<p>Rapid growth of energy consumption and greenhouse gas emissions from fossil fuels have promoted extensive research on biofuels. <a>Algal biofuels have been considered as a promising and environmentally friendly renewable energy source</a>. However, several limitations have inhibited the development of cost-effective biofuel production, which includes unstable cultivation caused by invading organisms and high cost of lipid extraction. This dissertation aims to investigate photochemical approaches to prevent culture collapse caused by invading organisms and biological approaches for the development of cost-effective lipid extraction methods.</p><p> </p><p>As a chemical-free water treatment technology, ultraviolet (UV) irradiation has been widely applied to inactivate pathogens but has not been used in algal cultivation to control invading organisms. To evaluate the potential of using UV irradiation to control invading algal species and minimize virus predation, <i>Tetraselmis sp. </i>and <i>Paramecium bursaria Chlorella virus 1</i> (PBCV-1) were examined as challenge organisms to evaluate effectiveness of UV disinfection. The concentration of viable (reproductively/infectively active) cells and viruses were quantified by a most probable number (MPN) assay and a plaque assay. A low-pressure collimated-beam reactor was used to investigate UV₂₅₄ dose-response behavior of both challenge organisms. A medium-pressure collimated-beam reactor equipped with a series of narrow bandpass optical filters was used to investigate the action spectra of both challenge organisms. Both challenge organisms showed roughly five log₁₀ units of inactivation for UV₂₅₄ doses over 120 mJ/cm². the most effective wavelengths for inactivation of <i>Tetraselmis</i> were from 254 nm to 280 nm, in which the inactivation was mainly attributed to UV-induced DNA damage. On the contrary, the most effective wavelength for inactivation of PBCV-1 was observed at 214 nm, where the loss of infectivity was mainly attributed to protein damage. These results provide important information for design of UV reactors to minimize the impact of invading organisms in algal cultivation systems.</p><p> </p><p>Additionally, a virus-assisted cell disruption method was developed for cost-effective lipid extraction from algal biomass. Detailed mechanistic studies were conducted to evaluate infection behavior of <i>Chlorovirus </i>PBCV-1 on <i>Chlorella sp.</i>, impact of infection on mechanical strength of algal cell wall, lipid yield, and lipid distribution. Viral disruption with multiplicity of infection (MOI) of 10^-8 completely disrupted concentrated algal biomass in six days. Viral disruption significantly reduced the mechanical strength of algal cells for lipid extraction. Lipid yield with viral disruption increased more than three times compared with no disruption control and was similar to that of ultrasonic disruption. Moreover, lipid composition analysis showed that the quality of extracted lipids was not affected by viral infection. The results showed that viral infection is a cost-effective process for lipid extraction from algal cells as extensive energy input and chemicals required by existing disruption methods are no longer needed.</p><p> </p><p>Overall, this dissertation provides innovative approaches for the development of cost-efficient algal biofuels. Application of UV disinfection and viral disruption significantly reduces chemical consumption and improves sustainability of algal biofuel production.<br></p>

A madeira é considerada um compósito natural de extrema complexidade, constituído basicamente por celulose, lignina, hemicelulose (polioses) e extrativos. Sua composição favorece ataques biológicos de diferentes espécies. Neste contexto, inúmeras técnicas vêm sendo estudadas e aplicadas para desinfestar e descontaminar obras de arte e bens culturais móveis elaborados em madeira e que sofreram ataques por fungos, bactérias ou insetos. A irradiação com raios gama, emitidos por isótopos instáveis como o cobalto-60, também vem sendo estudada como uma alternativa para esta finalidade. Este processo tem-se mostrado eficiente para eliminar infestação por insetos e microorganismos em objetos de madeira. Além disso, é rápido e não exige quarentena porque não gera resíduos tóxicos. Neste trabalho, objetivou-se avaliar os efeitos da radiação gama sobre algumas características das espécies de madeira Cedro Rosa e Imbuia. Para tanto, considerou-se que, tal como os demais métodos usualmente empregados na eliminação de microorganismos e insetos que atacam madeiras, o processamento por radiação não impede a reinfestação ou a recontaminação do objeto irradiado. Assim, foram aplicadas doses de radiação gama relativamente altas, até 100 kGy, de modo a investigar os efeitos da radiação, mesmo que as peças de madeira sejam submetidas repetidas vezes ao processo de irradiação, já que a dose absorvida é sempre cumulativa. Estudou-se a influência da radiação gama em propriedades físicas densidade aparente e retratibilidade, em propriedades mecânicas compressão paralela às fibras, flexão no módulo de elasticidade, dureza e cisalhamento e em propriedades térmicas estabilidade térmica das madeiras consideradas. Os resultados demonstraram que a radiação gama, na faixa de dose estudada, não promove alterações nas propriedades das espécies de madeiras investigadas. / Wood is considered a natural composite of extreme complexity, basically composed by cellulose, lignin, hemicelulose (polyosis) and extractives. Its composition favors biological attacks from different species. In this context, several techniques have been studied and applied for disinfecting and decontaminating wood-made works of art and cultural heritage, which have been damaged by fungi, bacteria and insects. Gamma radiation emitted by unstable isotopes, such as 60- cobalt, has also been studied as an alternative to the conventional wood preservatives. So, gamma rays treatment has been shown to be efficient to the removal of infestations by insects and microorganisms in wood-made artifacts, to be fast and not to require quarantine because it does not generate toxic waste. Similar to other techniques, this process does not prevent the irradiated material of re-infestation or recontamination. In this context, the effects of relatively high disinfestation gamma radiation doses (up to 100 kGy) on Cedro Rosa and Imbuia, two typical Brazilian wood species, are accompanied by the changes on the following attributes: apparent density, retracting, parallel compression to fibers, bending in the modulus of elasticity, hardness, shear and thermal stability. Results have shown that gamma radiation, in the studied dose range, does not promote alterations on properties of investigated wood species. In case of a re-infestation, these observations indicate that the wood species can be submitted to repeatedly irradiation processes without causing damage to their structure up to the studied dose range, in despite of radiation effects be always cumulative.

