Marine protist diversity in European coastal waters and sediments as revealed by high-throughput sequencing

Although protists are critical components of marine ecosystems, they are still poorly characterized. Here we analysed the taxonomic diversity of planktonic and benthic protist communities collected in six distant European coastal sites. Environmental deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from three size fractions (pico-, nano- and micro/mesoplankton), as well as from dissolved DNA and surface sediments were used as templates for tag pyrosequencing of the V4 region of the 18S ribosomal DNA. Beta-diversity analyses split the protist community structure into three main clusters: picoplankton-nanoplankton-dissolved DNA, micro/mesoplankton and sediments. Within each cluster, protist communities from the same site and time clustered together, while communities from the same site but different seasons were unrelated. Both DNA and RNA-based surveys provided similar relative abundances for most class-level taxonomic groups. Yet, particular groups were overrepresented in one of the two templates, such as marine alveolates (MALV)-I and MALV-II that were much more abundant in DNA surveys. Overall, the groups displaying the highest relative contribution were Dinophyceae, Diatomea, Ciliophora and Acantharia. Also, well represented were Mamiellophyceae, Cryptomonadales, marine alveolates and marine stramenopiles in the picoplankton, and Monadofilosa and basal Fungi in sediments. Our extensive and systematic sequencing of geographically separated sites provides the most comprehensive molecular description of coastal marine protist diversity to date.

An evaluation of surface micro- and mesoplastic pollution in pelagic ecosystems of the Western Mediterranean Sea

This study examines the distribution, abundance and characteristics of surface micro- and mesoplastic debris in the Western Mediterranean Sea. 41 samples were collected in 2011 (summer) and 2012 (summer). Results, firstly, revealed that micro- (<5mm) and mesoplastic debris were widely and uniformly distributed in this area with average concentrations of 130,000 parts/km(2) and 5700 parts/km(2), respectively. Importantly, a strong correlation between micro- and mesoplastic concentrations was identified. Secondly, a classification based on the shape and appearance of microplastics indicated the predominant presence of fragments (73 %) followed by thin films (14 %). Thirdly, the average mass ratio of microplastic to dry organic matter has been measured at 0.5, revealing a significant presence of microplastics in comparison to plankton. Finally, a correction method was applied in order to correct wind mixing effect on microplastics' vertical distribution. This data allows for a comprehensive view, for the first time, of the spatial distribution and nature of plastic debris in the Western Mediterranean Sea.

High diversity of skin-associated bacterial communities of marine fishes is promoted by their high variability among body parts, individuals and species

Animal-associated microbiotas form complex communities, which are suspected to play crucial functions for their host fitness. However, the biodiversity of these communities, including their differences between host species and individuals, has been scarcely studied, especially in case of skin-associated communities. In addition, the intraindividual variability (i.e. between body parts) has never been assessed to date. The objective of this study was to characterize skin bacterial communities of two teleostean fish species, namely the European seabass (Dicentrarchus labrax) and gilthead seabream (Sparus aurata), using a high-throughput DNA sequencing method. In order to focus on intrinsic factors of host-associated bacterial community variability, individuals of the two species were raised in controlled conditions. Bacterial diversity was assessed using a set of four complementary indices, describing the taxonomic and phylogenetic facets of biodiversity and their respective composition (based on presence/absence data) and structure (based on species relative abundances) components. Variability of bacterial diversity was quantified at the interspecific, interindividual and intraindividual scales. We demonstrated that fish surfaces host highly diverse bacterial communities, whose composition was very different from that of surrounding bacterioplankton. This high total biodiversity of skin-associated communities was supported by the important variability, between host species, individuals and the different body parts (dorsal, anal, pectoral and caudal fins).

Tracking changes to a microplankton community in a North Atlantic sea loch using the microplankton index PI(mp).

Microplankton plays a vital part in marine ecosystems and its importance has been recognised by the inclusion of microplankton community composition in regulatory frameworks such as the European Water Framework Directive and the Marine Strategy Framework Directive as an indicator of ecological status. Quantitative techniques are therefore required to assess the environmental status of the microplankton in a water body. Here we demonstrate the use of a method known as the Microplankton Index PI(mp) to evaluate changes in the microplankton community of the West coast Scottish Sea Loch Creran. Microplankton in this fjord has been studied since the 1970’s providing a data set spanning four decades. Our analysis compares an arbitrarily chosen reference period between 1979 and 1981 with a period between 2011 and 2013 and demonstrates that between these two periods community structure has changed considerably with a substantial drop in the numbers of observed diatoms accompanied by a rise in the number of autotrophic/mixotrophic dinoflagellates as well as an increase in the potentially toxin producing genus Pseudo-nitzschia and that these are related to changes in both the intensity and timing of local patterns of precipitation. The PI(mp) is shown to be a useful and robust method to visualise and quantify changes in the underlying structure of the microplankton community and is a powerful addition to the toolbox of techniques needed to determine the health of our seas.

