<p>The synthetic estrogen 17α-ethynylestradiol (EE2), an endocrine disruptor originating from birth control and hormone replacement therapy, is discharged in wastewater treatment plant (WWTP) effluents. The present study employed radio-labeled EE2 to examine the impact of temperature and salinity on the uptake of EE2 in male killifish (<em>Fundulus heteroclitus</em>), a model euryhaline teleost. Fish were exposed to a nominal concentration of 100 ng/L EE2 for 2 h. Actual concentrations were lower due to EE2 adsorption to the exposure system, but uptake rates were normalized to 100 ng/L. Oxygen consumption rates (MO₂), whole body EE2 uptake rates, and tissue-specific EE2 distribution were monitored. EE2 uptake by freshly killed fish was negligible. In killifish acclimated to 18^OC at 16 ppt (50 % seawater), MO₂ and EE2 uptake were both much lower after 24-h exposure to 10^OC and 4^OC, and increased after 24-h exposure to 26^OC. Transfer of killifish to fresh water for 24 h tended to lower EE2 uptake rate, and long-term acclimation to fresh water reduced it by about 70 %. Long-term acclimation to 100 % sea water (32 ppt) also reduced EE2 uptake rate by about 50 % relative to 16 ppt. However this was not seen in juvenile rainbow trout (<em>Oncorhynchus mykiss</em>) where uptake rates were the same in FW- and 16 ppt-acclimated trout. The tissue-specific accumulation of EE2 was found to be the highest (40-60 % of the total) in the liver plus gall bladder across all exposures, and the great majority of this was in the bile in killifish, regardless of temperature or salinity, whereas in trout accumulation was the highest in the carcass at 70 % of the total. The carcass was the next highest accumulator (30-40 %) in killifish, followed by the gut (10-20 %) with only small amounts in gills and spleen. Drinking rate, measured with radio-labeled polyethylene glycol-4000, was about 25-times greater in 16 ppt-acclimated killifish relative to freshwater-acclimated animals. However, drinking accounted for less than 30 % of gut accumulation, and therefore a negligible percentage of whole body EE2 uptake rates. In general, there were strong positive relationships between EE2 uptake rates and MO₂, suggesting similar pathways for uptake across the gills of these lipophilic molecules. These data will be useful in developing a predictive model of how variations in key environmental parameters (salinity, temperature, dissolved oxygen) affect EE2 uptake in estuarine fish, so as to determine optimal timing and location of WWTP discharges.</p> / Master of Science (MS)

Hormonaly active compounds in wastewaters represent nowdays a serious problem. Proceses currently used in watewater treatment plants (WWTP) are unefficient in removing these compounds from contaminated wastewaters. The compounds are supposed to sorb onto solid sludge elements and sediments. In this work seven endocrine disruptors were detected in the sludge samples from WWTPs. A new sensitive method for detection of seven selected endocrine disruptors (4-nonylphenol, bisphenol A, estriol, 17β-estradiol, estrone, 17α- ethynylestradiol, irgasan) was developed. The method is based on accelerated solvent extraction (ASE), gel permeation chromatography (GPC) and solid phased extraction. For final extract analysis, gas chromatography coupled with mass spectrometry (GC/MS) was used. The efficiency of this method was tested using artificially contaminated sludge and the method was used to analyse real samples from several WWTPs in Czech Republic. The effect of sludge age on detection of individual analytes was assessed as well. The concentrations of endocrine disruptors measured in the samples reached up to 1 µg/g. The results are comparable or higher then those reported in other works and they show the necessity of further research on endocrine disruptors in the environment.