Entre as principais dificuldades que os sistemas de tratamento de esgotos domésticos compostos por lagoas de estabilização enfrentam para atender as exigências impostas pela Resolução CONAMA 357/2005 podem se destacar as concentrações elevadas de nitrogênio amoniacal, fósforo e coliformes fecais e totais. Estes últimos, quando os sistemas utilizam corpos receptores classe 2 ou 3 geram a necessidade de desinfecção. Dentre as diversas tecnologias disponíveis para a desinfecção dos esgotos, a cloração é bastante atrativa pelo custo reduzido, em relação às outras tecnologias, bem como pela elevada experiência que as companhias e municipalidades possuem neste tipo de sistema, advinda dos sistemas de tratamento de água. Dentre os sistemas de cloração, o uso de hipoclorito de sódio torna-se interessante pela segurança, simplicidade das instalações e de operação. A desvantagem do uso deste tipo de sistema se dá na possibilidade da formação de subprodutos tóxicos, dentro os quais pode-se destacar os trihalometanos (THMs) e os ácidos haloacéticos (AHAs), porém esta formação pode ser teoricamente reduzida pela presença de elevadas concentrações de nitrogênio amoniacal nos efluentes destes sistemas, por sua reação preferencial com o cloro, na formação de cloraminas. Outras possibilidades de redução na formação é reduzir a concentração de precursores, reduzir a dosagem de cloro e utilizar método de descloração dos efluentes após a desinfecção. Este trabalho, ambientado no desenvolvimento das pesquisas do Edital 4 do PROSAB/FINEP (Programa Nacional de Pesquisa em Saneamento Básico), desenvolveu testes de desinfecção em efluentes de lagoas anaeróbias e facultativas utilizando como desinfetante hipoclorito de sódio, em dosagens de 8,0 mgCl/L e 12,0 mgCl/L. Desenvolveu também testes de desinfecção com efluentes anaeróbios e facultativos pós-tratados por meio de coagulação/floculação/sedimentação e coagulação/floculação/flotação com ar dissolvido, com dosagens de 2,0 mgCl/L e 4,0 mgCl/L. Para todos os testes de desinfecção foram realizados também testes de descloração das amostras. Desta forma foi possível estudar, dentro de uma faixa limitada, as possibilidades de reduzir a formação de subprodutos tóxicos na desinfecção de efluentes de lagoas de estabilização com hipoclorito de sódio. Nas amostras foram determinados por meio de cromatografia gasosa as concentrações dos THMs e AHAs. Por meio das metodologias do Standard Methods 20th Edition foram realizadas as análises físico-químicas dos parâmetros pH, temperatura, DQO, DBO, Ntotal, N-NH3, NTK, cloro livre, cloraminas, coliformes e outros microrganismos indicadores. A formação de THMs se mostrou bastante pequena, em média abaixo dos 20,0 µg/L em todos os testes, aparentemente devido às elevadas concentrações de N-NH3, da ordem de 30,0 mg/L. O mesmo não ocorreu com os AHAs. Em muitos testes a concentração encontrada de AHAs totais superou o padrão de 80,0 µg/L preconizados pela USEPA para água potável, e aqui adotado como padrão comparativo. A descloração reduziu em média 52% a concentração de AHAs nos testes, colocando quase todas as amostras dentro do padrão de 80,0 µg/L. Os bioensaios mostraram que a presença de cloro livre nas amostras eleva a toxicidade aguda em Daphnia similis provocada pelos efluentes e que a cloração seguida de descloração produz efluentes com toxicidade inferior à toxicidade dos efluentes apenas clorados e toxicidade também inferior àquela dos efluentes sem cloração. / The main difficulties that the stabilization ponds for sewage treatment systems face to meet the requirements imposed by CONAMA Resolution 357/2005 can highlight the high concentrations of ammonia nitrogen, phosphorus and fecal and total coliform. When the systems use receivers water bodies Class 2 or 3 generate the need for disinfection. Among the different technologies available for the disinfection of sewage, the chlorination is very attractive at reduced cost compared to other technologies, as well as the high experience that companies and municipalities have in this type of system, because the water treatment systems. About the chlorination technologies, the use of sodium hypochlorite becomes interesting for security, simplicity of installation and operation. The disadvantage of using this type of system is given the possibility of formation of toxic by-products, within which you can highlight the trihalomethanes (THMs) and haloacetic acids (AHAs), but this formation can theoretically be reduced by the presence of high concentrations of ammoniacal nitrogen in effluents of these systems, for its preferential reaction with the chlorine in the formation of cloraminas. Other possibilities for reducing the training is to reduce the concentration of precursors (organic matter), reduce the dosage of chlorine and use dechlorination method after disinfection. This work, developed in PROSAB / FINEP (National Program for Research on Sanitation) research announcement number 4, developed tests for disinfection of anaerobic and facultative ponds effluents using sodium hypochlorite as a disinfectant, in concentrations of 8.0 mgCl/L and 12.0 mgCl/L. Were also developed tests for disinfection with anaerobic and facultative ponds effluents post-treated by coagulation / flocculation / sedimentation and coagulation / flocculation / flotation with dissolved air, with concentrations of 2.0 mgCl/L and 4.0 mgCl/L. For all of disinfection tests were also conducted dechlorination tests. This made it possible to study the possibilities of reducing the formation of toxic byproducts of the stabilization ponds effluents disinfection with sodium hypochlorite. The THMs and AHAS concentrations were determined by gas chromatography. Through the methods of Standard Methods 20th Edition analyses were carried out physical-chemical parameters of pH, temperature, COD, BOD, total nitrogen, ammoniacal nitrogen, total Kjeldhal nitrogen, free chlorine, chloramines, coliforms and other indicator microorganisms. The formation of THMs was very low, on average below the 20.0 µg/L in all tests, apparently due to high concentrations of NH3-N, the order of 30.0 mg/L. The same did not occur with AHAs. In many tests found concentrations of AHAs exceeded the standard of 80.0 µg/L recommended by the USEPA for drinking water. Dechlorination reduced 52% of AHAs concentrations, in average. Bioassays showed that the presence of free chlorine in the samples increase the acute toxicity in Daphnia similes. Chlorination followed by dechlorination produces effluents with less toxicity than the toxicity of chlorinated and effluent without chlorination.

