Rates of microbial reactions are important in understanding groundwater chemistry and bioremediation. In aquifers, microbial rates depend on physicochemical and biological factors and also on how groundwater transport impacts microbial reactions at pore-scale. I numerically simulate microbial acetate consumption in a porous medium, focusing on how physical heterogeneity of the medium impacts the rates. My model is a 3-D cube, which represents a portion of a sandy aquifer. Acetate is supplied by groundwater flow through the cube, and microbes live on randomly distributed grain surfaces by oxidizing acetate. I simulated microbial acetate oxidation under a range of groundwater velocity, acetate concentrations, spatial heterogeneity, and other physicochemical conditions. The results demonstrate a significant gap in microbial kinetics between the pore-scale and continuum model. Specifically, microbial rates are larger in porous media of greater heterogeneity. For this reason, I propose that microbial parameters should not be applied directly to field-scale biogeochemical modeling.

The increasing cost of effluent treatment in the wool scouring industry is rapidly becoming a determining factor in the viability of existing scouring operations and new installations alike. This thesis details the development of an integrated effluent treatment process capable of treating the worst polluted effluent from a wool scour "heavy flow-down", to the point where it can either be economically discharged to local trade waste sewer, or directly discharged to river or ocean outfall with minimal environmental impact. The existing proprietary chemical flocculation process, Sirolan CF™, was improved by the addition of a bio-flocculation stage and turbidity monitoring and control, and the product from this process fed to an aerobic biological treatment system based upon the traditional activated sludge process. The biological treatment process was found to remove up to 98% of the BOD5 loading from the pre-treated liquor with a hydraulic residence time of at least 50 hours being required in the aerobic digestion vessels. A residual biorefractory COD of approximately 3,600mg/L was identified which could not be removed by biological treatment. When operating continuously, the biological process was observed to metabolically neutralise the pH 3.0 - 4.5 feed from the chemical flocculation system to pH &gt 7.0 without the need for supplemental addition of neutralising agents such as sodium hydroxide. This in itself provides a significant economic incentive for implementation of the process. Kinetic analysis of the biological process carried out under controlled laboratory conditions using a Bioflo 3000 continuous fermentor showed that the bio-chemical process followed substrate inhibition kinetics. An appropriate kinetic model was identified to represent the behaviour of the substrate degradation system, and modified by inclusion of a pseudo toxic concentration to account for the effect of pH inhibition upon the biological growth rate. The process was verified both at pilot plant scale and at demonstration plant scale at an operational wool scour. The demonstration plant was of sufficient size to handle the full heavy effluent flow-down from a small wool scour. At the time of publishing three full-scale effluent treatment systems based on this research had been sold to both domestic and international clients of ADM Group Ltd. who funded the research.

This thesis examined the use of two-phase partitioning bioreactors (TPPBs) for the biodegradation of poorly water-soluble compounds. TPPBs are stirred tank bioreactors composed of a biocatalyst-containing aqueous phase and an immiscible second phase containing large amounts of poorly water-soluble or toxic substrates. Degradation of the bioavailable substrate in the aqueous phase will result in equilibrium-driven partitioning of additional substrate from the immiscible phase into the aqueous phase, theoretically allowing for complete substrate degradation. Fundamental work was undertaken with the PCB-degrading organisms Burkholderia xenovorans LB400 in liquid-liquid and solid-liquid TPPBs. Initially biphenyl was used as the sole carbon source due to its hydrophobic nature and structural similarity to the environmentally relevant PCBs. The critical LogKO/W (octanol/water partitioning coefficient) of the organism was determined to be 5.5 and its growth kinetics on biphenyl were determined in a liquid-liquid TPPB. A polymer selection strategy for solid-liquid TPPBs was developed in the next chapter, and it was shown in the following chapter that biphenyl degradation in solid-liquid TPPBs was mass transfer limited, as described mathematically utilising the previously estimated microbial kinetics. The fundamental knowledge gained in the early chapters was then applied to the degradation of PCBs by the same organism. It was shown that the aqueous phase availability of PCBs is the rate-limiting step in biphasic bioreactors, and not the mass transfer rate. The low specific microbial degradation rates, resulting from substrate-limited growth were addressed with increased biomass concentrations; however, it was also found that an additional carbon source was required to maintain microbial activity over an extended period of time. Pyruvic acid was selected as a carbon source which, once added to actively PCB-degrading cells, maintained the cells’ activity towards PCBs and up to 85 % of 100 mg l-1 was degraded in 15 h. It was shown as the final contribution in this thesis that TPPBs can be combined with a PCB soil extraction step as a potential remediation scheme for PCB contaminated soil. PCBs were extracted from soil with polymer beads (up to 75 % removal), followed by biodegradation of the PCBs in a solid-liquid TPPB in which PCBs were delivered to the degrading organism from the same polymer. / Thesis (Ph.D, Chemical Engineering) -- Queen's University, 2007-08-07 16:11:00.494