Many chemicals present in the aquatic environment can interfere with physiological functions in fish. Exposure to chemicals can be revealed by the use of biomarkers. Induction of 7-ethoxyresorufin O-deethylase (EROD) activity is a commonly used biomarker for exposure to CYP1A inducers such as dioxins and polyaromatic hyrdrocarbons. Vitellogenin is a frequently used biomarker for estrogenic compounds in various fish species whereas a biomarker for androgens, spiggin, is only found in sticklebacks. The main objectives of this thesis were to evaluate gill EROD activity as a biomarker and the three-spined stickleback as a model species in ecotoxicological studies. EROD activities were measured in gill, liver and kidney in rainbow trout (Oncorhynchus mykiss) caged in urban areas in Sweden. EROD induction was most pronounced in the gill. Also in fish caged at reference sites, with an expected low level of known CYP1A inducers, a marked gill EROD induction was found. One suggested inducer in rural waters is humic substances (HS). To evaluate the EROD-inducing capacity of HS, three-spined sticklebacks (Gasterosteus aculeatus) were exposed to HS of natural or synthetic origin. Both kinds of HS caused significant EROD induction. Gill EROD activities were also induced in sticklebacks exposed to ethynylestradiol (EE2) and β-naphthoflavone (βNF), alone and in combinations. Production of vitellogenin was induced in sticklebacks exposed to ≥50 ng EE2/l and a significant decrease in spiggin production was observed in individuals exposed to 170 ng EE2/l. Results from this thesis further strengthen the contention that gill EROD activity is a very sensitive biomarker for CYP1A inducers and that the stickleback is a suitable biomonitoring species, especially for exposure to CYP1A inducers. The finding that not only classical CYP1A inducers but also HS and high EE2 concentrations stimulate gill EROD activity is of significance for the interpretation of biomonitoring data.

Neste trabalho foram realizados estudos de reações de bio-oxidação dos esteroides Progesterona e 17&alpha;-Etinilestradiol por fungos de ambiente marinho. Para a Progesterona foi feita uma triagem de reações com os caldos enzimáticos e as massas miceliais dos fungos Aspergillus sydowii CBMAI 934, Aspergillus sydowii CBMAI 935, Penicilium oxalicum CBMAI 1185 e Penicilium citrinum CBMAI 1186. Para as reações envolvendo os fungos A. sydowii CBMAI 935, P. oxalicum CBMAI 1185 e P. citrinum CBMAI 1186 os produtos majoritários observados foram os metabólitos Testosterona e Testololactona, produtos de oxidação da Progesterona promovida por enzimas do tipo Baeyer-Villiger monoxigenases. Para as reações envolvendo o fungo A. sydowii CBMAI 934 foram observados produtos de hidroxilação da Progesterona em carbonos do núcleo esteroidal não ativados eletronicamente, sendo produtos de oxidação da Progesterona promovida por enzimas do citocromo P-450. Da triagem foram selecionados os fungos A. sydowii CBMAI 934 e A. sydowii CBMAI 935 para executar reações de bio-oxidação em quintuplicatas (32ºC, 130 rpm, pH 7,4) utilizando o caldo enzimático e a massa micelial para isolamento e caracterizações espectroscópicas (RMN, EMAR e IV) e polarimétricas dos produtos majoritários. Após 14 dias de reação com o caldo enzimático do fungo A. sydowii CBMAI 934 foi isolado a 7,15-di-Hidroxiprogesterona com 13 % de rendimento em massa. Contudo não foi possível determinar com exatidão a configuração relativa dos grupos hidroxilas. Já para a reação com a massa micelial do fungo A. sydowii CBMAI 934 após 14 dias de reação foi isolado o produto 15&beta;-Hidroxiprogesterona com 58% de rendimento em massa. Após 7 dias de reação com o caldo enzimático do fungo