abstract: Reductive dechlorination by members of the bacterial genus Dehalococcoides is a common and cost-effective avenue for in situ bioremediation of sites contaminated with the chlorinated solvents, trichloroethene (TCE) and perchloroethene (PCE). The overarching goal of my research was to address some of the challenges associated with bioremediation timeframes by improving the rates of reductive dechlorination and the growth of Dehalococcoides in mixed communities. Biostimulation of contaminated sites or microcosms with electron donor fails to consistently promote dechlorination of PCE/TCE beyond cis-dichloroethene (cis-DCE), even when the presence of Dehalococcoides is confirmed. Supported by data from microcosm experiments, I showed that the stalling at cis-DCE is due a H2 competition in which components of the soil or sediment serve as electron acceptors for competing microorganisms. However, once competition was minimized by providing selective enrichment techniques, I illustrated how to obtain both fast rates and high-density Dehalococcoides using three distinct enrichment cultures. Having achieved a heightened awareness of the fierce competition for electron donor, I then identified bicarbonate (HCO3-) as a potential H2 sink for reductive dechlorination. HCO3- is the natural buffer in groundwater but also the electron acceptor for hydrogenotrophic methanogens and homoacetogens, two microbial groups commonly encountered with Dehalococcoides. By testing a range of concentrations in batch experiments, I showed that methanogens are favored at low HCO3 and homoacetogens at high HCO3-. The high HCO3- concentrations increased the H2 demand which negatively affected the rates and extent of dechlorination. By applying the gained knowledge on microbial community management, I ran the first successful continuous stirred-tank reactor (CSTR) at a 3-d hydraulic retention time for cultivation of dechlorinating cultures. I demonstrated that using carefully selected conditions in a CSTR, cultivation of Dehalococcoides at short retention times is feasible, resulting in robust cultures capable of fast dechlorination. Lastly, I provide a systematic insight into the effect of high ammonia on communities involved in dechlorination of chloroethenes. This work documents the potential use of landfill leachate as a substrate for dechlorination and an increased tolerance of Dehalococcoides to high ammonia concentrations (2 g L-1 NH4+-N) without loss of the ability to dechlorinate TCE to ethene. / Dissertation/Thesis / Ph.D. Microbiology 2013

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / A produÃÃo de camarÃo marinho à dominada pelo camarÃo branco Litopenaeus vannamei, sendo esta a espÃcie mais bem sucedida internacionalmente. A aquicultura à na maioria das vezes, vulnerÃvel a mudanÃas nas condiÃÃes ambientais que, podem gerar impactos negativos na produÃÃo. A maioria dos empreendimentos aquÃcolas realizam descarte inadequado de seus efluentes sem nenhuma aÃÃo para seu tratamento causando impacto ao meio ambiente. O presente trabalho teve como objetivo avaliar o desempenho zootÃcnico de pÃs-larvas de Litopenaeus vannamei cultivadas em Ãgua oriunda de um efluente tratado pela cianobactÃria Spirulina platensis na presenÃa e ausÃncia de probiÃtico. O experimento foi conduzido em uma carcinicultura localizada em Beberibe â CE, sendo este desenvolvido em duas fases, uma em Ãrea coberta e outra no ambiente externo com duraÃÃo total de 50 dias. O experimento constou de trÃs tratamentos, com cinco repetiÃÃes cada. No tratamento 1 a Ãgua utilizada seguiu o manejo de preparaÃÃo rotineiro dos prÃ-berÃÃrios com a adiÃÃo de bactÃrias do gÃnero Bacillus. O tratamento 2 contou com a Ãgua do efluente dos prÃ-berÃÃrios, tratada com S. platensis. O tratamento 3 contou com o efluente tratado com a adiÃÃo de bactÃrias. Foram alcanÃados percentuais mÃximos de remoÃÃo de acima de 99% para NAT e PO4- e NO3- no cultivo de S. platensis em efluente. Nos tratamentos testados foram detectadas diferenÃas a 5% de significÃncia estatÃstica de T2 e T3 em relaÃÃo a T1, em que no efluente tratado a concentraÃÃo de NAT e NH3 foi superior. A maior sobrevivÃncia foi encontrada no T2 (93%) seguida de 89% nos demais tratamentos. O maior peso mÃdio foi encontrado no T1 com 45 mg seguidos de T2 (42,6) e T3 (41,6), no entanto nÃo foram detectadas diferenÃas significativas em nenhum dos os parÃmetros zootÃcnicos testados. Diante do exposto conclui-se que o cultivo de pÃs-larvas de camarÃo marinho em Ãgua de efluente tratada por S. platensis, na presenÃa ou ausÃncia de probiÃtico, à uma alternativa viÃvel para a produÃÃo de camarÃes marinhos. / Marine shrimp production is dominated by the pacific white shrimp Litopenaeus vannamei, which is the species most successful internationally. Aquaculture is, most often, vulnerable to changes in