abstract: Widespread use of chlorinated solvents for commercial and industrial purposes makes co-occurring contamination by 1,1,1-trichloroethane (TCA), trichloroethene (TCE), and 1,4-dioxane (1,4-D) a serious problem for groundwater. TCE and TCA often are treated by reductive dechlorination, while 1,4-D resists reductive treatment. Aerobic bacteria are able to oxidize 1,4-D, but the biological oxidation of 1,4-D could be inhibited TCA, TCE, and their reductive transformation products. To overcome the challenges from co-occurring contamination, I propose a two-stage synergistic system. First, anaerobic reduction of the chlorinated hydrocarbons takes place in a H2-based hollow-fiber “X-film” (biofilm or catalyst-coated film) reactor (MXfR), where “X-film” can be a “bio-film” (MBfR) or an abiotic “palladium-film” (MPfR). Then, aerobic removal of 1,4-D and other organic compounds takes place in an O2-based MBfR. For the reductive part, I tested reductive bio-dechlorination of TCA and TCE simultaneously in an MBfR. I found that the community of anaerobic bacteria can rapidly reduce TCE to cis-dichloroethene (cis-DCE), but further reductions of cis-DCE to vinyl chloride (VC) and VC to ethene were inhibited by TCA. Also, it took months to grow a strong biofilm that could reduce TCA and TCE. Another problem with reductive dechlorination in the MBfR is that mono-chloroethane (MCA) was not reduced to ethane. In contrast, a film of palladium nano-particles (PdNPs), i.e., an MPfR, could the simultaneous reductions of TCA and TCE to mainly ethane, with only small amounts of intermediates: 1,1-dichloroethane (DCA) (~3% of total influent TCA and TCE) and MCA (~1%) in continuous operation. For aerobic oxidation, I enriched an ethanotrophic culture that could oxidize 1,4-D with ethane as the primary electron donor. An O2-based MBfR, inoculated with the enriched ethanotrophic culture, achieved over 99% 1,4-D removal with ethane as the primary electron donor in continuous operation. Finally, I evaluated two-stage treatment with a H2-based MPfR followed by an O2-MBfR. The two-stage system gave complete removal of TCA, TCE, and 1,4-D in continuous operation. / Dissertation/Thesis / Masters Thesis Civil, Environmental and Sustainable Engineering 2018

