Orientador: Liliane Lazzari Albertin / Resumo: A situação de escassez de recursos financeiros destinados ao tratamento e coleta de esgotos municipais, no Brasil, torna essencial a busca por tecnologias de tratamento que priorizem baixos custos de instalação e operação. Além da negligência que existe no tratamento de esgotos, os métodos utilizados, muitas vezes, não são capazes de remover os micropoluentes que estão presentes nos esgotos domésticos. Os micropoluentes podem ser desde fármacos, que são expelidos na urina ou descartados de maneira inadequada, a hormônios naturais ou sintéticos. Esse trabalho consistiu em realizar um processo de polimento do esgoto sanitário da cidade de Ilha Solteira – SP. O sistema de polimento é constituído por dois tanques, onde foi cultivada a macrófita Landoltia punctata. Foi também avaliado e feito a recirculação do efluente, no tratamento de esgoto. Neste sistema de polimento, foram avaliadas as eficiências de tratamento para os parâmetros de DBO, DQO, NT, NO3-, PT e ST. As maiores eficiências de remoção para DBO, DQO, NT, NO3-, PT e ST foram de 66,35%, 59,08%, 26,76%, 28,38%, 6,85% e 40,08%. Outro aspecto avaliado foi a taxa de crescimento da macrófita cultivada em esgoto sanitário. As maiores taxas de crescimento relativo foram de 3,84 e 3,17 g.m-2.d-1 MS. Considerando a presença de desreguladores endócrinos (DEs) no efluente da Estação de Tratamento de Esgoto (ETE) de Ilha Solteira, foi analisado a absorção dos estrógenos 17α-etinilestradiol (EE2) e o estriol (E3) pela macrófita, av... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The scarcity of financial resources for the treatment and collection of municipal wastewater in Brazil makes search for treatment technologies essential with low installation and operation costs. In addition, the methods used are often not able to remove the micropollutants that are present in domestic sewage. Micropollutants can range from drugs that are expelled in the urine or inappropriately discarded to natural or synthetic hormones. This work consisted of a process to polishing the sanitary sewage of the city of Ilha Solteira - SP. The treatment system consists of two tanks, where the macrophyte Landoltia punctata was grown. In one of the tanks, a recirculation system was performed. In this polishing system, the treatment efficiencies for the BOD, COD, TN, NO3-, TP and ST parameters were evaluated. The highest removal efficiencies for BOD, COD, TN, NO3-, TP and ST were 66,35%, 59,08%, 26,76%, 28,38%, 6,85% and 40,08%. Another aspect evaluated was the growth rate of the macrophyte. The highest relative growth rates were 3.84 and 3.17 g.m-2.d-1 MS. Considering the presence of endocrine disrupters (DEs) in the effluent from the Sewage Treatment Plant (ETE) in Ilha Solteira - SP, 17α-ethinyl estradiol (EE2) and estriol (E3) were analyzed on the plant to evaluate possible risks on the reuse of the biomass. The maximum achieved efficiencies for removal of E3 and EE2 were 83.63% and 83.50%, respectively. The recirculation tank was more effective at removing E3, while the non-r... (Complete abstract click electronic access below) / Mestre

