El objetivo de la Tesis es obtener una visión de la situación actual de la laguna costera del Mar Menor en relación con la contaminación por metales pesados procedentes de actividades mineras mediante el empleo de herramientas ecotoxicológicas. Comienza con el estudio de la entrada de residuos mineros en el ecosistema, su distribución en las aguas de la laguna así como los efectos tóxicos asociados. Continua valorando la biodisponibilidad de los metales contenidos en los sedimentos lagunares mediante pruebas de toxicidad y de bioacumulación; sus efectos en praderas de fanerógamas y comunidades de invertebrados asociados, para continuar determinando la potencial transferencia trófica de metales en las redes alimenticias lagunares y su posible biomagnificación. Finalmente, la Tesis concluye determinando la utilidad de indicadores propuestos para su aplicación en la Directiva Europea Marco del Agua cuando son aplicados a dos ecosistemas acuáticos contaminados por contaminación tóxica (metales pesados). / The objective of the Thesis is to determine the present situation of the Mar Menor coastal lagoon in relation with the contamination caused by heavy metals coming from old mining activities through the use of ecotoxicological tools. It begins with the study of the entrance of mining wastes in the lagoon ecosystem, its distribution in the lagoon waters as well as their toxic effects. It continues assessing the bioavailability of metals contained in sediments of the lagoon by means of bioaccumulation studies and toxicity tests; their possible deleterious effects on marine phanerogams and associated invertebrate communities. It also determines the potential trophic transfer of metals in trophic webs of the lagoon studying the possible biomagnification of metals. Finally, the Thesis concludes determining the utility of some indicators proposed for their application in the European Water Framework Directive when they are applied to aquatic ecosystems impacted by toxic contamination (heavy metals).

Pollution by toxic compounds is one of the most relevant environmental damages to ecosystems produced by human activity and, therefore, it must be considered in environmental protection and restoration of contaminated sites. According to this purposes, the main goal of this doctoral thesis has been to analyse the impact of several chlorophenols and heavy metals on the microbial communities of two typical Mediterranean soils. The ecological risk concentrations of each pollutant, which have been determined according to respirometric activity and changes in the microbial community composition, and the factors that influence on their effective toxic concentrations (bioavailable pollutants) have been analysed in order to predict their potential impact on different soil ecosystems and provide scientific data for the regulation of the soil protection policies. Moreover, resistant microorganisms with pollutant removal capacities have been isolated from artificially contaminated soil microcosms and tested in axenic cultures, to infer their potential usefulness for bioremediation. / La contaminació amb compostos tòxics antropogènics és un dels perjudicis ambientals més rellevants per als ecosistemes i, per tant, cal tenir-ho en compte en la protecció ambiental i la restauració de llocs contaminats. Amb aquests propòsits, l’objectiu principal d’aquesta tesi doctoral ha estat analitzar l’impacte de clorofenols i metalls pesants sobre la microbiota de dos sòls típicament mediterranis. S’han analitzat les concentracions de risc ecològic de cada contaminant, determinades segons activitat respiromètrica i canvis en la composició de la comunitat microbiana, i els factors que influeixen en les seves concentracions tòxiques efectives (contaminants biodisponibles), per predir l’impacte potencial sobre diferents ecosistemes edàfics i proporcionar evidències científiques per a les polítiques de protecció del sòl. D’altra banda, a partir de microcosmos edàfics contaminats artificialment, es van aïllar també microorganismes resistents amb capacitat per eliminar els contaminants, que van ser analitzats en cultius purs per avaluar-ne la utilitat potencial en bioremediació.