O cianeto é um composto tóxico, que pode ser encontrado no ambiente de maneira natural ou como resultado de atividades antropogênicas tais como a mineração de ouro e a indústria da galvanoplastia. Dentre as diferentes espécies do composto, o cianeto de hidrogênio (HCN) é considerado o mais tóxico, mesmo concentrações de 100ppm são letais para os seres humanos. Para a degradação destes compostos de cianeto a compostos menos tóxicos existem diferentes métodos de tratamento que podem ser químicos, físicos ou biológicos. O presente trabalho estudou a capacidade de degradação bacteriana de cianeto com cepas nativas isoladas de rejeito de mineração de ouro. Para isto, foram escolhidas três cepas dentro de um grupo de vinte cepas isoladas previamente. As cepas foram identificadas mediante as técnicas de MALDI-TOF e sequenciamento do gene 16s. Posteriormente realizou-se o processo de ativação e crescimento bacteriano no qual foram determinados os parâmetros de crescimento para cada uma das cepas, tais como pH, agitação, temperatura e meio de cultura. Após a etapa de ativação bacteriana, realizou-se a adaptação das três cepas em ambientes alcalinos. Nesta etapa, foram feitos ensaios em frascos agitados e avaliou-se o crescimento celular em função da formação de células viáveis para diferentes condições de pH (7; 8; 9,10 e 11). Com as cepas adaptadas ao pH 10 foram realizados ensaios de degradação bacteriana de cianeto em frascos agitados contendo 100mL de solução sintética de cianeto de potássio e 0,2mL de inoculo bacteriano. A concentração da solução de cianeto foi de 500mgL-1 e o pH de 10. Foram avaliadas três condições de temperatura (37, 32 e 27oC). Durante os ensaios foi estudado o crescimento bacteriano, o comportamento do pH e a degradação de cianeto. A quantificação do cianeto livre foi determinada pelo método polarográfico com eletrodo de mercúrio. Através dos resultados obtidos, foi possível identificar que as três cepas isoladas pertencem ás espécies Bacillus pumilus, Bacillus licheniformis e Bacillus subtilis. As cepas bacterianas apresentaram maior produção celular quando cultivadas em meio L.B a pH 7, velocidade de rotação de 190rpm e 37oC de temperatura. A máxima faixa de adaptação a ambientes alcalinos aconteceu em valores de pH 10. As melhores taxas de degradação de cianeto para a cepa B. subtilis e B. pumilus ocorreram em temperatura de 27oC e 65rpm, conseguindo degradar 100% do cianeto. A cepa B. licheniformis apresentou a melhor taxa de degradação de cianeto em temperatura de 32oC e 190rpm obtendo 99,5% de degradação. Através dos resultados obtidos no presente trabalho, foi possível avaliar o potencial de degradação de cianeto para as bactérias B. pumilus, B. licheniformis e B. subtilis as quais podem ser utilizadas como alternativa de tratamento em efluentes contaminados com cianeto. / Cyanide is a highly toxic compound that can be naturally found in the environment or as a result of anthropogenic activities such as gold mining and electroplating industry. Among the different species of the compound, hydrogen cyanide (HCN) is considered the most toxic, even concentrations of 100 ppm are lethal to humans. Degradation of this cyanide compound to less toxic compounds can be carried out through different methods such as chemical, physical and biological treatments. The present work investigated the bacterial degradation capacity of cyanide by isolated native strains from gold mining tailings. Three strains were selected from a group of twenty previously isolated strains which were identified using MALDI-TOF and 16s gene sequencing techniques. The bacterial activation and cellular growth were performed to determine the growth parameters for the strains, such as pH, temperature, rotation speed and culture medium. After the activation, the strains were adapted to grow in alkaline environments. During this phase, cellular growth was carried out in agitated flask at different pHs (7, 8, 9, 10 and 11). The adapted strains were used to perform cyanide degradation tests at pH 10 in agitated flask by using 100mL of a synthetic solution. The solution was composed of 500 ppm of potassium cyanide and 0,2mL of bacterial inoculum. During the experiments, three temperatures were evaluated (37, 32 and 27°C). Bacterial growth, cyanide degradation and pH behaviour were studied Free cyanide quantification was determined by polarographic method with a mercury electrode. It was found that the three isolated strains belong to Bacillus pumilus, Bacillus licheniformis and Bacillus subtilis groups. The highest bacterial growth was observed when the strains were cultivated in a L.B media at pH 7,0, 37°C by using 190 rpm of rotation velocity. The maximum range of adaptation to alkaline environments occurred in pH values of 10. The best cyanide degradation rate (100%) were achieved at 27°C for the strain B. suptilis and B. pumilus. The strain. B. licheniformis showed the best cyanide degradation rate (99,5%) at 37°C. The results obtained in the present work were able to evaluate the potential of cyanide degradation for the bacteria B. pumilus, B. licheniformis and B. subtilis. These results can be used as an alternative to treat wastewaters that are polluted with cyanide.