A. sydowii CBMAI 935 foram isolados os produtos Testosterona e Testololactona com rendimentos em massa de 24% e 36%, respectivamente. A Testololactona também foi isolada da reação com a massa micelial do fungo A. sydowii CBMAI 935 após 7 dias com rendimento em massa de 87%. Para o 17 &alpha;-Etinilestradiol foi feita uma triagem com os mesmos fungos utilizados na triagem das reações com a Progesterona, exceto com o fungo A. sydowii CBMAI 934. Após 7 dias de reação e nas mesmas condições das reações com a Progesterona, não foram observados produtos de biotransformação do 17 &alpha;-Etinilestradiol. Este estudo envolvendo a bio-oxidação da Progesterona mostra o potencial de biotransformação de fármacos esteroidais no meio ambiente. / In this work was performed bio-oxidation studies of steroids progesterone and 17&alpha;-ethynylestradiol by marine-derived fungi. For progesterone was performed a reactions screening with culture broth and micelia from Aspergillus sydowii CBMAI 934, Aspergillus sydowii CBMAI 935, Penicilium oxalicum CBMAI 1185 e Penicilium citrinum CBMAI 1186. For reactions using A. sydowii CBMAI 935, P. oxalicum CBMAI 1185 e P. citrinum CBMAI 1186 the majoritarian products obtained were testosterone and testololactone, products from progesterone oxidation by Baeyer-Villiger mono-oxygenases enzymes. For reactions using A. sydowii CBMAI 934 was observed hydroxylation products from progesterone in carbons of progesterone by eletronic factors. The enzymes than promoted the hydroxylation of the progesterone were of citochrome P-450 group. From screening was selected A. sydowii CBMAI 934 and A. sydowii CBMAI 935 for quintuplicate bio-oxydations (32ºC, 130 rpm, pH 7,4) using the culture broth and micelia for isolation and spectroscopicals caracterizations (NMR, HRMS and IR) and polarimetrics of majoritary products. After 14 days, the reaction with culture broth from A. sydowii CBMAI 934 was isolated 7,15-dihydroxyprogesterone with 13% yield. But, the configurations of hydroxy groups were not possible to determine. Already for reaction with micelia from A. sydowii CBMAI 934 was isolated 15&beta;-hydroxyprogesterone with 58% yield. After 7 days the reaction with culture broth from A. sydowii CBMAI 935 was isolated the testosterone and testololactone with 24% and 36% yields, respectively. The Testololactone also was isolated from reaction with micelia from A. sydowii CBMAI 935 after 7 days with 87% yield. For 17&alpha;-ethynylestradiol was performed a screening with same fungi used in reaction screening with Progesterone, except the A. sydowii CBMAI 934. After 7 days in same condictions of reactions with Progesterone, It was not observed products from 17&alpha;-ethynylestradiol. This study involving the bio-oxidation from Progesterone showed the potential of biontrasnformations steroidal drugs in environment.

Neste trabalho foram realizados estudos de reações de bio-oxidação dos esteroides Progesterona e 17&alpha;-Etinilestradiol por fungos de ambiente marinho. Para a Progesterona foi feita uma triagem de reações com os caldos enzimáticos e as massas miceliais dos fungos Aspergillus sydowii CBMAI 934, Aspergillus sydowii CBMAI 935, Penicilium oxalicum CBMAI 1185 e Penicilium citrinum CBMAI 1186. Para as reações envolvendo os fungos A. sydowii CBMAI 935, P. oxalicum CBMAI 1185 e P. citrinum CBMAI 1186 os produtos majoritários observados foram os metabólitos Testosterona e Testololactona, produtos de oxidação da Progesterona promovida por enzimas do tipo Baeyer-Villiger monoxigenases. Para as reações envolvendo o fungo A. sydowii CBMAI 934 foram observados produtos de hidroxilação da Progesterona em carbonos do núcleo esteroidal não ativados eletronicamente, sendo