A atrazina é um herbicida amplamente utilizado no Brasil e no mundo em diferentes culturas agrícolas, principalmente em culturas de milho, sorgo, soja e cana-de-açúcar, entretanto, pode se tornar um contaminante de águas superficiais e subterrâneas e do solo, gerando uma preocupação ambiental, pois desequilibra e interfere no ecossistema. A atrazina pode sofrer biodegradação, tornando o processo de biorremediação uma alternativa viável e ecologicamente aceitável para o tratamento de ambientes contaminados por esse herbicida. O microrganismo de referência nesse processo de biodegradação é a Pseudomonas sp. ADP que possui o plasmídio pADP-1, no qual se localizam os genes atzA, atzB, atzC, atzD, atzE e atzF, que codificam enzimas atuantes na via de degradação da atrazina. O presente estudo teve como objetivo o isolamento e a caracterização de bactérias do gênero Pseudomonas quanto à presença de genes de degradação da atrazina e quanto à capacidade de degradação desse herbicida. No presente trabalho foram isoladas 123 cepas de amostras de solo de diferentes regiões do Brasil, as quais foram caracterizadas através de provas bioquímicas e moleculares e identificadas como Pseudomonas aeruginosa (74,8%) e outras espécies (25,2%) pertencentes aos gêneros Pseudomonas, Achromobacter, Burkholderia, Cupriavidus e Rhizobium. A variabilidade genética dos isolados pertencentes ao gênero Pseudomonas foi analisada através da técnica de ERIC-PCR e demonstrou que todos os isolados apresentam alta diversidade genética (<80%). Os genes atzA, atzB, atzC, atzD, atzE e atzF foram detectados em seis isolados provenientes de amostras de solo das regiões Norte, Nordeste, Sudeste e Centro-Oeste do Brasil, sendo três deles da espécie Pseudomonas aeruginosa, um Cupriavidus pauculus, uma Burkholderia cepacea e um Rhizobium radiobacter. Apenas três isolados contendo os genes atz apresentaram plasmídios. Os isolados bacterianos que apresentaram os genes atz foram testados quanto à capacidade de crescimento utilizando a atrazina como única fonte de nitrogênio e quanto à capacidade de degradação desse herbicida. Todos os isolados testados apresentaram crescimento em meio ATZ-R, mas não foi possível observar a mineralização do herbicida, tanto em meio sólido quanto em meio líquido, todavia observou-se a degradação da atrazina por uma espécie de Pseudomonas aeruginosa (isolado P86), com redução de 44%, em meio líquido ATZ-R contendo 33 ppm de atrazina. / Atrazine - the herbicide widely used in Brazil and all over the world in different agricultural crops mainly in corn, sorghum, soy, and sugar cane can contaminate the superficial and subterraneous water, and soil creating an environmental concern due to imbalance and interference caused in ecosystem. Atrazine can suffer biodegradation, which can make the bioremediation process a viable and ecologically acceptable alternative for the treatment of the environment contaminated with this herbicide. The microorganism referred on that biodegradation process is Pseudomonas sp. ADP which has pADP-1 plasmid, where are the genes atzA, atzB, atzC, atzD, atzE, and atzF that codify active enzymes on atrazine degradation way. The aim of this present study was the isolation and the characterization of Pseudomonas genus bacteria due to the presence of the atrazine degradation genes, and the capacity of degradation of this herbicide. In the present work 123 samples of soil bacteria from different Brazilian regions were isolated, and characterized by biochemical and molecular tests, and identified as Pseudomonas aeruginosa (74.8%) and other species (25.2%) of Pseudomonas, Achromobacter, Burkholderia, Cupriavidus, and Rhizobium genus. Genetic variability of the isolated of Pseudomonas genus was analyzed by the ERIC-PCR technique, and demonstrate that all the isolated ones presented high genetic diversity (<80%). atzA, atzB, atzC, atzD, atzE, and atzF genes were detected in six isolated of soil samples from North, Northeast, Southeast, and Midwest of Brazil being 3 Pseudomonas aeruginosa, 1 Cupriavidus pauculus, 1 Burkholderia cepacea, and 1 Rhizobium radiobacter. Only 3 isolated with atz genes presented plasmids. Bacterial isolates that presented atz genes were tested for growth and degradation capacity using only atrazine as nitrogen source. All tested isolates presented growth on ATZ-R, but mineralization of this herbicide in solid or liquid media was not possible to observe. However, 1 specie Pseudomonas aeruginosa (isolate P86) presented atrazine degradation (reduction of 44%) in ATZ-R liquid media with 33 ppm of atrazine.

