Effect of ambient levels of ozone on photosynthetic components and radical scavenging system in leaves of African cowpea varieties

Tropospheric ozone (O3), a main component of photochemical oxidants, adversely affects not only human health but also vegetation. To clarify the long-term effects of ambient levels of tropospheric ozone (O3) on photosynthetic components and radical scavenging system in the leaves of cowpea ( Vigna unguiculata L.), two African varieties, Blackeye and Asontem, were grown in open-top chambers and exposed to filtered air (FA), non-filtered air (NF) or non-filtered air with additional O3 of approximately 50 nl l-1. Ambient levels of O3 significantly reduced chlorophyll concentration, quantum yield and activity of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco), thus contributing to the reduction in net photosynthetic rate at the reproductive growth stage of both varieties; with no significant variety difference in the sensitivity to O3. The O3-induced significant reduction in catalase activity was observed in Blackeye at vegetative and reproductive growth stages; and in Asontem at reproductive growth stage. On the other hand, exposure to O3 significantly increased ascorbate peroxidase activity in Blackeye at reproductive stage and did not significantly affect that in Blackeye at vegetative growth stage and that in Asontem at both growth stages. At reproductive growth stage, activities of monodehydroascorbate reductase and glutathione reductase were significantly increased by the exposure to O3 in both varieties. The results obtained in this study suggest that, although ascorbate peroxidase, monodehydroascorbate reductase and glutathione reductase played important roles in scavenging O3-induced reactive oxygen species in the leaves, radical scavenging ability of these enzymes is not sufficient to avoid detrimental effects of ambient levels of O3 on photosynthesis in both African cowpea varieties.

Light and dark carbon uptake by Dinophysis species in comparison to other photosynthetic and heterotrophic dinoflagellates

The marine dinoflagellate genus Dinophysis includes species that are the causative agents of diarrhetic shellfish poisoning (DSP). Recent findings indicate that some Dinophysis species are mixotrophic, i.e. capable of both autotrophic and heterotrophic nutrition. We investigated inorganic (and organic) carbon uptake by several species of Dinophysis in the Light and dark using the 'single-cell C-14 method', and compared uptake rates with those of photosynthetic Ceratium species and heterotrophic dinoflagellates in the genus Protoperidinium. Experiments were conducted with water from the Gullmar Fjord and from the Koster Strait (Swedish west coast). Nutrient-enriched phytoplankton from surface water samples were concentrated (20 to 70 mu m) and incubated at in situ temperature under artificial light conditions with high concentrations of inorganic C-14 (1 mu Ci ml(-1)). Individual cells of each desired species were manually isolated under a microscope and transferred to scintillation vials. C. tripes showed net C-14 uptake only during light periods, whereas both C. lineatum and C. furca showed C-14 uptake in the Light as well as uptake (and sometimes losses) in the dark. Dinophysis species had similar carbon fixation rates in Light compared to Ceratium species. For D. acuminata and D. norvegica, net carbon uptake occurred in both Light and dark periods. D. acuta showed a loss of carbon in the dark in one experiment, but in another, dark C uptake was significantly higher than uptake in Light. When exposed to Light, C. furca, D. norvegica and D. acuta had high specific carbon uptake rates. Growth rates for the different species were calculated from C-14 uptake by the cells during the first hours of incubation in light. D. acuminata and D. norvegica had similar maximum growth rates, 0.59 and 0.63 d(-1) (mu); the maximum growth rate of D. acuta was lower (0.41 d(-1)). The positive dark carbon uptake by Dinophysis may suggest a mixotrophic mode of nutrition. In one experiment, both D. norvegica and D. acuta showed a significantly higher carbon uptake in a dark bottle than in a Light bottle, which would be consistent with uptake of C-14-labeled organic matter by D. norvegica and D. acuta. Demonstration of direct uptake of dissolved and particulate organic matter would provide conclusive evidence of mixotrophy and this will require the development of new protocols for measuring organic matter uptake applicable to Dinophysis in the natural assemblages.