environmental conditions, that may have negative impacts on production. Most aquaculture ventures perform improper disposal of their effluents, without any treatment, causing harm to the environment. This study aim to evaluate the growth performance of post-larvae of Litopenaeus vannamei cultured in water originating from a effluent treated by the cyanobacterium Spirulina platensis in the presence or absence of probiotics. The experiment was conducted in a shrimp farm located in Beberibe - CE, which was developed in two phases: one in covered area and another in the external environment within a period of 50 days. The experiment consisted of three treatments with five replicates each. In treatment 1, the water used followed the routine preparation management of pre-nurseries with the addition of bacteria of the genus Bacillus. The treatment 2 had the water of the effluent of the shrimp nursery tank, treated with S. platensis. The treatment 3 had the effluent treated with the addition of bacteria. Maximum percentage of removals above 99% were achieved for TAN and PO4- and NO3- in cultivation of S. platensis in effluent. In the treatments tested were detected differences of 5% statistical significance of T2 and T3 compared to T1, with higher concentration of NAT and NH3 in the treated effluent. The higher survival was found in T2 (93%) followed by 89% in the other treatments. The highest average weight was found in T1 with 45 mg followed by T2 (42.6) and T3 (41.6), however no significant differences were detected in any of the zootechnical parameters tested. In view of what has been exposed here, it follows that the cultivation of marine shrimp post-larvae in effluent water treated with S. platensis, in the presence or absence of probiotics, is a viable alternative for the marine shrimp production.

Orientadores: Elias Basile Tambourgi, Galba Maria de Campos Takaki / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-09T16:12:47Z (GMT). No. of bitstreams: 1 Silva_ThayseAlvesdeLimae_M.pdf: 1548702 bytes, checksum: 41aadb70684cd66a5ae82bae01bd72ec (MD5) Previous issue date: 2007 / Resumo: O óleo bruto e seus destilados contêm quantidades significantes de compostos organossulfurados de baixo peso molecular, tais como tióis cicloalquilas e compostos aromáticos heterocíclicos baseados em tionefóis. Este último grupo de hidrocarbonetos aromáticos policíclicos sulfurados inclui benzotiofeno, dibenzotiofeno e seus derivados. Estes compostos têm sido objetos de interesse de estudos há décadas, pelo fato de constituírem a maior classe de contaminantes ambientais encontrados na água e no solo. Com o objetivo de consertar as conseqüências desta contaminação, tais como chuva ácida e poluição do ar, causadas pelo dióxido de enxofre liberado da combustão dos óleos, mais regulamentações na quantidade de enxofre no petróleo estão sendo estabelecidas. A biodessulfurização tem atraído a atenção dos pesquisadores pela sua aplicação na dessulfurização do petróleo, devido às condições do meio, como baixo consumo de energia e baixa emissão de dióxido de enxofre. Devido a esses fatores, a biodessulfurização, que opera em condições ambiente de temperatura e pressão, possui um grande potencial como uma técnica bastante promissora em alternativa à hidrodessulfurização. Neste trabalho foi estudada a cinética de crescimento de Pseudomonas fluorescens UCP 1514, que é uma linhagem selvagem do solo, em culturas em batelada contendo dibenzotiofeno, e seu potencial na biodessulfurização, para utilização na redução de enxofre contido nos combustíveis fósseis, de acordo com as regulamentações ambientais. O microrganismo foi capaz de metabolizar o composto e foram também identificadas através da amplificação do DNA, colônias mutantes fisiológicas, que cresceram na presença do dibenzotiofeno. Os resultados mostraram um decréscimo de 73% na concentração do composto, indicando que Pseudomonas fluorescens, seletivamente, remove enxofre do dibenzotiofeno para formar bifenil, indicando que este microrganismo oferece grande potencial na biodegradação de compostos sulfurados, podendo ser utilizado como catalisador biológico na desssulfurização e mineralização desses compostos, consequentemente, minimizando os danos ambientais causados pela poluição dessas substâncias / Abstract: Crude oil and its distillates contain significant amounts of low-molecular-mass organosulphur compounds such as alkyland cycloalkyl thiols, alkyl- and arylthioethers and aromatic heterocycles based on thiophene. This last group of polycyclic aromatic sulfurated hydrocarbons includes thiophene itself, benzothiophene, dibenzothiophene, and their alkylated derivatives. These compounds have been of concern for decades because they constitute a major class of ubiquitous environmental contaminants found in both air and sea areas. In order to mitigate the consequences