produtos de oxidação da Progesterona promovida por enzimas do citocromo P-450. Da triagem foram selecionados os fungos A. sydowii CBMAI 934 e A. sydowii CBMAI 935 para executar reações de bio-oxidação em quintuplicatas (32ºC, 130 rpm, pH 7,4) utilizando o caldo enzimático e a massa micelial para isolamento e caracterizações espectroscópicas (RMN, EMAR e IV) e polarimétricas dos produtos majoritários. Após 14 dias de reação com o caldo enzimático do fungo A. sydowii CBMAI 934 foi isolado a 7,15-di-Hidroxiprogesterona com 13 % de rendimento em massa. Contudo não foi possível determinar com exatidão a configuração relativa dos grupos hidroxilas. Já para a reação com a massa micelial do fungo A. sydowii CBMAI 934 após 14 dias de reação foi isolado o produto 15&beta;-Hidroxiprogesterona com 58% de rendimento em massa. Após 7 dias de reação com o caldo enzimático do fungo A. sydowii CBMAI 935 foram isolados os produtos Testosterona e Testololactona com rendimentos em massa de 24% e 36%, respectivamente. A Testololactona também foi isolada da reação com a massa micelial do fungo A. sydowii CBMAI 935 após 7 dias com rendimento em massa de 87%. Para o 17 &alpha;-Etinilestradiol foi feita uma triagem com os mesmos fungos utilizados na triagem das reações com a Progesterona, exceto com o fungo A. sydowii CBMAI 934. Após 7 dias de reação e nas mesmas condições das reações com a Progesterona, não foram observados produtos de biotransformação do 17 &alpha;-Etinilestradiol. Este estudo envolvendo a bio-oxidação da Progesterona mostra o potencial de biotransformação de fármacos esteroidais no meio ambiente. / In this work was performed bio-oxidation studies of steroids progesterone and 17&alpha;-ethynylestradiol by marine-derived fungi. For progesterone was performed a reactions screening with culture broth and micelia from Aspergillus sydowii CBMAI 934, Aspergillus sydowii CBMAI 935, Penicilium oxalicum CBMAI 1185 e Penicilium citrinum CBMAI 1186. For reactions using A. sydowii CBMAI 935, P. oxalicum CBMAI 1185 e P. citrinum CBMAI 1186 the majoritarian products obtained were testosterone and testololactone, products from progesterone oxidation by Baeyer-Villiger mono-oxygenases enzymes. For reactions using A. sydowii CBMAI 934 was observed hydroxylation products from progesterone in carbons of progesterone by eletronic factors. The enzymes than promoted the hydroxylation of the progesterone were of citochrome P-450 group. From screening was selected A. sydowii CBMAI 934 and A. sydowii CBMAI 935 for quintuplicate bio-oxydations (32ºC, 130 rpm, pH 7,4) using the culture broth and micelia for isolation and spectroscopicals caracterizations (NMR, HRMS and IR) and polarimetrics of majoritary products. After 14 days, the reaction with culture broth from A. sydowii CBMAI 934 was isolated 7,15-dihydroxyprogesterone with 13% yield. But, the configurations of hydroxy groups were not possible to determine. Already for reaction with micelia from A. sydowii CBMAI 934 was isolated 15&beta;-hydroxyprogesterone with 58% yield. After 7 days the reaction with culture broth from A. sydowii CBMAI 935 was isolated the testosterone and testololactone with 24% and 36% yields, respectively. The Testololactone also was isolated from reaction with micelia from A. sydowii CBMAI 935 after 7 days with 87% yield. For 17&alpha;-ethynylestradiol was performed a screening with same fungi used in reaction screening with Progesterone, except the A. sydowii CBMAI 934. After 7 days in same condictions of reactions with Progesterone, It was not observed products from 17&alpha;-ethynylestradiol. This study involving the bio-oxidation from Progesterone showed the potential of biontrasnformations steroidal drugs in environment.

