Orientador: Sarah Arana / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T12:36:44Z (GMT). No. of bitstreams: 1 Peterlini_MarianaFrem_M.pdf: 4643636 bytes, checksum: cd0d6e62df40edebf5ce8326f7c80cd0 (MD5) Previous issue date: 2012 / Resumo: A agricultura intensiva no Brasil tem aumentado o risco de contaminação dos recursos hídricos e dos organismos aquáticos por agroquímicos. Esse é o caso da atrazina, detectada em alta frequência em rios e córregos da região. Assim, neste trabalho, o pacu (Piaractus mesopotamicus) foi escolhido como bioindicador por sua importância econômica e ecológica em Bacias do Pantanal. As brânquias e o tegumento foram aqui avaliados como biomarcadores da toxicidade induzida por atrazina, uma vez que estes órgãos se encontram em contato direto com o ambiente aquático. Para avaliar a sensibilidade de alevinos de pacu e determinar a Concentração Letal Média (CL50) à atrazina foram conduzidos experimentos, em duplicata, em sistema estático por 96 horas, com as seguintes concentrações: 0,0; 13,2; 17,6; 22,0; 26,4; 30,8; 35,2; 39,6 mgL-1. A CL50 obtida para a atrazina foi de 28,58 mgL-1 (96h). Foram realizadas análises anatomopatológica, histopatológica qualitativa e semi-quantitativa para brânquias e análises anatomopatológica e histopatológica qualitativa para o epitélio tegumentar. Amostras de brânquia e de pele dorsal e ventral foram fixadas em Bouin e incluídas em parafina. Para o grupo controle não foi registrada qualquer alteração com os métodos empregados. Os alevinos expostos à atrazina apresentaram alterações comportamentais como: escurecimento da pele, boquejamento intenso e distúrbios motores, além disso, nesses aquários houve constante presença de muco na água; e, alterações anatomopatológicas, como: hiperemia na pele, dilatação da região ventral, presença de líquido no estômago e ascite sanguinolenta. Nas brânquias dos grupos tratados foram observadas as seguintes alterações morfológicas: edema e afastamento na região interlamelar, hipertrofia e hiperplasia na região interlamelar, destacamento do epitélio respiratório, hipertrofia de células do epitélio respiratório, congestão, telangiectasia e fusão de lamelas secundárias. Na pele dorsal e ventral dos grupos tratados não se observou alteração histopatológica. As alterações motoras observadas são provavelmente decorrentes do efeito neurotóxico da atrazina. As análises branquiais confirmam o comprometimento respiratório e de osmoregulação nesse órgão, sendo que a análise semi-quantitativa indicou a hipertrofia de células do epitélio respiratório como a lesão mais severa. Finalmente, as brânquias mostraram-se biomarcadores adequados na avaliação dos efeitos tóxicos da atrazina em pacu e o valor da CL50 sugere que a atrazina é levemente tóxica para o pacu, porém pelos demais resultados observados, se infere que só analisar a mortalidade não é o suficiente para determinar o dano causado por agrotóxicos em peixes, assim, recomenda-se o emprego de vários biomarcadores, como por exemplo, análise de vários outros órgãos. Os resultados obtidos reforçam a importância de estudos desta natureza, para embasar medidas ambientais preventivas e reguladoras quanto ao uso desse herbicida / Abstract: The intensive agriculture in Brazil has caused growing increase of pollution risks of hydric resources and the aquatic organisms by agrochemicals. This is the case of atrazine, detected in high frequency in the rivers and streams. Therefore, in this research, the pacu (Piaractus mesopotamicus) was chosen by bioindicator because of its great economic and ecological importance to the water ecosystem of the Pantanal region. The gills and skin were assessed here as biomarkers of induced toxicity by atrazine, considering that these organs are in direct contact with the aquatic environment. To evaluate the sensitivity of pacu fingerlings and determine the Median Lethal Concentration (LC50) by atrazine, experiments were conducted, performed in duplicate, in static system for 96 hours, with the following concentrations: 0.0; 13.2; 17.6; 22.0; 26.4; 30.8; 35.2; 39.6 mgL-1. The LC50 for pacu was 28.58 mgL-1 (96h). Anatomopathological, qualitative histopathological and semiquantitative histopathological analyses were performed for the gills, and anatomopathological and qualitative histopathological analysis were performed for the skin epithelium. Gills and dorsal and ventral skin samples were fixed in Bouin and processed for the inclusion in paraffin. The control group did not register any other alterations with the methods used. The fingerlings exposed to atrazine presented behavioral changes, such as: darkening of the skin, an increase in the frequency of gasping and motor disturbances. Furthermore, these aquariums presented the constant presence of mucus in the water; and, anatomopathological alterations, such as: skin hyperemia, dilation of the ventral region, the presence of liquid in the stomach and bloody ascites. In the gills of