O objetivo deste trabalho foi quantificar a formação de biofilme de Candida albicans e avaliar a integridade da superfície de uma resina acrílica termopolimerizável tratada com os sistemas de polimento líquido Biscover LV (Bisco, Schaumburg, USA) e Surface Coat (Kuraray, Tokyo, Japan), após 30 e 90 ciclos de desinfecção química. Um total de 280 corpos de prova (10 X 10 X 2 mm) foram confeccionados, esterilizados e aleatoriamente divididos em 20 grupos (n=14), de acordo com o tipo de polimento (mecânico de 0,3 ou 3 m, Surface Coat e Biscover LV), número de ciclos testados (0, 30 e 90) e solução utilizada: Nenhuma (controle negativo), Hipoclorito de sódio 1% (experimental) e água destilada (controle positivo). Após exposição às respectivas soluções de acordo com o protocolo adotado, as amostras foram imersas em saliva artificial por 2 h para a formação da película adquirida salivar e então inoculadas com 2 mL de suspensão de C. albicans (1.107 cel/mL) para fase de adesão durante 90 min. Após esta fase, as amostras foram lavadas e imersas em TSB estéril por 24 h para crescimento do biofilme em estufa a 37ºC sob agitação. Para quantificação do biofilme, foi utilizado o método de coloração por Cristal Violeta, em 240 espécimes divididos em 20 grupos de 12 amostras cada. Esta análise foi realizada por espectrofotômetro com comprimento de onda de 595 nm para mensuração do nível de absorbância do sobrenadante obtido após a descoloração do biofilme. Para análise qualitativa da integridade da camada de polimento liquido, 2 amostras de cada grupo (uma contaminada e outra não) tiveram suas superfícies fotografadas em seis campos distintos através do microscópio eletrônico de varredura (MEV). Os resultados foram submetidos o teste paramétrico ANOVA, seguido de teste de Tukey (p<0,05). Comparando os tipos de tratamentos de superfície avaliados dentro do mesmo tempo e soluções testadas não pode-se notar diferença estatisticamente significante entre os grupos de 0,3 m e Biscover LV ou 3 m e Surface Coat quando comparados entre si. Porém quando comparados os grupos de 0,3 m e Biscover LV aos grupos de 3 m e Surface Coat, estes apresentaram diferença estatisticamente significante. A análise em MEV dos corpos de prova colonizados demostrou a presença de C. albicans na superfície do corpo de prova sob a forma de leveduras e hifas em todos os grupos, enquanto que nos não colonizados foi observada uma degradação progressiva em todos os tipos de tratamento de superfície a medida que os números de ciclos e os desafios químicos eram aumentados. Apesar disso, o grupo Biscover LV apresentou uma quantidade de biofilme formada compatível com uma superfície lisa (0,3 m), enquanto que Surface Coat apresentou quantidade de biofilme formada compatível com a superfície rugosa (3,0 m). / The aim of this study was to quantify Candida albicans biofilm formation and evaluate the surface integrity of an denture base resin treated with liquid-polish Biscover LV (Bisco, Schaumburg, USA) and Surface Coat (Kuraray, Tokyo, Japan), after 30 and 90 cycles of chemical disinfection. A total of 280 specimens (10 x 10 x 2 mm) were prepared, sterilized and randomly divided into 20 groups (n = 14), according to the type of polishing (mechanical 0.3 or 3 m, Surface Coat and Biscover LV), number of cycles tested (0, 30, 90) and the solution used: none (negative-control), 1% sodium hypochlorite (experimental) and distilled water (positive-control). After exposure to the respective solutions according to the protocol adopted, the samples were immersed in artificial saliva for 2 h to form the acquired salivary pellicle and then inoculated with 2 mL of C. albicans (1.107 cel / mL) for 90 min to adhesion. After this, samples were washed and immersed in TSB for 24 h to biofilm growth at 37°C. For biofilm quantification, the method of crystal violet staining was used in 240 specimens, divided into 20 groups of 12 samples each. This analysis was performed by a spectrophotometer at a wavelength of 595 nm by measuring the level of supernatants absorbance obtained after the biofilms stain release. For qualitative analysis of the integrity of the liquid-polish layer, two samples from each group (one colonized and one not) had their surfaces photographed in six different fields through the scanning electron microscope (SEM). The results were submitted to ANOVA followed by Tukey\'s test (p<0.05). Comparing the types of surface treatments evaluated, within the same time and tested solutions, can not be noted statistically significant difference between groups of 0.3 m and Biscover LV or 3 m and Surface Coat. But when comparing the groups of 0.3 m and Biscover LV to groups of 3 mm and Surface Coat, there was a statistically significant difference. SEM analysis of the colonized specimens demonstrated the presence of C. albicans in the form of yeast and hyphae in all groups. In not colonized surfaces a progressive degradation was observed that was bigger when the number of cycles and chemical challenges were increased. Nevertheless, the group Biscover LV showed a amount of biofilm similar to a smooth surface (0.3 m), while Surface Coat presented an amount compatible with the roughened surface (3.0 m).