<p>Rezultati prikazani u ovom radu opisuju efekte in vivo primene atrazina (2-hloro-4-<br />etilamino-6-izopropilamino-s-triazin) na ex vivo steroidogenezu u Leydig-ovim ćelijama<br />peripubertalnih pacova (tretiranih sa 50 mg/kg i 200 mg/kg telesne mase od 23. do 50.<br />dana starosti). Dobijeni rezultati jasno ukazuju da 28-dnevna in vivo primena atrazina<br />snažno inhibira testikularnu steroidogenezu, smanjujući ekspresiju gena za steroidogene<br />enzime i druge regulatorne proteine uključene u kontrolu testikularne steroidogeneze u<br />Leydig-ovim ćelijama peripubertalnih pacova. Rezultati in vivo primene atrazina<br />pokazuju da atrazin snažno inhibira ekspresiju gena za luteinizirajući hormon receptor<br />(LHR), skevendžer receptor-B1 (SR-B1), steroidogeni faktor-1 (SF-1), steroidogeni<br />akutni regulatorni protein (StAR), translokator protein (TSPO), fosfodiesterazu 4B,<br />3&beta;&minus;hidroksisteroid dehidrogenazu (&Eta;SD), CYP17A1 i 17&beta;HSD. Rezultati u okviru ovih<br />istraživanja pokazuju da primena atrazina tokom prepubertalnog perioda razvoja mužjaka<br />pacova dovodi do dozno-zavisnog smanjenja nivoa cAMP i snažne inhibicije androgeneze<br />u prisustvu hCG. Obzirom na blokadu ekspresije LHR, prvog elementa u aktivaciji<br />cAMP-signalnog puta, moglo bi se predpostaviti da je to uzrok blokade androgeneze kod<br />atrazinom-tretiranih životinja. Takođe, rezultati ukazuju na inhibiciju supstrat-stimulisane<br />produkcije androgena paralelno sa redukcijom ekspresije steroidogenih enzima CYP17A1<br />i 17&beta;HSD. U drugom delu ove doktorske disertacije, ispitivan je efekat direktne in vitro<br />primene različitih doza atrazina (1 nM, 1 &mu;M, 20 &mu;M, 50 &mu;M) na ekspresiju i aktivnost<br />steroidogenih enzima u kulturi preči&scaron;ćenih Leydig-ovih ćelija testisa peripubertalnih<br />pacova, pri čemu je zabeleženo stimulatorno dejstvo pomenutog herbicida. Naime,<br />zabeleženo je povećanje bazalne i hCG-stimulisane produkcije testosterona praćeno<br />povećanim nivoom cAMP u medijumu tretiranih ćelija. Pri ispitivanju ekspresije gena za<br />steroidogene enzime i regulatorne proteine, zabeleženo je povećanje ekspresije gena za<br />SF-1, StAR, CYP17A1 i 17&beta;HSD u hCG-stimulisanim uslovima. Takođe, povećana je i<br />produkcija testosterona nakon dodavanja progesterona i androstendiona kao supstrata,<br />kod Leydig-ovih ćelija tretiranih sa atrazinom. Da bi poku&scaron;ali da objasnimo za&scaron;to postoje<br />razlike u efektu atrazina u zavisnosti od načina primene, postavili smo jednokratni in vivo<br />eksperiment sa atrazinom (tretman sa 50 mg/kg i 200 mg/kg telesne mase, životinje<br />tretirane 50. dana starosti). Rezultati ovih eksperimenata ukazali su na up-regulaciju<br />testikularne steroidogeneze, kao i na povećan nivo cAMP kod životinja tretiranih sa<br />atrazinom Stoga, nivo cAMP se pojavljuje kao karika koja povezuje sva tri kori&scaron;ćena<br />eksperimentalna pristupa. Međutim, ostaje otvoreno pitanje na koji način atrazin utiče na<br />modulaciju nivoa cAMP i to pitanje predstavlja motiv za dalja istraživanja. Sumarno,<br />dobijeni rezultati ukazuju da 24-časovni tretman atrazinom izaziva povećanje, a<br />prolongirani tretman snažno smanjenje steroidogenog kapaciteta Leydig-ovih ćelija<br />peripubertalnih pacova.</p> / <p> In the present study, we investigated the effects of oral dosing of atrazine (2-chloro-4-<br /> ethylamino-6-isopropylamino-s-triazine) to peripubertal male rats (50 mg/kg and 200<br /> mg/kg body weight daily from postnatal day 23 to 50) on ex vivo Leydig cell<br /> steroidogenesis. Leydig cells from treated rats were characterised by significant decline in<br /> mRNA transcripts of several genes responsible for steroidogenesis: luteinizing hormone<br /> receptor (LHR), scavenger receptor-B1, steroidogenic acute regulatory protein (StAR),<br /> translocator protein, steroidogenic factor-1 (SF-1), phosphodiesterase 4B,<br /> 3&beta;&minus;hydroxysteroid dehydrogenase (&Eta;SD), CYP17A1 and 17&beta;HSD. In the presence of<br /> human chorion gonadotropin, the dose-dependent decrease in extra cellular cAMP level<br /> and accordingly strong inhibition of androgenesis were obtained. The transcription of<br /> LHR gene in Leydig cells of atrazine-treated rats was down-regulated in a dose-dependent<br /> manner, which could be the reason for reduction in cAMP level and expression of cAMPdependent<br /> genes. The results also indicated inhibition of substrate-stimulated androgen<br /> production in parallel with reduced expression of steroidogenih enzymes CYP17A1 and<br /> 17&beta;HSD. In the second part of this study we examined direct 24 h in vitro effect of<br /> different doses of atrazine (1 nM, 1 &mu;M, 20 &mu;M, 50 &mu;M) on expression and activity of<br /> steroidogenic enzymes in purified Leydig cells obtained from peripubertal rats. Obtained<br /> results indicated that 24 h-incubation of peripubertal Leydig cells in the presence of<br /> atrazine increased steroidogenic capacity of that cells. Increased basal and hCGstimulated<br /> testosterone production were accompanied by increasing levels of