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A suite of biomarkers was developed using the crab Carcinus maenas and the mussel Mytilus edulis as test organisms. The ability of the biomarkers to differentiate amongst the major toxic components and to indicate the concentration of chemical mixtures was evaluated in the laboratory. Biomarkers were also applied in a field trial and their potential to monitor environmental water quality in a chemically contaminated estuary investigated. The results from the biomarker assays were compared with and validated against two commonly used toxicity tests (Tisbe battagliai LC-50, and Microtox®). Novel methods for recognising patterns of biomarker responses were developed and assessed. The most sensitive and reliable biomarker assays investigated were neutral red retention time in crabs and mussels and heart rate and glutathione-S-transferase activity in crabs. Effects were observed at environmentally realistic concentrations; for example lysosomal enlargement was observed in mussels exposed to a complex mixture containing chemicals at environmental quality standard concentrations. Exposure concentrations required to illicit biomarker responses were similar to toxicity test EC-50 values. The ease of interpretation and clarity of the results was enhanced when data from suites of biomarkers were pooled and analysed using multivariate statistical techniques (multidimensional scaling and cluster analysis). Multivariate analysis differentiated amongst mixtures containing solely organic chemicals, metals and metal and organic chemical mixtures. Exposure response relationships to complex mixtures were established for some of the individual biomarkers tested (crab heart rate and gill metallothionein) and also for suites of biomarkers when multivariate analysis was carried out. In the field biomarkers, in both transplanted and indigenous animals, were able to differentiate between clean and contaminated sites and indicate a pollution gradient along the Tees Estuary. This was not achieved using toxicity tests. The results were displayed clearly using multivariate analysis, enhancing the power of biomarkers as monitoring tools.

A poluição aquática em Santos é uma questão ambiental prioritária, interdisciplinar por natureza, devido aos seus efeitos ecológicos, de saúde pública e sócio-econômicos. Trata-se de um problema amplamente relatado, cuja evolução acompanhou o histórico de desenvolvimento econômico da região. A presente pesquisa partiu de análises pontuais de um único, porém representativo, canal de drenagem pluvial urbana e das praias receptoras de seu conteúdo, para uma análise da poluição aquática no município como um todo. Para isso, foram usadas diferentes metodologias: coletas de amostras de água do canal e do mar, seguidas de análises físicas, químicas e ecotoxicológicas; pesquisas sobre as políticas públicas relacionadas à poluição hídrica local e aplicação de entrevistas estruturadas em frequentadores das praias. Além disso, foi analisada uma série histórica de dados microbiológicos das águas costeiras santistas. Os resultados indicam toxicidade aguda eventual nas amostras do canal e toxicidade crônica nas amostras da água do mar, coletadas em 2007, e severa contaminação de origem fecal, ao menos nas últimas três décadas. A pesquisa sobre as políticas públicas constatou que algumas surtiram efeitos positivos, mas não foram suficientes para melhorar a qualidade da água e mantê-la adequada, seja por sua descontinuidade, seja pelo crescimento acelerado no município, tanto demográfico, quanto econômico. A maioria das ações tem despontado em resposta a situações críticas, sendo implantadas sem planejamento adequado, visando conter apenas as conseqüências ambientais, e não as reais causas dos problemas. Como as políticas públicas geralmente não surgiram de processos de gestão participativa, também foram analisados alguns instrumentos de participação pública vigentes em Santos existe um envolvimento ainda incipiente da população, mas com iniciativas interessantes e potencialmente viáveis. Em relação às entrevistas, foram analisadas as respostas de 83 pessoas, entre as quais 77% eram residentes e o restante turistas. A maioria dos entrevistados acredita que as praias estão poluídas e que a situação dos canais é ainda mais crítica, corroborando com os resultados encontrados nas análises ecotoxicológicas e com os dados de contaminação microbiológica. Grande parte dos entrevistados não se banha justamente pela qualidade da água duvidosa, mas não se sentem responsáveis pela situação, nem demonstram iniciativa para pressionar o poder público a revertê-la. Quando os dados da pesquisa são integrados, é possível constatar que a qualidade da água das praias de Santos é crítica, como resultado de um modelo insustentável de desenvolvimento, com falta de planejamento e políticas públicas inconsistentes. Essa situação é agravada pela passividade observada entre muitos dos frequentadores das praias, que, apesar de cientes da poluição, não se envolvem na busca de soluções. Espera-se que a pesquisa contribua para o controle da contaminação aquática local, através do estabelecimento de políticas e ações que considerem todas as dimensões do problema e especialmente as suas causas. O presente estudo demonstra que a integração de diferentes métodos de avaliação permite uma compreensão maior acerca dos problemas ambientais e consequentemente pode subsidiar soluções mais efetivas. Além disso, foi enfatizada a importância da interdisciplinaridade em estudos de questões ambientais. / The aquatic pollution in Santos is a priority environmental issue, which presents many dimensions, as the ecological, public health and socioeconomic ones, having thus an interdisciplinary nature. This question has been extensively studied and its evolution was strictly related to the history of the regional economic development. The first phase of the present survey consisted of punctual analyses, made during 2007, of a single (but representative) urban pluvial drainage channel and the receiving beaches which are directly influenced by its contribution (i.e., to its contaminants inputs), in order to estimate the water quality (according to the aquatic pollution) in the overall shore. To achieve that, different methods were employed: sampling of channel and sea waters, and physical, chemical and ecotoxicological analyses. The second phase consisted of an investigation on the public policies related to the local hydric pollution. Concomitantly, as the third part of this survey, we conducted a study on the public knowledge on the aquatic pollution, by the application of structured interviews to the beaches users. Complementarily, the historical series of microbiological data for the coastal waters of Santos was assessed, to provide additional data. The results indicated eventual acute toxicity of channel water samples and chronic toxicity of sea water samples, and a severe fecal contamination at least in the last three decades. The evalution of public policies indicated that even though some of them had positive effects, they were not sufficient to improve the water quality and to maintain it appropriate, whether due to their