treated groups were found the following morphological alterations: edema and spacing in the interlamellar region, hypertrophy and hyperplasia in the interlamellar region, epithelial lifting, hypertrophy of respiratory epithelium cells, congestion, telangiectasis and fusion of the secondary lamellae. For dorsal and ventral skin of treated groups no histological alterations were observed. The motor alterations observed are probably due to the neurotoxic effects of atrazine. The gills analysis confirm respiratory and osmoregulatory compromise in this organ, noting that semi-quantitative analysis indicated the hypertrophy of respiratory epithelium cells as the most severe lesion. Finally, the gills showed adequate biomarkers in the assessment of toxic atrazine effects in pacu and the LC50 value suggests that atrazine is lightly toxic to pacu. However, by other results observed, it is concluded that analyzing the mortality index alone is not enough to determine the damages caused by agrotoxics in fishes, so the use of other biomarkers is recommended, like for instance, analysis some other organs. The results obtained reinforce the importance of research of this nature, in order to establish preventive and regulatory environmental measures regarding the use of this herbicide / Mestrado / Histologia / Mestra em Biologia Celular e Estrutural

Qu’il s’agisse de crèmes solaires, d’additifs à carburant ou d’emballages alimentaires, ces produits d’utilisation courante contiennent des nanoparticules, et l’ensemble des processus de leur production, utilisation et vieillissement conduit à un relargage direct ou non dans les milieux aquatiques. Le rejet accru de nanoparticules dans les écosystèmes d’eau douce est donc à prendre en considération et soulève la question de leurs effets potentiels sur les organismes y vivant. Le programme collaboratif ANR P2N – MESONNET (2011 – 2015) s’est attaché à décrire le devenir des nanoparticules (transfert et transformation dans les milieux et dans les organismes vivants) ainsi qu’à déterminer leur écotoxicité. Le travail présenté dans ce manuscrit a pris part à cette évaluation en étudiant les effets de nanoparticules d’argent, de titane et de carbone (nanotubes) sur des Crustacés Amphipodes appartenant au genre Gammarus. L’approche expérimentale mise en œuvre a cherché à évaluer, en laboratoire, l’effet sur les gammares de l’exposition aux nanoparticules en focalisant sur des biomarqueurs mesurés au niveau (sub)individuel (cellulaire, physiologique, comportemental) et sur des paramètres ayant trait à leur rôle fonctionnel dans les écosystèmes. La démarche s’est voulue la plus proche possible des conditions naturelles en envisageant des scénarios de contamination environnementalement réalistes : exposition dans un milieu complexe (eau prélevée dans le milieu naturel) à de faibles concentrations en nanoparticules et en présence de nourriture. Dans ce contexte, l’étude des effets des nAg sur trois espèces de gammares a révélé une plus forte sensibilité de G. roeseli, chez lequel l’exposition a engendré une réduction de l’activité locomotrice et de la production de fines particules de matière organique (FPOM). Sur le plan physiologique, une augmentation de la consommation en oxygène a été observée chez des gammares exposés aux nAg, et ce, malgré une stabilité de réponse des marqueurs du métabolisme énergétique étudiés. La consommation en oxygène s’est avérée corrélée aux concentrations en argent mesurées dans les organismes, ainsi qu’à la taille des nanoparticules testées. L’influence de la forme des nanoparticules (nAg : triangulaires plates et sphériques ; nTiO2 : cubiques ou en bâtonnets) et de la présence de matière organique (NTC) a été étudiée en mésocosmes, en ciblant principalement des indicateurs cellulaires (réserve énergétique, réponse antioxydante, dommage...) et individuels (osmorégulation, ventilation et locomotion). Les résultats obtenus suggèrent des effets dépendants de la forme des nanoparticules. L’ensemble de ces travaux souligne l’intérêt d’une double approche, écologique et écotoxicologique, permettant de mieux appréhender les effets des nanoparticules sur les écosystèmes d’eau douce / Products incorporating nanoparticles are now ubiquitous in our everyday life. For example, sunscreens, fuel additives or food packaging materials contain nanoparticles which could be released in the aquatic environment during the entire life cycle of the nanoproduct, from its production to its use and aging. Therefore an increase of nanoparticle emissions in the aquatic ecosystems needs to be considered and raises the question about their potential effects on living organisms. The collaborative program ANR P2N – MESONNET (2011- 2015) was focused on the evaluation of the nanoparticle fate (transfer and transformation in media and living organisms) and on their ecotoxicity. The present work took part in this assessment by studying the effects of nanoparticles of silver, titanium