Os protocolos de Avaliação de Risco Ambiental (ERA) de Fármacos e Produtos de Cuidados Pessoais (FPCP) recomendam o uso de ensaios ecotoxicológicos tradicionais (por exemplo algas, bactérias, invertebrados, peixes) e a avaliação de efeitos em um único nível de organização biológica para a determinação dos efeitos potenciais dos FPCP à biota. Considerando que efeitos em nível de sub-indivíduo pode afetar igualmente a aptidão ecológica de organismos marinhos, e que os mesmos estão cronicamente expostos aos FPCP, o objetivo do presente estudo foi avaliar o risco ambiental de triclosan (TCS), ibuprofeno (IBU) e 17&alpha;-etinilestradiol (EE2) em sedimentos marinhos utilizando respostas de efeitos sub-individuais e populacionais. Por meio do HPLC-ESI-MS/MS, as concentrações ambientais de TCS e IBU foram quantificadas em sedimentos marinhos coletados no entorno do emissário submarino de esgoto de Santos (Baía de Santos, São Paulo - Brasil) com 15,14 e 49,0 ng.g-1, respectivamente, enquanto o EE2 não foi detectado (<33 ng.g-1). Uma bateria de ensaios de toxicidade crônica (desenvolvimento embriolarval) com ouriços-do-mar (Lytechinus variegatus) e bivalves (Perna perna) foi realizada (efeito a nível de indivíduo) após exposição a sedimentos contaminados com os FPCP. Além disso, foram analisados alguns biomarcadores de Fase I (etoxiresorufina O-deetilase EROD e dibenzilfluoresceína DBF), de Fase II (glutationa S-transferase GST) do metabolismo, do sistema antioxidante (glutationa peroxidase GPx), de neurotoxicidade (colinesterase ChE), de estresse oxidativo (peroxidação lipídica LPO e danos em DNA) e de citotoxicidade que foram selecionados para avaliação das respostas a nível de sub-indivíduo em mexilhões Mytella charruana. Todos os compostos analisados apresentaram efeitos sobre o desenvolvimento embriolarval de L. variegatus e P. perna em concentrações ambientalmente relevantes. Em nível de sub-indivíuo foi possível observar que o TCS causou efeitos cito-genotóxicos (diminuição da estabilidade da membrana lisossomal, peroxidação lipídica e danos em DNA) e neurotóxicos. O IBU causou efeitos citotóxicos e neurotóxicos, enquanto o EE2 apresentou efeitos citotóxicos e danos em DNA. Nesse sentido, mesmo em baixas concentrações os FPCP são potencialmente capazes de alterar os mecanismos de manutenção da homeostase. Os dados químicos e ecotoxicológicos foram integrados e os quocientes de risco estimados para TCS, IBU e EE2 apresentaram valores superiores a 1,0, indicando alto risco ambiental destes compostos em sedimentos marinhos. Estes são os primeiros dados de avaliação de risco ambiental de FPCP em sedimentos de uma zona costeira brasileira. Os resultados sugerem que a ERA de fármacos e produtos de cuidados pessoais deve contemplar, além dos ensaios de toxicidade tradicionais o uso de biomarcadores como indicadores dos primeiros sinais de efeitos e, assim, estabelecer uma avaliação de risco mais efetiva que assegure a proteção e funcionamento dos ecossistemas aquáticos. / The guidelines for the Environmental Risk Assessment (ERA) of pharmaceuticals and personal care products (PPCPs) usually recommend the use of standard ecotoxicity assays (e.g. algae, bacteria, invertebrate, fish) and the assessment of endpoints at individual level for the evaluation of potential effects of PPCPs on biota. Considering that effects at sub-individual level can also affect the ecological fitness of marine organisms, and that marine organisms are chronically exposed to PPCPs, the aim of the current study was to evaluate the environmental risk of triclosan (TCS), ibuprofen (IBU) and 17&alpha;-ethynylestradiol (EE2) in marine sediments using sub-individual and population endpoints. Using LC-ESI-MS/MS, the environmental levels of TCS and IBU were quantified in marine sediments from the vicinities of the Santos submarine sewage outfall (Bay of Santos, São Paulo, Brazil) at 15.14 and 49.0 ng g-1, respectively, while EE2 was not detected (<33ng g-1). A battery (n=3) of chronic bioassays (embryo-larval development) with a sea urchin (Lytechinus variegatus) and a bivalve (Perna perna) were performed at populational level after exposure to spiked sediment. Phases I (ethoxyresorufin O-deethylase EROD and dibenzylfluorescein dealkylase DBF) and II (glutathione S-transferase GST) of the metabolism, antioxidant system (glutathione peroxidase GPX), neurotoxicity (cholinesterase ChE), oxidative effects (lipid peroxidation LPO and DNA damage strand breaks) and cytotoxicity were selected to evaluate the sublethal responses in the bivalve Mytella charruana. These compounds showed developmental effects on L. variegatus and P. perna at environmentally relevant concentrations. At sub-individual level TCS induced cyto-genotoxic (reduction on stability of lysosome membrane, lipid peroxidation and DNA damage) and neurotoxic effects. IBU caused cyto and neurotoxic effect, while EE2 caused cytotoxic and DNA damage. Chemical and ecotoxicological data were integrated and the quotient risk estimated for TCS, IBU and EE2 showed values higher than 1.0, indicating high environmental risks of these compounds in sediments. These are the first data of risk assessment of pharmaceuticals and personal care products in sediments of a Brazilian coastal zone. The results suggests that the ERA of pharmaceuticals and personal care products must include, in addition to the standard toxicity tests, the use of biomarkers as indicators of the early warning signs and thus provide a more effective risk assessment to security the protection and functioning of aquatic ecosystems.

