Abrupt shifts in the Gironde fish community: an indicator of ecological changes in an estuarine ecosystem

For decades, global climate change has directly and indirectly affected the structure and function of ecosystems. Abrupt changes in biodiversity have been observed in response to linear or sudden modifications to the environment. These abrupt shifts can cause long-term reorganizations within ecosystems, with communities exhibiting new functional responses to environmental factors. Over the last 3 decades, the Gironde estuary in southwest France has experienced 2 abrupt shifts in both the physical and chemical environments and the pelagic community. Rather than describing these shifts and their origins, we focused on the 3 inter-shift periods, describing the structure of the fish community and its relationship with the environment during these periods. We described fish biodiversity using a limited set of descriptors, taking into account both species composition and relative species abundances. Inter-shift ecosystem states were defined based on the relationship between this description and the hydro-physico-chemical variables and climatic indices defining the main features of the environment. This relationship was described using generalized linear mixed models on the entire time series and for each inter-shift period. Our results indicate that (1) the fish community structure has been significantly modified, (2) environmental drivers influencing fish diversity have changed during these 3 periods, and (3) the fish-environment relationships have been modified over time. From this, we conclude a regime shift has occurred in the Gironde estuary. We also highlight that anthropogenic influences have increased, which re-emphasizes the importance of local management in maintaining fish diversity and associated goods and services within the context of climate change.

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

Successional dynamics in inland dune shrub communities drive changes in functional diversity

Communities can be defined as assemblages of species coexisting under particular environments. The relationship between environment and species are regulated by both environmental requirements –which ultimately determine the species capacity to establish and survive in a particular environment– and the ecological interactions occurring during assembly processes –which also determine community composition by conditioning species coexistence. In this context, plant functional traits are attributes that represent ecological strategies and determine how plants respond to environmental factors and interact with other species. Therefore, the analysis of how traits vary through the dynamics of communities, such as along successions, can give insights about how environmental requirements and species interactions may determine the composition and functional structure of these communities. The xerophytic shrub communities inhabiting inland sand dunes in SW Portugal are characterized by successional processes that are mainly driven by local (edaphic gradients and human disturbance) and regional (climate) processes. Therefore, they constitute an appropriate system for studying species interactions and environmentcommunity co-variations based on functional terms. Using these communities as a model, we evaluate the hypothesis that successional community changes in species composition of xerophytic shrub communities can result in concurrent changes in functional diversity

Characterization of a plankton community in a fish farm

Aim: The objective of this study was to analyze the composition of phytoplankton and zooplankton communities related to the dynamics of a fish farm; Methods: Samples were collected every other day, within a period of twenty consecutive days, during the dry and rainy seasons. Two samples were collected upstream from the fish pond (reservoir); the other four samples were collected in the fish farm area; Results: Rotifera and Chlorophyceae species were found in high densities at almost all sampling sites during both seasons under analysis. The higher phytoplankton species richness from site P3 to P6 was influenced by the management employed within the fish farm. The zooplankton community showed low values of density, species richness and equitability during the dry season; Conclusions: The continuous water flow and the addition of fertilizers (organic and inorganic) in the fish ponds had an influence on the plankton community, leading to a reduction in water quality and Cyanobacteria dominance.

Análise da estrutura e transporte do zooplâncton no estuário do Rio Caravelas (Bahia-Brasil) através do uso do zooscan