cAMP in the<br /> medium of treated cells. Also, in comparison to controls, gene expression revealed<br /> increased expression of SF-1, StAR, CYP17A1 and 17&beta;-HSD. When Leydig cells were<br /> challenged with progesterone and &Delta;4&ndash;androstenedione, testosterone production was<br /> increased in atrazine chalenged Leydig cells. To address these two opposite effects of<br /> atrazine we performed 24 h in vivo experiment in which peripubertal male rats (on<br /> postnatal day 50) were exposed to single atrazine treatment (50 mg/kg- and 200 mg/kgbody<br /> weight by gavage), and 24 h later, Leydig cells were isolated and testosterone levels<br /> in medium determined in basal and in hCG-stimulated conditions after 2 h-incubation<br /> period. Obtained results indicated that single in vivo exposure to atrazine was also<br /> accompanied 24 h later by up-regulation of Leydig cell androgenesis and increased cAMP<br /> level. According to the results obtained in this study, it seems that modulation of cAMP<br /> levels appear as a link that connects all three experimental approaches. However, the<br /> question of how atrazine affects the modulation of cAMP levels remains open, and<br /> present a motive for further research. In concluson, obtained results indicated that 24 h<br /> treatment with atrazine caused an increase, while prolonged treatment strongly reduce<br /> steroidogenic capacity of peripubertal Leydig cells.</p>

O estudo avaliou a efetividade de um sistema para tratamento de soluções aquosas contendo poluentes orgânicos emergentes (POE), denominados interferentes endócrinos (IE). Baseado nos processos oxidativos avançados (POA), empregando ozônio, radiação UV, peróxido de hidrogênio e variação de pH, o estudo possibilitou determinar a relação ideal oxidante/poluentes e o custo/benefício dos tratamentos com melhores desempenhos, de degradação do padrão contendo hormônios 17?-estradiol (E2) e 17?-etinilestradiol (EE2), e outra contendo atrazina (ATZ). Para os estudos, foi utilizado um planejamento fatorial 23, para identificar as melhores opções de degradação. Os tratamentos iniciaram com soluções em concentrações de 600 ?g L-1 de ATZ, em quadruplicatas, por períodos de até 10 minutos. Posteriormente a avaliação dos melhores tratamentos, foi estudada a cinética de mineralização de soluções a 5 mg L-1 de ATZ, em até 90 min. A concentração de estudo adotada para os hormônios foi de 1 mg L-1. A determinação da ATZ e dos hormônios (E2 e EE2) foi realizada por cromatografia em fase líquida (CLAE), utilizando a extração em fase sólida (EFS) quando necessário. Para o estudo foram construídos dois sistemas de tratamento paralelamente acoplados (S1 - escala laboratorial e S2 - escala piloto), com os respectivos volumes de 0,43 L e 20 L. O S2 possibilitou a avaliação da degradação e/ ou mineralização do fenol em solução a 50 mg L-1, porém em escala 60 vezes superior ao S1. As eficiências das mineralizações da ATZ e do fenol, foram avaliadas pela análise de Carbono Orgânico Total (COT). As condições experimentais pré-estabelecidas foram: (i) UV-C 254 nm, variando entre 0, 16 e 32 W; (ii) H2O2 entre 0, 50 e 100% do total teórico necessário para mineralização dos poluentes; (iii) O3 (4,6 g h-1) e (iv) pH 7; 9 e 11, previamente aplicadas às soluções estoque de ATZ. Após avaliação da melhor combinação para degradação, optou-se por trabalhar, nos demais estudos, apenas com soluções em pH 11,0. Na última etapa deste estudo, foram desenvolvidos dois métodos (direto e indireto) para avaliar a geração do gás ozônio em tempo real, possibilitando também realizar a medição do consumo de O3 durante os estudos. Para avaliar a eficiência dos métodos, os resultados analíticos foram convertidos em % de degradação ou mineralização e os custos dos tratamentos foram estimados. Comprovando a viabilidade desta proposta, resultados envolvendo o tratamento de solução padrão de atrazina (600 ?g L-1) apresentaram, nas melhores condições (pH 11, 30 mg L-1 de H2O2 e 4,6 g L-1 de O3), eficiência igual ou superior a 95% (<LQ) na degradação do herbicida, em apenas 2,5 minutos de tratamento e posteriormente soluções a 5 mg L-1, apresentando até 100% de mineralização em 90 minutos. Quanto aos hormônios foram obtidos poucos resultados acima do LD do equipamento, porém todos ficaram abaixo do LQ (50 ?g L-1). Quanto ao fenol (50 mg L-1), estudado no sistema piloto (S2), atingiu-se até 80% de mineralização em 24 horas e 100% de degradação. Os resultados obtidos demonstram que os sistemas apresentaram excelente eficácia conforme esperado / The study evaluated the effectiveness of a system for treatment of aqueous solutions containing emerging organic pollutants (EOP), called endocrine disruptors (ED). Based on advanced oxidation processes (AOP) using ozone, UV radiation, hydrogen peroxide and pH variation. This study allowed the determination of the ideal oxidant/pollutants ratio and the cost/benefit of treatments with better performance, regarding the degradation of the pattern containing hormones 17?-estradiol (E2) and 17?