As indústrias químicas são consideradas como o segmento industrial que gera os efluentes mais perigosos ao meio ambiente. Em virtude das concentrações expressivas de poluentes, tanto orgânicos quanto inorgânicos, os efluentes dessas indústrias podem interferir na atividade da biomassa de estações de tratamento de efluentes (E.T.E.), diminuindo sua eficiência e gerando efluentes tratados, porém em desacordo com a legislação pertinente. Para prevenir tais efeitos, o recebimento de efluentes industriais em E.T.E.s pode ser precedido por pré-tratamento, dentre os quais, o biológico, otimizado pela adição de microrganismos com capacidade de degradar poluentes. Foram isolados microrganismos com capacidade de degradar os compostos identificados como responsáveis pela toxicidade do efluente final da E.T.E.: benzeno, clorofórmio, 1,2-dicloroetano, pentaclorofenol, tricloroeteno, tolueno e p-xileno. Foram realizados testes de bioaumentação para pré tratar o efluente industrial, que foram avaliados através de ensaios físico-químicos e de toxicidade aguda para Vibrio fischeri e Daphnia similis. Os resultados obtidos demonstraram que o pré tratamento reduziu a toxicidade do efluente final da estação de tratamento. / Chemical industries are considered the industrial sector that generates the most dangerous effluents to the environment. Due to the high pollutant concentration, either organics or inorganics, the chemical industries effluents may interferer in the biomass activity in wastewater treatment plants (WWTP), what may reduce its efficiency and generate treated effluents in disagreement to the concerned law. In order to prevent such effects, the industrial effluents disposal in WWTPs can be preceded by biological pretreatment, which can be optimized by adding microorganisms capable of pollutants degradation. Microorganism strains that are able to degrade the compounds identified as the responsible for toxicity levels in the WWTP final effluent: benzene, chloroform, 1,2-dichloroethane, pentachlorophenol, trichloroethene, tolune and p-xylene were isolated. Bioaugmentation tests aiming the chemical effluent pretreatment were performed and they were evaluated through physical-chemical analysis and acute toxicity tests for Vibrio fischeri and Daphnia similis. The results showed that the industrial effluent pretreatment reduced the toxicity levels in the WWTP final effluent.

This study examined the effects of the presence of fungal growth on the phthalate ester content of plasticized polyvinyl chloride (pPVC) used as the vapor barrier component of metal building insulation and the impact the fungal growth may have on the resulting exposure potential to the workers in the building. Individual pieces of the pPVC material exhibiting fungal growth and individual pieces of the pPVC material exhibiting no fungal growth were collected from the building being investigated. Twenty-five pieces each of the pPVC material exhibiting fungal growth and of the pPVC material exhibiting no fungal growth were weighed, reduced to small pieces, and extracted with dichloromethane. The extracts were analyzed using a gas chromatograph coupled with a mass spectrometer. The phthalate content of four phthalate esters identified in the pPVC: diethyl phthalate (DEP), dibutyl phthalate (DBP), butyl benzyl phthalate (BBP), and di-(2-ethylhexyl) phthalate (DEHP), was quantified and the results extrapolated to concentration based on weight ([um]g/Kg) and area ([um]g/m2). A paired samples t test and independent samples t test was used to identify where significant differences occurred (α = 0.05) in the phthalate content between pieces. Using the mean concentration for each phthalate ester, a worst-case-scenario exposure was modeled. There was a statistically significant difference in the phthalate ester content between the pPVC exhibiting fungal growth and pPVC exhibiting no fungal growth for all four phthalate esters investigated. The mean phthalate ester concentration ranged from 13.74 [um]g/m2-34.94 [um]g/m2 in the pPVC exhibiting no fungal growth and 12.00 [um]g/m2-30.63 [um]g/m2 in the pPVC exhibiting fungal growth. The modeled exposure concentration in the building ranged from 0.39 [um]g/m3-0.98 [um]g/m3 and was generally lower than published exposure concentrations in similar settings.