of this contamination, such as acid rain and air pollution caused by sulphur dioxide released from the combustion of oils, more and more regulations on sulphur content in petroleum are being established. Biodesulfurization has attracted attention owing to its application to the desulfurization of petroleum due to its mild conditions, lower energy consumption and lower emission of sulphur dioxide. Thus biodesulfurization, which operates under room temperature and pressure conditions, is expected to be a complementary as well as promising alternative to hydrodesulfurization. In this work, we studied the growth kinetics of a bacterium of the strain Pseudomonas fluorescens UCP 1514, which is indigenous in soil, in batch cultures containing dibenzothiophene, a recalcitrant component of fossil fuels and its potential on the desulfurization, in order to use it for reducing the sulfur content of diesel oil in compliance with environmental regulations. The microorganism was capable to metabolize dibenzothiophene and we also identified the physiological mutant colonies that growing on dibenzothiopnhene by DNA amplification with polymerase chain reaction technique and the specific primer BOX.The results showed a decrease of 73% in dibenzothiophene content, indicating that Pseudomonas fluorescens selectively removes sulfur from dibenzothiophene to form biphenyl, indicating that this bacterium shows promising potential for decreasing the sulfur content of fossil fuels, can be used as biological catalyst, on desulfurization and mineralization these compounds, minimizing the problems caused by pollution of this substances / Mestrado / Sistemas de Processos Quimicos e Informatica / Mestre em Engenharia Química

Orientadores: Elias Basile Tambourgi, Ana Lucia Figueiredo Porto / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-16T02:11:19Z (GMT). No. of bitstreams: 1 Campos_EdgarSilveira_D.pdf: 8809758 bytes, checksum: 0005b20be087b28b91ebffb9ef5f140c (MD5) Previous issue date: 2010 / Resumo: Aproximadamente, 10.000 corantes e pigmentos diferentes são utilizados industrialmente, representando um consumo anual por volta de 7 x 105 toneladas mundialmente, com 26.500 só no Brasil. Em sua maioria, corantes são substâncias recalcitrantes e tóxicas, os quais são resistentes a degradação biológica. Na região metropolitana de São Paulo, três quatros dos efluentes industriais têxteis possuem resposta mutagênica positiva, o que torna a contaminação por corantes não só um problema ambiental, mas também um problema de saúde pública. Este trabalho tem como objetivo a seleção e avaliação de Pseudomonas para a descoloração de efluentes têxteis. Quatros espécies de Pseudomonas foram testadas, P. oleovorans, P. cepacia, P. putida e P. aeruginosa, com 14 corantes industriais, onde a P. oleovorans mostrou-se mais promissora, pois foi capaz de metabolizar 12 dos corantes testados. Posteriormente, a descoloração de corantes por P. oleovorans foi otimizada através de dois planejamentos experimentais consecutivos, onde foi possível reduzir o tempo de descoloração foi reduzido pela metade. As melhores condições para a P. oleovorans foram atingidas em condições de anoxia (zero RPM); pH inicial de 8,5; 32°C; uso de 25% do volume total do reator; tamanho da cultura inicial de 0,16 g l-1; e uma concentração de corantes de 50 mg l-1. Após a otimização, foi realizada descoloração de corante em cultura Fed-Batch, o que aumentou a massa de corante degradado em 10 vezes. Por conseguinte, a P. oleovorans foi imobilizada em espumas de poliuretano (PUF) e estas empacotadas em um reator anóxico para a descoloração de efluente sintético. A P. oleovorans foi capaz de descolorir efluente sintético contendo corantes em reator anóxico empacotado com PUF em apenas 24 horas, o que aumentou a produtividade em mais de seis vezes quando comparado a fermentação em Fed-Batch / Abstract: Approximately, different 10,000 dyes and pigments are for industrial use, representing an annual consumption of around 7 x 105 worldwide, 26,500 only in Brazil. In overall, dyes are toxic and recalcitrant substances, which are resistant to biological degradation. At the Sao Paulo's metropolitan region, three out of four textile industrial effluents have a positive mutagenic response, which makes dye contamination not only an environmental problem, but also a public health one. This work aims the selection and evaluation of Pseudomonas for textile effluent decolourization. Four Pseudomonas species were tested, P. oleovorans, P. cepacia, P. putida e P. aeruginosa, against 14 industrial dyes, where P. oleovorans showed to be the most promising one, it was capable to metabolize 12 dyes tested. Afterwards, P. oleovorans dye decolourization was optimized through two consecutive experimental designs, in which was possible to reduce the decolourization in half. The best