-ethinylestradiol (EE2), and another containing atrazine (ATZ). A 23 factorial design was used in the study to identify the best options for degradation. The treatments were started with solutions at concentrations of 600 ?g L-1 ATZ, in quadruplicate, for periods up to 10 min. Subsequently to the best treatment evaluation; the degradation of 5 mg L-1 ATZ solutions was evaluated, within 90 min. The adopted study concentration of the hormones was 0,05 mg L- 1 and 1,0 mg L- 1. The determination of the ATZ and hormones (E2 and EE2) was performed by liquid chromatography (HPLC) using solid phase extraction (SPE) when necessary. For this study, two parallel coupled processing systems were constructed (S1 - Laboratory scale and S2 - pilot scale) with the respective volumes of 0.43 L and 20 L. S2 system was developed to enable evaluation of the degradation of an already known substance, 50 mg L-1 phenol solution, but on a scale 60 times greater than S1. The efficiency of ATZ and phenol mineralization was evaluated by Total Organic Carbon (TOC). The pre-established experimental conditions were: (i) 254 nm UV-C, varying among 0, 16 and 32 W, (ii) H2O2 among 0, 50 and 100% of the theoretical value needed to complete mineralization of the pollutant, (iii) O3 (4,6 g h-1) and (iv) pH 7; 9 and 11 previously applied to all stock solutions. After assessing the best combination for degradation, it was chosen to work in the other experiments only solutions with pH 11.0. In the last stage of the study, two methods (an electric and a spectrophotometric) were developed to assess the ozone gas generation in real time and also allowing the measurement of O3 consumption during the studies. To evaluate the efficiency of the methods, analytical results were converted to % degradation or mineralization and treatment costs were estimated. Proving the feasibility of this proposal, preliminary results involving the treatment of atrazine standard solution (600 ?g L-1) had, in the best conditions (pH 11, 30 mg L-1 of H2O2 and 4,6 g L-1 of O3), efficiency equal or superior to 95% (<LQ) in degradation of the herbicide in just 2.5 minutes of treatment and then 5 mg L-1 solutions had 100% mineralization in 90 minutes. To hormones, few results above the DL of the equipment were obtained, but all were below the QL of the equipment (50 mg L-1). When phenol (50 mg L-1) was studied in the pilot system it reached up to 80% of mineralization within 24 hours. The obtained results shown that the systems developed reached high efficiency for treating the pollutants

Este trabalho tem como foco dois aspectos principais: i) O processamento e caracterização de elementos porosos vítreos, a partir de vidro reciclado e ii) A deposição de filmes de quitosana (CHI) e carboximetilcelulose (CMC). O objetivo é a avaliação da interação superficial desses filmes com o herbicida atrazina (ATZ) em meio aquoso. O processamento seguiu o princípio do preenchimento (filler principle), fazendo uso dos sais NaCl e o MgCO3 como fases formadoras de poros. A caracterização mostra que o NaCl, age como um elemento solúvel, inerte, exceto na interface NaCl-matriz vítrea, cuja estrutura cristalina final é a cristobalita. Contrariamente, o MgCO3 reage com a matriz introduzindo novas fases como o CaMg(SiO3)2. A estrutura final, de poros e da matriz, é distinta para cada um dos sais utilizados, principalmente quanto ao aspecto morfológico dos poros e as análises semiquantitativas mostraram que o cátion Na+ na interface vidro-NaCl e o íon Mg++ atuam como modificadores de cadeia. Medidas de porosimetria indicam que nos materiais processados com NaCl apresentam estruturas dos poros abertos com uniformidade na distribuição dos tamanhos e com certa regularidade de formatos quando comparados com os materiais processados com MgCO3. Com respeito à interação herbicida - materiais vítreos, esta foi avaliada por técnicas espectroscópicas, podendo-se inferir que há interação entre as superfícies ativadas quimicamente e a ATZ. A remoção do herbicida por filtragem simples através dos filmes de CHI e filmes de CHI+CMC depositados sobre as membranas foram inferiores numericamente aos valores obtidos pela ação da superfície vítrea ausente de filmes. Entretanto, os resultados indicam que ocorre uma melhor interação entre a CHI e o ATZ, quando ambos estão em solução a pH = 3,0. Por espectroscopia de fotoelétrons excitada por raios-X (XPS) houve o aumento das espécies O (1s), C (1s), N (1s) e Cl (2s) confirmando as interações com o herbicida, porém não sendo possível inferir se esta se dá por algum grupo proveniente da CHI ou por sítios livres disponíveis na superfície vítrea. Análises numéricas indicam remoções de ATZ na ordem de 10-12% com respeito às medidas realizadas em sistemas contendo uma única membrana. Avaliação complementar da remoção do metal (Cd) confirmam a vantagem das superfícies depositadas com CHI, para este tipo de interação, indicando que sistemas compostos podem ser vantajosos na remoção de diversos contaminantes. / This work is focused in two main aspects: i) The processing and characterization of porous vitreous pieces, produced from waste glass and ii) The deposition of chitosan (CHI) and carboximethilcelullose (CMC) on the vitreous surface. The evaluation