The present thesis aims to evaluate the use of antioxidant enzyme activities (AEA) of biofilm communities as biomarkers of metal pollution (mainly Zn) in fluvial ecosystems. In order to test AEA as biomarkers of metal pollution as well as their responses to environmental variables, several ecotoxicological experiments have been performed. A zoom from field studies (with high ecological realism) to microcosm experiments (under controlled conditions) has been done to fill the gap between both approaches and understand AEA responses under a multiple-stress (environmental and metallic) scenario. The field studies were carried out in the Riera d’Osor, a tributary of the Ter, located in the region of La Selva (Girona province) / L’objectiu d’aquesta tesi és avaluar l’ús de les activitats enzimàtiques antioxidants (AEA) dels biofilms com a biomarcadors de contaminació metàl·lica (principalment per zinc (Zn)) en els ecosistemes fluvials. Per tal d’estudiar la resposta de les AEA a la contaminació metàl•lica, així com també la resposta a altres canvis ambientals (situacions d’estrès múltiple), s’han realitzat diversos experiments ecotoxicològics fent un zoom des d’estudis de camp (amb un elevat realisme ecològic) fins a un estudi de laboratori utilitzant microcosmos (amb condicions controlades). Els estudis de camp s’han dut a terme a la riera d’Osor, afluent del Ter, situada a la comarca de la Selva (província de Girona)

Orientador: Denis Moledo de Souza Abessa / Resumo: Alguns contaminantes encontrados no ambiente costeiro incluem os princípios ativos de tintas anti-incrustantes, entre os quais se destaca o Irgarol. O Irgarol pode interagir com outros contaminantes comuns em áreas litorâneas antropizadas, tais como a amônia e os surfactantes, sendo que os efeitos dessas combinações são desconhecidos. O objetivo do presente trabalho foi avaliar a toxicidade de misturas binárias do biocida Irgarol com amônia e surfactante Alquil. Benzeno Sulfonado Linear - LAS, além das toxicidades dessas substâncias isoladas. Essa avaliação foi realizada em duas etapas. Na primeira etapa, náuplios de Artemia sp., copépodo Nitocra sp. e embriões de ouriço do mar Echinometra lucunter, foram expostos às concentrações isoladas de Irgarol, amônia e LAS, para definição dos níveis tóxicos dessas substâncias às espécies testadas. Na segunda etapa, náuplios de Artemia sp. foram expostos a uma mistura binária de contaminantes (i.é. Irgarol + amônia; ou Irgarol + LAS) a fim de se avaliar se as toxicidades das misturas seguem o padrão aditivo, sinérgico ou antagônico. Os dados obtidos da primeira etapa dos testes realizados com náuplios de Artemia sp. expostos à amônia, LAS e Irgarol apresentaram CL50-48h 122,85 mg/L, 21,21 mg/L, e 1,81 mg/L, respectivamente, e CEO de 100 mg/L, 81 mg/L e 10 mg/L, respectivamente. Para os testes realizados com os copépodos Nitocra sp., as respectivas CI50-7d foram estimadas em 27,89 mg/L, 4,61 mg/L, e 0,13 mg/L, enquanto as CEO foram 100 ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Contaminants found in the coastal environment include antifouling biocides, such as the Irgarol, ammonia and surfactants, among others. Since they can occur together in the environment, they can interact with each other and the resulting toxicities of their combinations are unknown. The aim this work was to evaluate the toxicity of binary mixtures of Irgarol with ammonia and the surfactant Linear Alkyl-Benzene Sulfonate - LAS, as well as the toxicity these single substances. This investigation was made in two steps. In the first step, nauplii of Artemia sp., the marine organisms Nitocra sp. (Copepoda), and embryos of the sea urchin Echinometra lucunter, were exposed to concentrations of Irgarol, ammonia and LAS, in order to define the toxic levels of each chemical to marine species. In the second step, nauplii of Artemia sp. were exposed to binary mixtures of the contaminants (Irgarol + ammonia; or Irgarol + LAS) in order to evaluate if the toxicities of the mixtures follow addictive, synergistic or antagonistic patterns. To nauplii Artemia sp. exposed to ammonia, LAS and Irgarol, the LC50-48h values were 122.85 mg/L, 21.21 mg/L, and 1.81 mg/L, respectively, and the LOECs were 100 mg/L, 81 mg/L and 10 mgl/L, respectively. The respective IC50-7d obtained in the tests with the copepod Nitocra sp., were 27.89 mg/L, 4.61 mg/L and 0.13 mg/L, while the LOECs were 100 mg/L, 8.1 mg/L and 10 mg/L. For the E. lucunter embryos, the EC50-42h of ammonia was 0.32 mg/L, while the LOEC was 0... (Complete abstract click electronic access below) / Mestre