conditions to P. oleovorans were achieved in anoxic conditions (0 RPM); initial pH of 8.5; 32°C; usage of 25% of total reactor; culture initial size of 0.16 g l-1; and dye concentration of 50 mg l-1. After the optimization, dye decolourization was performed in Fed-Batch, which increased the degraded dye mass 10-fold. At last, P. oleovorans was immobilized in Poly Urethane Foam (PUF), and packed in an anoxic reactor to the decolourization of a synthetic effluent. P. oleovorans was capable of decolourize the dye containing synthetic effluent in an anoxic reactor packed with PUF in only 24 hours, which increased the productivity in over than 6-fold when compared with Fed-Batch decolourization / Doutorado / Sistemas de Processos Quimicos e Informatica / Doutor em Engenharia Química

Submitted by Alisson Mota (alisson.davidbeckam@gmail.com) on 2015-07-06T19:29:41Z No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-06T19:38:38Z (GMT) No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-06T19:42:38Z (GMT) No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) / Made available in DSpace on 2015-07-06T19:42:38Z (GMT). No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) Previous issue date: 2014-04-11 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / The ever increasing consumption of fossil fuels puts oil as one of the most important products of human consumption. Undeniable technological and even social breakthroughs are coming era of "post-oil" . But this unbridled consumption has led to serious environmental aquatic ecosystems and land damage. Viable alternatives that promote the recovery of areas affected by these compounds are undoubtedly of utmost importance given the current scenario. Bioremediation has proven very promising in this area . And increasingly, new technology are aggregated to this field of research in an attempt to obtain more conclusive results, thus improving the overall understanding of how microorganisms interact in promoting bioremediation. This is precisely what has been proposed in this work. The use of Mass Spectrometry (MS) for identification of proteins of bacteria capable of growth in xenobiotics. Initially was selected near the Refinaria Isaac Sabbá (REMAN) species B. pumilus, with the potential ability to grow in petroleum and diesel. After conducting tests using oil and diesel separately with unique carbon sources, was proven capacity for growth of the species in these xenobiotics. Better growth/biomass production was recorded in crops having diesel as finte carbon, as well as the best protein yield. The proteomic profile in petroleum, diesel and yeast extract was determined. Comparisons between these profiles showed very marked difference between them. For analysis via mass spectrometry were selected 96 spots of B. pumilus were selected and identified by Peptide Mass Fingerprint (PMF) . Of this total 54 protein spots were identified. Some with potential involvement in direct bioremediation processes by B. pumilus. However, further studies are necessary to effective proof of proteins in the pathways of degradation of xenobiotics. / O consumo cada vez crescente dos combustíveis fósseis coloca o petróleo como um dos mais importantes produtos do consumo humano. Inegáveis avanços tecnológicos e até mesmo sociais são advindos da era “Pós-Petróleo”. Porém, esse consumo desenfreado tem gerado graves danos ambientais aos ecossistemas aquáticos e também terrestres. Alternativas viáveis capazes de promover a recuperação de áreas afetadas por esses compostos são sem dúvida de extrema importância diante do cenário atual. A biorremediação tem se mostrado bastante promissora nessa área. E cada vez mais novas tecnologia são agregadas a esse campo de investigação na tentativa de obter resultados mais conclusivos, melhorado assim, o entendimento global de como os microorganismos interagem na promoção da biorremediação. É justamente isso o que foi proposto nesse trabalho. O emprego da espectrometria (MS) de massa para identificação de proteínas de bactéria com capacidade de crescimento em xenobióticos. Inicialmente foi selecionada nas proximidades da Refinaria Isaac Sabbá (REMAN) a espécie B. pumilus, com potencial capacidade de crescer em petróleo e diesel. Após a realização de testes usando petróleo e diesel, separadamente, com fontes únicas de carbono, foi comprovada a capacidade de crescimento da espécie nesses xenobióticos. Melhor crescimento/produção de biomassa foi registrado nos cultivos tendo o diesel como finte de carbono, bem como o melhor rendimento protéico. O perfil proteômico em petróleo, diesel e extrato de levedura foi determinado. As comparações entre tais perfis demonstrou diferenças bastante acentuadas entre eles. Para análise via espectrometria massas foram selecionados 96 spots de B. pumilus e identificados por Peptide Mass Fingerprint (PMF). Desse total de spots, 54 proteínas foram identificadas. Algumas delas com potencial envolvimento nos processos diretos de biorremediação apresentada por B. pumilus. Porém, mais estudos são necessário para efetiva comprovação dessas proteínas nas vias de degradação de xenobióticos.