discontinuity, whether to the demographic and economic accelerated growth of the city. The majority of actions have been implemented as responses to critical situations, without proper and wider planning, aiming only to control the consequences, and not the real causes of the problems. The public policies generally were not a product of participative processes; some instruments of public participation in Santos also were analyzed the involvement of the population is still incipient. However there are interesting and potentially viable initiatives. Regarding to the interviews, 83 beach users were considered to this survey, 77% were local residents and the remaining were tourists. The most interviewees believe that beaches are polluted and that the channels situation is still more critic, being in accordance with the results of ecotoxicological and microbiological data. Many of the beach users do not have contact with sea water due to its doubtful quality, but they do not feel responsible for the situation and not show initiative to press the public governement to act. When the obtained data are integrated, it is possible to notice that the water quality in Santos beaches is critical, as a product of an unsustainable model of development, lacking of planning and inconsistent public policies. The situation is aggravated due to the passivity observed among many of the beach users, who are aware of the pollution, but do not involve themselves to find effective solutions. We hope that this research may contribute to the control of aquatic local contamination, by the establishment of policies, enforcements and actions which consider all the dimensions of the problem, in special its causes. Moreover this research demonstrated that the integration of the different assessment methods may provide a stronger comprehension of the environmental issues and which consequently can subsidize much more effectively their solutions. Therefore it is highlighted the importance of interdisciplinarity to the studies involving environmental issues.

For various reasons, existing methods for the assessment of aquatic pollution do not always adequately address the way in which contaminants affect receiving environments and their component ecosystems. The main advantage of biological assessment over the measurements of physical and chemical aspects of water quality is that biota provide an integrated response to all prevailing influences in their environment. Biological assessment protocols have been developed for a range of test organisms, from bacteria to mammals using measurement from molecular biomarkers to indicators at the population or community level of organisation. Macroinvertebrates in particular have been popular for ecological assessment of habitat and water quality because they are small and straight forward to sample and identify using relatively simple and inexpensive equipment and readily available taxonomic keys. However, various biological assessment techniques also have their limitations. Field-based assessment of biological communities does not provide direct evidence to determine underlying causal relationships, while laboratory or mesocosm toxicity tests are criticised for their limited ability to extrapolate to natural field conditions. To help bridge the gap, this thesis aims to investigate the efficacy of using caged macroinvertebrates in situ to assess the ecological condition of aquatic environments, and whether a causal relationship can be established when macroinvertebrates are deployed in situ at sites known to have impaired water quality. Endpoints employed in this thesis include survival, measurements of morphology (as a surrogate for growth) and condition and, for trials assessing sites that receive mine drainage, the tissue concentration of certain trace metals. Development of an in situ approach to water quality monitoring and assessment will potentially provide methods for use by resource managers, community groups and aquatic researchers that are less expensive and faster to run than existing methods and will complement other approaches employed in the assessment of water quality. In situ testing of water quality using macroinvertebrates requires the collection, handling, caging, deployment and retrieval of test organisms at sites of suspected pollutant impact. As such procedural factors may affect test organisms and potentially confound their responses, it is important to consider and understand as many of these factors as possible. Aquatic macroinvertebrates held in finer mesh cages had larger heads than in coarser mesh cages. This was likely due to increased substrate available for growth of epilithon and periphyton on which the caged organisms could graze. Caging density had no effect on amphipod mortality over the trial period, however, individuals held at higher densities increased in size (as indicated by longer dorsal lengths) more than those held at lower or intermediate densities. Temporary storage of test organisms in laboratory aquaria may facilitate the collection of abundances required for in situ trials, however, tanked individuals were smaller and had lower biomasses than individuals collected and deployed immediately. While this is likely to result from differences in feeding during the storage period, it is also possible that tank storage and the ?double handling? deleteriously affected them, or reduced their tolerance. The effects of transplanting macroinvertebrates between sites varied considerably depending on the characteristics of "source" and "transplant" sites. Certain taxa suffered marked mortality within 24 hours even at their source site, indicating an adverse effect of the caging itself, or perhaps via the change in food, shelter or microclimate which could potentially render them unsuitable as test organisms in caging studies. Other taxa did not differ in survival or body size when relocated between sites, with some evidence of increased growth at sites dissimilar from their source site. In general, organisms relocated to sites that are "similar" to their source environment performed less well at the transplant site. However, organisms transplanted to "dissimilar" sites were found to be bigger than those caged and deployed back to the source site. When employed to assess known pollution scenarios in and around Canberra, macroinvertebrate responses were, in some instances, able to be linked to specific environmental parameters or combinations thereof. In Case Study 1, findings varied in relation to the response endpoint being examined, and between test species, although concentrations of metals were significantly higher in tissue of macroinvertebrates deployed at the impact site downstream of the abandoned Captains Flat mine and increased with time exposed. In Case Study 2, freshwater shrimp suffered significant mortality within 24 hours of deployment at the impact sites, with larger individuals more susceptible at sites receiving urban stormwater runoff. While various biological effects were most closely correlated with ammonia concentrations at the site, different body size endpoints were affected in opposite ways. In Case Study 3, body size endpoints for one test organism varied consistently