The PhD thesis deals with determination of sulfonamides and tetracyclines in waste waters. Sulphonamides and tetracyclines are widely used in human and veterinary medicine. They are biologically active compounds and block biological processes in waste water treatment plants. These substances can influence microorganisms in the environment. Sulphonamides and tetracyclines induce resistance in low concentrations. These substances penetrate into the environment from industrial production, use in households, hospitals, veterinary clinics and fish farms. Tetracyclines and sulphonamides were determined in waste water from waste water treatment plants in Brno. Optimalization of extraction of these compounds was performed by solid phase extraction. Sulphonamides and tetracyclines were determined by liquid chromatography coupled with diode array detector or coupled with mass spectrometry. Optimalized method was used for determination tetracyclines and sulphonamides in waste water from waste water treatment plants in Brno in years 2007 – 2010.

Although wastewater treatment plants are a line of defense in minimizing indiscriminate output of microconstituents to natural waters, we do not possess a fundamental understanding of the mechanisms involved in microconstituent removal during wastewater treatment. With this in mind, experiments were designed to investigate the factors that can influence the fate of four microconstituents, carbamazepine (CBZ), 17alpha-ethinylestradiol (EE2), iopromide (IOP), and trimethoprim (TMP), during biological suspended culture treatment. Specifically, the role that various ecological members of biological treatment systems play in biotransforming these compounds was evaluated. Sorption assays were performed with inactivated biomass samples (ammonia oxidizing bacteria (AOB), laboratory enriched heterotrophic cultures free of active nitrifiers with low (Ox⁻) or high (Ox⁺) oxygenase activity, and a nitrifying activated sludge (NAS) from a full-scale wastewater treatment plant) to determine whether partitioning dictates removal of individual microconstituents. No microconstituents sorbed to the AOB culture. Neither CBZ nor IOP sorbed to Ox⁻, Ox⁺ and NAS cultures; however, EE2 and TMP sorbed to the Ox⁻, Ox⁺ and NAS biomass. Sorption was positively influenced by the presence of exopolymeric substances (EPS) associated with the cultures. The protein content of EPS affected EE2 and TMP sorption more appreciably than the polysaccharide content of EPS. Further experiments were performed to investigate microconstituent biodegradation by AOBs, Ox⁻ and Ox⁺ cultures. The influence of growth state and oxygenase activity on biotransformation by each culture was also evaluated. Results indicate that EE2 was the only microconstituent that was amenable to biotransformation by batch cultured AOB and heterotrophic cultures. EE2 was biotransformed but not mineralized by AOB chemostat and batch cultures. TMP was not transformed by AOB batch or chemostat cultures; however both EE2 and TMP were transformed by Ox⁻ and Ox⁺ chemostat cultures. Radiolabeled studies showed that EE2 was mineralized by this culture. Kinetically, AOBs dominated EE2 transformation to monohydroxylated metabolites; however, both Ox⁻ and Ox⁺ cultures further degraded and mineralized EE2 and metabolites generated by AOBs. These results indicate that biotransformation of EE2 by NAS may be limited by heterotrophic activity whereas TMP fate may be a function of heterotrophic activity only. Oxygenase activity did not limit EE2 or TMP biotransformation in chemostat cultures. Subsequent experiments that were performed to identify the factors that influence heterotrophic degradation of EE2 and TMP indicated that the presence of readily biodegradable substrates slows EE2 and TMP biotransformation. The impact of slowly biodegradable substrates like EPS on EE2 and TMP degradation was unclear. These results suggest that EE2 and TMP are most amenable to biodegradation in bioreactors where endogenous conditions dominate. / Ph. D.