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Os potenciais impactos ambientais provenientes da aquicultura intensiva tÃm aumentado a preocupaÃÃo da sociedade quanto à sustentabilidade da prÃpria atividade. Com o presente trabalho teve como objetivo avaliar, os parÃmetros zootÃcincos das tilÃpias do Nilo, qualidade de Ãgua, produÃÃo de biomassa algal e taxa de remoÃÃo dos compostos nitrogenados pela microalga Spirulina platensis em um cultivo integrado da tilÃpia do Nilo (Oreocrhomis niloticus) com a microalga S. platensis em um sistema fechado de circulaÃÃo de Ãgua. O experimento foi realizado no Centro de Biotecnologia Aplicada a Aquicultura (CEBIAQUA) do Departamento de Engenharia de Pesca da Universidade Federal do CearÃ. O cultivo das tilÃpias foi inteiramente casualizado e realizado em trÃs tratamentos com quatro repetiÃÃes nas densidades de 0,1; 0,15 e 0,20 juvenis L-1. As tilÃpias foram alimentadas ad libitum com raÃÃo comercial (0.8 mm), contendo 55% de proteÃna bruta (PB), fornecida Ãs 9, 13 e 17 h, sete dias por semana. Para o cultivo de Spirulina platensis, inicialmente, o inÃculo foi cultivado em meio quÃmico alternativo e posteriormente adaptado ao efluente de tilÃpia do Nilo e inoculado em 12 bacias de polietileno. Durante todo o experimento, a Ãgua das caixas dos peixes e das respectivas bacias das microalgas foi reutilizada no sistema de recirculaÃÃo fechado entre as duas culturas, havendo apenas reposiÃÃo de Ãgua por conta da evaporaÃÃo. ApÃs o completo desenvolvimento da S. platensis nas bacias, a coleta da microoalga foi realizada atravÃs de filtragem da Ãgua em telas de 60 Âm. Foram ainda monitorados o pH, temperatura, oxigÃnio dissolvido, bem como determinados os nÃveis de N-amÃnia, N-nitrato e N-nitrito durante o perÃodo experimental. Os resultados mostraram que a densidade de estocagem mais eficiente para o cultivo das tilÃpias foi 0,10 juvenis L-1, pois apesar de ter resultado na mesma biomassa final dos demais tratamentos, foi a que apresentou a menor taxa de conversÃo alimentar. A microalga S. platensis se adaptou facilmente e se desenvolveu muito bem no efluente das tilÃpias do Nilo, independente da densidade de estocagem dos peixes. Com relaÃÃo à qualidade de Ãgua, foi evidenciado que a integraÃÃo das culturas manteve os nÃveis de N-amÃnia (0,26  0,03 mg L-1), N-nitrato (2,55  0,71) e N-nitrito (0,19  0,01) dentro dos valores aceitÃveis para o cultivo da tilÃpia do Nilo. / The environmental potential impacts from intensive aquaculture have increased public concern about the sustainability of the activity itself. The present work aimed to evaluate the zootÃcincos parameters of Nile tilapia , water quality, algal biomass production and rate of nitrogen removal compounds by Spirulina platensis realize an integrated cultivation of Nile tilapia (Oreocrhomis niloticus) with Spirulina platensis in a closed system of water circulation. The experiment took place at the Biotechnology Aquaculture Applied Center (CEBIAQUA) of The Fishing Engineering Department of the Federal University of CearÃ. Tilapias culture was conducted in three treatments with four replicates at densities of 010, 0,15 and 0,20 juveniles L-1. The tilapias were fed ad libitum with a commercial ration (0,8 mm) containing 55% crude protein (CP), administered daily at 9, 13 and 17 h. For Spirulina platensis culture, initially, the inoculum was cultivated in an alternative chemical medium and later adapted to the Nile tilapia effluent and inoculated into 12 polyethylene basins. Throughout the experiment, the water of fishes boxes and of the respective microalgae basins was reused in the closed recirculation system between the two cultures, with only replacement of water due to evaporation. After the full development of S. platensis in the basins, the microalgae was collected using a 60 Âm screen. The pH, temperature, dissolved oxygen, and levels of N-ammonia, N-nitrate and N-nitrite were also monitored during all the experimental period. The results showed that the most efficient stocking density was 0,10 L-1, because despite having the same final biomass of the other treatments it presented the lowest feed conversion ratio. The microalgae S. platensis was easily adapted and developed very well in the Nile tilapiaâs effluent, regardless of fishes stocking density. With respect to water quality, it was evident that the cultures integration maintained the levels of N-ammonia (0,26  0,03 mg L-1), N-nitrates (2,55  0,71 mg L-1) and N- nitrites (0,19  0,01 mg L-1) within acceptable values for the Nile tilapia culture.