with respect to site and time factors, but none of the changes could be linked to any of the environmental data collected. Response variables for a different test species also indicated significant effects arising from both deployment site and time, however, each endpoint responded in a different way to the treatment factors, and aligned with different combinations of environmental data. In general, linking of macroinvertebrate responses with environmental data was difficult because of the high variability in the environmental data. However, it was further complicated by the mismatch in the level of replication between the two datasets. As a consequence of this, the macroinvertebrate data had to be collapsed to a lower level for comparison with the environmental data, resulting in a loss of natural variability and analytical power. Since only the strongest treatment effects, which could be detected above the background "noise", were detected and modelled against the environmental data, it is possible that other "cause" and "effect" relationships may have been overlooked. From these results, it is clear that many macroinvertebrate taxa are suitable for use as bioindicators in in situ trials, but that criteria used for selection of test species should definitely include more than just impact-sensitivity and abundance. However, there are several aspects associated with the experimental set up of field-based protocols involving caged macroinvertebrates that may limit their usefulness as a rapid and reliable bioassessment tool, and need to be considered when designing and undertaking these kinds of trials. It is also apparent that choice of endpoint can greatly influence conclusions, with detection of treatment effects reported in this thesis varying greatly depending on which morphological endpoint was examined. This study clearly demonstrated that there may be significant difficulties in establishing causal relationships between environmental data and biotic responses of macroinvertebrates deployed under field conditions. However, it has also shown that deployment of caged macroinvertebrates in situ may assist in the determination of biological effects arising from impaired water quality, which can then serve as the basis for more focussed laboratory or mesocosm studies in which environmental conditions can be more readily controlled or monitored.

Tämnaren är en slättsjö i östra Svealand där ammunition går att finna i bottensedimenten. Ammunitionen härstammar från övningar som Flygvapnet hade över sjön under 1950- och 1960-talen. Sjön är mycket grund och näringsrik och befinner sig i ett område som fortfarande påverkas starkt av landhöjningen orsakad av den senaste inlandsisens tillbakadragande. Detta innebär att Tämnaren växer igen i rask takt och beräknas vara i princip borta om ungefär 600 år. Sjön har ett skyddsvärde dels genom sin rikedom på olika fågelarter som håller till i närområdet men också ett rekreationsvärde för människor i området runt sjön. Detta gör att man försöker se vilka åtgärder som kan fungera för att hejda den snabba igenväxningen och muddring kan eventuellt bli aktuellt i sjön. Innan beslut om muddring tas bör halter och spridning av föroreningar från ammunitionen i sjöns sediment kartläggas.Muddring är en vanlig metod som används för att öka djupet i sjöar och vattendrag men den kan innebära vissa risker. Den här undersökningen har gjorts för att fastställa halter av tungmetallerna koppar, zink, kadmium och bly som finns i sjöns bottensediment. Vid eventuell muddring skulle dessa tungmetaller kunna spridas ut på ett större område i sjön och därför vill man veta metallhalterna och ifall sådana halter är skadliga. I den här undersökningen kunde både högre och lägre halter av tung-metaller detekteras, halterna för koppar och kadmium var de som var högst, i jämförelse med Natur-vårdsverkets, 2007, förväntade bakgrundshalter i svenska sediment. Det syntes också en tydlig trend i tungmetallshalterna då samtliga mätvärden visade upp högre halter i sydöstlig riktning i sjön. Även om inte alla halter var högre än väntat, med undantag för koppar och kadmium, så återstår risken att de kontaminerade sedimenten kan spridas vidare i sjön om man väljer att muddra den. Även om halterna är låga kan de fortfarande innebära risker att sprida dessa vidare till organismer och växtlighet i sjön.Miljöundersökningar är viktiga att göra innan man beslutar sig för att göra större ingrepp på naturen, såsom muddring, då det är viktigt att skydda den natur vi har. Tämnaren är en sjö som på många sätt är skyddad för att den har ett rikt djur- och naturliv som bör bevaras. Detta gör att det blir ännu mer aktuellt att göra ordentliga förundersökningar innan man tar sig för några större åtgärder. / Tämnaren is a lake in eastern Svealand, Sweden and it is a shallow, highly eutrophic lake that suffers from severe overgrowth and it will probably cease to exist in about 600 years due to land rise and sedimentation. The lake used to be the location for exercises for the Swedish Armed Forces during the years 1950 – 1960, and therefore old remnants of ammunition can be found on the lake bottom. There is a risk that these old remnants of ammunition could be leaking pollutants into the lake sediment and it is necessary to investigate the level of which these releases may be occurring. This thesis will focus on the four heavy metals copper, zink, cadmium and lead and analyses of these metals will be performed on the lake sediments. The goal is to determine the spatial distribution of the heavy metals and the content of them in the lake sediments since dredging might be necessary for the lake. The risks with dredging are many but in this thesis the dispersion of contaminated sediments are the primary consideration due to the conditions in the lake. The spatial distribution of heavy metals already dispersed is interesting to know because it is necessary to know if the ammunition is leaking pollutants through the lake.The results of the investigation showed some higher concentrations of heavy metals in the sediments, mainly for copper and cadmium. According to Naturvårdsverket, 2007, estimated background content of copper in sediment is 15 mg/kg and the highest measured value in Tämnaren is about ten times higher than that. For cadmium the estimated background value is 0,3 mg/kg and all measured values in this investigation were higher than that. For zink the estimated background value is 100 mg/kg and for lead 5 mg/kg and the measured values in this investigation showed lower values in general for both metals. It seems as some dispersion has already occurred, maybe due to wind and wave processes, causing resuspension of sediments. Overall the spatial distribution is showing a trend with higher heavy metal contents to south eastern direction in the investigated area which could be a consequence of wind and wave processes.Although the contents of heavy metals is a bit higher in comparison with estimated background contents in Swedish lake sediments only copper and cadmium are estimated to be higher than recom-mended for a safe environment. Due to the toxicity of copper and cadmium some caution is recommend-ed before the dredging process starts.