lt;p align="justify"gt; A água tem influência direta sobre a saúde, a qualidade de vida e o desenvolvimento do ser humano. Cerca de um quarto dos 4,8 bilhões de pessoas dos países em desenvolvimento não possuem acesso a fontes de água adequadas. Carvalho (1983), Silva e Araújo (2003), e outros, citam percentuais de contaminação por coliformes totais e fecais em água de cisterna que chegam a 91%. O presente estudo teve como objetivo, avaliar a eficiência do clorador simplificado por difusão na desinfecção da água utilizada para consumo humano, e para isso, fez-se análise microbiológica da água de 30 residências, correspondendo a 48% das existentes na bacia hidrográfica do Ribeirão do Laje, em Caratinga, MG localizadas à jusante do distrito de Santa Luzia, que utilizam água sem tratamento, captada de cisterna ou diretamente de nascente. Constatou-se que 90% das amostras analisadas estavam contaminadas com coliformes totais. Observou-se que a totalidade das amostras de água captadas de nascente estavam contaminadas por coliformes totais. Nenhuma captação de nascente está satisfatoriamente protegida. Quanto às cisternas, somente 28,6% possuem revestimento, 59% não possui tampa adequada, em 33% ocorre entrada de água de enxurrada no interior da cisterna, 43% estão distante de cursos d’água, e a média de distância das que estão próximas é de 12 metros, o que mostrou ser o mais provável fator de contaminação da água. Constatou-se que 70% dos entrevistados desconhecem normas básicas de construção e manutenção de cisternas. Segundo a percepção dos moradores entrevistados cuja escolaridade média de 3,2 anos de estudo, todos classificam a água consumida como sendo “excelente” ou “boa”, mas, contraditoriamente, 77% acreditam haver microorganismos patogênicos na água que consomem. Em 47% das residências pesquisadas não havia filtro doméstico para água. Implantou-se o clorador em 10 cisternas que apresentaram contaminação por coliformes, e observou-se que em três análises microbiológicas subseqüentes (10, 20 e 33 dias após implantação), não houve detecção de coliformes totais e fecais nas amostras. O teor de cloro ativo liberado pelo clorador ficou sempre abaixo de 0,2 mg L-1, e mesmos com teores menores que 0,1 mg L-1 não houve detecção de coliformes. Segundo a percepção dos moradores entrevistados, o clorador não interfere significativamente no paladar e cheiro da água, o que não desestimula seu uso. Concluiu-se neste estudo, que o clorador simplificado por difusão é um método eficiente, simples, de baixo custo, e que deve ter sua utilização estimulada junto à população que não tem acesso à água tratada, aspecto que em muito contribuirá para que se reduzam os riscos de transmissão de doenças de veiculação hídrica.lt;/pgt; / lt;p align="justify"gt; The present study it has as objective, to evaluate the efficiency of the clorador simplified for diffusion in the disinfection of the water used for human consumption, and for this, microbiological analysis of the water of 30 residences became, corresponding 48% of the existing residences in the hidrográfica basin of the Ribeirão do Laje, in Caratinga – MG located to the ebb tide of the district of Luzia Saint, that use water without treatment, caught of watering hole or spring. One evidenced that 90% of the analyzed samples were contaminated with total coliformes. It was observed that the totality of spring the caught water samples was contaminated by total coliformes. No apitation of spring satisfactorily is protected. How much to watering holes, 29% only they possess covering, 59% does not possess adjusted cover, in 33% occurs entered of torrent water in the interior of the watering hole, 43% are distant of courses d’água, and the distance average to that they are next is of 12 meters, what it showed to be the most likely factor of contamination of the water. One evidenced that 70% of the interviewed ones are unaware of basic norms of construction and maintenance of watering holes. According to perception of the interviewed inhabitants whose average escolaridade of 3,2 years of study, 100% classifies the consumed water as being “excellent” or “good”, but, contraditoriamente, 77% believe to have pathogenic microorganisms in the water that consume. In 47% of the searched residences it did not have domestic filter for water. The clorador in 10 watering holes was implanted that had presented contamination for coliformes, observed it that in three subsequent microbiological analyses (10, 20 and 33 days after implantation), did not have detention of total and fecais coliformes in the samples. The set free active chlorine text for the clorador was always below of 0,2 mg L-1, and same with 0,1 lesser texts that mg L-1 did not have detention of coliformes. According to perception of the interviewed inhabitants, the clorador does not intervene significantly with the palate and smells of the water, what it does not discourage its use. It was concluded in this study, that the clorador simplified for diffusion is an efficient, simple method, of low cost, and that it must have its use stimulated together to the population that does not have access to the treated water, aspect that in very will contribute so that the risks of transmission of illnesses of hídrica propagation are scrumbled.lt;/pgt;

A desinfecção dos canais radiculares figura como constante preocupação clínica. Esta pesquisa teve a proposta de verificar o nível de desinfecção alcançado pela instrumentação mecanizada utilizando o sistema K3 em relação ao uso desse mesmo sistema associado a um tratamento químico dentinário complementar. Foram utilizados 16 caninos inferiores unirradiculares, que foram aleatoriamente divididos em dois grupos. Foram realizados 2 ensaios em dias diferentes, cada ensaio formado por 8 dentes em cada grupo. As coroas dos dentes foram cortadas e o tamanho das raízes padronizado em 15 mm. Os canais foram esvaziados com auxilio de limas tipo K de números 10 ou 15 com hipoclorito de sódio a 1% seguido de 20 mL de tiosulfato de sódio a 5%. Seguiu-se então a odontometria. Os dentes foram impermeabilizados externamente por duas camadas de cianoacrilato de etila e montados em tubos Eppendorf a expensas de adesivo epóxico. Os conjuntos (raiz + Eppendorf) foram esterilizados em autoclave por 20 min a 134 °C. Os espécimes foram inoculados com uma da suspensão correspondente à concentração bacteriana 0,5 da escala de Mc Farland (1,5 x 108 UFC). A primeira coleta microbiológica foi realizada imediatamente após o tempo de incubação para estabelecer o número de unidades formadoras de colônias. O preparo químicocirúrgico dos canais radiculares do grupo 1 foi realizado apenas por