Submitted by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-12-05T15:45:54Z No. of bitstreams: 1 Tese - João M. S. Lima.pdf: 3551790 bytes, checksum: 602e786b4a54ff5164341a8a64252e5b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-12-05T15:46:11Z (GMT) No. of bitstreams: 1 Tese - João M. S. Lima.pdf: 3551790 bytes, checksum: 602e786b4a54ff5164341a8a64252e5b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-12-05T15:46:42Z (GMT) No. of bitstreams: 1 Tese - João M. S. Lima.pdf: 3551790 bytes, checksum: 602e786b4a54ff5164341a8a64252e5b (MD5) / Made available in DSpace on 2016-12-05T15:46:42Z (GMT). No. of bitstreams: 1 Tese - João M. S. Lima.pdf: 3551790 bytes, checksum: 602e786b4a54ff5164341a8a64252e5b (MD5) Previous issue date: 2016-06-30 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Biosurfactants are compounds produced by fungi and bacteriain different behavioral and environmental conditions, and can be used as an option for the bioremediation of environments contaminated by oil and its derivatives. The selection of fungi, endophytic and epiphytic bacteria, with potential biosurfactant activity, was evaluated through the following tests: Biodegradability using discoloration of 2,6 Dichloro Phenol Indophenol (DCPIP); oil drop-collapse; and emulsification, through surface tension evaluation with Du Nouy ring and the resulting production curve of the most promising microorganisms. Eight fungal samples were effective in the biodegradability test, since they promoted discoloration of indophenol in 24 and 48 hours: S31 Phoma sp, S36 Phoma sp, S24 Rhizopus oryzae, S46 Fusarium sp., S42 Fusarium sp., S32 Fusarium sp, S33 Fusarium sp. and S51 Fusarium sp. All of these fungal isolates produced laccase, pectinase, amylase and lipase. Of these, the S31 Phoma sp. isolate presented a diesel emulsion of 1.5 cm, or 52%, to reduce water surface tension from 72.43 mN/m to 51.03 mN/m. Among the bacteria selected, the M87 Microbacterium sp. isolate presented the best correlation between the tests. This isolate produced 3.0 g/L of biosurfactants, and the fractions promoted reduction of water surface tension below 40 mN/m. The best fraction (F1) was analyzed through FT-IR infrared UFLC-MS and 1H NMR which demonstrated the presence of compounds probably related to fatty acids, a type of non-ionic biosurfactant, that could be used in the cosmetic industry or as an emulsifier in bioremediation processes. No toxic activity was detected either in the bacterial lyophilized aqueous extract or the fungal aqueous extract. Further studies are required in order to: Promote the growth of these, pure or consortium, cultures; identify the best carbon source for the production of biosurfactants; and perform a more thorough chemical characterization of the different biosurfactants and bioemulsifiers from the fungi and bacteria selected. / Os biossurfactantes são compostos produzidos por fungos e bactérias em diferentes condições ambientais e comportamentais e podem ser usados como alternativa para a biorremediação de ambientes contaminados por petróleo e seus derivados. A seleção de fungos e bactérias endofíticas e epifíticas, com provável atividade biossurfactante, foi realizada por meio dos seguintes testes: biodegradabilidade, utilizando descoloração do 2,6 Dicloro Fenol Indofenol (DCPIP); colapso da gota de petróleo; emulsificação, avaliação da tensão superficial pelo método do anel de Du Noüy, e curva de produção dos micro-organismos mais promissores. Amostras fúngicas em um total de oito mostraram-se eficientes no teste de biodegradabilidade, pois promoveram a descoloração do indofenol em 24 e 48h, estas foram identificadas como: S31 Phoma sp., S36 Phoma sp., S24 Rhizopus oryzae, S46 Fusarium sp., S42 Fusarium sp., S32 Fusarium sp., S33 Fusarium sp. e S51 Fusarium sp. Destes, o isolado S31 Phoma sp. apresentou emulsão do diesel de 1,5 cm ou 52% com redução da tensão superficial da água de 72,43 mN/m para 51,03 mN/m, além disso, os oito fungos selecionados apresentaram atividade na produção de enzimas: lacase, pectinase, amilase e lipase. Dentre as bactérias, o isolado M87 Microbacterium sp., mostrou melhor correlação entre os testes realizados. Esse isolado produziu 3,0 g/L de biossurfactantes e suas frações promoveram a redução da tensão superficial da água abaixo de 40 mN/m. A melhor fração (F1) foi analisada por FT-IR infravermelho, UFLC-MS e RMN 1H. As diferentes técnicas mostraram a presença de compostos provavelmente relacionados a ácidos graxos, um tipo de biossurfactante não iônico que pode ser explorado futuramente na indústria de cosméticos ou como emulsificante em processos de biorremediação. O teste para avaliar a citotoxicidade do extrato liofilizado e aquoso da bactéria, e o de toxicidade do extrato aquoso do fungo, não mostraram atividade tóxica. Novos estudos fazem-se necessários no sentido de promover o crescimento dessas culturas, puras ou em consórcio; utilizando diferentes fontes de carbono com o intuito de identificar a melhor fonte para a produção de biossurfactantes, assim como aprofundar os estudos de caracterização química dos diferentes biossurfactantes e bioemulsificantes produzidos, tanto pelo fungo como pela bactéria selecionada.