and carbon (nanotube) on the Crustaceans Amphipod Gammarus sp.. The experimental approach was designed to assess, in laboratory conditions, the effect of nanoparticles on the gammarids, focusing on biomarker measurements at the (sub)individual level (cellular, physiological, behavioral) and on some parameters related to their functional roles. Experiments were designed as close as possible to natural conditions using environmentally realistic contamination scenario: exposure in a complex media (natural water from river) to low nanoparticle concentrations and with food. In this context, the effects of nAg were investigated in three gammarids species and results revealed a higher sensibility of G. roeseli, for which exposure induced a decrease of the locomotor activity and of the production of fine particles of organic matter (FPOM). At the physiological level, an increase of the oxygen consumption was observed in gammarids exposed to nAg, despite a stability observed for the studied markers of energetic metabolism. The oxygen consumption was correlated with the silver concentration measured in the organisms, and also with the tested nanoparticle sizes. The influence of the nanoparticle forms (nAg: plate triangular and spherical; nTiO2: cubic and rod-shaped) and of the organic matter (CNT) was studied in mesocosms by focusing mainly on cellular indicators (energetic storage, antioxidant response, damage…) and individual ones (osmoregulation, ventilation and locomotion). The obtained results suggest form-dependent effects of nanoparticles. This work shows the interest of a coupled approach using ecological and ecotoxicological endpoints to better understand the effects of nanoparticles on freshwater ecosystems

Les polychlorobiphényles (PCB) et les polybromodiphényléthers (PBDE) sont deux familles de composés hydrophobes partageant certaines propriétés physico-chimiques. Des travaux antérieurs ont montré que ces composés pouvaient affecter les traits d’histoire de vie des poissons. Il s’avère cependant que les études approchant des situations environnementales sont rares. Cette thèse se divise en trois parties. La première partie s’est attachée à évaluer les effets d’une exposition à un mélange réaliste de PCB et de PBDE sur les traits d’histoire de vie du poisson-zèbre. Les résultats ont montré une taille asymptotique plus élevée, une probabilité de ponte retardée chez les individus exposés et une faible survie des larves F1 en situation de jeûne issues des pontes précoces des individus exposés. La seconde partie a cherché à identifier, sur la base d’un modèle bioénergétique (DEB), les modes d’action physiologique (PMoA) de PCB et PBDE. Deux PMoAs probables ont été révélés: le premier est lié à l’augmentation de la fraction d’énergie allouée à la maintenance et la croissance somatique et le deuxième est lié à l’augmentation des coûts de production d’un œuf. La troisième partie de la thèse s’est intéressée à évaluer les conséquences de la contamination sur la dynamique de population de poissons. Les résultats ont montré une réduction de l’abondance numérique, une augmentation de la biomasse et un grand risque de surexploitation chez la population exposée, Les résultats de cette thèse pourraient constituer un outil d’aide à une gestion durable des stocks halieutiques qui prennent en compte les effets de multiples contaminations qui se surajoutent à la pression de pêche. / Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) are two families of hydrophobic compounds sharing some similar environmental properties. Previous studies have demonstrated that these compounds could affect fish life-history traits. However, few studies have focused on environmental situations. This thesis is divided into three parts. The first part aimed to evaluate the effects of a realistic mixture of PCBs and PBDEs on the life-history traits of zebrafish. The results showed a slower growth, but to a larger asymptotic length, and delayed spawning probability in exposed fish. In addition, offspring issued from early spawning events of exposed fish exhibited a lower larval survival under starvation condition. The second part aimed to identify, on the basis of a bioenergetic model (DEB), the physiological modes of action (PMoA) of PCBs and PBDEs. Two potential PMoAs have been revealed: the first one was through an increase of the fraction of energy allocated to somatic maintenance and growth and the second one was through an increase of the cost of production of an egg. The third part focused on the population dynamical consequences of the individual life-history effects of PCBs and PBDEs. The results showed a lower abundance, a higher biomass and a higher risk of overexploitation in exposed population compared to control population. The results of this thesis could be a helpful tool for sustainable management of fish stocks that take into account the effects of multiple contaminations that are added to the fishing pressure.