A paracoccidioidomicose (PCM), doença sistêmica de caráter granulomatoso, causada pelo fungo termodimórfico Paracoccidioides brasiliensis. A PCM é endêmica na América Latina e atinge principalmente indivíduos do sexo masculino com atividades econômica ligada a agricultura. Os pacientes com PCM exigem tratamento a base de sulfametoxazol/trimetoprim, anfotericina B, e derivados azólicos por longos períodos. A gp43, possui 416 aminoácidos, onde um trecho específico de 15 aminoácidos (QTLIAIHTLAIRYAN) designado como (P10), é reconhecido pelos linfócitos T de camundongos e humanos. No presente estudo avaliamos o efeito aditivo da imunização do P10 com às drogas antifúngicas utilizadas no tratamento da PCM. Nossos resultados indicam um efeito aditivo entre a imunização com P10 e o tratamento medicamentoso em camundongos Balb/c infectados. Associado a redução significativa da carga fúngica no pulmão, baço e fígado desses animais, detectamos aumento dos níveis de IL-12 e IFN-? e diminuição de IL-4 e IL-10. / Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides brasiliensis, a thermal dimorphic fungus. PCM is endemic in Latin America affecting mainly rural workers. The patients with PCM demand treatment the base of sulfametoxazole/trimethoprim, amphotericin B and derivatives azolic. The gp43 has 416-mer and mice and human T lymphocytes are stimulated by a 15-mer peptide designated as P10 (QTLIAIHTLAIRYAN). In the present work we evaluated the additive effect of P10 immunization and antifungal drugs utilizing in PCM treatment. Ours results showed an additive protective effect between immunization with P10 and drugs treatment with infected Balb/c mice. Associated to the reduction of fungal burden in the lung, spleen and liver we observed increase of levels of IL-12 and IFN-? and reduction of IL-4 and IL-10.

Stenotrophomonas maltophilia é um bacilo Gram-negativo, não fermentador, considerado um microorganismo pouco virulento, relacionado principalmente a infecções associadas à assistência a saúde. A S. maltophilia apresenta um padrão de resistência intrínseca à maioria das classes de antibióticos. A droga de escolha para o tratamento das infecções por S. maltophilia é a sulfametoxazol/ trimetoprima (SMX/TMP). Entretanto, estudos atuais relatam o aumento da resistência a esse antibiótico, o que limita assim as opções para terapia efetiva. Outras opções de tratamento são o levofloxacino e a tigeciclina, porém, faltam estudos clínicos e in vitro dessas drogas. A proposta deste estudo foi avaliar os possíveis mecanismos de resistência a SMX/TMP e as quinolonas em isolados clínicos de pacientes internados no Instituto Central do Hospital das Clínicas e do Hospital A.C. Camargo. Foram avaliadas 106 amostras de S. maltophilia isoladas de pacientes adultos com infecção relacionada à assistência a saúde, internados no Instituto Central do Hospital das Clínicas da FMUSP e no Hospital de Câncer A.C Camargo durante o período de dezembro de 2008 a dezembro de 2010. A sensibilidade à SMX/TMP foi de 78,3%, para levofloxacino de 82% e 14,2% para cirpofloxacino, para minociclina de 100% e tigeciclina 91,6%. Foi realizado PCR para detecção dos genes sul1, sul2 e dfrA1 para avaliar a resistência à SMX/TMP, os genes int1 e iscr2 para avaliação da presença de elementos genéticos móveis e os genes gyrA, qnr, smeD, smeT e aac(6)-Ib-cr para avaliação da resistência as quinolonas. Quatorze amostras (13,2%) foram positivas para o gene sul1. Desses isolados, nove amostras apresentavam resistência ao SMX/TMP com CIM50 de 8 g/mL e CIM90 de 128 g/mL. Cinco amostras positivas para o gene sul1 foram sensíveis a SMX/TMP com CIM50 de 1 g/mL e CIM90 de 1 g/mL. A sequencia do integron1 da amostra com CIM >125 g/mL mostrou um tamanho aproximado de 4000 pb contendo os genes cassetes aac4 e aadA1 e a região qac/sul1. Uma amostra resistente a SMX/TMP foi positiva para o gene sul2 localizado na transposase-like ISCR 2. Observamos a presença de quatro novos qnr em cepas de S. maltophilia e a presença da enzima aac(6)-ib-cr em 4 amostras. 