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2018-06-27T11:56:06Z No. of bitstreams: 1 PDF - Liliane Silva Câmara de Oliveira.pdf: 25636983 bytes, checksum: 0b12e9559d0756d268a1438a25de32ed (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2018-07-03T17:48:57Z (GMT) No. of bitstreams: 1 PDF - Liliane Silva Câmara de Oliveira.pdf: 25636983 bytes, checksum: 0b12e9559d0756d268a1438a25de32ed (MD5) / Made available in DSpace on 2018-07-03T17:48:57Z (GMT). No. of bitstreams: 1 PDF - Liliane Silva Câmara de Oliveira.pdf: 25636983 bytes, checksum: 0b12e9559d0756d268a1438a25de32ed (MD5) Previous issue date: 2017-03-23 / The water is one of the most important resources for maintaining life on planet earth, and only a small part of the whole that is available is capable of favoring human life: is called fresh water. It is found in rivers, glaciers, lakes and subterranean vegetation with low salinity, and in these days he has been suffering it is suffering from anthropic actions related to urbanization, industrialization, bathing, fish farming, agronomy and agriculture. These actions cause contamination of these waters and degradation of the natural environment, putting at risk the maintenance of life. This research was carried out in the Municipality of Pureza, located 59 km from the capital of Rio Grande do Norte, Where is the source of the Maxaranguape River, considered one of the most important of the State, that was object of observation of this study during the year 2016. Its nascent and its margins are used as seaside resort and it has been showing signs of degradation from these actions. This environmental scenario motivated the accomplishment of this study, whose objective was to investigate the occurrence of possible genotoxic effects of the water pollution of the Maxaranguape River in fish of the genus Tilapia from an environmental perspective. For that was used as method observatory records with annotations and photos; listening to the surrounding residents and utilities of the river were performed, as well as microbiological, physical-chemical and Micronucleus (MN) tests from blood red blood cells of Tilapia fish. The results showed that the river is in the process of degradation and may still be a source of public health problems. It presents sandbanks and changes in its course, microbiological and physico-chemical changes in its waters and the extinction of species such as Tilapia, which made genotoxic study impossible. The river is being disrespected by city dwellers and public authorities, being used in many wrong ways, dying for lack of inspection, maintenance and preservation. It is an urgent need for control over the city's administrators, a serious job of cleaning, maintenance, inspection and conservation. As it is also necessary an educational intervention work with the environment residents and visitors who use the River and its source as recreational points. / A água é um dos recursos mais importantes para a manutenção da vida no planeta terra e apenas uma pequena parte do todo que está disponível é capaz de favorece a vida humana: é a chamada água doce. Ela é encontrada em Rios, geleiras, lagos e lenções subterrâneos com salinidade baixa, e nos dias atuais vem sofrendo com as ações antrópicas ligadas a urbanização, industrialização, balneabilidade, piscicultura, agronomia e agropecuária. Essas ações provocam a contaminação dessas águas e degradação do ambiente natural, colocando em risco a manutenção da vida. Essa pesquisa foi realizada no Município de Pureza, localizado a 59 km da capital do Rio Grande do Norte, onde está a nascente do Rio Maxaranguape, considerado um dos mais importantes do Estado, objeto de observação desse estudo durante o ano de 2016. Sua nascente e suas margens são utilizadas como balneário e ele vem apresentando sinais de degradação provenientes dessa ação. Este cenário ambiental motivou a realização desta pesquisa, cujo objetivo é investigar a ocorrência de possíveis efeitos genotóxicos da poluição aquática do Rio Maxaranguape em peixes do gênero Tilapia em uma perspectiva ambiental. Para tanto, foi utilizado como método de coleta de dados registros observatórios com anotações e fotos; escuta dos moradores do entorno e utilitários do rio foram realizados, além de testes microbiológicos, físico-químicos e do Micronúcleo (MN) a partir de eritrócitos sanguíneos de peixes Tilapia. Os resultados mostraram que o rio encontra-se em processo de degradação, podendo ainda ser fonte de problemas de saúde pública. Ele apresenta bancos de areia e modificações em seu curso, alterações microbiológicas e físico-químicas em suas águas e a extinção de espécies como a Tilapia, o que impossibilitou o estudo genotóxico. O Rio está sendo desrespeitado pelos moradores da cidade e autoridades públicas, sendo utilizado de muitas maneiras incorretas, morrendo pela falta de fiscalização, manutenção e preservação. É urgente a realização de um sério trabalho de limpeza, manutenção, fiscalização e conservação, assim como é necessário um trabalho de intervenção educacional com os moradores do entorno e visitantes que utilizam o Rio e sua nascente como pontos recreativos.