No presente estudo, duas espécies nativas, Hydra viridissima Pallas, 1766 e Hydra salmacidis Silveira, Gomes e Silva, 1997 foram estudadas visando obter informações sobre a biologia, a ecologia e a potencialidade destas espécies como organismos-teste em estudos ecotoxicológicos. Para isso, foram avaliados alguns aspectos de seus ciclos de vida, como as taxas de crescimento individual e populacional por meio do cultivo das hidras em laboratório, e foram investigados seus potenciais predadores e presas preferenciais dentre os principais componentes da comunidade de invertebrados aquáticos, presentes em corpos de água doce similares aos ambientes onde as espécies ocorrem naturalmente. Avaliou-se também o efeito de diferentes agentes tóxicos, tais como metais, sais, fármacos e surfactantes, às espécies estudadas, bem como seus efeitos sobre a capacidade de regeneração destas hidras. Além disso, avaliou-se a toxicidade de cianotoxinas e de amostras ambientais de água e sedimento aos organismos. Os resultados indicaram que macroinvertebrados, como as ninfas de Odonata, as larvas do Diptera Chaoborus sp. e os Copepoda Cyclopoida, além do peixe Poecilia reticulata, usuais predadores de invertebrados em águas doces, não consumiram as hidras, e que H. viridissima tem como presas preferenciais as fases jovens dos copépodos calanóides e pequenos cladóceros, enquanto que H. salmacidis, além destas presas, também tem preferência pelos organismos adultos dos copépodos calanóides. Os tempos de geração obtidos para as espécies de Hydra não diferiram significativamente entre si (5,2 ± 1,14 e 5,2 ± 0,79 dias para H. viridissima e H. salmacidis, respectivamente) e foram relativamente curtos, enquanto que H. viridissima apresentou um tempo de duplicação da população (6,38 ± 2,40 dias) significativamente maior do que H. salmacidis (4,11 ± 0,21 dias). Em relação à sensibilidade das hidras aos agentes tóxicos, as duas espécies foram bastante sensíveis quando comparadas a espécies de outros grupos de invertebrados rotineiramente utilizados como organismos-teste, e ora uma, ora outra espécie teve maior sensibilidade aos agentes tóxicos testados. Estas espécies nativas de Hydra são bastante adequadas para os estudos ecotoxicológicos por serem facilmente cultivadas, com baixo custo, e por responderem por meio de modificações morfológicas graduais às condições de crescente toxicidade, evidenciando, além das concentrações letais, as concentrações subletais de agentes tóxicos ou amostras ambientais. São, portanto, excelentes organismos-teste e devido à sua capacidade de regeneração podem ser utilizadas como indicadores de toxicidade. / In the present study, two native species, Hydra viridissima Pallas, 1766 and Hydra salmacidis Silveira, Gomes and Silva, 1997 were studied regarding their biology, ecology and performance as test organisms for ecotoxicological studies. Aspects of their life cycle such as individual and population growth rates were assessed by culturing them in the laboratory, and their potential predators and preferential preys were investigated among the main components of communities from water bodies similar to those in which the hydras naturally occur. Several acute ecotoxicological tests were performed to assess the effect of different compounds such as metals, salts, pharmaceuticals and surfactants to Hydra species. The effect of these toxicants on Hydra regeneration ability was also assessed. Besides that, tests were performed with cianotoxins and environmental samples of water and sediments. The results indicated that macroinvertebrates such as Odonata nymphs, Chaoborus sp. larvae (Diptera), Cyclopoida copepods, and the guppy Poecilia reticulata, usually predators of invertebrates in freshwaters, did not consume the hydras. It was also found that H. viridissima has positive food selection for the nauplii and copepodites of calanoid copepods and small cladocerans, whereas H. salmacidis, besides these preys, also have preference for adult calanoid copepods. Generation times did not significantly differ between the species of Hydra (5.2 ± 1.14 and 5.2 ± 0.79 days for H. viridissima and H. salmacidis, respectively) and were relatively short. H. viridissima had a population doubling time (6.38 ± 2.40 days) significantly longer than that of H. salmacidis (4.11 ± 0.21 days). Both species of Hydra had high sensitivity to the toxicant compounds tested when compared to other species of invertebrate groups routinely used as test organisms. Regarding the sensitivity to the compounds tested, one or the other species can be more sensitive depending on the compound. These native species of Hydra are suitable for as test organisms because: they can be easily and low costing laboratory cultured, and they respond to increasing toxicity by gradually changing their morphology showing lethal or sublethal effects of toxicants or contaminated environmental samples. They are therefore excellent test organisms and due to their ability to regenerate they can be used as toxicity indicators.