100% das cepas foram positivas para o gene do sistema de efluxo smeDEF e 12/38 amostras tiveram o gene smeT do sistema de efluxo smeDEF, ,porém não foi observada mutação nesse gene. Na sequencia de aminoácidos da girase A de 15 amostras resistentes a levofloxacino não observamos mutações relacionadas à resistência a quinolonas. As cepas resistentes a SMX/TMP apresentaram um padrão policlonal. Dezoito amostras resistentes ao levofloxacino apresentaram 14 perfis clonais, distribuídos em 10 clusters. S. maltophilia exibe múltiplos mecanismos de resistência, nesse estudo observamos um grande número de cepas com elementos genéticos móveis carregando o gene sul1 e outros genes de resistência. A S. maltophilia pode ser um importante reservatório de transmissão de genes de resistência / Stenotrophomonas maltophilia is a gram-negative, non-fermenter, considered a low virulent organism, mainly related to healthcare associated infections. S. maltophilia shows a pattern of intrinsic resistance to many classes of antibiotics. The drug of choice for the treatment of infections caused by S. maltophilia is SMX/TMP, however, current studies have reported increased resistance to this antibiotic, thus limiting the options for effective therapy. Among the treatment options appear tigecycline and levofloxacin, but clinical trials and studies in vitro to such drugs are lacking. The purpose of this study was to evaluate the possible mechanisms of resistance to SMX/TMP and quinolones in clinical isolates from patients admitted to the Institute\'s Central Clinical Hospital and the Hospital A.C Camargo. We evaluated 106 strains of S. maltophilia isolated from adult patients with healthcare associated infections, at the Instituto Central do Hospital das Clínicas and the Cancer Hospital AC Camargo in the period of December 2008 to December 2010. The sensitivity to SMX/TMP was 78.3%, 82% for levofloxacin, 14,2% for ciprofloxacin, minocycline 100% and for tigecycline 91.6%. PCR was performed for detection of gene sul1, sul2 and dfrA1 to evaluate the resistance to SMX/TMP, genes iscr2 int1 was performed to evaluate the presence of mobile genetic elements and genes gyrA, qnr, smeD , smeT and aac (6 \')-Ib-cr for evaluation of resistance to quinolones. Fourteen samples (13.2%) were positive for the gene sul1. In these isolates, nine samples showed resistance to SMX/TMP with MIC50 of 8 g/ml and MIC90 of 128 g/mL. Five strains were positive for sul1 gene and were susceptible to SMX/TMP with MIC50 of 1 g/ml and MIC90 of 1 g/mL. The sequence of the integron class 1 strain with an MIC> 125 g/mL showed an approximate size of 4000 bp containing the gene cassettes aadA1, aac4 and qac/sul1. A strain resistant to SMX/TMP was positive for the gene sul2 located on ISCR2 a transposase-like. We observed the presence of four new qnr in strains of S. maltophilia and the presence of the enzyme aac (6 \')-ib-cr in 4 samples. 100% of the strains were positive for the gene of the efflux system smeDEF and 12/38 samples had the gene smeT repressor of smeDEF efflux system, however there was no mutation in this gene. In the amino acid sequence of gyrase A of 15 strains resistant to levofloxacin did not observe mutations related to resistance to quinolones. Strains resistant to SMX/TMP had a polyclonal PFGE pattern. Eighteen strains resistant to levofloxacin showed 14 clonal profiles 14 divided into 10 clusters S. maltophilia displays multiple mechanisms of resistance. In this study, we observed a large number of strains with mobile genetic elements carrying the sul1 gene and other resistance genes. S. maltophilia is may be an important source for transmission of genes of resistance

Stenotrophomonas maltophilia é um bacilo Gram-negativo, não fermentador, considerado um microorganismo pouco virulento, relacionado principalmente a infecções associadas à assistência a saúde. A S. maltophilia apresenta um padrão de resistência intrínseca à maioria das classes de antibióticos. A droga de escolha para o tratamento das infecções por S. maltophilia é a sulfametoxazol/ trimetoprima (SMX/TMP). Entretanto, estudos atuais relatam o aumento da resistência a esse antibiótico, o que limita assim as opções para terapia efetiva. Outras opções de tratamento são o levofloxacino e a tigeciclina, porém, faltam estudos clínicos e in vitro dessas drogas. A proposta deste estudo foi avaliar os possíveis mecanismos de resistência a SMX/TMP e as quinolonas em isolados clínicos de pacientes internados no Instituto Central do Hospital das Clínicas e do Hospital A.C. Camargo. Foram avaliadas 106 amostras de S. maltophilia isoladas de pacientes adultos com infecção relacionada à assistência a saúde, internados no Instituto Central do Hospital das Clínicas da FMUSP e no Hospital de Câncer A.C Camargo durante o período de dezembro de 2008 a dezembro de 2010. A sensibilidade à SMX/TMP foi de 78,3%, para levofloxacino de 82% e 14,2% para