of the active aspects aiming at interactions with the herbicide atrazine (ATZ) was realized in aqueous medium. The processing follows the filler principle making use of NaCl and MgCO3 as porous phases formation. Characterization showed that NaCl acts as a soluble, inert phase, with interaction over NaCl-Matrix interface, resulting in cristobalite phase as final structure. Conversely, the MgCO3 reacts along the matrix generating new phases such as CaMg(SiO3)2. The final porous and matrix structure also differs to each used salt, mainly concerning morphological aspects of the porous where semiquantitive analysis point to the Na+ in glass-NaCl interface and to Mg++ as the main chain modifiers. Measurements by porosimetry has showed that in the materials processed with NaCl the porous structure are typically open with uniform size distribution and present a certain regularity of forms when compared with the membranes processed with MgCO3. Concerning an herbicide interaction, which was evaluated by spectroscopic techniques inferring interaction between chemically active surfaces and ATZ. The herbicide removal through CHI and CHI+CMC deposited films resulted numerically lower than those values attained to glass surface absent of films. Nevertheless, the results point that to a better interaction between CHI and ATZ when both are dissolved at pH 3,0. By XPS scanning it was possible to follow the variation of the surface concentration with increasing of the elements O (1s), C (1s), N (1s) e Cl (2s) confirming surface interaction, despite not being feasible to define what functional groups take place in the interaction. Numerical analysis presents herbicide removal in the order of 10-12% concerning measure performed over a single membrane. Complementary tests of metal removal (Cd) confirmed the advantage of CHI surface in this type of interaction, making evident that composed filtration system could be ideal in the removal of distinct contaminants.

O estudo avaliou a efetividade de um sistema para tratamento de soluções aquosas contendo poluentes orgânicos emergentes (POE), denominados interferentes endócrinos (IE). Baseado nos processos oxidativos avançados (POA), empregando ozônio, radiação UV, peróxido de hidrogênio e variação de pH, o estudo possibilitou determinar a relação ideal oxidante/poluentes e o custo/benefício dos tratamentos com melhores desempenhos, de degradação do padrão contendo hormônios 17?-estradiol (E2) e 17?-etinilestradiol (EE2), e outra contendo atrazina (ATZ). Para os estudos, foi utilizado um planejamento fatorial 23, para identificar as melhores opções de degradação. Os tratamentos iniciaram com soluções em concentrações de 600 ?g L-1 de ATZ, em quadruplicatas, por períodos de até 10 minutos. Posteriormente a avaliação dos melhores tratamentos, foi estudada a cinética de mineralização de soluções a 5 mg L-1 de ATZ, em até 90 min. A concentração de estudo adotada para os hormônios foi de 1 mg L-1. A determinação da ATZ e dos hormônios (E2 e EE2) foi realizada por cromatografia em fase líquida (CLAE), utilizando a extração em fase sólida (EFS) quando necessário. Para o estudo foram construídos dois sistemas de tratamento paralelamente acoplados (S1 - escala laboratorial e S2 - escala piloto), com os respectivos volumes de 0,43 L e 20 L. O S2 possibilitou a avaliação da degradação e/ ou mineralização do fenol em solução a 50 mg L-1, porém em escala 60 vezes superior ao S1. As eficiências das mineralizações da ATZ e do fenol, foram avaliadas pela análise de Carbono Orgânico Total (COT). As condições experimentais pré-estabelecidas foram: (i) UV-C 254 nm, variando entre 0, 16 e 32 W; (ii) H2O2 entre 0, 50 e 100% do total teórico necessário para mineralização dos poluentes; (iii) O3 (4,6 g h-1) e (iv) pH 7; 9 e 11, previamente aplicadas às soluções estoque de ATZ. Após avaliação da melhor combinação para degradação, optou-se por trabalhar, nos demais estudos, apenas com soluções em pH 11,0. Na última etapa deste estudo, foram desenvolvidos dois métodos (direto e indireto) para avaliar a geração do gás ozônio em tempo real, possibilitando também realizar a medição do consumo de O3 durante os estudos. Para avaliar a eficiência dos métodos, os resultados analíticos foram convertidos em % de degradação ou mineralização e os custos dos tratamentos foram estimados. Comprovando a viabilidade desta proposta, resultados envolvendo o tratamento de solução padrão de atrazina (600 ?g L-1) apresentaram, nas melhores condições (pH 11, 30 mg L-1 de H2O2 e 4,6 g L-1 de O3), eficiência igual ou superior a 95% (<LQ) na degradação do herbicida, em apenas 2,5 minutos de tratamento e posteriormente soluções a 5 mg L-1, apresentando até 100% de mineralização em 90 minutos. Quanto aos hormônios foram obtidos poucos resultados acima do LD do equipamento, porém todos ficaram abaixo do LQ (50 ?g L-1). Quanto ao fenol (50 mg L-1), estudado no sistema piloto (S2), atingiu-se até 80% de mineralização em 24 horas e 100% de degradação. Os resultados obtidos demonstram que os sistemas apresentaram excelente eficácia conforme esperado / The study evaluated the effectiveness of a system for treatment of aqueous solutions containing emerging organic pollutants (EOP), called endocrine disruptors (ED). Based on advanced oxidation processes (AOP) using ozone, UV radiation, hydrogen peroxide and pH variation. This study allowed the determination of the ideal oxidant/pollutants ratio and the cost/benefit of treatments with better performance, regarding the degradation of the pattern containing hormones 17?