Nos últimos anos o aumento do consumo de agrotóxicos em todo o mundo tem preocupado agências nacionais e internacionais em decorrência dos diversos impactos ambientais e das sérias implicações à saúde humana. Dentre os inúmeros produtos utilizados na agricultura destaca-se o agrotóxico Vertimec 18CE, indicado para 42 usos distintos, compreendendo 24 culturas, visando o combate de 17 diferentes pragas. Considerando os riscos ambientais associados ao uso de agrotóxicos, se faz necessária a realização de estudos mais abrangentes para melhor compreender as inter-relações existentes no processo de contaminação e os reais efeitos sobre a biota aquática. Nesse sentido, procurou-se, na presente pesquisa, avaliar os efeitos do Vertimec 18CE sobre Danio rerio (Cypriniformes, Cyprinidae) por meio de experimentos in situ, a partir da simulação da contaminação do ambiente aquático por água proveniente do carreamento superficial de parcelas de solo contaminadas e não contaminadas (controle), além da aplicação direta do contaminante na coluna d\'água; bem como por experimentos laboratoriais com diferentes diluições do runoff (simulando o aporte diferenciado quantitativamente), com amostras ambientais dos mesocosmos e pela avaliação da sensibilidade ao produto comercial Vertimec 18CE. Para isso foram utilizadas duas parcelas de solo com 8 \'M POT.2\' cada e inclinação de 6%, das quais uma recebeu a contaminação segundo a dose recomendada para o cultivo de morango (0,125 litros de calda por \'M POT.2\') e a outra somente aspersão com água, servindo como controle. Posteriormente foi simulada uma chuva de 20 a 30 mm e coletada a água de runoff para a execução dos experimentos in situ e laboratoriais com as diluições de runoff. Simultaneamente foram realizadas análises físicas e químicas da água de runoff e dos mesocosmos, além da caracterização física e química do sedimento do experimento in situ e das parcelas de solo utilizadas. As análises limnológicas do experimento in situ demonstraram o aumento de nutrientes (nitrogênio e fósforo), material em suspensão e turbidez, com efeitos sobre a concentração de clorofila a, feofitina, condutividade e oxigênio dissolvido. Os resultados obtidos nos testes de toxicidade indicaram diferenças entre os testes laboratoriais (com condições mais controladas) e in situ (condições mais próximas ao sistema natural), registrando-se toxicidade crônica apenas no experimento in situ, para o tratamento com contaminação direta com o agrotóxico. Não foram observados efeitos agudos ou crônicos nos experimentos laboratoriais com as diluições de runoff e nem com as amostras de água dos mesocosmos. Os efeitos tóxicos registrados nos testes de toxicidade crônica de curta duração indicam maior tempo de permanência da formulação comercial em relação ao seu princípio ativo isolado, a abamectina, demonstrando os riscos potenciais do uso do produto comercial sem estudos prévios. Quanto à sensibilidade os valores de CL50, 48h estiveram entre 3,88 e 6,13 \'mü\'l/L de Vertimec 18CE (média de 4,74 \'mü\'l/L), enquanto que os valores da CE50, 48h estiveram entre 2,46 e 5,63 \'mü\'l/L (média de 3,66 \'mü\'l/L). No teste crônico de curta duração obteve-se valores de 3,0 \'mü\'l/L; 2,0 \'mü\'l/L e 2,5 \'mü\'l/L para CEO, CENO e VC, respectivamente. / In recent years the increased of pesticides use in the world has been concerned national and international agencies as a result of various environmental impacts and the serious implications to human health. Among the many products used in agriculture, stands out the pesticide Vertimec 18CE, nominated for 42 different uses, comprising 24 cultures, aimed to combat 17 different pests. Considering the environmental risks associated with pesticide use, it is necessary to carry out further studies to better understand the interrelationships within the process of contamination and the actual effects on aquatic biota. In this sense, this study aimed to evaluate the effects of Vertimec 18CE on Danio rerio (Cypriniformes, Cyprinidae) through in situ experiments, by simulating the contamination of the aquatic environment by water from the surface runoff of contaminated and uncontaminated (control) soil, beyond the direct application of the contaminant in the water column, as well laboratory experiments with different dilutions of the runoff (simulating the contribution quantitatively different), with samples of the mesocoms and the assessment of the sensitivity to the commercial product Vertimec 18CE. For this we used two parcels of land with 8 \'M POT.2\' each and incline of 6%, one received a recommended dose contamination for the cultivation of strawberries (0.125 L per \'M POT. 