Quantification of microbial communities in subsurface marine sediments of the Black Sea and off Namibia

Organic-rich subsurface marine sediments were taken by gravity coring up to a depth of 10 m below seafloor at six stations from the anoxic Black Sea and the Benguela upwelling system off Namibia during the research cruises Meteor 72-5 and 76-1, respectively. The quantitative microbial community composition at various sediment depths was analyzed using total cell counting, catalyzed reporter deposition fluorescence in situ hybridization (CARD FISH) and quantitative real-time PCR (Q-PCR). Total cell counts decreased with depths from 10(9) to 10(10) cells/mL at the sediment surface to 10(7)-10(9) cells/mL below one meter depth. Based on CARD FISH and Q-PCR analyses overall similar proportions of Bacteria and Archaea were found. The down-core distribution of prokaryotic and eukaryotic small subunit ribosomal RNA genes (16S and 18S rRNA) as well as functional genes involved in different biogeochemical processes was quantified using Q-PCR. Crenarchaeota and the bacterial candidate division JS-1 as well as the classes Anaerolineae and Caldilineae of the phylum Chloroflexi were highly abundant. Less abundant but detectable in most of the samples were Eukarya as well as the metal and sulfate-reducing Geobacteraceae (only in the Benguela upwelling influenced sediments). The functional genes cbbL, encoding for the large subunit of RuBisCO, the genes dsrA and aprA, indicative of sulfate-reducers as well as the mcrA gene of methanogens were detected in the Benguela upwelling and Black Sea sediments. Overall, the high organic carbon content of the sediments goes along with high cell counts and high gene copy numbers, as well as an equal abundance of Bacteria and Archaea.

Anaerobic oxidation of methane at a marine methane seep in a forearc sediment basin off Sumatra, Indian Ocean

A cold methane seep was discovered in a forearc sediment basin off the island Sumatra, exhibiting a methane-seep adapted microbial community. A defined seep center of activity, like in mud volcanoes, was not discovered. The seep area was rather characterized by a patchy distribution of active spots. The relevance of anaerobic oxidation of methane (AOM) was reflected by C-13-depleted isotopic signatures of dissolved inorganic carbon. The anaerobic conversion of methane to CO2 was confirmed in a C-13-labeling experiment. Methane fueled a vital microbial community with cell numbers of up to 4 x 10(9) cells cm(-3) sediment. The microbial community was analyzed by total cell counting, catalyzed reporter deposition fluorescence in situ hybridization (CARD FISH), quantitative real-time PCR (qPCR), and denaturing gradient gel electrophoresis (DGGE). CARD FISH cell counts and qPCR measurements showed the presence of Bacteria and Archaea, but only small numbers of Eukarya. The archaeal community comprised largely members of ANME-1 and ANME-2. Furthermore, members of the Crenarchaeota were frequently detected in the DGGE analysis. Three major bacterial phylogenetic groups (delta-Proteobacteria, candidate division OP9, and Anaerolineaceae) were abundant across the study area. Several of these sequences were closely related to the genus Desulfococcus of the family Desulfobacteraceae, which is in good agreement with previously described AOM sites. In conclusion, the majority of the microbial community at the seep consisted of AOM-related microorganisms, while the relevance of higher hydrocarbons as microbial substrates was negligible.

When males are more inclined to stay at home :Insights into the partial migration of a pelagic seabird provided by geolocators and isotopes

Partial migration has never been studied in pelagic seabirds, but investigating old unresolved questions in new contexts can provide useful fresh insights. We used geolocators and stable isotopes to investigate this phenomenon in a migratory pelagic seabird, the Cory’s shearwater (Calonectris diomedea). Although most birds migrated to the southern hemisphere, 8.1% of studied birds (N = 172) remained close to the breeding colony (Selvagem Grande, Madeira, Portugal), foraging within the Canary current. Almost all resident birds were males, while age or body size did not predict migratory status. Despite displaying a high repeatability (R = 0.72) in the choice of wintering area, residency was not a fixed strategy and individuals could switch between migrating and staying in the Canary current in different years. The predictions resulting from the “body size” and the “social dominance” hypotheses, in which larger individuals or dominant individuals, respectively, remain closer to the breeding areas, were not supported by our data. Resident males were able to occupy the nesting burrows much earlier than migratory males and arrival time in this species is known to affect the probability of engaging in a reproductive attempt. The selective pressure to arrive early at the colony is therefore the most likely explanation for the maintenance of this partial migration system.

