and became sympatric in the studied area.

Egg production rates and/or hatching success in the copepods Acartia clausi, Calanus helgolandicus and Temora longicornis were negatively affected by a late spring (May-June 2003) phytoplankton bloom in the North Adriatic Sea, dominated mainly by the large diatom Cerataulina pelagica. Highest total concentrations of 3.3- 104 cells-ml(-1) were located in the vicinity of the Po River, which also corresponded to the area where the highest numbers of phaeophorbides were measured (0.779, 0.528 and 0.419 mu g.l(-1), respectively, compared to an average of the remaining stations of 0.183 +/- 0.049 SD), suggesting some grazing on the bloom. Phytoplankton biomass in terms of carbon was dominated by diatoms, representing on average 42% of total phytoplankton carbon and more than 80% at several stations. Cerataulina pelagica, Cyclotella spp., Chaetoceros spp. and small unidentified centric diatoms dominated the diatom community numerically but C. pelagica was by far the dominant diatom in terms of carbon due to its large cell size. This species represented more than 60% of the diatom biomass at nine of the 14 stations sampled, and was absent only at one station, which was the most offshore station sampled during the cruise. Although polyunsaturated aldehydes (PUAs) were not detected, other oxylipins which are hydroxy and keto derivatives of eicosapentaenoic and docosahexaenoic acids that affect copepod reproduction were found in these samples. Hence, we can attribute the negative impact of diatoms not only to PUAs, as previously believed, but also to these compounds. This is the first direct evidence of the presence of oxylipins other than PUAs in marine blooms dominated by diatoms.

Photophysiological parameters provide useful insights into the effects of environmental forcings on phytoplankton physiology. We present data on the short-term photoacclimative responses of phytoplankton throughout the water column during a diel sampling (every 1.5–3 h for 33 h) at a coastal station in the Gulf of Naples (Italy) in November 1996. Liposoluble pigments (high performance liquid chromatography), variable fluorescence (Prim-Prod probe) and picoplankton cell counts and autofluorescence (flow cytometry) were investigated every 1.5–3 h over a period of 33 h. The phytoplankton was phased to the alternation of light and dark and also showed acclimation to the different light intensities. Photoprotective pigments were synthesized during the day at the surface (0–20 m) and were significantly correlated with light intensity changes, as well as with the effective quantum yield of fluorescence. At night, recovery of photosystems from excess light was observed as was redistribution of nutrients and algae due to vertical convective motions caused by thermal dissipation. Equations linking photobiological parameters and time or light evolution were inferred to obtain kinetic coefficients. These were then used as biological tracers of vertical mixing whose velocity in the surface layer was estimated to be ,0.05 cm s 21 .

Bioactive polyunsaturated aldehydes (PUAs) are produced by several marine phytoplankton (mainly diatoms) and have been shown to have a detrimental effect on a wide variety of organisms, including phytoplankton and invertebrates. However, their potential impact on marine bacteria has been largely neglected. We assess here the effect of three PUAs produced by marine diatoms: 2E,4E-decadienal, 2E,4E-octadienal and 2E,4E-heptadienal, on the growth of 33 marine bacterial strains, including 16 strains isolated during a bloom of the PUA-producing diatom Skeletonema marinoi in the Northern Adriatic Sea. A concentration-dependent growth reduction was observed for 19 bacterial strains at concentrations ranging from 3 to 145 micromolL(-1). Surprisingly, Eudora adriatica strain MOLA358 (Flavobacteriaceae) and Alteromonas hispanica strain MOLA151 (Alteromonadaceae) showed growth stimulation upon exposure to PUAs at concentrations between 13 and 18 micromolL(-1). The remaining 12 strains were unaffected by even very high PUA concentrations. Strains isolated during the diatom bloom showed remarkable resistance to PUA exposures, with only two out of 16 strains showing growth inhibition at PUA concentrations below 106, 130, and 145 micromolL(-1) for 2E,4E-decadienal, 2E,4E-octadienal and 2E,4E-heptadienal, respectively. No correlation between taxonomical position and sensitivity to PUA was observed. Considering that many bacteria thrive in close vicinity of diatom cells, it is likely that these compounds may shape the structure of associated bacterial communities by representing a selection force. This is even more relevant during the final stages of blooms, when senescence and nutrient limitation increase the potential production and release of aldehydes.