A poluição aquática em Santos é uma questão ambiental prioritária, interdisciplinar por natureza, devido aos seus efeitos ecológicos, de saúde pública e sócio-econômicos. Trata-se de um problema amplamente relatado, cuja evolução acompanhou o histórico de desenvolvimento econômico da região. A presente pesquisa partiu de análises pontuais de um único, porém representativo, canal de drenagem pluvial urbana e das praias receptoras de seu conteúdo, para uma análise da poluição aquática no município como um todo. Para isso, foram usadas diferentes metodologias: coletas de amostras de água do canal e do mar, seguidas de análises físicas, químicas e ecotoxicológicas; pesquisas sobre as políticas públicas relacionadas à poluição hídrica local e aplicação de entrevistas estruturadas em frequentadores das praias. Além disso, foi analisada uma série histórica de dados microbiológicos das águas costeiras santistas. Os resultados indicam toxicidade aguda eventual nas amostras do canal e toxicidade crônica nas amostras da água do mar, coletadas em 2007, e severa contaminação de origem fecal, ao menos nas últimas três décadas. A pesquisa sobre as políticas públicas constatou que algumas surtiram efeitos positivos, mas não foram suficientes para melhorar a qualidade da água e mantê-la adequada, seja por sua descontinuidade, seja pelo crescimento acelerado no município, tanto demográfico, quanto econômico. A maioria das ações tem despontado em resposta a situações críticas, sendo implantadas sem planejamento adequado, visando conter apenas as conseqüências ambientais, e não as reais causas dos problemas. Como as políticas públicas geralmente não surgiram de processos de gestão participativa, também foram analisados alguns instrumentos de participação pública vigentes em Santos existe um envolvimento ainda incipiente da população, mas com iniciativas interessantes e potencialmente viáveis. Em relação às entrevistas, foram analisadas as respostas de 83 pessoas, entre as quais 77% eram residentes e o restante turistas. A maioria dos entrevistados acredita que as praias estão poluídas e que a situação dos canais é ainda mais crítica, corroborando com os resultados encontrados nas análises ecotoxicológicas e com os dados de contaminação microbiológica. Grande parte dos entrevistados não se banha justamente pela qualidade da água duvidosa, mas não se sentem responsáveis pela situação, nem demonstram iniciativa para pressionar o poder público a revertê-la. Quando os dados da pesquisa são integrados, é possível constatar que a qualidade da água das praias de Santos é crítica, como resultado de um modelo insustentável de desenvolvimento, com falta de planejamento e políticas públicas inconsistentes. Essa situação é agravada pela passividade observada entre muitos dos frequentadores das praias, que, apesar de cientes da poluição, não se envolvem na busca de soluções. Espera-se que a pesquisa contribua para o controle da contaminação aquática local, através do estabelecimento de políticas e ações que considerem todas as dimensões do problema e especialmente as suas causas. O presente estudo demonstra que a integração de diferentes métodos de avaliação permite uma compreensão maior acerca dos problemas ambientais e consequentemente pode subsidiar soluções mais efetivas. Além disso, foi enfatizada a importância da interdisciplinaridade em estudos de questões ambientais. / The aquatic pollution in Santos is a priority environmental issue, which presents many dimensions, as the ecological, public health and socioeconomic ones, having thus an interdisciplinary nature. This question has been extensively studied and its evolution was strictly related to the history of the regional economic development. The first phase of the present survey consisted of punctual analyses, made during 2007, of a single (but representative) urban pluvial drainage channel and the receiving beaches which are directly influenced by its contribution (i.e., to its contaminants inputs), in order to estimate the water quality (according to the aquatic pollution) in the overall shore. To achieve that, different methods were employed: sampling of channel and sea waters, and physical, chemical and ecotoxicological analyses. The second phase consisted of an investigation on the public policies related to the local hydric pollution. Concomitantly, as the third part of this survey, we conducted a study on the public knowledge on the aquatic pollution, by the application of structured interviews to the beaches users. Complementarily, the historical series of microbiological data for the coastal waters of Santos was assessed, to provide additional data. The results indicated eventual acute toxicity of channel water samples and chronic toxicity of sea water samples, and a severe fecal contamination at least in the last three decades. The evalution of public policies indicated that even though some of them had positive effects, they were not sufficient to improve the water quality and to maintain it appropriate, whether due to their discontinuity, whether to the demographic and economic accelerated growth of the city. The majority of actions have been implemented as responses to critical situations, without proper and wider planning, aiming only to control the consequences, and not the real causes of the problems. The public policies generally were not a product of participative processes; some instruments of public participation in Santos also were analyzed the involvement of the population is still incipient. However there are interesting and potentially viable initiatives. Regarding to the interviews, 83 beach users were considered to this survey, 77% were local residents and the remaining were tourists. The most interviewees believe that beaches are polluted and that the channels situation is still more critic, being in accordance with the results of ecotoxicological and microbiological data. Many of the beach users do not have contact with sea water due to its doubtful quality, but they do not feel responsible for the situation and not show initiative to press the public governement to act. When the obtained data are integrated, it is possible to notice that the water quality in Santos beaches is critical, as a product of an unsustainable model of development, lacking of planning and inconsistent public policies. The situation is aggravated due to the passivity observed among many of the beach users, who are aware of the pollution, but do not involve themselves to find effective solutions. We hope that this research may contribute to the control of aquatic local contamination, by the establishment of policies, enforcements and actions which consider all the dimensions of the problem, in special its causes. Moreover this research demonstrated that the integration of the different assessment methods may provide a stronger comprehension of the environmental issues and which consequently can subsidize much more effectively their solutions. Therefore it is highlighted the importance of interdisciplinarity to the studies involving environmental issues.