2\') and the other only water spraying, as a control. Subsequently we simulated a 20 to 30 mm of rain and runoff water was collected for the addition of in situ and laboratory experiments with dilutions of runoff. Were performed physical and chemical analysis of runoff water and the mesocosms, as well as physical and chemical characterization of sediment of in situ experiment and the parcels of soil used. Limnological analyses of in situ experiment showed an increase of nutrients (nitrogen and phosphorus), suspended solids and turbidity, with effects on the concentration of chlorophyll, pheophytin, conductivity and dissolved oxygen. The results obtained in toxicity tests indicated differences between the laboratory tests (with more controlled conditions) and in situ (conditions closer to the natural system), registering only partial chronic toxicity in the in situ experiment, to treatment with direct pesticide contamination. There were no acute or chronic effects in laboratory experiments with dilutions or with runoff and water samples from mesocosms. Toxic effects reported in chronic toxicity tests indicated higher persistence of the commercial formulation in relation to its isolated active ingredient, abamectin, demonstrating the potential risks of the use of commercial product without prior studies. In relation to the sensitivity of LC50, 48h values were between 3.88 and 6.13 \'mü\'l/L Vertimec 18CE (medium value of 4.74 \'mü\'l/L), whereas the EC50, 48h values were among 2.46 and 5.63 \'mü\'l/L (medium value of 3.66 \'mü\'l/L). In the partial chronic test we obtained a values of 3.0 \'mü\'l/L, 2.0 and 2.5 \'mü\'l/L for LOEC, NOEC and CV, respectively.

This thesis is focused on attestation of ecotoxicity of octachlorstyrene (OCS), persistent organic pollutant (POP), by biological tests of toxicity. The toxicity of OCS was determined by using two kinds of alternative tests applied on fresh water organisms. First of them, Thamnotoxkit, is based on observation of mobility, immobilization or even killing of crustacean Thamnocephalus platyurus. The second one, Daphnotoxkit, applies crustacean Daphnia magna. For purposes of comparison, a standard test of toxicity – so called test of root growth inhibition of Sinapis alba and two fytotests – test of growth inhibition of Lemna minor and test of root growth inhibition of Allium cepa were performed as well.

Introduction: Prostate cancer is the most common type of cancer in Sweden and is often treated using antiandrogenic drug therapy. Two substances belonging to this class of pharmaceuticals are bicalutamide and flutamide. After excretion from the human body, the drug molecules enter the wastewater treatment plant (WWTP). The WWTPs are not effective enough to completely remove pharmaceutical residues, why presence of both bicalutamide and flutamide can be detected in WWTP effluent water. Previous findings: Antiandrogens have been reported to affect reproduction in adult fish, but studies regarding possible effects on the embryonic development of fish are few. Aim: The present study sought to investigate if exposure to bicalutamide or flutamide cause toxicity in the early developmental stages of zebrafish embryos, and whether negative effects occur within concentrations relevant to measured environmental levels. Method: A modified OECD FET-test was used, where additional sublethal endpoints were included and the time period for assessment extended to 144 hours post fertilization (hpf). In addition, a locomotor activity assay was performed at 144 hpf in order to observe any sub-lethal swimming behavioral effects. Results: High doses (10 mg/L) of flutamide led to 100% lethality of the zebrafish embryos but the results suggest no acute toxic effects in the high dose treatment group of bicalutamide, or of either flutamide or bicalutamide within in the low (0.1 mg/L) or intermediate (1 mg/L) treatment groups. Neither did the locomotor activity assay result in statistically significant results, although the pattern of swimming activity in the low dose groups suggests that behavioral developmental effects could be present. Conclusions: High doses of flutamide caused mortality of the embryos, but no lethal or sublethal effects were present at environmentally relevant concentrations. The modest outcome of present study however suggests that further investigation of behavioral developmental effects of antiandrogens could be of future relevance. Analysis of the expression of genes related to neuronal growth, memory and other cognitive behaviors associated with behavioral changes, would then be of interest for further studies.

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