-estradiol (E2) and 17?-ethinylestradiol (EE2), and another containing atrazine (ATZ). A 23 factorial design was used in the study to identify the best options for degradation. The treatments were started with solutions at concentrations of 600 ?g L-1 ATZ, in quadruplicate, for periods up to 10 min. Subsequently to the best treatment evaluation; the degradation of 5 mg L-1 ATZ solutions was evaluated, within 90 min. The adopted study concentration of the hormones was 0,05 mg L- 1 and 1,0 mg L- 1. The determination of the ATZ and hormones (E2 and EE2) was performed by liquid chromatography (HPLC) using solid phase extraction (SPE) when necessary. For this study, two parallel coupled processing systems were constructed (S1 - Laboratory scale and S2 - pilot scale) with the respective volumes of 0.43 L and 20 L. S2 system was developed to enable evaluation of the degradation of an already known substance, 50 mg L-1 phenol solution, but on a scale 60 times greater than S1. The efficiency of ATZ and phenol mineralization was evaluated by Total Organic Carbon (TOC). The pre-established experimental conditions were: (i) 254 nm UV-C, varying among 0, 16 and 32 W, (ii) H2O2 among 0, 50 and 100% of the theoretical value needed to complete mineralization of the pollutant, (iii) O3 (4,6 g h-1) and (iv) pH 7; 9 and 11 previously applied to all stock solutions. After assessing the best combination for degradation, it was chosen to work in the other experiments only solutions with pH 11.0. In the last stage of the study, two methods (an electric and a spectrophotometric) were developed to assess the ozone gas generation in real time and also allowing the measurement of O3 consumption during the studies. To evaluate the efficiency of the methods, analytical results were converted to % degradation or mineralization and treatment costs were estimated. Proving the feasibility of this proposal, preliminary results involving the treatment of atrazine standard solution (600 ?g L-1) had, in the best conditions (pH 11, 30 mg L-1 of H2O2 and 4,6 g L-1 of O3), efficiency equal or superior to 95% (<LQ) in degradation of the herbicide in just 2.5 minutes of treatment and then 5 mg L-1 solutions had 100% mineralization in 90 minutes. To hormones, few results above the DL of the equipment were obtained, but all were below the QL of the equipment (50 mg L-1). When phenol (50 mg L-1) was studied in the pilot system it reached up to 80% of mineralization within 24 hours. The obtained results shown that the systems developed reached high efficiency for treating the pollutants

A atrazina é um herbicida amplamente utilizado no Brasil e no mundo em diferentes culturas agrícolas, principalmente em culturas de milho, sorgo, soja e cana-de-açúcar, entretanto, pode se tornar um contaminante de águas superficiais e subterrâneas e do solo, gerando uma preocupação ambiental, pois desequilibra e interfere no ecossistema. A atrazina pode sofrer biodegradação, tornando o processo de biorremediação uma alternativa viável e ecologicamente aceitável para o tratamento de ambientes contaminados por esse herbicida. O microrganismo de referência nesse processo de biodegradação é a Pseudomonas sp. ADP que possui o plasmídio pADP-1, no qual se localizam os genes atzA, atzB, atzC, atzD, atzE e atzF, que codificam enzimas atuantes na via de degradação da atrazina. O presente estudo teve como objetivo o isolamento e a caracterização de bactérias do gênero Pseudomonas quanto à presença de genes de degradação da atrazina e quanto à capacidade de degradação desse herbicida. No presente trabalho foram isoladas 123 cepas de amostras de solo de diferentes regiões do Brasil, as quais foram caracterizadas através de provas bioquímicas e moleculares e identificadas como Pseudomonas aeruginosa (74,8%) e outras espécies (25,2%) pertencentes aos gêneros Pseudomonas, Achromobacter, Burkholderia, Cupriavidus e Rhizobium. A variabilidade genética dos isolados pertencentes ao gênero Pseudomonas foi analisada através da técnica de ERIC-PCR e demonstrou que todos os isolados apresentam alta diversidade genética (<80%). Os genes atzA, atzB, atzC, atzD, atzE e atzF foram detectados em seis isolados provenientes de amostras de solo das regiões Norte, Nordeste, Sudeste e Centro-Oeste do Brasil, sendo três deles da espécie Pseudomonas aeruginosa, um Cupriavidus pauculus, uma Burkholderia cepacea e um Rhizobium radiobacter. Apenas três isolados contendo os genes atz apresentaram plasmídios. Os isolados bacterianos que apresentaram os genes atz foram testados quanto à capacidade de crescimento utilizando a atrazina como única fonte de nitrogênio e quanto à capacidade de degradação desse herbicida. Todos os isolados testados apresentaram crescimento em meio ATZ-R, mas não foi possível