Report of the Working Group on Introductions and Transfers of Marine Organisms (WGITMO). 16-18 March 2016 Olbia, Italy

The 42th meeting of the ICES Working Group on Introductions and Transfers of Marine Organisms (WGITMO) was held in Olbia, Italy, 16–18 March 2016, with Anna Occhipinti-Ambrogi as host and Henn Ojaveer as chairperson. Representatives from 19 countries participated in the meeting. Attendants were from Belgium, Canada, Dennark, Estonia, Finland, France, Germany, Ireland, Israel, Italy, Lithuania, Norway, Poland, Portugal, Russia, Spain, Sweden, United Kingdom and United States. Sweden contributed by cor-respondence. The objectives of the meeting were to update information and discuss several aspects related to the introductions and transfers of non-indigenous aquatic species. Data and information management were two of the discussion topics of the meeting, with special focus on the better exploitation of the ‘Information system on aquatic non-indigenous and cryptogenic species’ (AquaNIS). The WGITMO also dedicated time for addressing the MSFD D2 issues: indicator on new non-indigenous species introduced by human activities, and opportunities and problems related to cross-regional comparison of non-indigenous species indicators. Preparation of the manuscript of the alert report on sea squirt Didemnum vexillum, which is to be published in ICES CRR series, was discussed and the steps to be taken to finalise the report were agreed. As usual, adequate time was devoted to discuss national reports, to exchange of information on the management of NIS and to review ongoing and planned research activities. The approach taken during the meeting facilitated presentations and discussions on the issues of relevance related to the Terms of References as well as on a few generic and strategically-important issues of general relevance to bioinvasions. The meeting began with a full-day joint meeting with the Working Group on Ballast and Other Ship Vectors (WGBOSV), which provided an opportunity to discuss and address issues of common interest, such as shipping and biofouling as introduction vectors. The proposed ICES demonstration advice on ‘Risk management of non-indigenous species associated with shipping in the Arctic’ was discussed, and edits were suggested for both the orientation of the demonstration advice as well as for the exact questions to be asked. Both working groups agreed that the practice of conducting back-to-back meetings with one joint day is useful and will continue in 2017. All Terms of References to be addressed for 2016 were discussed. For some Terms of Ref-erence, more detailed presentations were given, and a short overview of the information and subsequent discussion is provided herein at the end of each section. This report is structured so that each Term of Reference is dealt with in sequential order. The main body of the report contains summaries of the presentations and discussions with the more detailed documents being contained in the Annexes. WGITMO progressed each of the Terms of Reference by either completing the task or clearly identifying and agreeing on the inter-sessional activities required to still finalise the work in 2016. From 2017, WGITMO will be shifted to multi-annual management.

Rare sex or out of reach equilibrium? The dynamics of F-IS in partially clonal organisms

Background: Partially clonal organisms are very common in nature, yet the influence of partial asexuality on the temporal dynamics of genetic diversity remains poorly understood. Mathematical models accounting for clonality predict deviations only for extremely rare sex and only towards mean inbreeding coefficient (F-IS) over bar < 0. Yet in partially clonal species, both F-IS < 0 and F-IS > 0 are frequently observed also in populations where there is evidence for a significant amount of sexual reproduction. Here, we studied the joint effects of partial clonality, mutation and genetic drift with a state-and-time discrete Markov chain model to describe the dynamics of F-IS over time under increasing rates of clonality. Results: Results of the mathematical model and simulations show that partial clonality slows down the asymptotic convergence to F-IS = 0. Thus, although clonality alone does not lead to departures from Hardy-Weinberg expectations once reached the final equilibrium state, both negative and positive F-IS values can arise transiently even at intermediate rates of clonality. More importantly, such "transient" departures from Hardy Weinberg proportions may last long as clonality tunes up the temporal variation of F-IS and reduces its rate of change over time, leading to a hyperbolic increase of the maximal time needed to reach the final mean (F-IS,F-infinity) over bar value expected at equilibrium. Conclusion: Our results argue for a dynamical interpretation of F-IS in clonal populations. Negative values cannot be interpreted as unequivocal evidence for extremely scarce sex but also as intermediate rates of clonality in finite populations. Complementary observations (e.g. frequency distribution of multiloci genotypes, population history) or time series data may help to discriminate between different possible conclusions on the extent of clonality when mean (F-IS) over bar values deviating from zero and/or a large variation of F-IS over loci are observed.