O pesticida Vertimec® 18CE é amplamente utilizado no controle de pestes na agricultura como inseticida, nematicida e acaricida. Embora aplicado sobre plantações agrícolas, ele pode atingir os sistemas aquáticos via escoamento superficial e afetar muitas espécies não-alvo. Um experimento simulando o escoamento sobre um campo contaminado consistiu em duas áreas separadas por uma distância de 6,0 metros, uma não-contaminada servindo como controle, e o outra contaminada com 0,125 L/\'M POT.2\' de Vertimec por meio de um pulverizador manual. As amostras do lixiviado oriundo do escoamento foram coletadas na parte inferior das áreas experimentais em declive. Uma fração das amostras foi utilizada para análises físicas e químicas e para realização de testes de toxicidade aguda em laboratório, e o restante da amostra foi inoculado em 12 câmaras cilíndricas, imersas em um tanque preenchido com a água da represa do Lobo, localizado em um mesocosmo construído em área próxima. Além da realização de um ensaio crônico in situ, também investigamos os efeitos de amostras ambientais oriundas dos mesocosmos a parir de ensaios agudos e crônicos realizados em laboratório com diferentes espécies de invertebrados aquáticos. Três espécies de invertebrados foram utilizadas como organismos teste neste estudo: Chironomus sancticaroli (Insecta Diptera), Hydra viridissima (Cnidaria, Hydridae) e Girardia tigrina (Plathyelminthes, Turbellaria). Os valores de LC50 96h para o pesticida Vertimec® 18EC foram: 0,14 mL/L (2,52 mg/L de abamectina) para H. viridissima, 0,28 mL/L (494,0 ug/L de abamectina) para G. tigrina e 0,54 \'10 POT.-5\' mL/L (97,0 ng/L de abamectina) para C. sancticaroli. Estes resultados indicam que C. sancticaroli tem uma sensitividade muito mais elevada a este pesticida do que as outras espécies testadas. No experimento em mesocosmos foi observado que no tratamento onde foi inoculado o lixiviado oriundo da área contaminada, o comprimento e a largura do corpo e da cápsula cefálica das larvas sobreviventes foram menores quando comparadas a dos indivíduos que foram expostos aos controles, evidenciando que a exposição crônica ao Vertimec via lixiviado oriundo da área contaminada afetou o crescimento das larvas de C. sancticaroli. Além disso, algumas larvas no mesocosmo contaminado (via runoff) tiveram deformidades no mento (4,2%). As análises referentes à comunidade de macroinvertebrados bentônicos total nos mesocosmos ao final do experimento mostraram que no tratamento com aplicação direta do Vertimec houve 100% de mortalidade de C. sancticaroli e um decréscimo de quase 80% na riqueza de táxons e na densidade total da comunidade bentônica quando comparada com a comunidade no controle. Nenhum táxon pertencente à Classe Insecta foi registrado no tratamento contaminado diretamente pelo contaminante utilizado corroborando a elevada toxicidade deste inseticida. Considerando todos os resultados, podemos concluir que o Vertimec® 18EC é altamente tóxico à comunidade de macroinvertebrados bentônicos e que os ecossistemas aquáticos contaminados com o escoamento superficial de áreas agrícolas com aplicação deste pesticida serão adversamente afetados. / The pesticide Vertimec® 18EC is largely used to control pests in agriculture acting as insecticide, nematicide and acaricide. Although applied in agricultural lands it can reach the aquatic systems via surface runoff and affect many non-target species. A field experiment simulating the runoff over a field plot contaminated which consisted of two 6 m apart agricultural plots, one non-contaminated, acting as control, and the other contaminated with 0.125 L/\'M POT.2\' of Vertimec using a backpack sprayer. Runoff samples were collected downstream the plots. One fraction was used for physical and chemical analysis and laboratory acute toxicity tests and the remaining sample was inoculated in 12 cylindrical chambers, immersed in a water tank filled with Lobo Reservoir water, in a nearby constructed mesocosm. Three species of freshwater invertebrates were used as test-organisms in this study: Chironomus sancticaroli (Insecta, Diptera), Hydra viridissima (Cnidaria, Hydridae) and Girardia tigrina (Plathyelminthes, Turbellaria). The values of LC50 96 h for Vertimec® 18EC were: 0.14 mL/L (2.52 mg/L of abamectin) for H. viridissima, 0.028 mL/L (494 ug/L of abamectin) for G. tigrina and 0.54 \'10 POT.-5\' mL/L (97 ng/L of abamectin) para C. sancticaroli. These results indicate that C. sancticaroli has a much higher sensitivity to this pesticide than the other species tested. In the experiments in mesocosms it was found that in the highest concentration tested, the length and width of the body and cephalic capsules of the larvae in the contaminated runoff were smaller than those of the larvae from both, the control and non-contaminated runoff, evidencing that the chronic exposure to the Vertimec contaminated runoff affected adversely the growth of C. sancticaroli larvae. In addition some larvae in the contaminated mesocosm had mentum deformities (4.2%). The analyses of the whole community in the mesocosms at the end of the experiment showed that in the treatment with direct application of Vertimec, there were 100% mortality of all C. sancticaroli inoculated larvae, and almost 80% decrease in the richness of taxa and in the total density of benthic organisms when compared with the community in the control. No taxa belonging to the Class Insecta were recorded in the contaminated treatment corroborating the strong toxicity of this insecticide. From all results obtained we conclude that Vertimec® 18EC is highly toxic to benthic macro-invertebrates community and that aquatic ecosystems contaminated by agricultural runoff with this pesticide will be adversely affected.