Ce travail concerne l'étude de la dégradation enzymatique de micropolluants pharmaceutiques récalcitrants présents dans l'eau. Tout d’abord, les efficacités de trois laccases différentes issues respectivement de : Pycnoporus sanguineus CS43, Trametes versicolor (Tv) et Myceliophtora thermophila ont été comparés lors d’essais de dépollution de solutions modèles renfermant trois antibiotiques (amoxicilline, ciprofloxacine et sulfaméthoxazole) et un antiépileptique (carbamazépine). Les essais ont été réalisés avec les laccases libres en présence ou non de médiateurs redox. L'impact de plusieurs paramètres opératoires sur les performances des enzymes a également été étudié. Puis, une nouvelle méthode d’immobilisation des laccases impliquant l’activation du support (microparticules à base de silice commerciales) par du glutaraldéhyde en phase vapeur a été mise au point et optimisée en utilisant la méthodologie de plans d’expériences. Après immobilisation, la laccase Tv s’est avérée être la plus active. Des essais de dégradation en présence de médiateurs redox ont confirmé l’efficacité de l’enzyme immobilisée et sa possible réutilisation lors de cycles successifs. La toxicité des solutions après traitement a été évaluée par des tests Microtox®. La laccase Tv a également été immobilisée sur des nanoparticules non commerciales à base de silice ou d’argile ainsi que sur des composites à base de silice et d’argile. La laccase Tv immobilisée sur les supports composites riches en silice a montré une plus grande réactivité et de meilleures performances pour l'élimination des composés cibles. / This work is focused on the study of the enzymatic depletion of recalcitrant pharmaceutical micropollutants in water. The potential degradation of three antibiotics (amoxicillin, ciprofloxacin and sulfamethoxazole) and one anti-epileptic (carbamazepine) was studied with three laccases: Pycnoporus sanguineus CS43, Trametes versicolor (Tv) and Myceliophtora thermophila. Free laccase systems were evaluated for pharmaceuticals depletion on model solutions in the presence or absence of redox mediators and the impact of several parameters on the performance of laccases for degradation were studied. The enzymes were then immobilized on different solid supports: commercial silica, laboratory synthetized nano-silica and clay based composite nanomaterials and used for degradation tests. A novel methodology for the covalent binding of laccases onto carriers was developed by using glutaraldehyde in vapour phase and the best immobilization conditions were determined through a 23 full factorial design. The immobilized Tv shown the highest activity and was tested in presence of redox mediators. Moreover, the reusability was evaluated in several degradation cycles and the toxicity of the solutions after treatment was assessed with the Microtox® test. In comparison to laccase immobilized on commercial silica, the Tv supported on laboratory synthetized materials showed higher activity and a better performance for the removal of target compounds.

Le delta du Rhône est une vaste plaine alluviale recouverte d’étangs et de lagunes, parsemée de cultures diverses, principalement du riz (21’100 ha en 2009, Centre Français du Riz, 2010). Des herbicides répandus en avril (pré levée, prétilachlor) et en juin (poste levée, bentazone ; MCPA et dichlorprop) sont utilisés dans la riziculture pour lutter contre les mauvaises herbes. L’étang de Vaccarès, qui est classé «réserve de biosphère» par l’UNESCO pour sa grande richesse biologique reçoit des pesticides directement issus des rizières par les canaux de drainage. D’un point de vue scientifique, peu d’études sont consacrées au devenir des pesticides dans les eaux estuariennes. En outre, la plupart de ces études ont été focalisées sur la photochimie bien que la biodégradation soit impliqué dans l’atténuation des pesticides. Alors le but de ce travail de thèse est de comprendre le devenir des herbicides de riziculture en évaluant le rôle de la photochimie direct et indirect et de la biodégradation. Nous avons développé donc un modèle capable de décrire la transformation de molécules organique par la voie photochimique dans les eaux estuariennes. Enfin nous avons déterminé les processus principaux impliqués dans l'atténuation rapide des herbicides de riziculture en Camargue. / The Ile de Camargue basin is the central part of the Rhône Delta in the south of France included between the two branches of the river. The higher parts of this area are agricultural lands, mainly rice fields, whereas the lower parts are lagoons and natural wetlands of international importance for biodiversity preservation.Bentazone, dichlorprop, and MCPA are post-emerging herbicides applied in rice fields from mid April to mid June. Rice fields grown under flooded conditions represent an environment where these conditions increase the potential for pesticides to transport in runoff from rice fields to non- target surface waters.To predict their fate in the surface waters of the Camargue and to asses their risk, it is necessary to understand their chemical reaction under environmental conditions.Reactive photoinduced species (RPS, hydroxyl radical HO., singlet oxygen 1O2 and dissolved organic matter triplet state) in estuarine waters (Vaccarès lagoon) were characterized experimentally to estimate the role of photochemistry in rice paddy. We then developed a kinetic model describing the solar photo-transformation of xenobiotic organic compounds induced by the three different photooxidants RPS. Sunlight photodegradation of bentazone, dichlorprop (DCPP), MCPA and alachlor (used as a surrogate for the similar Pretilachlor, pre-emergence herbicide) and microbial transformation were conducted experimentally to estimate near-surface half-lives in surface water. Parallel to the experimental study, an assessment of the elimination rates of those herbicides by sorption and sedimentation, and volatilization were obtained by the model MASAS (Modelling of Anthropogenic Substances in Aquatic Systems).The half-lives of the herbicides obtained experimentally were then compared with those obtained from field data. This allowed to assessing the main processes involved in the attenuation of herbicide concentrations.