Today a lot of attention is focused on compounds called endocrine disrupters (EDs) among substances released to environment by humans. They are a group of substances which can disturb function of hormonal system of organisms including humans. Their poor removal at wastewater treatment plants (WwTP) were shown at various studies, thus they can reach the environment in water. A prospective way for the degradation of EDs at WwTP can be their removal by ligninolytic fungi. They are able to degrade lots of lignin-like aromatic substances because of their highly nonspecific enzymes. In this work growth and enzyme production capability of four ligninolytic fungal strains were monitored on three solid substrates (straw pellets, poplar sawdust mixed with straw pellets, oak sawdust with straw pellets), which may be suitable substrates for fungal growth in bioreactors for wastewater treatment. Ability of these enzymes to degrade EDs were tested in in-vitro degradation experiment. Trametes versicolor was found as best degrading strain with 20 μg/ml of bisphenol A, 17 α- ethynylestradiol and nonylphenol degraded below a quantification limit within 24 hours. Fungal strains degraded EDs well on all of the three substrates but wood sawdust seemed to be a better substrate for fungal growth because straw pellets...

Os protocolos de Avaliação de Risco Ambiental (ERA) de Fármacos e Produtos de Cuidados Pessoais (FPCP) recomendam o uso de ensaios ecotoxicológicos tradicionais (por exemplo algas, bactérias, invertebrados, peixes) e a avaliação de efeitos em um único nível de organização biológica para a determinação dos efeitos potenciais dos FPCP à biota. Considerando que efeitos em nível de sub-indivíduo pode afetar igualmente a aptidão ecológica de organismos marinhos, e que os mesmos estão cronicamente expostos aos FPCP, o objetivo do presente estudo foi avaliar o risco ambiental de triclosan (TCS), ibuprofeno (IBU) e 17&alpha;-etinilestradiol (EE2) em sedimentos marinhos utilizando respostas de efeitos sub-individuais e populacionais. Por meio do HPLC-ESI-MS/MS, as concentrações ambientais de TCS e IBU foram quantificadas em sedimentos marinhos coletados no entorno do emissário submarino de esgoto de Santos (Baía de Santos, São Paulo - Brasil) com 15,14 e 49,0 ng.g-1, respectivamente, enquanto o EE2 não foi detectado (<33 ng.g-1). Uma bateria de ensaios de toxicidade crônica (desenvolvimento embriolarval) com ouriços-do-mar (Lytechinus variegatus) e bivalves (Perna perna) foi realizada (efeito a nível de indivíduo) após exposição a sedimentos contaminados com os FPCP. Além disso, foram analisados alguns biomarcadores de Fase I (etoxiresorufina O-deetilase EROD e dibenzilfluoresceína DBF), de Fase II (glutationa S-transferase GST) do metabolismo, do sistema antioxidante (glutationa peroxidase GPx), de neurotoxicidade (colinesterase ChE), de estresse oxidativo (peroxidação lipídica LPO e danos em DNA) e de citotoxicidade que foram selecionados para avaliação das respostas a nível de sub-indivíduo em mexilhões Mytella charruana. Todos os compostos analisados apresentaram efeitos sobre o desenvolvimento embriolarval de L. variegatus e P. perna em concentrações ambientalmente relevantes. Em nível de sub-indivíuo foi possível observar que o TCS causou efeitos cito-genotóxicos (diminuição da estabilidade da membrana lisossomal, peroxidação lipídica e danos em DNA) e neurotóxicos. O IBU causou efeitos citotóxicos e