Peripheral communities of the Eastern Lau Spreading Center and Valu Fa Ridge: community composition, temporal change and comparison to near-vent communities

Western Pacific hydrothermal vents will soon be subjected to deep-sea mining and peripheral sites are considered the most practical targets. The limited information on community dynamics and temporal change in these communities makes it difficult to anticipate the impact of mining activities and recovery trajectories. We studied community composition of peripheral communities along a cline in hydrothermal chemistry on the Eastern Lau Spreading Center and Valu Fa Ridge (ELSC-VFR) and also studied patterns of temporal change. Peripheral communities located in the northern vent fields of the ELSC-VFR are significantly different from those in the southern vent fields. Higher abundances of zoanthids and anemones were found in northern peripheral sites and the symbiont-containing mussel Bathymodiolus brevior, brisingid seastars and polynoids were only present in the northern peripheral sites. By contrast, certain faunal groups were seen only in the southern peripheral sites, such as lollipop sponges, pycnogonids and ophiuroids. Taxonomic richness of the peripheral communities was similar to that of active vent communities, due to the presence of non-vent endemic species that balanced the absence of species found in areas of active venting. The communities present at waning active sites resemble those of peripheral sites, indicating that peripheral species can colonize previously active vent sites in addition to settling in the periphery of areas of venting. Growth and mortality were observed in a number of the normally slow-growing cladorhizid stick sponges, indicating that these animals may exhibit life history strategies in the vicinity of vents that differ from those previously recorded. A novel facultative association between polynoids and anemones is proposed based on their correlated distributions.

First metabolomic approach of the epiphytic bacteria-marine diatom Haslea ostrearia relationships

The marine diatom Haslea ostrearia [1] produces a water-soluble blue-pigment named marennine [2] of economic interest. But the lack of knowledge of the ecological conditions, under which this microalga develops in its natural ecosystem, more especially bacteria H. ostrearia interactions, prevents any optimization of its culture in well-controlled conditions. The structure of the bacterial community was analyzed by PCR-TTGE before and after the isolation of H. ostrearia cells recovered from 4 localities, to distinguish the relative part of the biotope and the biocenose and eventually to describe the temporal dynamic of the structure of the bacterial community at two time-scales. The differences in genetic fingerprints, more especially high between two H. ostrearia isolates (HO-R and HO-BM) showed also the highest differences in the bacterial structure [3] as the result of specific metabolomics profiles. The non-targeted metabolomic investigation showed that these profiles were more distinct in case of bacteria-alga associations than for the H. ostrearia monoculture Here we present a Q-TOF LC/MS metabolomic fingerprinting approach [3]: - to investigate differential metabolites of axenic versus non axenic H. ostrearia cultures. - to focus on the specific metabolites of a bacterial surrounding associated with the activation or inhibition of the microalga growing. The Agilent suite of data processing software makes feature finding, statistical analysis, and identification easier. This enables rapid transformation of complex raw data into biologically relevant metabolite information.

Preliminary metabolomic approach on the bacterial structure community of Haslea ostrearia

The marine diatom Haslea ostrearia [1] produces a water-soluble blue-pigment named marennine [2] of economic interest. But the lack of knowledge of the ecological conditions, under which this microalga develops in its natural ecosystem, more especially bacteria H. ostrearia interactions, prevents any optimization of its culture in well-controlled conditions. The structure of the bacterial community was analyzed by PCR-TTGE before and after the isolation of H. ostrearia cells recovered from 4 localities, to distinguish the relative part of the biotope and the biocenose and eventually to describe the temporal dynamic of the structure of the bacterial community at two time-scales. The differences in genetic fingerprints, more especially high between two H. ostrearia isolates (HO-R and HO-BM) showed also the highest differences in the bacterial structure [3] as the result of specific metabolomics profiles. The non-targeted metabolomic investigation showed that these profiles were more distinct in case of bacteria-alga associations than for the H. ostrearia monoculture Here we present a Q-TOF LC/MS metabolomic fingerprinting approach [3]: - to investigate differential metabolites of axenic versus non axenic H. ostrearia cultures. - to focus on the specific metabolites of a bacterial surrounding associated with the activation or inhibition of the microalga growing. The Agilent suite of data processing software makes feature finding, statistical analysis, and identification easier. This enables rapid transformation of complex raw data into biologically relevant metabolite information.