cirpofloxacino, para minociclina de 100% e tigeciclina 91,6%. Foi realizado PCR para detecção dos genes sul1, sul2 e dfrA1 para avaliar a resistência à SMX/TMP, os genes int1 e iscr2 para avaliação da presença de elementos genéticos móveis e os genes gyrA, qnr, smeD, smeT e aac(6)-Ib-cr para avaliação da resistência as quinolonas. Quatorze amostras (13,2%) foram positivas para o gene sul1. Desses isolados, nove amostras apresentavam resistência ao SMX/TMP com CIM50 de 8 g/mL e CIM90 de 128 g/mL. Cinco amostras positivas para o gene sul1 foram sensíveis a SMX/TMP com CIM50 de 1 g/mL e CIM90 de 1 g/mL. A sequencia do integron1 da amostra com CIM >125 g/mL mostrou um tamanho aproximado de 4000 pb contendo os genes cassetes aac4 e aadA1 e a região qac/sul1. Uma amostra resistente a SMX/TMP foi positiva para o gene sul2 localizado na transposase-like ISCR 2. Observamos a presença de quatro novos qnr em cepas de S. maltophilia e a presença da enzima aac(6)-ib-cr em 4 amostras. 100% das cepas foram positivas para o gene do sistema de efluxo smeDEF e 12/38 amostras tiveram o gene smeT do sistema de efluxo smeDEF, ,porém não foi observada mutação nesse gene. Na sequencia de aminoácidos da girase A de 15 amostras resistentes a levofloxacino não observamos mutações relacionadas à resistência a quinolonas. As cepas resistentes a SMX/TMP apresentaram um padrão policlonal. Dezoito amostras resistentes ao levofloxacino apresentaram 14 perfis clonais, distribuídos em 10 clusters. S. maltophilia exibe múltiplos mecanismos de resistência, nesse estudo observamos um grande número de cepas com elementos genéticos móveis carregando o gene sul1 e outros genes de resistência. A S. maltophilia pode ser um importante reservatório de transmissão de genes de resistência / Stenotrophomonas maltophilia is a gram-negative, non-fermenter, considered a low virulent organism, mainly related to healthcare associated infections. S. maltophilia shows a pattern of intrinsic resistance to many classes of antibiotics. The drug of choice for the treatment of infections caused by S. maltophilia is SMX/TMP, however, current studies have reported increased resistance to this antibiotic, thus limiting the options for effective therapy. Among the treatment options appear tigecycline and levofloxacin, but clinical trials and studies in vitro to such drugs are lacking. The purpose of this study was to evaluate the possible mechanisms of resistance to SMX/TMP and quinolones in clinical isolates from patients admitted to the Institute\'s Central Clinical Hospital and the Hospital A.C Camargo. We evaluated 106 strains of S. maltophilia isolated from adult patients with healthcare associated infections, at the Instituto Central do Hospital das Clínicas and the Cancer Hospital AC Camargo in the period of December 2008 to December 2010. The sensitivity to SMX/TMP was 78.3%, 82% for levofloxacin, 14,2% for ciprofloxacin, minocycline 100% and for tigecycline 91.6%. PCR was performed for detection of gene sul1, sul2 and dfrA1 to evaluate the resistance to SMX/TMP, genes iscr2 int1 was performed to evaluate the presence of mobile genetic elements and genes gyrA, qnr, smeD , smeT and aac (6 \')-Ib-cr for evaluation of resistance to quinolones. Fourteen samples (13.2%) were positive for the gene sul1. In these isolates, nine samples showed resistance to SMX/TMP with MIC50 of 8 g/ml and MIC90 of 128 g/mL. Five strains were positive for sul1 gene and were susceptible to SMX/TMP with MIC50 of 1 g/ml and MIC90 of 1 g/mL. The sequence of the integron class 1 strain with an MIC> 125 g/mL showed an approximate size of 4000 bp containing the gene cassettes aadA1, aac4 and qac/sul1. A strain resistant to SMX/TMP was positive for the gene sul2 located on ISCR2 a transposase-like. We observed the presence of four new qnr in strains of S. maltophilia and the presence of the enzyme aac (6 \')-ib-cr in 4 samples. 100% of the strains were positive for the gene of the efflux system smeDEF and 12/38 samples had the gene smeT repressor of smeDEF efflux system, however there was no mutation in this gene. In the amino acid sequence of gyrase A of 15 strains resistant to levofloxacin did not observe mutations related to resistance to quinolones. Strains resistant to SMX/TMP had a polyclonal PFGE pattern. Eighteen strains resistant to levofloxacin showed 14 clonal profiles 14 divided into 10 clusters S. maltophilia displays multiple mechanisms of resistance. In this study, we observed a large number of strains with mobile genetic elements carrying the sul1 gene and other resistance genes. S. maltophilia is may be an important source for transmission of genes of resistance