Ecotoxicology of Nano-TiO2 An Evaluation of its Toxicity to Organisms of Aquatic Ecosystems

The production and use of synthetic nanoparticles is growing rapidly, and therefore the presence of these materials in the environment seems inevitable. Titanium dioxide (TiO2) presents various possible uses in industry, cosmetics, and even in the treatment of contaminated environments. Studies about the potential ecotoxicological risks of TiO2 nanoparticles (nano-TiO2) have been published but their results are still inconclusive. It should be noted that the properties of the diverse nano-TiO2 must be considered in order to establish experimental models to study their toxicity to environmentally relevant species. Moreover, the lack of descriptions and characterization of nanoparticles, as well as differences in the experimental conditions employed, have been a compromising factor in the comparison of results obtained in various studies. Therefore, the purpose of this paper is to make a simple review of the principal properties of TiO2, especially in nanoparticulate form, which should be considered in aquatic toxicology studies, and a compilation of the works that have been published on the subject.

Abundance of sedentary consumers and sessile organisms along the wave exposure gradient of subtropical rocky shores of the south-west Atlantic

Sedentary consumers play an important role on populations of prey and, hence, their patterns of abundance, distribution and coexistence on shores are important to evaluate their potential influence on ecosystem dynamics. Here, we aimed to describe their spatio-temporal distribution and abundance in relation to wave exposure in the intertidal rocky shores of the south-west Atlantic to provide a basis for further understanding of ecological processes in this system. The abundance and composition of the functional groups of sessile organisms and sedentary consumers were taken by sampling the intertidal of sheltered and moderately exposed shores during a period of one year. The sublittoral fringe of sheltered areas was dominated by macroalgae, while the low midlittoral was dominated by bare rock and barnacles. In contrast, filter-feeding animals prevailed at exposed shores, probably explaining the higher abundance of the predator Stramonita haemastoma at these locations. Limpets were more abundant at the midlittoral zone of all shores while sea urchins were exclusively found at the sublittoral fringe of moderately exposed shores, therefore, adding grazing pressure on these areas. The results showed patterns of coexistence, distribution and abundance of those organisms in this subtropical area, presumably as a result of wave action, competition and prey availability. It also brought insights on the influence of top-down and bottom-up processes in this area.

Toxicity of the therapeutic potassium permanganate to tilapia Oreochromis niloticus and to non-target organisms Ceriodaphnia dubia (microcrustacean cladocera) and Pseudokirchneriella subcapitata (green microalgae)

Potassium permanganate is a chemical compound widely used in aquaculture for the control and removal of parasites, and in the prevention of diseases caused by bacteria and fungi. However, this compound can be toxic to fish, being a strong oxidant. Moreover, there is no consistent information in the literature about its toxicity to non-target organisms. The purpose of this study was to evaluate the acute toxicity (LC50;96h) of potassium permanganate for tilapia, Oreochromis niloticus, and to determine its toxic effects on nontarget organisms using ecotoxicological assays performed with the microcrustacean Ceriodaphnia dubia and with the green microalgae Pseudokirchneriella subcapitata. The results showed that the concentration of 1.81 mg L-1 of potassium permanganate caused acute toxic effect in tilapia fingerlings. The ecotoxicological assays demonstrated that concentrations above 0.12 mg L-1 can cause chronic toxic effects on non-target organisms, indicating possible deleterious effects on the food chain of the aquatic ecosystem that may receive the discharge of effluents released by fish cultures treated with this chemotherapy. All toxic concentrations determined in this study were below those recommended in the literature for the use of this chemotherapy in fish cultures, demonstrating that this type of therapy should be more carefully considered in order to avoid damage to the treated fish and to the environment. (C) 2011 Elsevier B.V. All rights reserved.