>Magister Scientiae - MSc / Current water treatment technologies do not remove many endocrine disruptor compounds (EDCs) such as 17α-ethynylestradiol (EE2) in its entirety, and the amount of these pollutants that continues to enter the aquatic environment through wastewater effluents is still capable of causing harmful health effects. Therefore the development of simpler and more sensitive biosensor system for detection of EE2 must be developed which have high responsiveness, low cost and easy handling. Therefore the aim of this study was to work towards the development of rapid test system of polyaniline-laccase on gold enzyme nanobiosensor (PANI-PSSA/Lac/Glu) for water estrogens. Preliminary studies were first done on the materials used in this study: estrogens, laccase, gold nanoparticles (AuNPs), and electropolymerized PANI-PSSA. Laccase was shown to be active towards EE2 and the enzyme could be stored for over three months. EE2 solution also could be used for over three months. Buffer used in this study was found to be suitable. Phosphoric acid (H3PO4) was a suitable electrolyte than hydrochloric acid (HCl) to be used for the electropolymerization of aniline and was used because it has same ions as the McIlvaine buffer (McIlB) which the post-deposition CVs indicated the formation of electrochemically very stable film. AuNPs were successfully synthesized and its size was identified to be less than 22 nm. McIlB used for testing electrochemical properties of AuNP. CVs of GC/PANI-PSSA and GC/PANIPSSA/ Au showed no difference before and after exposure to aq. EE2 solution, an indication of being re-usable and could also serve as stable immobilising platform in laccase biosensor. When interrogating with electrochemical impedance spectroscopy (EIS), the charge transfer resistance (Rct) of both GC/PANI-PSSA and GC/PANI-PSSA/Lac/Glu showed an average increase by about 2.4% and 21% before and after exposure of EE2, respectively. This shows that the GC/PANI-PSSA/Lac/Glu was a functional EE2 biosensor and showing a positive step towards achieving a re-usable biosensor for EE2 as a model water estrogen. Future work Page | vi will focus on exploring different ways of improving the biosensor’s surface regeneration and its sensitivity to EE2.

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2017-12-04T13:25:24Z No. of bitstreams: 2 Dissertação - Fernanda Lisboa Martins - 2017.pdf: 4877141 bytes, checksum: 9bbe680537dbf0bf672e46c97951bc77 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-12-06T11:35:00Z (GMT) No. of bitstreams: 2 Dissertação - Fernanda Lisboa Martins - 2017.pdf: 4877141 bytes, checksum: 9bbe680537dbf0bf672e46c97951bc77 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-12-06T11:35:00Z (GMT). No. of bitstreams: 2 Dissertação - Fernanda Lisboa Martins - 2017.pdf: 4877141 bytes, checksum: 9bbe680537dbf0bf672e46c97951bc77 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-09-22 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / This work compared the efficiency of removing the 17-α-ethynylestradiol (EE2) through advanced oxidative processes (POA) and granular activated carbon adsorption (CAG). EE2 is a synthetic female hormone common in domestic sewage and effluent from pharmaceutical industries. This hormone is a type of emerging pollutant that causes endocrine disruption in living beings even in concentrations of nanograms order. The EE2 removal was analyzed in a prototype of a 10 L reactor, with upward flow CAG fluidized bed, using four lamps emitting ultraviolet radiation, with batch time of 2 hours in each process. Photolysis showed 5% of efficiency, while photocatalysis removed 87% of EE2. The adsorption processes in CAG and titanium dioxide immobilized at CAG removed 82 and 89%, respectively. / Este trabalho comparou a eficiência de remoção de 17-α-etinilestradiol (EE2) por meio de processos oxidativos avançados (POA) e adsorção em carvão ativado granular (CAG). O EE2 é um hormônio feminino sintético comum no esgoto doméstico e no efluente de indústrias farmacêuticas. Esse hormônio é um tipo de poluente emergente que causa desregulação endócrina nos seres vivos mesmo em concentrações da ordem de nanogramas. Então foi analisada a remoção do EE2 em um protótipo de reator de 10 L, com leito fluidizado de CAG, de fluxo ascendente, utilizando quatro lâmpadas que emitem radiação ultravioleta, com tempo de batelada de 2 horas, cada processo. A fotólise apresentou 5% de eficiência, enquanto a fotocatálise removeu 87% de EE2, e os processos de adsorção em CAG e em dióxido de titânio imobilizado em CAG removera, 82 e 89%, respectivamente.