Atualmente a classe dos antibióticos se destaca pelo grande consumo e também pelo risco à saúde quando administrado de forma equivocada, esse aumento deve-se ao destaque cada vez maior da indústria de produtos farmacêuticos. Outra questão a ser levantada é a contaminação do meio ambiente por essa classe substâncias, pois após o uso pelo ser humano ocorre a eliminação natural de parte da concentração administrada, assim sendo liberadas nos esgotos e, posteriormente, contaminando os corpos d\'água, a fauna e flora local. Assim, foi proposto um métodos diferente para a degradação desses compostos, utilizando a eletrodos de difusão gasosa (EDG) capazes de gerar peróxido de hidrogênio in situ e em meio ácido, precursores dos radicais hidroxila, responsáveis pela degradação. Foram estudados EDG\'s não catalisados e catalisados, com diferentes porcentagens de ftalocianina de ferro II, com relação à quantidade de peróxido produzido, melhor potencial de produção e cinética do processo. Os resultados revelaram que a incorporação de 0,5% de Ft-Fe no eletrodo apresentou os melhores resultados. Esse eletrodo foi escolhido então para realizar as degradações dos antibióticos sulfametoxazol e trimetoprim. Na célula eletroquímica utilizou-se o processo Fenton, as reações ocorreram em uma faixa de potencial (-0,4 V ≤ E ≤ -1,4 V) e no reator foi utilizado um potencial fixo (-1,75 V) mas utilizando processos de Fenton e Foto-Fenton. As amostras degradadas foram avaliadas por técnicas analíticas de espectroscopia no ultravioleta (UV), cromatografia líquida de alta eficiência (HPLC) e teor de carbono orgânico total (TOC). Os resultados mostraram que, na célula eletroquímica, o melhor potencial de degradação foi de -1,1 V, com uma taxa de redução de 25,5% para o trimetoprim e 96,0% do sulfametoxazol e uma diminuição do teor de carbono orgânico total de 10,4%. Para o reator o melhor resultado foi obtido para o processo de Foto-Fenton onde houve uma redução de 16,9% do teor de carbono orgânico total e uma redução de 99,7% do sulfametoxazol e 11,3% do trimetoprim, em um potencial de -1,75 V. Considerando a formação de subprodutos foi elaborada uma rota de degradação com os possíveis compostos formados. / Currently the class of antibiotics is notable for the large consumption and also the risk to health when administered in error, this increase is due to the growing prominence of the pharmaceutical industry. Another issue to be addressed is the environmental contamination by substances that class, because after use by humans part of the concentration administered is naturally eliminated, thus being released into sewers and subsequently contaminating water bodies, the local fauna and flora. Thus, different methods has been proposed for the degradation of these compounds, using gas diffusion electrodes (GDE) capable of generating hydrogen peroxide in situ and in acid medium, precursors of hydroxyl radicals, responsible for degradation. We studied GDE\'s not catalyzed and catalyzed with different percentages of iron phthalocyanine II, with respect to the amount of peroxide produced better yield potential and kinetic process. The results revealed that the incorporation of 0.5% of Ft-Fe in the electrode showed the best results. This electrode was then chosen to perform the degradation of the antibiotic sulfamethoxazole and trimethoprim. In the electrochemical cell used in the Fenton process, the reactions occurred in a potential range (-0.4 V ≤ E ≤ -1.4 V) and in the reactor was used a fixed potential (-1.75 V) but using Fenton and photo-Fenton processes. Degraded samples were analyzed by analytical techniques, ultraviolet spectroscopy (UV), high performance liquid chromatography (HPLC) and total organic carbon content (TOC). The results showed that in the electrochemical cell, the best degradation potential was -1.1 V, with a reduction rate of 25.5% for trimethoprim and 96.0% for sulfamethoxazole and a decreased carbon content total of 10.4%. For the reactor the best result was obtained for the photo-Fenton process where there was a reduction of 16.9% of the total organic carbon content and a reduction of 99.7% of sulfamethoxazole and 11.3% of trimethoprim, in a potential of -1.75 V. Considering the formation of byproducts was drafted a route with the possible degradation compounds formed.