Analyse du compartiment mésozooplanctonique et écologie alimentaire printanière de la sardine, Sardina pilchardus (Walbaum, 1782), et de l’anchois, Engraulis encrasicolus (Linné, 1758) adultes dans le Golfe de Gascogne

Dietary studies of marine species constitute an important key to improve the understanding of its biology and of its role in the ecosystem. Thus, prey-predator relationships structure and determine population dynamics and the trophic network at the ecosystem scale. Among the major study sites, the marine ecosystem is submitted to natural and anthropogenic constraints. In the North-Eastern part of the Atlantic Ocean, the Bay of Biscay is a large open area surrounded South by Spain and East by France. This bay is an historic place of intense fishery activities for which the main small pelagic species targeted are the pilchard, Sardina pilchardus and the anchovy, Engraulis encrasicolus. The aim of this work is to analyze the trophic ecology of these two small pelagic fish in spring in the Bay of Biscay. To do this, a first section is devoted to their prey composed by the mesozooplanktonic compartment, through a two-fold approach: the characterization of their spatio-temporal dynamics during the decade 2003-2013 and the measurement of their energetic content in spring. For this season, it appears that all prey types are not worth energetically and that the Bay of Biscay represents a mosaic of dietary habitat. Moreover, the spring mesozooplankton community presents a strong spatial structuration, a temporal evolution marked by a major change in abundance and a control by the microphytoplankton biomass. The second section of this work is relative to a methodological approach of the trophic ecology of S. pilchardus and E. encrasicolus. Three different trophic tracers have been used: isotopic ratios of carbon and nitrogen, parasitological fauna and mercury contamination levels. To improve the use of the first of these trophic tracers, an experimental approach has been conducted with S. pilchardus to determine a trophic discrimination factor. Finally, it appears that the use of these three trophic tracers has always been permitted to highlight a temporal variability of the relative trophic ecology of these fish. However, no spatial dynamics could be identified through these three trophic tracers.

Evaluation of the antioxidant activity and antitumor activity of marine invertebrates extracts

Dissertação de Mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologias, Universidade do Algarve, 2014

White-faced storm-petrels Pelagodroma marina predated by gulls as biological monitors of plastic pollution in the pelagic subtropical northeast atlantic

Marine plastic pollution is rapidly growing and is a source of major concern. Seabirds often ingest plastic debris and are increasingly used as biological monitors of plastic pollution. However, virtually no studies have assessed plastics in seabirds in the deep subtropical North Atlantic. We investigated whether remains of white-faced storm-petrels (WFSP) present in gull pellets could be used for biomonitoring. We analysed 263 pellets and 79.0% of these contained plastic debris originating in the digestive tract of WFSP. Pellets with no bird prey did not contain plastics. Most debris were fragments (83.6%) with fewer plastic pellets (8.2%). Light-coloured plastics predominated (71.0%) and the most frequent polymer was HDPE (73.0%). Stable isotopes in toe-nails of WFSP containing many versus no plastics did not differ, indicating no individual specialisation leading to differential plastic ingestion. We suggest WFSP in pellets are highly suitable to monitor the little known pelagic subtropical Northeast Atlantic.

Effects of hook and bait on targeted and bycatch fishes in an equatorial atlantic pelagic longline fishery

We examined the effects of different hook style and bait type combinations on the catches of targeted, bycatch, and discarded fishes in equatorial Atlantic waters. In total, 221 longline sets (>305,000 hooks) were deployed from Portuguese pelagic longline vessels (SELECT -PAL Project) during the February–October fishing season. Three different hook styles and two bait types were tested: the traditional J-hook was compared to two circle hooks (one non-offset and one with 10° offset), and squid bait was compared to mackerel. Catch per unit effort (CPU Es) were calculated and compared between the different hook style and bait type combinations, which indicated that the effects of hook style and bait on the CPU Es were species-specific. For example, swordfish CPU Es were higher with J-hooks baited with squid, while for targeted tunas and blue shark only the bait effect was significant, but with opposite effect (i.e., higher catches of tuna with squid bait and higher catches of blue shark with mackerel bait). For the discarded species, at-haulback mortality was also species-specific. Proportions of alive vs dead specimens at time of fishing gear retrieval did not vary significantly by hook style or bait type combinations. The total retained catch was analyzed in value per unit effort (VPU E), and indicated losses in fishery revenue when mackerel was used instead of squid, but not when circle hooks were used instead of J-hooks.