Spatial-temporal dynamics of zooplankton in the Caravelas river estuary (Bahia, Brazil). The survey was conducted in order to describe the zooplankton community of the estuary Caravelas (Bahia, Brazil), to quantify and relate the patterns of horizontal and vertical transport with the type of tide (neap and spring) and tidal phase (flood and ebb). Zooplankton samples were collected with the aid of a suction pump (300L), filtered in plankton nets (300μm) and fixed in saline formalin 4%. Samples were collected at a fixed point (A1), near the mouth of the estuary, with samples taken at neap tides and spring tides during the dry and rainy seasons. Samples were collected for 13 hours, at intervals of 1 hour in 3 depths: surface, middle and bottom. Simultaneous collection of biological, we measured the current velocity, temperature and salinity of the water through CTD. In the laboratory, samples were selected for analysis in estereomicroscope, with 25 groups identified, with Copepoda getting the highest number of species. The 168 samples obtained from temporal samples were subsampled and processed on equipment ZooScan, with the aid of software ZooProcess at the end were generated 458.997 vingnettes. 8 taxa were identified automatically, with 16 classified as a semi-automatic. The group Copepoda, despite the limited taxonomic refinement ZooScan, obtained 2 genera and 1 species identified automatically. Among the seasons dry and wet groups Brachyura (zoea), Chaetognatha, and the Calanoid copepods (others), Temora spp., Oithona spp. and Euterpina acutifrons were those who had higher frequency of occurrence, appearing in more than 70% of the samples. Copepoda group showed the largest percentage of relative abundance in both seasons. There was no seasonal variation of total zooplankton, with an average density of 7826±4219 org.m-3 in the dry season, and 7959±3675 org.m-3 in the rainy season, neither between the types and phases of the tides, but seasonal differences were significant recorded for the main zooplankton groups. Vertical stratification was seen for the major zooplankton groups (Brachyura, Chaetognatha, Calanoida (other), Oithona spp, Temora spp. e Euterpina acutifrons). The scale of this stratification varied with the type (square or tide) and tidal phase (flood or ebb). The instantaneous transport was more influenced by current velocity, with higher values observed in spring tides to the total zooplankton, however, there was a variation of this pattern depending on the zooplankton group. According to the data import and export of total zooplankton, the outflow of organisms of the estuary was higher than the input. The results suggest that the estuary of Caravelas may influence the dynamics of organic matter to the adjacent coast, with possible consequences in National Marine Park of Abrolhos

Environmental Dynamics of Dissolved Black Carbon in Aquatic Ecosystems

Black carbon (BC), the incomplete combustion product from biomass and fossil fuel burning, is ubiquitously found in soils, sediments, ice, water and atmosphere. Because of its polyaromatic molecular characteristic, BC is believed to contribute significantly to the global carbon budget as a slow-cycling, refractory carbon pool. However, the mass balance between global BC generation and accumulation does not match, suggesting a removal mechanism of BC to the active carbon pool, most probable in a dissolved form. The presence of BC in waters as part of the dissolved organic matter (DOM) pool was recently confirmed via ultrahigh resolution mass spectrometry, and dissolved black carbon (DBC), a degradation product of charcoal, was found in marine and coastal environments. However, information on the loadings of DBC in freshwater environments and its global riverine flux from terrestrial systems to the oceans remained unclear. The main objectives of this study were to quantify DBC in diverse aquatic ecosystems and to determine its environmental dynamics. Surface water samples were collected from aquatic environments with a spatially significant global distribution, and DBC concentrations were determined by a chemical oxidation method coupled with HPLC detection. While it was clear that biomass burning was the main sources of BC, the translocation mechanism of BC to the dissolved phase was not well understood. Data from the regional studies and the developed global model revealed a strong positive correlation between DBC and dissolved organic carbon (DOC) dynamics, indicating a co-generation and co-translocation between soil OC and BC. In addition, a DOC-assistant DBC translocation mechanism was identified. Taking advantage of the DOC-DBC correlation model, a global riverine DBC flux to oceans on the order of 26.5 Mt C yr-1 (1 Mt = 1012 g) was determined, accounting for 10.6% of the global DOC flux. The results not only indicated that DOC was an important environmental intermediate for BC transfer and storage, but also provided an estimate of a major missing link in the global BC budget. The ever increasing DBC export caused by global warming will change the marine DOM quality and may have important consequences for carbon cycling in marine ecosystem.