Atualmente a classe dos antibióticos se destaca pelo grande consumo e também pelo risco à saúde quando administrado de forma equivocada, esse aumento deve-se ao destaque cada vez maior da indústria de produtos farmacêuticos. Outra questão a ser levantada é a contaminação do meio ambiente por essa classe substâncias, pois após o uso pelo ser humano ocorre a eliminação natural de parte da concentração administrada, assim sendo liberadas nos esgotos e, posteriormente, contaminando os corpos d\'água, a fauna e flora local. Assim, foi proposto um métodos diferente para a degradação desses compostos, utilizando a eletrodos de difusão gasosa (EDG) capazes de gerar peróxido de hidrogênio in situ e em meio ácido, precursores dos radicais hidroxila, responsáveis pela degradação. Foram estudados EDG\'s não catalisados e catalisados, com diferentes porcentagens de ftalocianina de ferro II, com relação à quantidade de peróxido produzido, melhor potencial de produção e cinética do processo. Os resultados revelaram que a incorporação de 0,5% de Ft-Fe no eletrodo apresentou os melhores resultados. Esse eletrodo foi escolhido então para realizar as degradações dos antibióticos sulfametoxazol e trimetoprim. Na célula eletroquímica utilizou-se o processo Fenton, as reações ocorreram em uma faixa de potencial (-0,4 V ≤ E ≤ -1,4 V) e no reator foi utilizado um potencial fixo (-1,75 V) mas utilizando processos de Fenton e Foto-Fenton. As amostras degradadas foram avaliadas por técnicas analíticas de espectroscopia no ultravioleta (UV), cromatografia líquida de alta eficiência (HPLC) e teor de carbono orgânico total (TOC). Os resultados mostraram que, na célula eletroquímica, o melhor potencial de degradação foi de -1,1 V, com uma taxa de redução de 25,5% para o trimetoprim e 96,0% do sulfametoxazol e uma diminuição do teor de carbono orgânico total de 10,4%. Para o reator o melhor resultado foi obtido para o processo de Foto-Fenton onde houve uma redução de 16,9% do teor de carbono orgânico total e uma redução de 99,7% do sulfametoxazol e 11,3% do trimetoprim, em um potencial de -1,75 V. Considerando a formação de subprodutos foi elaborada uma rota de degradação com os possíveis compostos formados. / Currently the class of antibiotics is notable for the large consumption and also the risk to health when administered in error, this increase is due to the growing prominence of the pharmaceutical industry. Another issue to be addressed is the environmental contamination by substances that class, because after use by humans part of the concentration administered is naturally eliminated, thus being released into sewers and subsequently contaminating water bodies, the local fauna and flora. Thus, different methods has been proposed for the degradation of these compounds, using gas diffusion electrodes (GDE) capable of generating hydrogen peroxide in situ and in acid medium, precursors of hydroxyl radicals, responsible for degradation. We studied GDE\'s not catalyzed and catalyzed with different percentages of iron phthalocyanine II, with respect to the amount of peroxide produced better yield potential and kinetic process. The results revealed that the incorporation of 0.5% of Ft-Fe in the electrode showed the best results. This electrode was then chosen to perform the degradation of the antibiotic sulfamethoxazole and trimethoprim. In the electrochemical cell used in the Fenton process, the reactions occurred in a potential range (-0.4 V ≤ E ≤ -1.4 V) and in the reactor was used a fixed potential (-1.75 V) but using Fenton and photo-Fenton processes. Degraded samples were analyzed by analytical techniques, ultraviolet spectroscopy (UV), high performance liquid chromatography (HPLC) and total organic carbon content (TOC). The results showed that in the electrochemical cell, the best degradation potential was -1.1 V, with a reduction rate of 25.5% for trimethoprim and 96.0% for sulfamethoxazole and a decreased carbon content total of 10.4%. For the reactor the best result was obtained for the photo-Fenton process where there was a reduction of 16.9% of the total organic carbon content and a reduction of 99.7% of sulfamethoxazole and 11.3% of trimethoprim, in a potential of -1.75 V. Considering the formation of byproducts was drafted a route with the possible degradation compounds formed.