Tools For Spatiotemporally Specific Proteomic Analysis In Multicellular Organisms

The emergence of mass spectrometry-based proteomics has revolutionized the study of proteins and their abundances, functions, interactions, and modifications. However, in a multicellular organism, it is difficult to monitor dynamic changes in protein synthesis in a specific cell type within its native environment. In this thesis, we describe methods that enable the metabolic labeling, purification, and analysis of proteins in specific cell types and during defined periods in live animals. We first engineered a eukaryotic phenylalanyl-tRNA synthetase (PheRS) to selectively recognize the unnatural L-phenylalanine analog p-azido-L-phenylalanine (Azf). Using Caenorhabditis elegans, we expressed the engineered PheRS in a cell type of choice (i.e. body wall muscles, intestinal epithelial cells, neurons, pharyngeal muscles), permitting proteins in those cells -- and only those cells -- to be labeled with azides. Labeled proteins are therefore subject to "click" conjugation to cyclooctyne-functionalized affnity probes, separation from the rest of the protein pool and identification by mass spectrometry. By coupling our methodology with heavy isotopic labeling, we successfully identified proteins -- including proteins with previously unknown expression patterns -- expressed in targeted subsets of cells. While cell types like body wall or pharyngeal muscles can be targeted with a single promoter, many cells cannot; spatiotemporal selectivity typically results from the combinatorial action of multiple regulators. To enhance spatiotemporal selectivity, we next developed a two-component system to drive overlapping -- but not identical -- patterns of expression of engineered PheRS, restricting labeling to cells that express both elements. Specifically, we developed a split-intein-based split-PheRS system for highly efficient PheRS-reconstitution through protein splicing. Together, these tools represent a powerful approach for unbiased discovery of proteins uniquely expressed in a subset of cells at specific developmental stages.

Refugium for surface life on Snowball Earth in a nearly enclosed sea? A numerical solution for sea-glacier invation through a narrow strait

Abstract Where photosynthetic eukaryotic organisms survived during the Snowball Earth events of the Neoproterozoic remains unclear. Our previous research tested whether a narrow arm of the ocean, similar to the modern Red Sea, could have been a refugium for photosynthetic eukaryotes during the Snowball Earth. Using an analytical ice-flow model, we demonstrated that a limited range of climate conditions could restrict sea-glacier flow sufficiently to allow an arm of the sea to remain partially free from sea-glacier penetration, a necessary condition for it to act as a refugium. Here we expand on the previous study, using a numerical ice-flow model, with the ability to capture additional physics, to calculate sea-glacier penetration, and to explore the effect of a channel with a narrow entrance. The climatic conditions are made selfconsistent by linking sublimation rate to surface temperature. As expected, the narrow entrance allows parts of the nearly enclosed sea to remain safe from sea-glacier penetration for a wider range of climate conditions.

Influence of Baculovirus anticarsia on the growth rate and survival of some nontarget aquatic organisms.

Recently, microbial pest control agents (MPCAs) have been worldwide used to reduce chemical pesticide use and to diminish the high risk of those compounds in the environment. Among various MPCAs, the nuclear polyhedrosis virus Baculovirus anticarsia is widely used in Brazil in the biological control of the velvet bean caterpillar. Although literature data do not show adverse effects of baculoviruses to nontarget organisms, it is necessary to evaluate their toxicity or patogenicity in order to study th environmental risk of those products and to register the formulations in the Brazilian Environmental Regularory Agency - IBAMA. In the presente work, the influence of a Baculovirus anticarsia formulation was evaluted to measure the consequences in the growth rateof the green algae Selenastrum capricornutum, the duckweed Lemna valdiviana and the microcrustacean Daphnia similis. The survival of the fish Hyphessobrycon scholzei exposed during 28 days was also evaluated. No significative adverse effects (P > 0.05) were observed in the test organisms which were exposed to 1-1000 times the maximum calculated pesticide concentration following a direct application to 15 cm layer of water.

Prediction of a low-risk concentration of diflubenzuron to aquatic organisms and evaluation of clay and gravel in reducing the toxicity.