observar a mineralização do herbicida, tanto em meio sólido quanto em meio líquido, todavia observou-se a degradação da atrazina por uma espécie de Pseudomonas aeruginosa (isolado P86), com redução de 44%, em meio líquido ATZ-R contendo 33 ppm de atrazina. / Atrazine - the herbicide widely used in Brazil and all over the world in different agricultural crops mainly in corn, sorghum, soy, and sugar cane can contaminate the superficial and subterraneous water, and soil creating an environmental concern due to imbalance and interference caused in ecosystem. Atrazine can suffer biodegradation, which can make the bioremediation process a viable and ecologically acceptable alternative for the treatment of the environment contaminated with this herbicide. The microorganism referred on that biodegradation process is Pseudomonas sp. ADP which has pADP-1 plasmid, where are the genes atzA, atzB, atzC, atzD, atzE, and atzF that codify active enzymes on atrazine degradation way. The aim of this present study was the isolation and the characterization of Pseudomonas genus bacteria due to the presence of the atrazine degradation genes, and the capacity of degradation of this herbicide. In the present work 123 samples of soil bacteria from different Brazilian regions were isolated, and characterized by biochemical and molecular tests, and identified as Pseudomonas aeruginosa (74.8%) and other species (25.2%) of Pseudomonas, Achromobacter, Burkholderia, Cupriavidus, and Rhizobium genus. Genetic variability of the isolated of Pseudomonas genus was analyzed by the ERIC-PCR technique, and demonstrate that all the isolated ones presented high genetic diversity (<80%). atzA, atzB, atzC, atzD, atzE, and atzF genes were detected in six isolated of soil samples from North, Northeast, Southeast, and Midwest of Brazil being 3 Pseudomonas aeruginosa, 1 Cupriavidus pauculus, 1 Burkholderia cepacea, and 1 Rhizobium radiobacter. Only 3 isolated with atz genes presented plasmids. Bacterial isolates that presented atz genes were tested for growth and degradation capacity using only atrazine as nitrogen source. All tested isolates presented growth on ATZ-R, but mineralization of this herbicide in solid or liquid media was not possible to observe. However, 1 specie Pseudomonas aeruginosa (isolate P86) presented atrazine degradation (reduction of 44%) in ATZ-R liquid media with 33 ppm of atrazine.

Esta tese descreve o desenvolvimento de metodologias eletroanalíticas, em especial a voltametria de onda quadrada com o eletrodo de gota pendente de Hg, para determinação de atrazina em águas de rios do Estados de São Paulo e em amostras de solo. A exatidão das metodologias foi avaliada por comparação com HPLC e estudos de recuperação. Inicialmente desenvolveu-se o método em modo estático, no qual a maior sensibilidade foi obtida em pH 2 e freqüência de 400 Hz, com limites de detecção e determinação de 2 e 10 microgramas L-1, respectivamente. O estudo do comportamento eletroquímico revelou que a atrazina comporta-se reversivelmente em altas freqüências de onda quadrada. Dentre os metabólitos da atrazina, a desetilatrazina e desiosopropilatrazina, também são eletroativas nas condições estudadas, sendo possíveis interferentes, ao contrário da hidroxiatrazina e didealquilatrazina. Sistemas em fluxo contínuo e de injeção seqüencial foram desenvolvidos para determinação de atrazina em extrato de solo e águas naturais, bem como em estudos de adsorção. Em águas, os limites de detecção e determinação obtidos por injeção seqüencial foram de 4,5 e 15 microgramas L-1. Estudos de adsorção revelaram que atrazina liga-se mais fortemente a solos em meios ácidos. As metodologias em fluxo permitiram um significativo aumento da velocidade analítica e economia de reagentes, automatizando as determinações. / This thesis describes the development of electroanalytical methodologies, especially the square wave voltammetry with the hanging mercury drop electrode, for determination of the herbicide atrazine in river waters of the São Paulo state and soil samples. The accuracy of the methodologies was evaluated by comparison with HPLC and recovery studies. Initially, a stationary method was developed, in which the highest sensitivity was attained at pH 2 and at frequency of 400 Hz, with detection and quantification limits of 2 and 10 micrograms L-1, respectively. The electrochemical behavior of atrazine was reversible at the higher square wave frequencies. Among the atrazine metabolites, deethylatrazine and deisopropylatrazine are also electroactive under the studied conditions, being possible interferences, contrary to hydroxyatrazine and didealkylatrazine. Continuous flow and sequential injection systems were developed for determination of atrazine in soil extracts and natural waters, as well as in adsorption studies. In waters, the detection and quantification limits attained by sequential injection were 4.5 and 15 micrograms L-1. Adsorption studies showed that atrazine binds more strongly to soil in acidic media. The flow analysis methodologies lead to significant increase in the sampling throughput and reagent saving, automating the determinations.