Submitted by Luciana Sebin (lusebin@ufscar.br) on 2016-09-27T13:41:48Z No. of bitstreams: 1 TeseASM.pdf: 6858970 bytes, checksum: 83be7f6c0d85cb91801e150418917482 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-27T19:51:56Z (GMT) No. of bitstreams: 1 TeseASM.pdf: 6858970 bytes, checksum: 83be7f6c0d85cb91801e150418917482 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-27T19:52:06Z (GMT) No. of bitstreams: 1 TeseASM.pdf: 6858970 bytes, checksum: 83be7f6c0d85cb91801e150418917482 (MD5) / Made available in DSpace on 2016-09-27T19:52:15Z (GMT). No. of bitstreams: 1 TeseASM.pdf: 6858970 bytes, checksum: 83be7f6c0d85cb91801e150418917482 (MD5) Previous issue date: 2016-03-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / The aim of this study was to evaluate the effects of diuron and carbofuran, isolated and mixed, on Raphidocelis subcapitata, Paramecium caudatum and Ceriodaphnia silvestrii and carry out a preliminary risk assessment of these pesticides for Brazilian water bodies. For this, acute and chronic toxicity tests were performed with individual compounds and in combination, in its standard form and commercial formulations. The results showed that in individual action, these pesticides significantly inhibited the R. subcapitata population growth and caused physiological (chlorophyll a content) and morphological (complexity and cell size) changes in the cells. For this alga, the toxicity of standard active ingredients and their commercial formulations showed no significant differences. For the protozoan P. caudatum, acute toxicity tests evidenced an increase of mortality with increasing exposure time for the selected compounds. Moreover, these pesticides caused a significant decrease in population growth, number of generations and biomass of P. caudatum. The toxicity of standard diuron and its commercial formulation was similar for this protozoan, while commercial carbofuran was more toxic than its standard form, suggesting a synergistic interaction with “inert” ingredients. In contrast, for the cladoceran C. silvestrii, acute toxicity tests showed that diuron commercial product was 7.2 times more toxic than its standard active ingredient, whereas for carbofuran, commercial product was 1.5 times less toxic than its standard active ingredient. Regarding chronic toxicity of diuron herbicide, both standard and commercial form caused hormetic effects on fertility of C. silvestrii female. Carbofuran significantly reduced the reproduction of C. silvestrii at concentrations from 0.38 μg L-1 onwards. The tropical species C. silvestrii was more sensitive to these pesticides than cladocerans commonly used in temperate regions, and in addition, for carbofuran, this species was the most sensitive organism already tested (EC50-48h = 0,86 μg L-1). According to the toxicity tests results of diuron and carbofuran mixtures, acute and chronic exposures on all selected test organisms caused significant deviations from the toxicity predicted by Concentration Addition and Independent Action reference models. For P. subcapitata, data indicated the occurrence of synergism in the mixture of these compounds, especially when diuron is the dominant chemical in combination. For P. caudatum, dose ratio dependent deviation (synergism caused by carbofuran and antagonism caused by diuron) was observed in acute exposure and deviation for antagonism was verified for population growth at sublethal concentrations of the standard compounds mixtures. For C. silvestrii, antagonism was observed in low doses and synergism at high doses, predominating synergism in acute exposures of standard pesticides mixtures, while for reproduction the deviation for antagonism was found. The commercial product mixtures of diuron and carbofuran showed significantly different effects and more severe than the standard compounds mixtures for P. caudatum and C. silvestrii. Therefore, diuron and carbofuran mixture can interact and cause toxic responses different from those predicted for individual compounds. According to the concentrations found in water bodies in Brazil, these compounds can cause direct and indirect toxic effects on communities of algae, protozoa and cladocerans and may change the structure and functioning of aquatic ecosystems. In a preliminary risk assessment of these pesticides, risk quotients were larger than 1, indicating that diuron and carbofuran represent potential ecological risks to Brazilian water bodies. Given the above, it is concluded that the pesticides studied alone and in combination, caused toxicity to test organisms and represent potential ecological risk for aquatic environments, which