Novel insights into the dynamics of green turtle fibropapillomatosis

Outbreaks of fibropapillomatosis (FP), a neoplastic infectious disease of marine turtles, have occurred worldwide since the 1980s. Its most likely aetiological agent is a virus, but disease expression depends on external factors, typically associated with altered environments. The scarcity of robust long-term data on disease prevalence has limited interpretations on the impacts of FP on turtle populations. Here we model the dynamics of FP at 2 green turtle foraging aggregations in Puerto Rico, through 18 yr of capture-mark-recapture data (1997−2014). We observed spatiotemporal variation in FP prevalence, potentially modulated via individual site-fidelity. FP ex pression was residency dependent, and FP-free individuals developed tumours after 1.8 ± 0.8 yr (mean ± SD) in the infected area. Recovery from the disease was likely, with complete tumour regression occurring in 2.7 ± 0.7 yr (mean ± SD). FP does not currently seem to be a major threat to marine turtle populations; however, disease prevalence is yet unknown in many areas. Systematic monitoring is highly advisable as human-induced stressors can lead to deviations in host− pathogen relationships and disease virulence. Finally, data collection should be standardized for a global assessment of FP dynamics and impacts.

Composition and temporal dynamics of a temperate rocky cryptobenthic fish assemblage

Temporal variation in a temperate cryptobenthic fish assemblage at the Arrabida Marine Park (Portugal) was assessed by visual Surveys during 2002 and 2003. A total of 9596 fish from 11 families and 30 species was recorded. There were no changes in structure or density at the assemblage level between years, whereas diversity changed significantly due to a higher number of abundant species in the second year. A similar seasonal trend was found between years, with a significant overall density increase in autumn. This is partially explained by the arrival of new recruits of some of the most abundant species in the assemblage. Assemblage diversity and structure also changed across seasons. A group of species encompassing Gobius xanthocephalus, Tripterygion delaisi, Parablennius pilicornis, Gobius paganellus, Lepadogaster candollii and Lepadogaster spp. were analysed in detail. The temporal patterns of two of the most abundant species, G. xanthocephalus and T delaisi, mimicked the overall temporal patterns of the assemblage. We suggest that the inter-annual stability in density of this subtidal fish assemblage may be similar to what has been reported for the intertidal and that strong post-settlement processes are probably shaping this assemblage.

Transboundary dimensions of marine spatial planning: Fostering inter-jurisdictional relations and governance

There is broad agreement that marine spatial planning (MSP) should incorporate transboundary considerations, reflecting the cross-border nature of marine and coastal ecosystem dynamics and maritime resources and activities. This is recognised in the European Union's recent legislation on MSP, and experience in transboundary approaches is developing through official processes and pilot studies.

Development of molecular approaches to assess the diversity and abundance of marine nematodes assemblages for assessment of Good Environmental Status of the estuarine and marine ecosystems