Background: Evidence from experimental studies suggests that atrazine and its analytes alter the timing of puberty in laboratory animals. Such associations have not been investigated in humans. Objective: To determine the association between in utero exposure to atrazine analytes and earlier menarche attainment in a nested case-control study of the population-based Avon Longitudinal Study of Parents and Children. Methods: Cases were girls who reported menarche before 11.5 years while controls were girls who reported menarche at or after 11.5 years. Seven atrazine analyte concentrations were measured in maternal gestational urine samples (sample gestation week median (IQR): 12 (8–17)) during the period 1991–1992, for 174 cases and 195 controls using high performance liquid chromatography-tandem mass spectrometry. We evaluated the study association using multivariate logistic regression, adjusting for potential confounders. We used multiple imputation to impute missing confounder data for 29% of the study participants. Results: Diaminochlorotriazine (DACT) was the most frequently detected analyte (58%>limit of detection [LOD]) followed by desethyl atrazine (6%), desethyl atrazine mercapturate (3%), atrazine mercapturate (1%), hydroxyl atrazine (1%), atrazine (1%) and desisopropyl atrazine (0.5%). Because of low detection of other analytes, only DACT was included in the exposure–outcome analyses. The adjusted odds of early menarche for girls with DACT exposures≥median was 1.13 (95% Confidence Interval [95% CI]:0.82, 1.55) and exposure Conclusions: This study is the first to examine the association between timing of menarche and atrazine analytes. We found a weak, non-significant association between in-utero exposure to atrazine metabolite DACT and early menarche, though the association was significant in the subset of girls with complete confounder information. Further exploration of the role of these exposures in female reproduction in other cohorts is needed.

Background : Atrazine is an agricultural herbicide used extensively in corn production worldwide. Atrazine is an endocrine disruptor and has been linked to many other deleterious health outcomes. Exposure assessment studies have been carried out in Iowa among farm and non-farm populations. However, commercial pesticide applicators have been left out of those studies. Atrazine is also used in developing countries in grain production. In developing countries there is great concern about acute pesticide poisonings, but chronic exposure to less acutely toxic pesticides has not been studied extensively. This study assessed the in-home contamination of atrazine among commercial pesticide applicators in Iowa and then quantitatively analyzed these results with results from similar studies. Occupational inhalation exposure to atrazine was also assessed in Honduras. Methods : Dust samples were taken from 29 commercial pesticide applicator households in four different locations. This sampling was done once during the atrazine application season and again six months later during winter months to assess atrazine persistence. Occupational and household characteristics were analyzed for associations with atrazine dust levels. Data from two previous studies that analyzed farm and non-farm household dust samples for atrazine were combined with data from the commercial applicator's homes. This new and larger dataset was analyzed to identify which population has the greatest risk for take-home atrazine exposure and what determinants were associated with in-home atrazine dust levels. Lastly, corn production practices in Honduras were evaluated and personal air samples were taken from pesticide applicators during atrazine application to assess inhalation exposure. Results : The first study found that atrazine levels persist into the winter months in the homes of commercial applicators. Atrazine handling (days, pounds, and acres sprayed) were all positively associated with in-home atrazine levels. Commercial applicators that change their shoes inside had higher atrazine levels. More frequent floor cleaning was associated with lower atrazine levels. The second study identified commercial applicators' homes as the most contaminated compared with farmers who apply atrazine to their own land, farmers who hire out atrazine application, and non-farm homes. Farmers that apply their own atrazine also had significantly higher atrazine levels in their homes. The association between atrazine handling and household atrazine levels was highly significant in this study (p < 0.001). In Honduras, atrazine is applied to corn fields with tractor/boom equipment and manual backpack sprayers. Despite applying about one-fifteenth the amount of atrazine, backpack sprayers are exposed to nearly equal amounts of atrazine via inhalation exposure and likely have greater exposure via the dermal route. Among backpack sprayers, which type of spray nozzle used is associated with inhalation exposure. Among tractor/boom applicators, tractor drivers have much lower inhalation exposure than workers who operate and observe the boom. Conclusions : The amount of atrazine handled is the most important determinant for predicting in-home atrazine levels in Iowa. Ubiquitous atrazine contamination and its distribution within homes and among household type provide strong evidence for the take-home pathway. While some improvements have been made in Honduras regarding pesticide application, poor farm workers and small farmers still use antiquated pesticide application techniques which leads to a higher risk of inhalation and dermal exposure.

Surface water quality of the Liverpool Plains (NSW), a series of floodplains comprising the floodplain of the Mooki River, is poor despite the introduction of conservation tillage that has reduced soil erosion and, with it, the transport of sediment, nutrients and chemicals to streams.The aim of this thesis was to provide a basis for recommending the possible wider use of biofilters, by determining their effectiveness in reducing pollutant (mainly atrazine) transport at multiple scales in the Liverpool Plains, quantifying the importance of relevant processes, including the capacity of soil biofilters to degrade the retained pollutants, and evaluating the effect of pollutants on the biofilter vegetation. These objectives were explored in two sub-catchments within the Liverpool Plains, namely Big Jacks Creek and the Blackville.Soils are predominantly vertosols that crack deeply on drying, resulting in initially high infiltration rates on wetting and high water holding capacity. Field monitoring revealed that biofilters such as grassed waterways, natural grasslands or vegetated filter strips (VFS) positioned at multiple catchment scales can significantly reduce pollutant concentrations in runoff. The use of biofilters is discussed, along with other best management practices that will be needed to manage pesticide loads both at source and in the transport pathway. / Doctor of Philosophy (PhD)