Xanthates, key compounds used in the mining industry for the separation of metals from ores, may have a significant impact on the ecosystems once discarded into the tailings. The aims of this study were to : characterize chemical and microbiological conditions in a tailings lagoon (Brown Creek gold mine); examine the feasability of using indigenous bacteria to degrade xanthate under laboratory conditions; and, optimize the biodegradation process by employing biofilm technology and plant-microbe associations. To achieve these, a simple but accurate HPLC method was developed. The chemical and biological constitution of the tailings lagoon was monitored over a 2 year period from March 1995 to February 1997. Xanthate degradative performance was improved by Phragmites australis and its root associated bacterial community. Cumulative amounts of xanthate degraded by this plant-microbe association were significantly higher than any other treatment, specifically glass-attached and free-living bacterial populations. An increased bacterial number in the extensively developed root, the presence of extracellular layer as revealed from scanning electron microscope and significance of plant removal via uptake were thought to be the possible explanations for the enhanced biodegradation activity observed. In view of the above, use of glass-attached bacteria and plant-microbe associations might provide a feasible solution to degrade xanthate in the tailings lagoon. The results obtained from this study suggest that the plant-microbe associations could provide a cost effective method to degrade/remove xanthate from the tailings lagoon runoff. This approach offers several ecological advantages. Firstly, it offers the advantage of a solar-powered treatment system that is aesthetically pleasing. Secondly, bioremediation can be done on site, thereby eliminating waste transportation costs. Finally, it has the added attraction of a higher potential for public acceptance / Doctor of Philosophy (PhD)

“In the decomposition of organic/botanic materiality, decayed and decaying exotic weeds are printed and imprinted on the host vessel: The surviving trace becomes a code - a sign - a semiotic map = disjecta membra: being there ... then destroyed... but still remaining.” THE BODY OF VISUAL AND WRITTEN RESEARCH, 'DECOMPOSE', is a cross-disciplinary interrogation, interpreting overlapping meanings in the Botanical Biodegradation of Weeds through Visual Art/Science practices and processes expressed as Text +Paintings. DECOMPOSE validates the Act of Art, Botanical Biodegradation of Weeds, as both: I. An expression of the Subjective, Expansive and Ephemeral nature of Art, Artist and Materials and II. An incarnation of the nature of Time and Sublime Beauty, that articulates, and expands perceptions of Art, Artist and Materials as Text + Paintings. The 'equation': DECAY + WEEDS = BEAUTY expands to encompass key elements in the DECOMPOSE body of research: BOTANICAL BIODEGRADATION + AUSTRALIAN EXOTIC, FERAL and NOXIOUS WEED SPECIES + ARTIST + MATERIALS + ART + SCIENCE + TIME = DECAY-PAINTINGS = RESEARCH = SUBLIME BEAUTY Argued by quantitative and qualitative example, DECOMPOSE is at once: I. Subjective: a conceptual and translative process expressed through the personal vision of the artist. II. Expansive: an interrogation of a single process, at once finite and infinite in meanings and extractions. III. Ephemeral: investigations and results signifying the specific and universal decay of all things. / Doctor of Philosophy (PhD)

[Typescript] Includes bibliography.

Bibliography: leaves 177-191.