suggests the implementation by the regulatory agencies of more stringent measures for the use of pesticides to improve the protection of aquatic biota. / O objetivo desta pesquisa foi avaliar os efeitos do diuron e do carbofuran, isolados e em mistura, sobre as espécies Raphidocelis subcapitata, Paramecium caudatum e Ceriodaphnia silvestrii e realizar uma avaliação de risco preliminar destes agrotóxicos para os corpos d’água brasileiros. Para isso, testes de toxicidade aguda e crônica foram realizados com os compostos isolados e em mistura, nas suas formas padrão e comercial. Os resultados mostraram que, em ação individual, estes agrotóxicos inibiram significativamente o crescimento populacional de R. subcapitata e causaram alterações fisiológicas (conteúdo de clorofila a) e morfológicas (complexidade e tamanho celular) nas células. Para esta espécie de alga, a toxicidade dos ingredientes ativos nas formas padrão e comercial não apresentaram diferenças significativas. Para o protozoário P. caudatum, os testes de toxicidade aguda evidenciaram um aumento da mortalidade com o aumento do tempo de exposição aos compostos selecionados. Além disso, estes agrotóxicos causaram uma diminuição significativa no crescimento populacional, no número de gerações e na biomassa de P. caudatum. A toxicidade do diuron padrão e de sua formulação comercial foi similar para este protozoário, enquanto que o carbofuran comercial foi mais tóxico do que a sua forma padrão, sugerindo a ocorrência de interações sinérgicas com os ingredientes “inertes”. Diferentemente, para o cladócero C. silvestrii, os testes de toxicidade aguda mostraram que o produto comercial de diuron foi 7,2 vezes mais tóxico do que o seu ingrediente ativo padrão, enquanto que para o carbofuran, o produto comercial foi 1,5 vezes menos tóxico do que o seu ingrediente ativo padrão. Em relação à toxicidade crônica, o herbicida diuron, tanto na forma padrão como comercial, causou efeitos horméticos sobre a fertilidade das fêmeas de C. silvestrii. Já o carbofuran reduziu significativamente a reprodução de C. silvestrii em concentrações a partir de 0,38 μg L-1. A espécie tropical C. silvestrii foi mais sensível a estes agrotóxicos do que cladóceros comumente utilizados em regiões temperadas, e além disso, para o carbofuran, esta espécie foi o organismo-teste mais sensível já registrado na literatura (CE50-48h = 0,86 μg L-1). De acordo com os resultados dos testes de toxicidade das misturas de diuron e carbofuran, exposições agudas e crônicas sobre todos os organismos-teste selecionados causaram desvios significativos da toxicidade prevista pelos modelos de referência de Adição de Concentração e Ação Independente. Para a alga R. subcapitata, os dados evidenciaram a ocorrência do sinergismo na mistura destes compostos, principalmente quando o diuron é o produto químico dominante na mistura. Para o P. caudatum, o desvio dependente da proporção da dose (sinergismo causado pelo carbofuran e antagonismo causado pelo diuron) foi observado em exposições agudas e desvio para o antagonismo foi verificado para o crescimento populacional em concentrações subletais das misturas dos compostos padrões. Para a C. silvestrii, observou-se o antagonismo em baixas doses e o sinergismo em altas doses, prevalecendo o sinergismo nas exposições agudas das misturas dos agrotóxicos padrões, enquanto que para a reprodução foi encontrado o desvio para o antagonismo. As misturas dos produtos comerciais de diuron e carbofuran apresentaram efeitos significativamente diferentes e mais graves do que as misturas dos compostos padrões para o P. caudatum e a C. silvestrii. Portanto, diuron e carbofuran em mistura podem interagir e causar respostas tóxicas diferentes das previstas para os compostos individuais. De acordo com as concentrações encontradas nos corpos d’água brasileiros, estes compostos podem causar efeitos tóxicos diretos e indiretos sobre as comunidades de algas, protozoários e cladóceros, podendo alterar a estrutura e o funcionamento dos ecossistemas aquáticos. Na avaliação de risco preliminar destes agrotóxicos, os quocientes de risco foram maiores do que 1, indicando que o diuron e o carbofuran representam riscos ecológicos potenciais para os corpos d’água brasileiros. Diante do exposto, conclui-se que os agrotóxicos estudados, isolados e em mistura, causaram toxicidade aos organismos avaliados e representam riscos ecológicos potenciais para os ambientes aquáticos, o que sugere a implantação pelas agências reguladoras de medidas mais restritivas para o uso destes agrotóxicos, visando uma melhor proteção da biota aquática.