Plastificantes podem ser definidos como aditivos de baixa volatilidade utilizados para aumentar a processabilidade, flexibilidade ou diminuir a dureza de materiais poliméricos. Os ftalatos e adipatos utilizados como plastificantes, por sua baixa solubilidade em água e pelo alto coeficiente de partição octanol/água, tendem a se acumular no solo e sedimentos. Estes compostos são considerados potencialmente carcinogênicos, teratogênicos e disruptores endócrinos. A presente pesquisa compreendeu a biorremediação ex-situ do solo contaminado com resíduos de uma unidade industrial de plastificantes, utilizando reatores aeróbios, com microrganismos indígenas e exógenos adaptados através da adição de inóculo retirado da Estação de Tratamento de Efluentes por Lodos Ativados desta indústria. Foram avaliados os plastificantes: DIBP (Di-isobutilftalato), DBP (Dibutilftalato), DOP (Dioctilftalato),DIDP (Di-isodecilftalato), DIAP (Di-isoamilftalato) e DOA (dioctiladipato). Após a caracterização geotécnica do solo da área de plastificantes em 10 diferentes pontos, foram retiradas as quantidades para a biorremediação em 8 diferentes pontos (100kg/ponto) com os teores totais de plastificantes compreendidos entre 17 mg/kg solo a 6222 mg/kg solo. Análises mineralógicas, físicas e químicas foram realizadas posteriormente e indicaram que possivelmente a capacidade de troca catiônica do solo era devida aos plastificantes. Na biorremediação, os teores iniciais de plastificantes no solo, variaram de 85 mg/kg a 1688 mg/kg e após 120 dias de biodegradação em reatores aeróbios, as eficiências de remoção foram de 75 a 97%. Conforme as análises de fingerprint da comunidade bacteriana, ao final do processo, as bactérias presentes no solo eram originárias do lodo e do solo inicial e as análises de CGMS identificaram os metabólitos MEHP e os sub-produtos finais da biodegradação. / Plasticizers are low volatility compounds that offer flexibility and processability to resins. The phthalates and adipates, used as plasticizers, have low water solubility e high partition octanol/water(Kow) and accumulate in soil and sediments. This compounds are considered teratogenics, carcinogenics and as endocrine disruptors. This study evaluated the bioremediation of tropical soil contaminated with plasticizers process wastes, in aerobic conditions, with and without introduction of acclimated bacteria. After geological analysis of soil, considering ten differents points on the factory area, it was selected the soil for biodegradation of eight points (100kg/point) representing 17mg total plasticizers/kg soil to 6222mg total plasticizers/kg soil. Mineralogical, physical and chemical analysis were done and the results showed that perhaps the cationic change capacity was due to plasticizers. The plasticizers contents in soil were 85-1688mg/kg and after 120 days of biodegradation in eight aerobic reactors, the removal efficiencies were 75-97%. The fingerprint analysis showed that the final bacteria present in reactors originated from soil and sludge and the CGMS analysis identified the metabolic MEHP, and showed the sub-products and final products of biodegradation.

This study investigates the impacts of selected Leguminous Tree Species (LTS) and kaolinite pre-amendment on oil-contaminated soil. It covered assessment of different levels of contamination (0, 25, 50, 75 and 100 ml in 4000 g soil; which represents the degree of light crude oil spillage concentration as 0.0, 0.63, 1.25, 1.88 and 2.5 %v/w) on the growth performance of Albizia adianthifolia, Albizia odoratissima, Bauhinia monandra, Delonix regia, Peltophorum pterocarpum and Tetrapleura tetraptera LTS investigated. Percentage germination, seedling height, seedling girth, number of leaves and number of nodules decreased as the concentrations of crude oil in soil samples increased. LTS affected soil physicochemical properties. Soil acidity decreased; soil organic matter, carbon content and exchangeable ions increased. N, P and K were altered in the LTS planted soil as compared to controls, but there were no significant (P > 0.05) differences. There were increased microbial counts in the crude oil-contaminated soil planted with LTS as compared with non-LTS planted soils. Hydrocarbon removal was significantly higher (P < 0.05, n = 3) in LTS planted soil than in non-planted soil. D. regia planted soils had most hydrocarbon removal and had significantly more growth in terms of plant height, girth and leaf production in the field. Kaolinite (10 and 20 g samples) applications were suitable and effective sorbent agents for oil-contamination at the different oil concentrations. The sorption potential of kaolinite increased with the increase in kaolinite to 20 g. The potential re-usability of kaolinite after the initial use for oil sorption was analysed and 10 g of burnt kaolinite sorbed 43.62%, while 20 g sorbed 58.90%. The rate of oil sorption by fresh kaolinite was significantly higher than burnt kaolinite. Results show the considerable potential of phytoremediation protocols with LTS and kaolinite as combined remediating agents for oil spill remediation in the humid tropics.