Resumo: Predição da concentração de baixo risco de diflubenzuron para organismos aquáticos e avaliação da argila e brita na redução da toxicidade. O diflubenzuron é um inseticida que além de ser usado agricultura, tem sido amplamente empregado na piscicultura, apesar do seu uso ser proibido nesta atividade. Este composto não consta na lista da legislação brasileira que estabelece limites máximos permissíveis em corpos de água para a proteção das comunidades aquáticas. No presente trabalho, a partir da toxicidade do diflubenzuron em organismos não-alvo, foi calculada a concentração de risco para somente 5% das espécies (HC5). O valor deste parâmetro foi estimado em aproximadamente 7 x 10-6 mg L-1 . Este baixo valor é devido à extremamente alta toxicidade do diflubenzuron para dafnídeos e à grande variação de sensibilidade entre as espécies testadas. Dois matérias de relativamente baixo custo se mostraram eficientes na remoção da toxicidade do diflubenzuron de soluções contendo este composto. Dentre esses materiais, a argila expandida promoveu a redução em aproximadamente 50% da toxicidade de uma solução contendo diflubenzuron. Os resultados podem contribuir para políticas públicas no Brasil relacionadas ao estabelecimento de limites máximos permissíveis de xenobióticos no compartimento aquático. Também, para a pesquisa de matérias inertes e de baixo custo com potencial de remoção de xenobióticos presentes em efluentes da aquicultura ou da agricultura. Abstract: Diflubenzuron is an insecticide that, besides being used in the agriculture, has been widely used in fish farming. However, its use is prohibited in this activity. Diflubenzuron is not in the list of Brazilian legislation establishing maximum permissible limits in water bodies for the protection of aquatic communities. In this paper, according toxicity data of diflubenzuron in non-target organisms, it was calculated an hazardous concentration for only 5% of the species (HC5) of the aquatic community. This parameter value was estimated to be about 7 x 10 -6 mg L -1 . The low value is due to the extreme high toxicity of diflubenzuron to daphnids and to the large variation in sensitivity among the species tested. Two relatively low cost and inert materials were efficient in removing the diflubenzuron from solutions containing this compound. Among these materials, expanded clay shown to promote reduction of approximately 50% of the toxicity of a solution containing diflubenzuron. The results may contribute to the establishment of public policies in Brazil associated to the definition of maximum permissible limits of xenobiotics in the aquatic compartment. This study is also relevant to the search of low cost and inert materials for xenobiotics removal from aquaculture or agricultural effluents.

Relationship between phenotype and environment in marine calcifying organisms: exploring growth and shell properties of different mollusks species in past and modern scenario

Coastal ecosystems represent an inestimable source of biodiversity, being among the most productive areas on the planet. Despite the great ecological and economic value of those environments, many threats endanger the species living in this ecosystem, like the rapid warming and the sea acidification, among many other. Benthic calcifying organisms (e.g. mollusks, corals and echinoderms) in particular, are among the most exposed to those hazards. These organisms use calcium carbonate as a structural and protective material through the biomineralization process, biologically controlled by the organism, but nevertheless, strongly influenced by the environmental surroundings. Evaluating how a changing environment can influence the process of biomineralization is critical to understand how those species of great ecological and economic importance will face the ongoing climate change. This thesis investigates the mechanism of biomineralization in different mollusks’ species of the Adriatic Sea, providing detailed descriptions of shells skeletal, biometric and growth parameters. Applying a multidisciplinary and multi-scale research approach, the influence of external environmental factors on the process of shell formation has been investigated. To achieve this purpose analysis were conducted both on current populations and on fossil remain, which allows to investigate ecological responses to past climate transitions. Mollusks’ shells in fact are one of the best tools to understand climate change in the past, present and future, since they record the environmental conditions prevailed during their life, reflected on the geochemical properties, microstructure and growth of the shell. This approach allowed to overcome the time scale limit imposed by field and laboratory survey, and better understand species long term adaptive response to changing environment, a crucial issue to define proper conservation and management strategies. Furthermore, the investigation of fossil record of mollusks assemblages offered the opportunity to evaluate the long-term biotic response to anthropogenic stressors in the north Adriatic Sea.