Many exogenous and endogenous compounds are referred to as endocrine disruptors (EDCs), as they interfere with natural synthesis, signaling and metabolism of endogenous hormones. Common exogenous endocrine disruptors are benzo(a)pyrene (BaP) and 17α-ethinylestradiol (EE2). Endogenous endocrine disruptor 17β-estradiol (E2) is frequently present in the environment as well. In this thesis, the effect of the mentioned EDCs and their combinations on gene and protein expression of CYP1B1, 3A1 and 3A2 in rat liver, kidney and lung was determined. Protein expression was studied using Western blot method and specific antibodies; gene expression was assessed by quantitative PCR. Moreover, the effect of tested EDCs and their combinations on BaP metabolism and CYP3A specific activity (measured as testosterone 6β-hydroxylation) were studied in liver microsomal samples. It was confirmed, that BaP significantly increases CYP1B1 expression in rat liver and lung both alone and together with EE2 or E2. Pretreatment of rat with E2 and BaP increases the ability of BaP to induce CYP1B1 expression. On the contrary, EE2, E2 and their combination decrease the CYP1B1gene expression. The rate of BaP metabolites formed in liver microsomal samples increases in rats pretreated with BaP and its combinations. In liver, there was...

Organic contaminants from industrial and/or domestic effluents may be harmful to humans directly or indirectly by degrading the quality of the aquatic environment. Consequently these contaminants must be reduced to levels that are not harmful to humans and the environment before disposal. Chemical, physical and biological methods exist for the removal of these pollutants from effluents. Among the available chemical methods, heterogeneous photocatalytic oxidation has been found particularly effective in removing a large number of persistent organics in water. In this study, photocatalytic degradation was explored for the removal of reactive azo-dye (textile dye), triclocarban (disinfectant), clopyralid (herbicide) and three endocrine disrupting compounds (EDCs) (estrone, 17ß-estradiol and 17α-ethinylestradiol) from synthetic effluents. The major factors affecting the photocatalytic processes including the initial concentration of the target compounds, the amount of catalyst, the light intensity, the type of catalyst, the electron acceptor, the irradiation time and the pH were studied. Other oxidation techniques including (O3, H2O2, UV) were also studied. Generally UV light is used in combination with titanium dioxide, as photocatalyst, to generate photoinduced charge separation leading to the creation of electron-hole pairs. The holes act as electron acceptors hence the oxidation of organics occur at these sites. These holes can also lead to the formation of hydroxyl radicals which are also effective oxidants capable of degrading the organics. The results obtained in this study indicated that photolysis (i.e. UV only) was found to have no effect on the degradation of reactive azo-dye (RO16). However, complete photocatalytic degradation of 20 mg/L (3.24×10-2 mM) RO16 was achieved in 20 minutes in the presence of 1g/L TiO2 Degussa P25 at pH 5.5. Comparison between various types of catalysts (i.e. Degussa P25, VP Aeroperl, Hombifine N) gave varied results but Degussa P25 was the most effective photocatalyst hence it was selected for this study. For RO16 the optimum catalyst concentration was 0.5 g/L TiO2 with initial concentration of 20 mg/L RO16. It was found that the disappearance of RO16 satisfactorily followed the pseudo first-order kinetics according to Langmuir-Hinshelwood (L-H) model. The rate constant was k= 0.0928 mol/min. Photodegradation of TCC was studied in 70%v acetonitrile: 30%v water solutions. UV light degraded TCC effectively and the reaction rates increased with decreasing initial concentration of TCC. UV/TiO2 gave unsatisfactory degradation of triclocarban (TCC) since only 36% were removed in 60 minutes with initial concentration of TCC 20 mg/L. The degradation of clopyralid and the EDCs was studied using three oxidation systems UV/TiO2, UV/H2O2 and O3. Complete degradation of clopyralid (3,6-DCP) was achieved with UV/TiO2 in about 90 minutes at an optimum catalyst concentration of 1g/L. Zero-order kinetics was found to describe the first stage of the photocatalytic reaction in the concentration range 0.078-0.521 mM. At pH 5 the rate constant was 2.09×10-6-4.32×10-7 M.s-1.Complete degradation of all the three EDCs was achieved with UV/H2O2 in 60 minutes at catalyst concentration of (2.94×10-2 M). On the other hand complete degradation of the EDCs was achieved in just 2 minutes with ozonation. For high concentration EDCs, TiO2/UV gave low efficiency of degradation as compared with ozone and H2O2/UV. First-order kinetics was found to describe the photocatalytic reaction of the EDCs. / Education Service Department of the Libyan Government