meio do sistema rotatório K3. Cones de papel absorvente esterilizados foram inseridos no canal radicular para nova coleta. Os dentes do Grupo 2 também tiveram os canais radiculares preparados com sistema rotatório K3 até a fase de irrigação-aspiração final com hipoclorito de sódio a 0,5% e EDTA-T a 17%. Após essa etapa foi realizado o tratamento químico da dentina que correspondeu à inserção de solução de hipoclorito de sódio a 1% no interior da cavidade pulpar e com uma lima tipo K de número 25 a solução foi agitada durante um minuto. Esse procedimento foi repetido 5 vezes num total de 10 mL dessa substância e somando um tempo de 10 minutos. A irrigação final foi efetuada com 10 mL de solução de hipoclorito de sódio a 0,5% seguidas de 20 mL de solução de EDTA- T a 17% (pH 7,0). Nova coleta para exame microbiológico foi realizada. Essas suspensões sofreram diluições seriadas de 10-1 a 10-7 em água peptonada antes do preparo químico-cirúrgico e de10-1 a 10-5 após o mesmo As diluições foram então semeadas em triplicata em placas contendo TSA. Após o período de 24 h de incubação as placas que apresentaram crescimento bacteriano tiveram o número de unidades formadoras de colônias (UFCs) determinado. Embora houvesse redução da população bacteriana em ambos os grupos no que se refere ao pré e pós-operatório, os resultados demonstraram que não houve diferença estatística significante entre os grupos de estudo (p>0,05). / The effective disinfection of root canals represents a constant clinical concern. This research aimed at verifying the level of disinfection achieved by mechanical instrumentation using the K3 system when compared to the associated use of the same system and a complementary chemical dentinal treatment. Sixteen singlerooted lower canines were randomly divided into two groups. Two experiments, each using 8 teeth per group, were conducted on different days. The dental crowns were sectioned and the root length was standardized to 15 mm. The canals were instrumented using #10 or 15 K-files with 1% sodium hypochlorite, followed by 5% sodium thiosulfate. The root canals were measured. The teeth were made externally impermeable by two layers of ethyl cyanoacrylate and placed in Eppendorf tubes using epoxy resin. The compounds (root + Eppendorf) were sterilized in an autoclave for 20 min at 134oC. A suspension that corresponds to the bacterial concentration of 0.5 on the McFarland scale (1.5 x 108 CFU) was inoculated onto the specimens. The first microbiological sample collection was done immediately after incubation in order to determine the number of colony-forming units. Chemical preparation of the root canals of group 1 was done using the K3 rotatory system exclusively. Sterile paper cones were inserted into the canal for a new collection. Teeth from group 2 also had their root canals prepared using the K3 rotatory system up to the phase of final irrigation with 0.5% sodium hypochlorite and 17% EDTA-T. Next, chemical dentinal treatment was done by inserting a 1% sodium hypochlorite solution into the pulp cavity and agitating for one minute with the aid of a #25 K-file. This step was repeated 5 times using a total of 10 ml of the solution for a total duration of 10 minutes. Final irrigation was done using 10 ml of 0.5% sodium hypochlorite solution followed by 20 ml of 17% EDTA-T (pH 7.0). A new collection for microbiological examination was done. These suspensions underwent serial dilutions from 10-1 to 10- 7 using peptone water before preparation and from 10-1 to 10-5 afterwards. The dilutions were then inoculated thrice onto TSA plates. After an incubation period of 24h, the number of colony-forming units (CFU) was determined for the plates which presented with bacterial growth. Although there was a reduction of the bacterial population in both experimental groups when pre and post-operative counts were compared, results demonstrated that there was no statistically significant difference between the two study groups (p>0.05).

Neste estudo foi avaliado o potencial tóxico de diferentes desinfetantes (cloro, ácido peracético, radiação ultravioleta e ozônio) utilizados na desinfecção de esgotos sanitários. Para tanto, foram realizados ensaios de desinfecção (em diversas concentrações e tempos de contato) com o esgoto doméstico originário da estação de tratamento de esgotos da cidade de Araraquara (SP) e, posteriormente, ensaios de toxicidade a fim de verificar possíveis efeitos agudos e crônicos nos seguintes organismos-teste: Daphnia similis, Ceriodaphnia silvestrii, Chironomus xanthus, Danio rerio e Allium cepa. Todos os desinfetantes, nas condições experimentais testadas, foram capazes de produzir efeitos deletérios aos organismos-teste utilizados nesta pesquisa. O cloro foi considerado o desinfetante mais tóxico, sendo seguido pelo ozônio, ácido peracético e radiação UV. Verificou-se ainda que quando o efluente não desinfetado foi tóxico aos organismos-alvo, sua toxidez foi potencializada com a adição dos diferentes agentes desinfetantes. Os resultados obtidos neste estudo sugerem que a utilização do cloro para desinfecção de esgotos sanitários, sem prévia descloração, deve ser revista, em face da eficiência satisfatória de inativação de bactérias proporcionada por outros agentes de desinfecção potencialmente menos tóxicos (tais como o ácido peracético e a radiação UV). / In this study, it was evaluated the toxic potential of different disinfectant agents (chlorine, peracetic acid, ultraviolet radiation and ozone) used in the disinfection of urban wastewater. For so much, disinfection assays were accomplished (in several concentrations and contact times) with the domestic sewage from Araraquara city (SP) and toxicity bioassays were developed in order to verify possible acute and chronic effects in the following test-organisms: Daphnia similis, Ceriodaphnia silvestrii, Chironomus xanthus, Danio rerio and Allium cepa. All the disinfectants, in the tested experimental conditions, were capable to produce harmful effects to the test-organisms used in this research. Chlorine was considered the most toxic disinfectant, being followed by ozone, peracetic acid and UV radiation. It was noticed that when the effluent, by itself, was toxic to the test-organism, its toxicity was increased with the different disinfecting agents\' addition. The results obtained in this study suggest that the use of chlorine as a wastewater disinfectant, without previous dechlorination, should be reviewed, because it was observed that other disinfection agents (such as peracetic acid and UV radiation) were able to promote satisfactory levels of bacteria inactivation for potentially less toxicity.