Submitted by Guacira de Figueiredo Eufrasio Pauly null (guafe@yahoo.com.br) on 2018-03-22T19:42:28Z No. of bitstreams: 1 Avaliação da toxicidade de misturas Irgarol e contaminantes ambientais comumente observados em áreas costeiras.pdf: 1773405 bytes, checksum: d022473735b82f51e6340d000b00e009 (MD5) / Approved for entry into archive by Disleide Silvia Valerio Gounella null (disleide@clp.unesp.br) on 2018-03-23T18:54:19Z (GMT) No. of bitstreams: 1 pauly_gfe_me_svic.pdf: 1773405 bytes, checksum: d022473735b82f51e6340d000b00e009 (MD5) / Made available in DSpace on 2018-03-23T18:54:19Z (GMT). No. of bitstreams: 1 pauly_gfe_me_svic.pdf: 1773405 bytes, checksum: d022473735b82f51e6340d000b00e009 (MD5) Previous issue date: 2018-02-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Alguns contaminantes encontrados no ambiente costeiro incluem os princípios ativos de tintas anti-incrustantes, entre os quais se destaca o Irgarol. O Irgarol pode interagir com outros contaminantes comuns em áreas litorâneas antropizadas, tais como a amônia e os surfactantes, sendo que os efeitos dessas combinações são desconhecidos. O objetivo do presente trabalho foi avaliar a toxicidade de misturas binárias do biocida Irgarol com amônia e surfactante Alquil. Benzeno Sulfonado Linear - LAS, além das toxicidades dessas substâncias isoladas. Essa avaliação foi realizada em duas etapas. Na primeira etapa, náuplios de Artemia sp., copépodo Nitocra sp. e embriões de ouriço do mar Echinometra lucunter, foram expostos às concentrações isoladas de Irgarol, amônia e LAS, para definição dos níveis tóxicos dessas substâncias às espécies testadas. Na segunda etapa, náuplios de Artemia sp. foram expostos a uma mistura binária de contaminantes (i.é. Irgarol + amônia; ou Irgarol + LAS) a fim de se avaliar se as toxicidades das misturas seguem o padrão aditivo, sinérgico ou antagônico. Os dados obtidos da primeira etapa dos testes realizados com náuplios de Artemia sp. expostos à amônia, LAS e Irgarol apresentaram CL50-48h 122,85 mg/L, 21,21 mg/L, e 1,81 mg/L, respectivamente, e CEO de 100 mg/L, 81 mg/L e 10 mg/L, respectivamente. Para os testes realizados com os copépodos Nitocra sp., as respectivas CI50-7d foram estimadas em 27,89 mg/L, 4,61 mg/L, e 0,13 mg/L, enquanto as CEO foram 100 mg/L, 8,1 mg/L, e 10 mg/L. A CE50-42h obtida nos testes com embriões de Echinometra lucunter expostos à amônia foi de 0,32 mg/L, enquanto a CEO foi de 0,1 mg/L. Todas as concentrações de exposição ao LAS foram tóxicas para os embriões de Echinometra lucunter, e não foi observada toxicidade do Irgarol. Ambas as misturas binárias de Irgarol e amônia e Irgarol e LAS apresentaram melhor ajuste para o modelo IA (ação independente), com efeitos antagônicos e sinérgicos, respectivamente. As ferramentas utilizadas foram úteis para prever o padrão de respostas das misturas químicas sobre as espécies. / Contaminants found in the coastal environment include antifouling biocides, such as the Irgarol, ammonia and surfactants, among others. Since they can occur together in the environment, they can interact with each other and the resulting toxicities of their combinations are unknown. The aim this work was to evaluate the toxicity of binary mixtures of Irgarol with ammonia and the surfactant Linear Alkyl-Benzene Sulfonate - LAS, as well as the toxicity these single substances. This investigation was made in two steps. In the first step, nauplii of Artemia sp., the marine organisms Nitocra sp. (Copepoda), and embryos of the sea urchin Echinometra lucunter, were exposed to concentrations of Irgarol, ammonia and LAS, in order to define the toxic levels of each chemical to marine species. In the second step, nauplii of Artemia sp. were exposed to binary mixtures of the contaminants (Irgarol + ammonia; or Irgarol + LAS) in order