O pesticida Vertimec® 18CE é amplamente utilizado no controle de pestes na agricultura como inseticida, nematicida e acaricida. Embora aplicado sobre plantações agrícolas, ele pode atingir os sistemas aquáticos via escoamento superficial e afetar muitas espécies não-alvo. Um experimento simulando o escoamento sobre um campo contaminado consistiu em duas áreas separadas por uma distância de 6,0 metros, uma não-contaminada servindo como controle, e o outra contaminada com 0,125 L/\'M POT.2\' de Vertimec por meio de um pulverizador manual. As amostras do lixiviado oriundo do escoamento foram coletadas na parte inferior das áreas experimentais em declive. Uma fração das amostras foi utilizada para análises físicas e químicas e para realização de testes de toxicidade aguda em laboratório, e o restante da amostra foi inoculado em 12 câmaras cilíndricas, imersas em um tanque preenchido com a água da represa do Lobo, localizado em um mesocosmo construído em área próxima. Além da realização de um ensaio crônico in situ, também investigamos os efeitos de amostras ambientais oriundas dos mesocosmos a parir de ensaios agudos e crônicos realizados em laboratório com diferentes espécies de invertebrados aquáticos. Três espécies de invertebrados foram utilizadas como organismos teste neste estudo: Chironomus sancticaroli (Insecta Diptera), Hydra viridissima (Cnidaria, Hydridae) e Girardia tigrina (Plathyelminthes, Turbellaria). Os valores de LC50 96h para o pesticida Vertimec® 18EC foram: 0,14 mL/L (2,52 mg/L de abamectina) para H. viridissima, 0,28 mL/L (494,0 ug/L de abamectina) para G. tigrina e 0,54 \'10 POT.-5\' mL/L (97,0 ng/L de abamectina) para C. sancticaroli. Estes resultados indicam que C. sancticaroli tem uma sensitividade muito mais elevada a este pesticida do que as outras espécies testadas. No experimento em mesocosmos foi observado que no tratamento onde foi inoculado o lixiviado oriundo da área contaminada, o comprimento e a largura do corpo e da cápsula cefálica das larvas sobreviventes foram menores quando comparadas a dos indivíduos que foram expostos aos controles, evidenciando que a exposição crônica ao Vertimec via lixiviado oriundo da área contaminada afetou o crescimento das larvas de C. sancticaroli. Além disso, algumas larvas no mesocosmo contaminado (via runoff) tiveram deformidades no mento (4,2%). As análises referentes à comunidade de macroinvertebrados bentônicos total nos mesocosmos ao final do experimento mostraram que no tratamento com aplicação direta do Vertimec houve 100% de mortalidade de C. sancticaroli e um decréscimo de quase 80% na riqueza de táxons e na densidade total da comunidade bentônica quando comparada com a comunidade no controle. Nenhum táxon pertencente à Classe Insecta foi registrado no tratamento contaminado diretamente pelo contaminante utilizado corroborando a elevada toxicidade deste inseticida. Considerando todos os resultados, podemos concluir que o Vertimec® 18EC é altamente tóxico à comunidade de macroinvertebrados bentônicos e que os ecossistemas aquáticos contaminados com o escoamento superficial de áreas agrícolas com aplicação deste pesticida serão adversamente afetados. / The pesticide Vertimec® 18EC is largely used to control pests in agriculture acting as insecticide, nematicide and acaricide. Although applied in agricultural lands it can reach the aquatic systems via surface runoff and affect many non-target species. A field experiment simulating the runoff over a field plot contaminated which consisted of two 6 m apart agricultural plots, one non-contaminated, acting as control, and the other contaminated with 0.125 L/\'M POT.2\' of Vertimec using a backpack sprayer. Runoff samples were collected downstream the plots. One fraction was used for physical and chemical analysis and laboratory acute toxicity tests and the remaining sample was inoculated in 12 cylindrical chambers, immersed in a water tank filled with Lobo Reservoir water, in a nearby constructed mesocosm. Three species of freshwater invertebrates were used as test-organisms in this study: Chironomus sancticaroli (Insecta, Diptera), Hydra viridissima (Cnidaria, Hydridae) and Girardia tigrina (Plathyelminthes, Turbellaria). The values of LC50 96 h for Vertimec® 18EC were: 0.14 mL/L (2.52 mg/L of abamectin) for H. viridissima, 0.028 mL/L (494 ug/L of abamectin) for G. tigrina and 0.54 \'10 POT.