neurotóxicos, enquanto o EE2 apresentou efeitos citotóxicos e danos em DNA. Nesse sentido, mesmo em baixas concentrações os FPCP são potencialmente capazes de alterar os mecanismos de manutenção da homeostase. Os dados químicos e ecotoxicológicos foram integrados e os quocientes de risco estimados para TCS, IBU e EE2 apresentaram valores superiores a 1,0, indicando alto risco ambiental destes compostos em sedimentos marinhos. Estes são os primeiros dados de avaliação de risco ambiental de FPCP em sedimentos de uma zona costeira brasileira. Os resultados sugerem que a ERA de fármacos e produtos de cuidados pessoais deve contemplar, além dos ensaios de toxicidade tradicionais o uso de biomarcadores como indicadores dos primeiros sinais de efeitos e, assim, estabelecer uma avaliação de risco mais efetiva que assegure a proteção e funcionamento dos ecossistemas aquáticos. / The guidelines for the Environmental Risk Assessment (ERA) of pharmaceuticals and personal care products (PPCPs) usually recommend the use of standard ecotoxicity assays (e.g. algae, bacteria, invertebrate, fish) and the assessment of endpoints at individual level for the evaluation of potential effects of PPCPs on biota. Considering that effects at sub-individual level can also affect the ecological fitness of marine organisms, and that marine organisms are chronically exposed to PPCPs, the aim of the current study was to evaluate the environmental risk of triclosan (TCS), ibuprofen (IBU) and 17&alpha;-ethynylestradiol (EE2) in marine sediments using sub-individual and population endpoints. Using LC-ESI-MS/MS, the environmental levels of TCS and IBU were quantified in marine sediments from the vicinities of the Santos submarine sewage outfall (Bay of Santos, São Paulo, Brazil) at 15.14 and 49.0 ng g-1, respectively, while EE2 was not detected (<33ng g-1). A battery (n=3) of chronic bioassays (embryo-larval development) with a sea urchin (Lytechinus variegatus) and a bivalve (Perna perna) were performed at populational level after exposure to spiked sediment. Phases I (ethoxyresorufin O-deethylase EROD and dibenzylfluorescein dealkylase DBF) and II (glutathione S-transferase GST) of the metabolism, antioxidant system (glutathione peroxidase GPX), neurotoxicity (cholinesterase ChE), oxidative effects (lipid peroxidation LPO and DNA damage strand breaks) and cytotoxicity were selected to evaluate the sublethal responses in the bivalve Mytella charruana. These compounds showed developmental effects on L. variegatus and P. perna at environmentally relevant concentrations. At sub-individual level TCS induced cyto-genotoxic (reduction on stability of lysosome membrane, lipid peroxidation and DNA damage) and neurotoxic effects. IBU caused cyto and neurotoxic effect, while EE2 caused cytotoxic and DNA damage. Chemical and ecotoxicological data were integrated and the quotient risk estimated for TCS, IBU and EE2 showed values higher than 1.0, indicating high environmental risks of these compounds in sediments. These are the first data of risk assessment of pharmaceuticals and personal care products in sediments of a Brazilian coastal zone. The results suggests that the ERA of pharmaceuticals and personal care products must include, in addition to the standard toxicity tests, the use of biomarkers as indicators of the early warning signs and thus provide a more effective risk assessment to security the protection and functioning of aquatic ecosystems.