to evaluate if the toxicities of the mixtures follow addictive, synergistic or antagonistic patterns. To nauplii Artemia sp. exposed to ammonia, LAS and Irgarol, the LC50-48h values were 122.85 mg/L, 21.21 mg/L, and 1.81 mg/L, respectively, and the LOECs were 100 mg/L, 81 mg/L and 10 mgl/L, respectively. The respective IC50-7d obtained in the tests with the copepod Nitocra sp., were 27.89 mg/L, 4.61 mg/L and 0.13 mg/L, while the LOECs were 100 mg/L, 8.1 mg/L and 10 mg/L. For the E. lucunter embryos, the EC50-42h of ammonia was 0.32 mg/L, while the LOEC was 0.1 mg/L. All the tested concentrations of LAS were toxic to the embryos of E. lucunter, where Irgarol did not caused toxicity to these organisms. Both binary mixtures of Irgarol and ammonia and Irgarol and LAS showed better fit to the model IA (Independent Action), with antagonistic and synergistic effects, respectively. The used tools were useful to predict the toxicities of chemical mixtures to marine tropical species. / 4693520

Algumas atividades antrópicas podem provocar alterações nos recursos naturais, seja ela no rio, no solo ou no mar. Os efeitos destas alterações podem ser observados na biota, na qualidade das águas, na disponibilidade de nutrientes, interferindo em todos os elementos que compõem o ecossistema, influenciando-o em maior ou menor grau. O presente estudo procurou determinar o efeito dos poluentes sulfato de zinco, cloreto de amônia, dodecilsulfato de sódio e benzeno, numa espécie bioindicadora marinha, el larvas de mexilhão (Perna perna), além de acrescentar dados sobre a biologia e analisar quimicamente a presença de hidrocarbonetos nos tecidos dos animais adultos e das larvas. Para tanto foram utilizados testes de toxicidade, estudos sobre a bioenergética (consumo de oxigênio e excreção de amônia), enzimas biomarcadoras e excreção de fósforo e nitrogênio. Com relação à biologia, foram encontrados indivíduos sexualmente maduros a partir de 26,1 e 27 mm de comprimento para fêmeas e machos, respectivamente, e há diferença, ainda que esta não seja estatisticamente significativa, entre ovócitos e larvas submetidos a ação de poluentes. A análise de hidrocarbonetos indicou que existe uma contribuição biogênica e petrogênica no litoral norte do Estado de São Paulo. Para as demais análises verificou-se que, quando comparados aos grupos controles, as larvas foram sensíveis e responderam aos diferentes poluentes, geralmente com inibição da atividade. / Some antropic activities may be the cause fo alterations in natural resources, being it river, soil or sea. These alteration effects may be observed in the biota, water quality, nutrients disponibility, interfering with all elements that are part of a ecosystem, with greater or lesser influence degree. The present study was directed to determine the effect of pollutants, such as zinc sulphate, ammonia chlorate, sodium dodecilsulphate and benzene, acting over larvae of a marine bioindicator, the mussel (Perna perna), besides adding data on biology and chemically analysing hidrocarbon presence in larvae and adult animal tissues. In order to obtain these results, toxicity tests were used and bioenergetic (oxygen consumption and ammonia excretion), biomarkers, phosphorus and nitrogen excretion, were studied. As for biology, individuals sexually mature were found starting with 26,1 and 27 mm length females and males, respectively, and there is difference between larvae and ovocites, although statistically not significant, submitted to pollutants action. The hydrocarbon analysis indicates a biogenic and petrogenic contribution to north coast of São Paulo State. For the other analyses the observed results, when compared to control groups, showed that larvae are sensitive, and responded to different pollutants, generally with activity inhibition.