A poluição dos corpos de água é cada vez maior devido ao lançamento de efluentes, esgotamento sanitário e disposição inadequada de resíduos sólidos. Esse fato causa grande preocupação à comunidade científica, pois à poluição associam-se riscos à saúde causado por poluentes bioativos, compostos químicos orgânicos persistentes e outros compostos que poderiam estar presentes na água. Dentre as substâncias que causam risco à saúde se destacam os desreguladores endócrinos, que são compostos químicos exógenos que interferem na atividade hormonal. Os sistemas de tratamento de água convencionais não removem alguns compostos, tais como pesticidas e hormônios, os quais deveriam ser removidos. O uso do ozônio e cloro vem sendo reportado para tal finalidade, pois atuam como oxidantes que são capazes de oxidar compostos orgânicos, além de serem poderosos desinfetantes. Portanto, o objetivo deste estudo foi avaliar a eliminação e/ou redução dos desreguladores endócrinos - DE em estações de tratamento de água com o uso do ozônio e do cloro. Aliado ao estudo, avaliar qual será a eficiência de remoção de organismos indicadores nas doses utilizadas para oxidação de DE (Coliformes Totais e E.coli) e avaliar a ecotoxicidade em peixes de amostras cloradas contendo 17\'beta\'-estradiol. O cloro e o ozônio foram capazes de remover eficientemente estrona (E1) e 17\'beta\'-estradiol (E2) de águas, contudo a eficiência de remoção diminui em concentrações na ordem de ng/L, necessitando-se de altas doses de ozônio e cloro para a completa remoção dos hormônios estudados. Tanto o cloro como o ozônio foram eficientes na inativação de organismos indicadores nas doses comumente utilizadas em ETAs e nas doses requeridas para a remoção dos contaminantes estudados. Amostras contendo E2 após serem cloradas apresentaram toxicidade crônica a peixes da espécie Danio rerio, o efeito estrogênico foi observado principalmente em machos. Foi necessário uma dose de ozônio de 0,95 e 3,8 mg/L para a remoção de 93 e 98% de E2 e E1 respectivamente (Co = 100 ng/L). Considerando o cloro como oxidante após a aplicação de 1,5 mg/L com tempo de contato de 24 h chegou-se a uma remoção de 99,6 e 97,5% para o E2 e E1 respectivamente (Co = 500 ng/L). Tão importante quanto remover o estrona e o 17\'beta\'-estradiol foi verificar e identificar quais foram os subprodutos formados após a oxidação dos mesmos. Foram identificados vários subprodutos tanto utilizando a degradação por meio do cloro como do ozônio. Alguns destes compostos identificados foram persistentes e recalcitrantes. Para o ozônio a completa remoção foi atingida após 10 mg/L de \'O IND.3\' e 16 mg/L de \'O IND.3\' para os subprodutos do E1 e E2 respectivamente. Considerando o cloro mesmo após um tempo de contato de 16 dias, havia ainda subprodutos da degradação na água. / The pollution of water bodies is increasing due to the release of effluents, sewage and inadequate solid waste disposal. This fact causes great concern to the scientific community, because pollution are associated with health risks caused by pollutants of bioactive compounds and other persistent organic compounds that could be present in water. Among the substances that can cause health risks are the endocrine disrupters, which are exogenous chemicals that interfere with hormonal activity. The conventional water treatment systems does not remove certain compounds, such as pesticides and hormones, which should be removed. The use of ozone and chlorine has been reported for this purpose because it acts as an oxidant that can oxidize organic compounds, and addicionally are powerful disinfectants. Therefore, the aim of this study was the elimination and/or reduction of endocrine disrupters during the water treatment using ozone and chlorine. Allied to the study, evaluate the efficiency of indicator organisms removal in the doses used for the oxidation of endocrine disrupters (coliform and E.coli) and assess the ecotoxicity of fish in chlorinated samples containing 17\'beta\'- estradiol. Chlorine and ozone were able to efficiently remove estrone (E1) and 17\'beta\'-estradiol (E2) of water, but the removal efficiency decreases at concentrations in the ng/L. Complete removal of the hormones was reach with high doses of ozone and chlorine. Both chlorine and ozone were effective in the inactivation of indicator organisms at commonly doses used in water treatment plants and in the doses required for the removal of the pollutants studied. Samples containing chlorinated E2 had chronic toxicity to fish Danio rerio, the estrogenic effect was observed mainly in males, females in some groups estrogenic effect was not observed. The dose of ozone necessary for the removal of 93 and 98% of E1 and E2 (Co = 100 ng/L) were 0.95 and 3.8 mg/L, respectively. The application of 1.5 mg/L of chlorine with a contact time of 24 h achieved removal of 99.6 and 97.5% for E1 and E2 respectively (Co = 500 ng/L) . As important as removing estrone and 17\'beta\'-estradiol was to verify and identify which were the byproducts formed after oxidation. Several byproducts were identified by the degradation using chlorine and ozone. Some of these compounds were identified to be persistent and recalcitrant. The ozone dose required to the complete removal of the byproducts was achieved after 10 mg/L \'O IND.3\' and 16 mg/L \'O IND.3\' for the byproducts of E1 and E2 respectively. The byproducts of chlorine even after a contact time of 16 days persisted in the water.