Several marine diatoms produce polyunsaturated aldehydes (PUAs) that have been shown to be toxic to a wide variety of model organisms, from bacteria to invertebrates. However, very little information is available on their effect on phytoplankton. Here, we expand previous studies to six species of marine phytoplankton, belonging to different taxonomic groups that are well represented in marine plankton. The effect of three PUAs, 2 E ,4 E -decadienal, 2 E ,4 E -octadienal and 2 E ,4 E -heptadienal, was assessed on growth, cell membrane permeability, flow cytometric properties and morphology. A concentration-dependent reduction in the growth rate was observed for all cultures exposed to PUAs with longer-chained aldehydes having stronger effects on growth than shorter-chained aldehydes. Clear differences were observed among the different species. The prymnesiophyte Isochrysis galbana was the most sensitive species to PUA exposure with a lower threshold for an observed effect triggered by mean concentrations of 0.10 μmol L −1 for 2 E ,4 E -decadienal, 1.86 μmol L −1 for 2 E ,4 E -octadienal and 3.06 μmol L −1 for 2 E ,4 E -heptadienal, and a 50% growth inhibition (EC 50 ) with respect to the control at 0.99, 2.25 and 5.90 μmol L −1 for the three PUAs, respectively. Alternatively, the chlorophyte Tetraselmis suecica and the diatom Skeletonema marinoi (formerly S. costatum ) were the most resistant species with 50% growth inhibition occurring at concentrations at least two to three times higher than I. galbana . In all species, the three PUAs caused changes in flow cytometric measures of cell size and cell granulosity and increased membrane permeability, assessed using the viability stain SYTOX Green. For example, after 48 h 51.6 ± 2.6% of I. galbana cells and 15.0 ± 1.8% of S. marinoi cells were not viable. Chromatin fragmentation was observed in the dinoflagellate Amphidinium carterae while clear DNA degradation was observed in the chlorophyte Dunaliella tertiolecta . Concentrations used are in a significant range for affecting growth and performance of phytoplankton living in close vicinity of PUA-producing algae. Thus, PUAs may act as allelochemicals by mediating interactions among planktonic organisms.

Phytoplankton pigment diversity and photoacclimation during the natural day-night cycle was investigated at a fixed location in the Strait of Sicily in July 1997 using HPLC pigment analysis on fractionated samples ( 3 µm) and flow cytometry. Picophytoplankton dominated phytoplankton biomass in terms of chl a with an average value of 57% and was mainly represented by prokaryotes, prymnesiophytes and pelagophytes. Prochlorococcus and picoeukaryotes con- tributed equally to the picophytoplankton in terms of chl a, but Prochlorococcus were numerically more abundant and were represented by 2 ecotypes, one replacing the other according to depth. Larger phytoplankton were dominated by prymnesiophytes and diatoms. Photoacclimation was evi- dent from changes in pigment content and strongly increased with depth. The deep chlorophyll max- imum (DCM), present between 75 and 90 m, showed a diverse and rich phytoplankton community with the 2 size classes almost equally represented. Growth rates of Prochlorococcus and Synechococ- cus, estimated from cell cycle measurements, were 0.67 and 0.41 d -1 , respectively, at 75 m. Only Prochlorococcus was able to sustain a good growth rate of 0.43 d -1 at the base of the DCM (90 m) where only 0.5% of incident light was available. Light-shift experiments using onboard incubated natural seawater showed much faster kinetic coefficients for acclimation in picophytoplankton than in larger algae. In general, the data describe the dynamics of picophytoplankton and its light adapta- tion through the water column and in the DCM, and can be considered representative of stable sum- mer conditions in the Mediterranean Sea.