-5\' mL/L (97 ng/L of abamectin) para C. sancticaroli. These results indicate that C. sancticaroli has a much higher sensitivity to this pesticide than the other species tested. In the experiments in mesocosms it was found that in the highest concentration tested, the length and width of the body and cephalic capsules of the larvae in the contaminated runoff were smaller than those of the larvae from both, the control and non-contaminated runoff, evidencing that the chronic exposure to the Vertimec contaminated runoff affected adversely the growth of C. sancticaroli larvae. In addition some larvae in the contaminated mesocosm had mentum deformities (4.2%). The analyses of the whole community in the mesocosms at the end of the experiment showed that in the treatment with direct application of Vertimec, there were 100% mortality of all C. sancticaroli inoculated larvae, and almost 80% decrease in the richness of taxa and in the total density of benthic organisms when compared with the community in the control. No taxa belonging to the Class Insecta were recorded in the contaminated treatment corroborating the strong toxicity of this insecticide. From all results obtained we conclude that Vertimec® 18EC is highly toxic to benthic macro-invertebrates community and that aquatic ecosystems contaminated by agricultural runoff with this pesticide will be adversely affected.

La pollution croissante des milieux aquatiques nécessite la mise au point de nouvelles technologies permettant d’optimiser la surveillance de la qualité de l’eau. Dans ce contexte, nous avons développé un biomarqueur de susceptibilité du degré de la pollution globale des milieux aquatiques intitulé « Sentinelle ». Le principe du biomarqueur Sentinelle est basé sur le niveau de coexpression de deux protéines « Multidrug Resistance » (MDR), la protéine ABCG2-like et la P-gp, dans les érythrocytes de la truite Salmo trutta fario. Le biomarqueur sentinelle a été validé en conditions in vitro grâce au développement des cultures primaires d’érythrocytes de truite. Après l’exposition des globules rouges de truites à des concentrations croissantes d’un polluant modèle, le Benzo-a-pyrène, l’expression de la protéine ABCG2-like et de la P-gp augmente d’une manière dose dépendante. Le biomarqueur Sentinelle a ensuite été validé en milieu naturel sur des truites fario en provenance de différents cours d’eau d’Auvergne. En milieu naturel, les deux protéines MDR sont exprimées différemment dans les érythrocytes de truites fario selon le degré de contamination du cours d’eau. En effet, dans une rivière où la pollution est faible voire nulle, seule la protéine ABCG2-like est exprimée, alors que dans une rivière présentant une contamination plus importante, la P-gp et l’ABCG2-like sont toutes les deux coexprimées par une réponse de type relais. Les expériences menées en conditions in vitro et en milieu naturel, laissent supposer que la protéine ABCG2-like assure une fonction de garde alors que la P-gp assurerait une fonction de protection défensive. En conséquence, selon le niveau d’expression de la protéine de garde et de la protéine de défense, le degré de contamination de la rivière pourrait être évalué. L’intérêt de l’utilisation du biomarqueur Sentinelle a aussi été validé sur des Salmonidés en provenance de pisciculture. Ce nouvel outil biologique apporte des informations plus intégratives et plus précoces sur la qualité des milieux aquatiques, informations essentielles pour une meilleure gestion des ressources en eau. / Increasing aquatic pollution requires the development of new technologies for to optimize the monitoring of water quality. In this context, we have developed a “biomarker of susceptibility” designating the degree of global pollution in aquatic medium, entitled "Sentinel". The Sentinel biomarker is based on the co-expression level of two major "Multidrug resistance" (MDR) proteins, such as ABCG2-like protein and P-gp, in erythrocytes of brown trout’s in response to aquatic pollution. After developing a primary erythrocyte culture, the Sentinel biomarker was validated in a controlled medium. Trout erythrocytes exposure to increasing concentrations of Benzo-a-pyrene, a model pollutant, induced an increase expression of ABCG2-like protein and P-gp by a dose-dependent response. The Sentinel biomarker was then developed in a natural environment, using the erythrocytes of brown trout collected from the various rivers located in the Auvergne region of France. In the natural environment, both MDR proteins are differentially expressed in the erythrocytes of brown trout depending on the degree of contamination of rivers. Indeed, wild brown trout erythrocytes in an uncontaminated river, expressed only the ABCG2-like protein, whereas, in a river presenting a higher contamination, P-gp and ABCG2-like were both coexpressed with a relay response. Experiments in vitro conditions and natural environment, suggest that ABCG2-like protein acts as a vanguard protective protein, in complement to P-gp which acts as a “defensive” protective protein. Consequently, according to the expression level of the vanguard and defense proteins, the degree of contamination of the river could be evaluated. The use advantage of the Sentinel biomarker has also been validated on Salmonidae erythrocytes form farmed fish. This new tool provides biological information more early and integrative on the quality of aquatic environments. These informations are essential for better management of water resources.