水資源缺乏引起的一系列問題在世界上已建得到廣泛關注，因此，確保提供潔淨衛生的水在保護人類健康和環境方面起著重要作用。近來，光催化作為頗有前景的替代方法被廣泛應用殺菌除污。二氧化鈦是目前研究最多應用最廣的光催化劑。基於紫外光譜照射，催化劑表面產生活性氧化物種，這些物種具有強氧化性能殺滅細胞。 / 本文首次研究了母體菌種大腸桿菌BW25113和它的同源單基因缺陷變異體對光催化殺菌的靈敏度差異。母體菌種和變異菌種表現出不同的耐受性。基於此，能幫助發掘出重要的變種。通過生物化學方法，可以檢測出不同菌種的生理性特徵。結合其他方法，可以進一步揭示光催化殺菌的生理性機理。 / 首先，我們篩選出了兩種重要的變異體。一種是大腸桿菌JW1081，即脂肪酸變異體，該菌種缺乏脂肪酸合成調節的關鍵基因。一種是大腸桿菌JW3942，即乙酰輔酶A變異體，該菌種缺乏乙酰輔酶A合成調控得到關鍵激酶。我們發現脂肪酸變異體對光催化處理的耐受性稍低，而乙酰輔酶A變異體則耐受性較高。 同時發現，溫度可以調節細胞膜的不飽和酸和飽和酸的比例。因此，我們提出脂肪酸和乙酰輔酶A是光催化殺菌中的重要影響因子。 / 更進一步研究發掘了細胞內酶和光催化產生的活性氧物種間的關係。大腸桿菌JW3914，即過氧化氫酶變異體，是發現的另一個重要的變異體。通過對母體和變異體的淬滅劑實驗，主要的殺菌活性氧物種是光催化產生的雙氧水，而不是羥基自由基。細胞體內的過氧化氫酶誘導在母體菌體內發現，而未在變異體內檢測到。 / 本課題採用母體/單基因變異體的研究方法，為全面深刻理解光催化殺菌的深層機理提供一種全新的研究思路。 / Many problems associated with the lack of clean, fresh water worldwide are well known. Developing countries will particularly be affected by water availability problems and there will be further pressure on water demand resulting from economic development and population growth. Therefore, the provision of safe and clean water plays a key role in protecting human health and the environment. Recently, photocatalytic oxidation (PCO) has been widely accepted as a promising alternative method of water disinfection. Titanium dioxide (TiO₂) has been investigated extensively and is the most widely used photocatalyst. Upon the irradiation of UVA lamp, reactive charged and oxidative species are generated on TiO₂ surface and can inactive the bacterial cells. / In this study, the photocatalytic performances of a parental strain (E.coli BW25113) and its isogenic single-gene deletion mutant strains have been investigated for the first time. These bacterial strains exhibited different sensitivies towards photocalytic inactivation. Based on this, it can help reveal some important mechanism from the mutations. Biotic factors were confirmed by physiological biochemical measurement. / Firstly, we screened out the potential mutation fabF⁻ mutant (E. coli JW1081, carrying the mutation of fabF759(del)::kan) and coaA⁻ mutant (E. coli JW3942, carrying the mutation of coaA755(del)::kan). The isogenic fabF⁻ mutant is slightly more susceptible, and coaA⁻ mutant is less susceptible to photocatalytic inactivation. Through conditioning temperature, it adjusts the ratio of unsaturated to saturated fatty acid (FA) of cell membrane. We propose that FA profile and coenzyme A level significantly affect photocatalytic inactivation of bacteria. Moreover, we show photogenerated electrons (e⁻) can directly inactivate the cells of E. coli. / Furthermore, we report the relationship between the bacterial intracellular enzyme and the reactive charged and oxidative species (ROSs) generated during photocataltic inactivation. The katG⁻ mutant, E. coli JW3914, carrying the mutation of katG729(del)::kan is another important mutation. The parental and katG⁻ mutant strains reveal that photogenerated H₂O₂ but not OH[subscript free] is another important reactive oxygen species to inactivate bacteria. The inducible catalase (CAT) corresponding to H₂O₂can be detected in parental strain but not in katG⁻ mutant. / The research methodology using parental/single-gene deletion mutant strains is expected to shed light on fully understanding of the fundamental mechanism of photocatalytic inactivation of E. coli. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Gao, Minghui. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 130-177). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Acknowledgements --- p.i / Abstract --- p.v / Table of contents --- p.ix / List of Figures --- p.xiii / List of Plates --- p.xvii / List of Tables --- p.xviii / List of Equations --- p.xix / Abbreviations --- p.xxi / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Water crisis --- p.1 / Chapter 1.2 --- Traditional disinfection methods --- p.3 / Chapter 1.2.1 --- Chlorination --- p.4 / Chapter 1.2.2 --- Ozonation --- p.6 / Chapter 1.2.3 --- Ultraviolet irradiation --- p.8 / Chapter 1.2.4 --- Multiple disinfectants --- p.10 / Chapter 1.3 --- Advanced oxidation process (AOPs) --- p.10 / Chapter 1.3.1 --- Hydrogen Peroxide/Ozone (H₂O₂/O₃) --- p.11 / Chapter 1.3.2 --- Ozone/Ultraviolet Irradiation (O₃/UV) --- p.12 / Chapter 1.3.3 --- Hydrogen Peroxide/ Ultraviolet Irradiation (H₂O₂/UV) --- p.12 / Chapter 1.3.4 --- Fenton's --- p.Reaction / Chapter 1.4 --- Solar photocatalytic disinfection (SPC-DIS) --- p.14 / Chapter 1.4.1 --- Photocatalyst-TiO₂ --- p.31 / Chapter 1.4.2 --- Irradiation sources --- p.35 / Chapter 1.4.3 --- TiO₂ photocatalytic process --- p.35 / Chapter 1.4.4 --- The role of photogenerated reactive charged and oxidative species (ROSs) --- p.38 / Chapter 1.5 --- Bacteria --- p.40 / Chapter 1.5.1 --- E. coli BW25113 --- p.40 / Chapter 1.5.2 --- E. coli Keio Collection --- p.41 / Chapter 1.5.3 --- Bacterial defense mechanism towards oxidative stresses --- p.44 / Chapter 1.6 --- Photocalytic applications --- p.53 / Chapter 