Antibiotic resistance challenges the practice and development of modern medicine. The aim of this thesis was to test the hypothesis that antibiotic resistance is reversible once the selection pressure of an antibiotic is removed. A decisive reduction (85%) in trimethoprim and trimethoprim-sulfamethoxazole over 24 months in Kronoberg County, Sweden, is described. The resistance baseline prior to the intervention and the effects of the intervention on resistance levels, trimethoprim resistance genes (dfr-genes) and population structure in Escherichia coli were studied. The effects of different algorithms for excluding patient duplicate isolates were small but systematic. An identical algorithm was used throughout. The drastic decrease in the use of trimethoprim containing drugs did not result in a corresponding decrease in trimethoprim resistance. This was true both for total trimethoprim resistance and for trimethoprim mono-resistance. The distributions of E. coli phenotypes, dfr-genes and E. coli sequence types were stable. The marginal effect on resistance rates was explained by a low fitness cost of trimethoprim resistance observed in vitro and the high levels of associated resistance in trimethoprim resistant isolates. Trimethoprim resistance was, although widespread in the E. coli population, more common in certain E. coli sequence types. The distributions of dfr-genes were different in E. coli and K. pneumoniae and between different E. coli sequence types. These results indicate mechanisms related to the genetic back-bone of E coli to be important for the acquisition and persistence of antibiotic resistance. The findings of this thesis indicates that, at least for some classes of antibiotics, we may have overestimated the usefulness of a strategy for reversing antimicrobial resistance based on the fitness cost of resistance. We have equally underestimated the conserving effects of associated resistance. The stability of the dfr-genes and E. coli sequence types underlines the importance of associated resistance and successful lineages in the spread and maintenance of antibiotic resistance in E. coli.

A paracoccidioidomicose (PCM), doença sistêmica de caráter granulomatoso, causada pelo fungo termodimórfico Paracoccidioides brasiliensis. A PCM é endêmica na América Latina e atinge principalmente indivíduos do sexo masculino com atividades econômica ligada a agricultura. Os pacientes com PCM exigem tratamento a base de sulfametoxazol/trimetoprim, anfotericina B, e derivados azólicos por longos períodos. A gp43, possui 416 aminoácidos, onde um trecho específico de 15 aminoácidos (QTLIAIHTLAIRYAN) designado como (P10), é reconhecido pelos linfócitos T de camundongos e humanos. No presente estudo avaliamos o efeito aditivo da imunização do P10 com às drogas antifúngicas utilizadas no tratamento da PCM. Nossos resultados indicam um efeito aditivo entre a imunização com P10 e o tratamento medicamentoso em camundongos Balb/c infectados. Associado a redução significativa da carga fúngica no pulmão, baço e fígado desses animais, detectamos aumento dos níveis de IL-12 e IFN-? e diminuição de IL-4 e IL-10. / Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides brasiliensis, a thermal dimorphic fungus. PCM is endemic in Latin America affecting mainly rural workers. The patients with PCM demand treatment the base of sulfametoxazole/trimethoprim, amphotericin B and derivatives azolic. The gp43 has 416-mer and mice and human T lymphocytes are stimulated by a 15-mer peptide designated as P10 (QTLIAIHTLAIRYAN). In the present work we evaluated the additive effect of P10 immunization and antifungal drugs utilizing in PCM treatment. Ours results showed an additive protective effect between immunization with P10 and drugs treatment with infected Balb/c mice. Associated to the reduction of fungal burden in the lung, spleen and liver we observed increase of levels of IL-12 and IFN-? and reduction of IL-4 and IL-10.

The aim of the diploma thesis was to monitor seasonal concentration changes of 7 antibiotics norfloxacin, levofloxacin, ciprofloxacin, azithromycin, erythromycin, trimethoprim and sulfamethoxazole in wastewater influent and (cleaned) water effluent in the Sewage Treatment Plant (STP) České Budějovice. Time-proportional 24 hours pooled samples of wastewater were collected every month from March 2011 to February 2012 in the influent and effluent pof the STP. The concentrations of target compounds were determined by using in line SPE/LC-MS/MS analysis. The highest average concentration in the influent was detected in case of norfloxacin (0.563 microgram/l) and ciprofloxacin (0.406 microgram/l). The highest average concentration in the effluent was detected in the case of trimethoprim (0.255 microgram/l) and erythromycin (0.117 microgram/l). Higher concentration of antibiotics was measured in the colder periods of the year. It was connected with increased use of antibiotics and with less cleaning efficiency of the STP in this season. The highest removal efficiency was determined for norfloxacin and ciprofloxacin, the lowest for erythromycin. The influence of the season on the removal efficiency of antibiotics was found esp. for azithromycin,trimethoprim and sulfamethoxazole.