In Europe, the concerns with the status of marine ecosystems have increased, and the Marine Directive has as main goal the achievement of Good Environmental Status (GES) of EU marine waters by 2020. Molecular tools are seen as promising and emerging approaches to improve ecosystem monitoring, and have led ecology into a new era, representing perhaps the most source of innovation in marine monitoring techniques. Benthic nematodes are considered ideal organisms to be used as biological indicator of natural and anthropogenic disturbances in aquatic ecosystems underpinning monitoring programmes on the ecological quality of marine ecosystems, very useful to assess the GES of the marine environment. dT-RFLP (directed Terminal-Restriction Fragment Length Polymorphism) allows to assess the diversity of nematode communities, but also allows studying the functioning of the ecosystem, and combined with relative real-time PCR (qPCR), provides a high-throughput semi-quantitative characterization of nematode communities. These characteristics make the two molecular tools good descriptors for the good environmental status assessment. The main aim of this study is to develop and optimize the dT-RFLP and qPCR in Mira estuary (SW coast, Portugal). A molecular phylogenetic analysis of marine and estuarine nematodes is being performed combining morphological and molecular analysis to evaluate the diversity of free-living marine nematodes in Mira estuary. After morphological identification, barcoding of 18S rDNA and COI genes are being determined for each nematode species morphologically identified. So far we generated 40 new sequences belonging to 32 different genus and 17 families, and the study has shown a good degree of concordance between traditional morphology-based identification and DNA sequences. These results will improve the assessment of marine nematode diversity and contribute to a more robust nematode taxonomy. The DNA sequences are being used to develop the dT-RFLP with the ability to easily process large sample numbers (hundreds and thousands), rather than typical of classical taxonomic or low throughput molecular analyses. A preliminary study showed that the digest enzymes used in dT-RFLP for terrestrial assemblages separated poorly the marine nematodes at taxonomic level for functional group analysis. A new digest combination was designed using the software tool DRAT (Directed Terminal Restriction Analysis Tool) to distinguished marine nematode taxa. Several solutions were provided by DRAT and tested empirically to select the solution that cuts most efficiently. A combination of three enzymes and a single digest showed to be the best solution to separate the different clusters. Parallel to this, another tool is being developed to estimate the population size (qPCR). An improvement in qPCR estimation of gene copy number using an artificial reference is being performed for marine nematodes communities to quantify the abundance. Once developed, it is proposed to validate both methodologies by determining the spatial and temporal variability of benthic nematodes assemblages across different environments. The application of these high-throughput molecular approaches for benthic nematodes will improve sample throughput and their implementation more efficient and faster as indicator of ecological status of marine ecosystems.

Federative dynamics in the EU under the influence of EU citizenship rights in times of crisis

The federative debate has been tied to the history of the EU integration from the start. Even though the EU is a polity in formation, the comparison with the state model has limited usefulness in assessing the evolution of federative dynamics in the EU. In the context of the EU, federalism should not be regarded as a static arrangement, but rather as a process that has been unfolding despite or because of EU integration setbacks. By the same token, EU citizenship is necessarily distinct from national citizenship. Our purpose in this article is to understand the federative dynamics of the EU in relation to the emergence of a “community of rights”. We also aim to establish if, and how, the on-going crisis has triggered a change in the EU federative process as a result of the weakening/strengthening of citizenship rights.

Impact of anthropogenic stressors on the microbial communities in marine holobionts and ecosystems

Free-living or host-associated marine microbiomes play a determinant role in supporting the functioning and biodiversity of marine ecosystems, providing essential ecological services, and promoting the health of the entire biosphere. Currently, the fast and restless increase of World’s human population strongly impacts life on Earth in the forms of ocean pollution, coastal zone destruction, overexploitation of marine resources, and climate change. Thanks to their phylogenetic, metabolic, and functional diversity, marine microbiomes represent the Earth’s biggest reservoir of solutions against the major threats that are now impacting marine ecosystems, possibly providing valuable insights for biotechnological applications to preserve the health of the ocean ecosystems. Microbial-based mitigation strategies heavily rely on the available knowledge on the specific role and composition of holobionts associated microbial communities, thus highlighting the importance of pioneer studies on microbial-mediated adaptive mechanisms in the marine habitats. In this context, we propose different models representing ecologically important, widely distributed, and habitat-forming organisms, to further investigate the ability of marine holobionts to dynamically adapt to natural environmental variations, as well as to anthropogenic stress factors. In this PhD thesis, we were able to supply the characterization of the microbial community associated with the model anthozoan cnidaria Corynactis viridis throughout a seasonal gradient, to provide critical insights into microbiome-host interactions in a biomonitoring perspective. We also dissected in details the microbial-derived mitigation strategies implemented by the benthonic anthozoan Anemonia viridis and the gastropod Patella caerulea as models of adaptation to anthropogenic stressors, in the context of bioremediation of human-impacted habitats and for the monitoring and preservation of coastal marine ecosystems, respectively. Finally, we provided a functional model of adaptation to future ocean acidification conditions by characterizing the microbial community associated with the temperate coral Balanophyllia europaea naturally living at low pH conditions, to implement microbial based actions to mitigate climate change.