The wood-degrading basidiomycete, Phanerochaete chrysosporium, has been studied as a model organism in elucidating the mechanisms and pathways enabling this white-rot fungus to degrade recalcitrant lignin. These same mechanisms are implicated in the mineralisation of environmentally persistent, toxic phenolic chemicals. For this reason, P. chrysosporium has been exploited in a number of environmentally sound technologies, including the degradation of the indigestible lignin component in agricultural waste for the generation of digestible animal feedstocks or high sugar content raw materials for ethanol production; brightening processes in the pulp and paper industry; the detoxification and decolourisation of industrial effluents; and the bioremediation of hazardous waste sites. The improvement of these technologies is dependant on ongoing research involving strain selection, strain development using genetic engineering approaches and process development. Strain improvement using non-recombinant methods is beneficial in that it does not limit the inherent robustness observed amongst natural variants. In this research, through a breeding programme, ten P.chrysosporium sibling strains were screened for variable ligninase activities and pollutant degradation capabilities in order to further describe previously identified differences between these organisms. A conventional stationary liquid culture technique was effectively miniaturised from 10 ml flask cultures to a 96-well microtitre plate format, for the assessment of multigenic traits amongst sibling strains. Using the 96-well microtitre plate method, the relationships between P. chrysosporium growth kinetics, peroxidase production, pollutant sensitivity and pollutant degradation was explored. Significant correlations were primarily associated with P. chrysosporium growth [P < 0.05]. Percentage p-cresol removal and tannic acid tolerance were both correlated with a shorter lag phase in growth [tannic acid: r = 0.7698, P < 0.05; p-cresol: r = 0.7584, P < 0.05] and lower stationary phase biomass levels [tannic acid: r = 0.8177, P < 0.05; p-cresol: r = 0.7803, P < 0.05]. A significant correlation (linear relationship) was also detected between percentage Poly-R478 decolourisation and time of onset of MnP [r = 0.9689, P < 0.001]. No correlation was observed between dye decolourisation, p-cresol degradation, lignin degradation and lignin peroxidase (LiP) or manganese peroxidase (MnP) activities [P > 0.05]. These results imply that differences in the biosynthetic pathways for biomass accumulation in sibling strains play a significant role in the intraspecific variation observed in pollutant sensitivity, pollutant degradation, and enzyme production. Categorical analysis of intraspecific differences was assessed according to four criterions. These included growth, extracellular peroxidase activities, tolerance to toxic pollutants and the biodegradation of model pollutants. Sibling strains showing the most variable responses in three or more of the selective criterion were recommended for further studies. These strains include P. chrysosporium ME446, BS 2.52, BS 13, BS 17, BS 18, and BS 24. Interestingly, BS 2.52 (a dikaryotic strain generating from the crossing of two haploid progeny) showed significantly lower degradation capabilities than the wildtype parent strain ME446. The inherited variability observed between sibling strains is to be further explored through proteome and transcriptome analysis and genetic linkage studies aimed at describing the mechanisms or pathways conferring tolerance to or degradation of environmental pollutants. In examining fewer organisms at this next level, the number of replicates examined can be increased and thus the power of detection of experimental procedures improved, enabling the detection of multigenic traits amongst genetically related organisms. Growth was shown to play a significant role in the intraspecific differences detected in pollutant sensitivity and degradation between sibling strains. Little is known about the mechanism of growth and differentiation, or the role of differentiation in regulating the lignolytic activity in this organism. The membrane gradostat bioreactor and a unique plug-flow membrane bioreactor were evaluated as novel tools with which to further explore the relationship between secondary metabolism, pollutant degradation and biofilm development in sibling strains. High yield MnP production at levels as high as 1478.8 U.l-1 was achieved using a laboratory scale membrane gradostat bioreactor. Furthermore, extensive mycelial differentiation and tissue formation are reported for P. chrysosporium in both the membrane gradostat bioreactor and plug-flow membrane bioreactor. Intraspecific differences in the extent of this differentiation were observed in strains ME446, BS 13, BS 17 and BS 26 cultured using the membrane gradostat bioreactor, highlighting the potential of these techniques as a platform for future strain improvement strategies.