This work is aimed at the study of the effect of 17α-ethinylestradiol (EE2) on the production and characteristics of ligninolytic enzymes (laccase, Mn-dependent peroxidase and lignin peroxidase) in I. lacteus, T. versicolor, P. chrysosporium and P. ostreatus cultures grown on two types of liquid media. Enzyme activity production in fungal cultures was affected by the composition of culture medium. In the case of P. chrysosporium, the addition of EE2 to the complex- medium cultures led to a MnP activity stimulation and simultaneously LiP production was partially repressed in these cultures. In the mineral MM medium, no effect of EE2 on enzyme production by P. chrysosporium was observed. In EE2 treated MM cultures of P. ostreatus lower MnP activities were found when compared to biotic controls. In the case of T. versicolor cultures, the addition of EE2 to the complex medium caused laccase and LiP stimulation in the cultures. In the MM medium, however, only laccase production was affected by EE2. I. lacteus MnP production was partially repressed by EE2 in MM medium. In contrast to that, significantly higher MnP activities were detected in complex- medium I. lacteus cultures after the treatment with EE2. Further EE2 degradation by the fungal cultures was studied. The highest degradation effeciency was...

The effects of cypermethrin, copper and glyphosate on the endocrine system and subsequent response to female pheromones were investigated in mature male brown trout (Salmo trutta) parr.  Responses measured were the amount of strippable milt, blood plasma levels of both an androgen (11-ketotestosterone (11-KT)) and a progestin (17α,20β-dihydroxy-4-pregnen-3-one (17,20b-P)), and behavioural changes. This was done in a two phased investigation where parr were exposed to one of the following via ambient water: 1) 0.1 or 1.0 μg L-1 cypermethrin, 2) 10 or 100 μg L-1 copper (Cu2+), or 3) 150 μg L-1 glyphosate for a 96 hour period.  Phase one was a priming experiment exposing parr to a treatment followed by priming with PGF2α or ovarian fluid (OVF). Atlantic salmon (Salmo salar) parr were, also exposed to glyphosate during phase I. The second phase was centered on behavioural observations.  Exposed parr were placed in a 35,000 L stream aquarium together with two ovulated females and four anadromous males. After the experiments a blood sample was taken, milt volumes measured and testes weighed.  The plasma was analyzed for 11-KT and 17,20b-P concentrations using radioimmunoassay (RIA). Results from phase I-priming: 1.0 μg L-1 cypermethrin exposure lowered 17,20b-P and 11-KT; Copper exposure lowered milt volumes; glyphosate exposure lowered 11-KT in salmon and raised 17,20b-P in trout.  Results from phase II-behaviour: 1.0 μg L-1 cypermethrin exposure lowered 11-KT, milt and spawning behaviour; copper exposure lowered spawning behaviour and raised 11-KT; Glyphosate exposure lowered 11KT; continuous cypermethrin exposure raised 17,20b-P, 11-KT and gave a tendency towards increased aggression. It is concluded that low concentration exposure to the compounds examined can induce negative effects on male salmonid endocrine systems, either through a disruption in the olfactory system or through a direct effect. / Extern doktorand från Södertörns högskola

A synthetic estrogen 17α-ethinylestradiol (EE2) is the main active component of the hormonal contraceptive pills. The rise of consumption of hormonal contraceptives has increased the risk of the back negative effects of EE2 to aquatic organisms. EE2 belongs to the endocrine disruptive compounds known for mimicking natural hormones. A more detailed examination of the transformation of this compound in vivo and in vitro can contribute to a better understanding of its negative effects. This master thesis is therefore devoted to the study of the metabolism of EE2 in two selected model organisms. The ligninolytic fungus Pleurotus ostreatus is the type of fungi with promising biodegradation ability to a lot of pollutants. These properties have led to numerous studies of the degradation potential of P. ostreatus towards EE2, with the possibility of removing this compound from the environment. EE2 has been degraded by the fungus P. ostreatus in vivo resulting in one hydroxylated metabolite, which estrogenic activity is in need for further study. In vitro studies were carried out with a microsomal fraction isolated from the mycelium of this fungus. The conversion of EE2 in vitro via CYPs dependent on NADPH has not been demonstrated, however using KHP as a cofactor, there was one metabolite of EE2 found,...