1.7 --- Significance of the project --- p.55 / Chapter 2. --- Objectives --- p.58 / Chapter 3. --- Genetic studies of the roles of fatty acid and coenzyme A in photocatalytic inactivation of Escherichia coli --- p.61 / Chapter 3.1 --- Introduction --- p.61 / Chapter 3.2 --- Materials and methods --- p.65 / Chapter 3.2.1 --- Photocatalyst --- p.65 / Chapter 3.2.2 --- Bacterial nutrient --- p.66 / Chapter 3.2.3 --- Bacterial strains --- p.67 / Chapter 3.2.4 --- Photocatalytic inactivation --- p.69 / Chapter 3.2.5 --- Fatty acid profile --- p.72 / Chapter 3.2.6 --- Fluorescent measurement of bacterial coenzyme A content --- p.74 / Chapter 3.2.7 --- The role of photogenerated electrons (e⁻) to bacterial inactivation --- p.74 / Chapter 3.2.8 --- Transmission Electron Microscopic (TEM) --- p.75 / Chapter 3.2.9 --- Photoelectrochemical measurement --- p.77 / Chapter 3.3 --- Results --- p.77 / Chapter 3.3.1 --- Photocatalytic inactivation --- p.77 / Chapter 3.3.2 --- Effects of pre-incubation at different temperatures --- p.80 / Chapter 3.3.3 --- Fatty acid profile --- p.83 / Chapter 3.3.4 --- Fluorescent measurement of bacterial coenzyme A content --- p.84 / Chapter 3.3.5 --- The role of electron (e⁻) in photocataytic inactivation --- p.84 / Chapter 3.3.6 --- Transmission electron microscopy (TEM) --- p.89 / Chapter 3.3.7 --- Photocurrent measurement --- p.90 / Chapter 3.4 --- Discussion --- p.90 / Chapter 3.5 --- Conclusions --- p.96 / Chapter 4 --- Genetic and physiological studies of the role of Catalase and H₂O₂ in photocatalytic inactivation of E. coli --- p.98 / Chapter 4.1 --- Introduction --- p.98 / Chapter 4.2 --- Materials and methods --- p.101 / Chapter 4.2.1 --- Bacterial strains --- p.101 / Chapter 4.2.2 --- Photocatalytic performance --- p.102 / Chapter 4.2.3 --- Scavenger studies --- p.103 / Chapter 4.2.4 --- Effects of different pHs on photocatalytic inactivation --- p.104 / Chapter 4.2.5 --- Measurement of bacterial catalase activity and H₂O₂ --- p.104 / Chapter 4.2.6 --- Transmission electron microscopy (TEM) --- p.105 / Chapter 4.2.7 --- Atomic absorption spectrophotometer (AAS) --- p.105 / Chapter 4.3 --- Results and discussion --- p.106 / Chapter 4.3.1 --- Photocatalytic performance --- p.106 / Chapter 4.3.2 --- Scavenger studies --- p.108 / Chapter 4.3.3 --- Contribution of hydrogen peroxide (H₂O₂) --- p.111 / Chapter 4.3.4 --- Effects of different pHs on photocatalytic inactivation --- p.114 / Chapter 4.3.5 --- Bacterial catalase (CAT) activity --- p.116 / Chapter 4.3.6 --- Destruction model of bacterial cells --- p.118 / Chapter 4.4 --- Conclusions --- p.120 / Chapter 5. --- General conclusions --- p.122 / Chapter 6. --- Prospectives --- p.125 / Chapter 7. --- Appendix --- p.127 / Chapter 8. --- References --- p.130

The emergence of pathogens that are more difficult to inactivate than bacteria, such as C. parvum and G. lamblia, has led to the enactment of more stringent drinking water regulations. The Long Term 2 Enhanced Surface Water Treatment Rule (LT2ESWTR), promulgated in January 2006, requires increased inactivation of C. parvum. However, increasing the disinfectant dose to enhance inactivation, especially when using chlorine, increases production of carcinogenic disinfection by-products (DBPs) such as trihalomethanes (THMs). As a result of the risks posed by DBPs, the Stage 2 Disinfectants and Disinfection By-Products Rule (Stage 2 D/DBP) was promulgated to limit exposure to DBPs by requiring systems to monitor concentrations at the worst cases locations in the distribution system. The purpose of this study was to evaluate sonication as an alternate disinfection strategy to reduce THM formation. Prior research has demonstrated the inactivation kinetics of sonication. Therefore, if sonication also reduces THM formation, this disinfection technology could help water utilities simultaneously comply with the Stage 2 D/DBP Rule and the LT2ESWTR. Water samples were prepared with varying concentrations of natural organic matter (NOM). THM formation potential reduction was evaluated by treating the water samples with sonication at 20 kHz for 0 seconds, 30 seconds, 60 seconds, 5 minutes and 10 minutes. After treatment, the samples were chlorinated and incubated at 20„aC to form THMs. After incubation times from 1 to 7 days, THMs were extracted, and gas chromatography with electron capture detection was used to quantify THM concentrations in treated and control samples. For experimental water with an NOM concentration of 1 mg/L that was dosed with 6 mg/L of NaOCl, the average THM formation potential reduction was 40% for sonication times of 30 seconds, 5 minutes and 10 minutes. The data for 60 seconds of treatment do not follow the same trends as the other data. Additional study is necessary to increase precision of the experimental data; however, this study supports sonication as a potential method of THM reduction.