PCBs are among the most problematic marine contaminants. Converging towards the oceans via the rivers and the atmosphere, they concentrate in sediments where they become a permanent threat to organisms living at their contact. PCBs are extremely resistant, bioaccumulated and some congeners are considered as highly toxic. The North Sea is considered as a highly contaminated area ;however little information is available regarding the impact of PCBs on key benthic organisms of this region.<p>Ubiquist, abundant and generally recognized as a good bioindicator species, the common NE Atlantic sea star Asterias rubens (L.) is an ecosystem-structuring species in the North Sea and was chosen as an experimental model. The present study focused on the characterization of PCB bioaccumulation in A. rubens exposed through different routes (seawater, food, sediments) and on subsequent biological responses, at immune and sucellular levels. The considered responses were respectively (i) the production of reactive oxyggen species (ROS) by sea stars amoebocytes, which constitutes the main line of defence of echinoderms against pathogenic challenges and (ii) the induction of a cytochrome P450 immunopositive protein (CYP1A IPP) which, in vertebrates, is involved in PCB detoxification.<p>Experimental exposures carried out have shown that A. rubens efficiently accumulates PCBs. Exposure concentrations were always adjusted to match those encountered in the field. PCB concentrations reached in sea stars during the experiments matched the values reported in field studies ;therefore our experimental protocol was found to accurately simulate actual field situations. Uptake kinetics were related to the planar conformation of the considered congeners :non-coplanar PCB uptake was described using saturation models, whereas coplanar PCBs (c-PCBs) were bioaccumulated according to bell-shaped kinetics. Non-coplanar congeners generally reached saturation concentrations whithin a few days or a few weeks, which means that sea stars can be used to pinpoint PCB contamination shortly after occurrence. On the other hand, c-PCB concentrations reached a peak followed by a sudden drop, indicating the probable occurrence of c-PCB-targeted metabolization processes in sea stars. Our experimental studies also demonstrated that seawater was by far the most efficient route for PCB uptake in sea stars and that even if PCB levels in seawater are extremely low compared to sediment-associated concentrations, seawater constitutes a non-negligible route for PCB uptake in marine invertebrates. Among the different body compartments, bodywall displayed the highest bioaccumulative potency and can therefore be considered as particularly interesting for field biomonitoring applications. Rectal caeca, which play a central role in digestion and excretion processes in sea stars, have also rised particular interest as results suggest these organs could be involved in the elimination of PCB 77 degradation products. <p>The field work carried out during the present study showed that PCB concentrations measured in A. rubens tissues reflect environmental levels of certain congeners. As it was the case in experimental conditions, A. rubens differentially accumulated PCB congeners according to their planarity. Strong relationships were found between concentrations measured in sediments and those determined in sea stars body wall for certain non-coplanar congeners (e.g. 118 and 138), thus allowing to consider A. rubens as a suitable bioindicator species for medium-chlorinated PCB congeners. On the other hand, sea stars appeared to be able to regulate -to a certain extent- their content in coplanar PCBs. This implies that (i) A. rubens cannot be strictly considered as an indicator organism for c-PCBs and (ii) c-PCBs probably affect essential aspects of sea star biology, potentially leading to deleterious effects.<p>The present study addressed effects of PCB exposure on A. rubens biology, in both experimental and field conditions. In experimental conditions, PCBs were found to significantly alter ROS production by sea stars amoebocytes. This alteration also occurred in a congener-specific way :c-PCBs were found to significantly affect, and probably impair sea stars immune system, whereas non-coplanar congeners had no effect. In the field, the PCB contribution to immunotoxicity could not be determined because none of our studies considered ROS production along with c-PCB concentration measurements. However, the levels of ROS production by sea stars amoebocytes measured in field and experimental conditions were found to potentially lead to altered immunity, and therefore to impair sea stars defence against pathogenic agents. <p>A specially designed ELISA was used to measure CYP1A IPP in experimental and field conditions. Experimental work has shown that the induction of this protein was related to PCB exposure in a congener-specific fashion :c-PCBs alone were found to strongly induce the production of CYP1A IPP according to a dose-dependent relationship. These results have highlighted many similarities between the dioxin-like responsiveness of CYP1A IPP induction in sea stars and that occurring in vertebrates. This strongly suggests similarities in the toxicity-triggering mechanism of dioxins and c-PCBs. In the field, CYP1A IPP induction was found to be significantly related to PCB levels determined in bottom sediments. It can thus be considered as a valuable biomarker. Further research is however needed to better characterize the influence of physico-chemical and physiological parameters on CYP1A induction to refine the interpretation of the information gathered