Polyunsaturated aldehydes (PUA) have recently been shown to induce reactive oxygen species (ROS) and possibly reactive nitrogen species (RNS, e.g., peroxynitrite) in the diatom Skeletonema marinoi (S. marinoi), which produces high amounts of PUA. We now are attempting to acquire better understanding of which reactive molecular species are involved in the oxidative response of S. marinoi to PUA. We used flow cytometry, the dye dihydrorhodamine 123 (DHR) as the main indicator of ROS (but which is also known to partially detect RNS), and different scavengers and inhibitors of both nitric oxide (NO) synthesis and superoxide dismutase activity (SOD). Both the scavengers Tempol (for ROS) and uric acid (UA, for peroxynitrite) induced a lower DHR-derived green fluorescence in S. marinoi cells exposed to the PUA, suggesting that both reactive species were produced. When PUA-exposed S. marinoi cells were treated with the NO scavenger 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), an opposite response was observed, with an increase in DHR-derived green fluorescence. A higher DHR-derived green fluorescence was also observed in the presence of sodium tungstate (ST), an inhibitor of NO production via nitrate reductase. In addition, two different SOD inhibitors, 2-methoxyestradiol (2ME) and sodium diethyldithiocarbamate trihydrate (DETC), had an effect, with DETC inducing the strongest inhibition after 20 min. These results indicate the involvement of O2 (•) generation and SOD activity in H2 O2 formation (with downstream ROS generation dependent from H2 O2 ) in response to PUA exposure. This is relevant as it refines the biological impact of PUA and identifies the specific molecules involved in the response. It is speculated that in PUA-exposed S. marinoi cells, beyond a certain threshold of PUA, the intracellular antioxidant system is no longer able to cope with the excess of ROS, thus resulting in the observed accumulation of both O2 (•-) and H2 O2 . This might be particularly relevant for population dynamics at sea, during blooms, when cell lysis increases and PUA are released. It can be envisioned that in the final stages of blooms, higher local PUA concentrations accumulate, which in turn induces intracellular ROS generation that ultimately leads to cell death and bloom decay.

Villar, Emilie ... et. al.-- 11 pages, 8 figures, supplementary materials www.sciencemag.org/content/348/6237/1261447/suppl/DC1, Agulhas rings provide the principal route for ocean waters to circulate from the Indo-Pacific to the Atlantic basin. Their influence on global ocean circulation is well known, but their role in plankton transport is largely unexplored. We show that, although the coarse taxonomic structure of plankton communities is continuous across the Agulhas choke point, South Atlantic plankton diversity is altered compared with Indian Ocean source populations. Modeling and in situ sampling of a young Agulhas ring indicate that strong vertical mixing drives complex nitrogen cycling, shaping community metabolism and biogeochemical signatures as the ring and associated plankton transit westward. The peculiar local environment inside Agulhas rings may provide a selective mechanism contributing to the limited dispersal of Indian Ocean plankton populations into the Atlantic. © 2015, American Association for the Advancement of Science. All rights reserved, We appreciate the commitment of the following people and sponsors: CNRS (in particular, Groupement de Recherche GDR3280); European Molecular Biology Laboratory (EMBL); Genoscope/CEA; VIB; Stazione Zoologica Anton Dohrn; UNIMIB; Fund for Scientific Research–Flanders; Rega Institute, KU Leuven; the French Ministry of Research; the French government Investissements d’Avenir programmes OCEANOMICS (ANR-11-BTBR-0008), FRANCE GENOMIQUE (ANR-10-INBS-09-08), MEMO LIFE (ANR-10-LABX-54), PSL* Research University (ANR-11-IDEX-0001-02), and ANR (projects POSEIDON/ ANR-09-BLAN- 0348, PHYTBACK/ANR-2010-1709-01, PROMETHEUS/ANR-09-PCS-GENM-217, SAMOSA/ANR-13-ADAP-0010, and TARAGIRUS/ANR-09-PCS-GENM-218); European Union FP7 (MicroB3/No.287589, IHMS/HEALTH-F4-2010-261376, and MaCuMBA/No.311975); ERC Advanced Grant Award to C.B. (Diatomite: 294823); Gordon and Betty Moore Foundation grant (no. 3790) to M.B.S.; Spanish Ministry of Science and Innovation grant CGL2011-26848/BOS MicroOcean PANGENOMICS to S.G.A.; TANIT (CONES 2010-0036) from the Agència de Gestió d´Ajusts Universitaris i Reserca to S.G.A.; JSPS KAKENHI grant no. 26430184 to H.O.; NASA Ocean Biology and Biogeochemistry program (NNX11AQ14G and NNX09AU43G) to E.B.; The Italian Research for the Sea (Flagship Project RITMARE) to D.I.; and FWO, BIO5, and Biosphere 2 to M.B.S.We also appreciate the support and commitment of Agnès b. and Etienne Bourgois, the Veolia Environment Foundation, Region Bretagne, Lorient Agglomeration, World Courier, Illumina, the EDF Foundation, FRB, the Prince Albert II de Monaco Foundation, and the Tara schooner and its captains and crew. We thank MERCATOR-CORIOLIS and ACRI-ST for providing daily satellite data during the expedition. We are also grateful to the French Ministry of Foreign Affairs for supporting the expedition and to the countries that graciously granted sampling permissions. Tara Oceans would not exist without continuous support from 23 institutes (http://oceans.taraexpeditions.org)