Die weltweit mehr als 7500 Amphibienarten sind durch anthropogene Ursachen wie Habitatzerstörung, Krankheitsverbreitung, Klimawandel und Umweltverschmutzung in ihrem Bestand bedroht. Einige der Ursachen sind kaum erforscht, so die Verschmutzung aquatischer Ökosysteme durch endokrine Disruptoren (EDs), Substanzen, die mit dem Hormonsystem interagieren. Ausgehend von neuen molekularen Markern, welche die Ermittlung des genetischen Geschlechts erstmals bei einigen Hyliden und Bufoniden erlauben, wurde in der vorliegenden Arbeit auf die Wirkung des synthetischen Östrogens 17α- Ethinylestradiol (EE2)und des Weichmachers Bisphenol A (BPA) fokussiert. Für drei Bufonidenarten wurde zunächst die Geschlechtsgebundenheit von Mikrosatelliten getestet und ein XX/XY-System nachgewiesen. Diese und bereits etablierte Marker wurden anschließend in ein neu entwickeltes Versuchsdesign für ED-Studien integriert: Nach gleichzeitiger Aufzucht von Modell- (Xenopus laevis) und Nicht-Modell-Arten (Hyla arborea, Bufo viridis) unter EE2- bzw. BPA- Exposition wurde das genetische Geschlecht bestimmt und mit dem anatomisch und histologisch ermittelten phänotypischen Geschlecht erglichen. Die drei Anuren zeigten starke Empfindlichkeitsunterschiede gegenüber beiden EDs. Umweltrelevante Konzentrationen beeinflussten die somatische Entwicklung und führten zu artspezifischen Gonaden-Fehlbildungen. EE2 bewirkte zahlreiche partielle und komplette Geschlechtsumwandlungen, mit stärkeren Effekten bei X. laevis. Diese Arbeit zeigt somit, dass bereits niedrige EE2- und BPA-Konzentrationen zu starken Schädigungen führen können und die Substanzen aufgrund ihrer erheblichen aquatischen Präsenz als ernstzunehmende Faktoren der Amphibienkrise anzusehen sind. Die Ermittlung des genetischen Geschlechts wird als wichtig eingestuft, um verlässliche Aussagen über ED-Effekte zu treffen. Zudem sollten an der Modell-Art X. laevis gewonnene Erkenntnisse nicht vorbehaltlos auf andere Amphibienarten extrapoliert werden. / The more than 7500 known amphibian species are globally threatened, mainly due to anthropogenic causes like habitat destruction, dispersing diseases, climate change and environmental pollution. Some of the causes are barely investigated, e.g. the pollution of aquatic ecosystems with endocrine disrupting compounds (EDCs), substances that interfere with the hormone system. Based on new molecular markers, for the first time allowing genetic sexing in some hylids and bufonids, this thesis focused on the effects of the synthetic estrogen 7α-ethinylestradiol(EE2) and the plasticizer bisphenol A (BPA). Initially, several microsatellite markers were tested for sex-linkage in three bufonid species, and an XX/XY system could be revealed. Subsequently, these and other established markers were integrated into a newly developed experimental design for EDC-research: after simultaneous exposure of model (Xenopus laevis) and non-model species (Hyla arborea, Bufo viridis) to EE2 or BPA, metamorphs were genetically sexed. Anatomically and histologically determined phenotypic sexes were directly compared with the genetic sex of each individual. The three anurans showed striking differences in their susceptibilities in both EDCexperiments. Environmentally relevant concentrations affected the somatic development and led to species-specific gonadal anomalies. In addition, EE2 provoked high numbers of mixed sex and completely sex-reversed individuals, with more pronounced effects in X. laevis than in the two non-model species. This work shows that low concentrations of EE2 and BPA lead to severe damages. Due to their widespread presence in the aquatic environment, these substances might contribute to the worldwide amphibian crisis. To produce reliable results in EDC-studies, genetic sexing is considered important. Furthermore, findings gained with the model species X. laevis should not unreservedly be extrapolated to other amphibian species.

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.