via this biomarker. <p>Results obtained in our study have lead to questionning international regulations applying to PCB biomonitoring in the marine environment. For instance, we strongly suggest that the selection of congeners to be systematically considered should be revised to include c-PCBs. Indeed, in our experiments PCB toxicity was almost always attributable to the sole c-congeners. Historically, determination of c-PCB concentrations was extremely difficult due to analytical limitations ;however, nowadays, these problems have been overcome and do no more justify their exclusion from monitoring studies. <p>Although A. rubens appeared to be quite resistant to PCB contamination, levels measured in sea stars from the southern North Sea can possibly affect their immune and endocrine systems in a subtle way, but with relatively low risk for this species at the short-term. However, this does not mean that other species in this region undergo similarly low risks, or that sea star-structured ecosystems may not become affected in the long-term<p> / Doctorat en sciences, Spécialisation biologie animale / info:eu-repo/semantics/nonPublished

De nombreux cours d’eau sont exposés aux polluants métalliques. Dans ces milieux, les communautés microbiennes structurées sous forme de biofilms peuvent être affectées par ces polluants. Or, une des questions prégnantes en écotoxicologie concerne l’évaluation des impacts toxiques dans le cas de combinaisons multi-stress. Dans ce contexte, l’objectif principal de ce travail était d’évaluer l’influence de la température sur la réponse du biofilm à une exposition au cuivre (Cu). Cet objectif a été abordé en considérant l’influence de la température, d’une part, sur la sensibilité des communautés au Cu, et d’autre part, sur l’exposition des communautés de biofilm à ce métal. Ces travaux ont été réalisés en microcosmes de laboratoires et menés sur des biofilms prélevés à différentes saisons. Nous avons tout d’abord optimisé les protocoles utilisés pour mesurer i) les niveaux de tolérance des communautés au Cu et ii) la bioaccumulation du Cu dans les biofilms. Les résultats des expérimentations ont révélé que la température pouvait moduler l’impact structural et fonctionnel du Cu sur les biofilms, du fait notamment d’une diminution de la bioaccumulation aux plus fortes températures. En complément, les résultats ont mis en évidence l’influence d’une hausse de la température sur la sensibilité des communautés au Cu, qu’elles aient été préalablement exposées (PICT) ou non à ce métal. Cependant, nous avons observé que les effets de la température sur la réponse du biofilm étaient variables d’une étude à l’autre, suggérant une influence de nombreux paramètres, tels que la composition initiale de la communauté, les niveaux de stress appliqués (température et Cu), le compartiment microbien étudié (phototrophe ou hétérotrophe), ou les fonctions considérées / Many streams are impacted by metallic pollution. In such ecosystems, microbial communities, which grow preferentially as biofilms, can be affected by these pollutants. However, there is a need to better assess the impact of toxic substances under multi-stress interactions. In this context, the main aim of this work was to evaluate the effect of temperature on biofilm response to copper (Cu) exposure. Accordingly, we addressed the influence of temperature on microbial community sensitivity to Cu on the one hand, and its influence on biofilm exposure to this metal on the other hand. Microcosm experiments were performed under laboratory conditions using biofilms sampled in winter or summer. This work allowed us to set up new methodological developments, which were applied to improve the protocols used to measure (i) Cu tolerance levels of biofilm microbial communities and (ii) Cu bioaccumulation in biofilms. Results showed that temperature could modulate structural and functional effects of Cu on biofilms, notably due to a decrease in Cu bioaccumulation at higher temperatures. In addition, our findings indicate that temperature increase could influence the sensitivity of microbial communities to Cu, whether they have been previously chronically exposed (PICT) or not to this metal. Nonetheless, temperature effects on biofilms varied among experiments, revealing that the influence of temperature on the effects of Cu on microbial biofilms depends on many parameters, such as the initial composition of communities, the level of the applied stresses (temperature and Cu), the microbial compartment studied (i.e. phototrophic or heterotrophic), or the measured functional parameters

To determine the ecotoxicity of analgetics, first the individual ecotoxicity values of individual analgetics are determined and then a mixture of two analgetics is tested. To determine the toxicity, both standard and alternative toxicity tests are used (daphnia magna, sinapsis alba, scenedesmus subspicatum, vibrio fischeri, thamnotoxkit FTM a daphnotoxkit FTM magna). The analgetics being whish tested are the commonly used medicines ibuprofen, ASA, diclofenac and paracetamol, which are the most frequently used medicines in the Czech Republic and whole Europe. To determine the ecotoxicity of the polymers, I‘m using an indirect method of examination, in which I determine the antagonistic or synergistic effects of the mixture of monomers from which the polymer is prepared and into which it slowly decomposes in nature. For the determination both standard and alternative toxicity tests are used. The polymers the toxicity of which is being determined are the habitually used polymer PET and the formaldehyde resine known as bakelite in Eastern Europe.