Agulhas rings provide the principal route for ocean waters to circulate from the Indo-Pacific to the Atlantic basin. Their influence on global ocean circulation is well known, but their role in plankton transport is largely unexplored. We show that, although the coarse taxonomic structure of plankton communities is continuous across the Agulhas choke point, South Atlantic plankton diversity is altered compared with Indian Ocean source populations. Modeling and in situ sampling of a young Agulhas ring indicate that strong vertical mixing drives complex nitrogen cycling, shaping community metabolism and biogeochemical signatures as the ring and associated plankton transit westward. The peculiar local environment inside Agulhas rings may provide a selective mechanism contributing to the limited dispersal of Indian Ocean plankton populations into the Atlantic. ispartof: Science vol:348 issue:6237 ispartof: location:United States status: published

High-throughput sequencing of Prasinovirus DNA polymerase and host green algal (Mamiellophyceae) ribosomal RNA genes was used to analyse the diversity and distribution of these taxa over a ∼10 000 km latitudinal section of the Indian Ocean. New viral and host groups were identified among the different trophic conditions observed, and highlighted that although unknown prasinoviruses are diverse, the cosmopolitan algal genera Bathycoccus, Micromonas and Ostreococcus represent a large proportion of the host diversity. While Prasinovirus communities were correlated to both the geography and the environment, host communities were not, perhaps because the genetic marker used lacked sufficient resolution. Nevertheless, analysis of single environmental variables showed that eutrophic conditions strongly influence the distributions of both hosts and viruses. Moreover, these communities were not correlated, in their composition or specific richness. These observations could result from antagonistic dynamics, such as that illustrated in a prey-predator model, and/or because hosts might be under a complex set of selective pressures. Both of these reasons must be considered to interpret environmental surveys of viruses and hosts, because covariation does not always imply interaction.

Most diatom species produce at least two classes of oxylipins, polyunsaturated aldehydes (PUAs) and non-volatile oxylipins (NVOs). These compounds can act as grazer deterrents by reducing the reproductive success of copepod grazers. The aim of the present study was to better understand the impact of late-winter diatom blooms and of diatom-derived PUAs and NVOs on the reproduction of two dominant copepod species in the northern Adriatic Sea, in two consecutive years. Higher diatom abundance in 2005 compared to 2004 coincided with higher egg-production rates in both Acartia clausi and Calanus helgolandicus. The increased contribution of diatoms to the phytoplankton assemblage in 2005 also led to a higher phytoplankton production of NVOs, whereas PUA levels were low in both years. These three-times-higher oxylipin levels correlated with a decrease in average hatching success in both A. clausi and C helgolandicus from similar to 80% in 2004 to similar to 60% in 2005, indicating that the observed negative effects on copepod recruitment in the latter year were related to the greater production of diatom NVOs. Our findings reveal the importance of these metabolites and offer an explanation as to why diatom blooms may be more toxic for copepods in some years compared to others. (C) 2015 Elsevier B.V. All rights reserved.