This thesis examined the biological treatment of gas streams containing benzene, toluene, ethylbenzene and o-xylene (BTEX) using solid-liquid two-phase partitioning bioscrubbers (SL-TPPBs). SL-TPPBs consist of a cell containing aqueous phase and a polymeric solid phase that sequesters poorly water soluble and/or toxic substrates, mitigating substrate toxicity in the aqueous phase and improving the gas mass transfer during treatment of VOC contaminated gases. An initial investigation of oxygen transport determined that the polymers in a stirred-tank SL-TPPB enhance gas-liquid mass transfer. In addition, a study on biodegradation kinetics of BTEX by a bacterial consortium identified and quantified substrate interactions such as inhibition, enhancement and cometabolism. The stirred-tank SL-TPPB was then experimentally investigated for treatment of BTEX gas streams during steady-state and dynamic step-change operation to determine performance of the system relative to other biotreatment methods. A mathematical model was developed to predict system performance, which included the microbial kinetic model structure and parameters estimated during kinetic and oxygen mass transfer studies. As a less energy intensive alternative, an airlift SL-TPPB was operated and characterized. The airlift SL-TPPB was compared to an airlift liquid-liquid TPPB (silicone oil as sequestering phase) and a single phase airlift over dynamic step-change loadings, which showed that the airlift SL-TPPB outperformed the single phase airlift by >30% and had similar performance to the liquid-liquid airlift. However, the airlift SL-TPPB performance was lower relative to the stirred-tank SL-TPPB by >15%. Steady-state operation of the airlift SL-TPPB identified a range of operating conditions that provided maximum performance and conditions that were not oxygen limited. This prompted a study of oxygen mass transfer and hydrodynamics in the airlift system, which identified that the addition of polymers to an airlift does not cause physical enhancement of the gas-liquid mass transfer coefficient, but improves aqueous phase mixing and enhances overall oxygen transfer rate. A tanks-in-series mathematical model was formulated to predict performance of the airlift SL-TPPB, wherein the number of tanks-in-series to describe mixing in the airlift was obtained from a residence time distribution analysis of the airlift system completed during the hydrodynamic investigation. This thesis contributes a low-energy solution for the effective treatment of gases contaminated with BTEX. / Thesis (Ph.D, Chemical Engineering) -- Queen's University, 2009-08-18 16:16:22.598

Submitted by Programa de Pós-graduação em Biotecnologia (mebiotec.ufba@gmail.com) on 2017-04-05T12:59:59Z No. of bitstreams: 1 Dissertacao Final - Diego Montes.pdf: 3792184 bytes, checksum: 932c7d285674a48aa5a90cb88c53dd85 (MD5) / Approved for entry into archive by Delba Rosa (delba@ufba.br) on 2017-07-03T15:41:00Z (GMT) No. of bitstreams: 1 Dissertacao Final - Diego Montes.pdf: 3792184 bytes, checksum: 932c7d285674a48aa5a90cb88c53dd85 (MD5) / Made available in DSpace on 2017-07-03T15:41:00Z (GMT). No. of bitstreams: 1 Dissertacao Final - Diego Montes.pdf: 3792184 bytes, checksum: 932c7d285674a48aa5a90cb88c53dd85 (MD5) / CAPES / As Bactérias redutoras de sulfato (BRS) são de enorme importância do ponto de vista industrial e ambiental. O sulfeto produzido pelas BRS constitui um grande problema para o setor petrolífero por causar da biocorrosão nas instalações e por ser tóxico ao ser humano. O monitoramento dessas bactérias é realizado com frequência a fim de qualificar o tratamento para prevenir a produção de sulfeto. O tratamento comumente utilizado é a injeção de biocidas cuja utilização deve ser proporcional a densidade de BRS a fim de minimizar danos econômicos e ambientais. O objetivo deste trabalho foi o de desenvolver um método indireto, baseado no consumo de aceptores de elétrons (sulfato) para monitorar populações de BRS. A metodologia consistiu, a princípio, no desenvolvimento de um método de análise por cromatográfia de íons que, além do sulfato e nitrato, pode também separar outros 15 ânions como: tungstato, sulfeto, fosfato, molibdato, oxalato, nitrito, cloreto, piruvato, butirato, propionato, acetato, fluoreto, formiato, succinato e citrato. O método de avaliação foi testado para se ajustar aos padrões regidos pelo INMETRO e ANVISA (órgãos brasileiros de regulamentação). Adicionalmente foi realizada a adaptação de um método espectrofotométrico para determinação de sulfato para assim auxiliar o processo cromatográfico. O tempo de corrida no cromatografo foi de 45 minutos que foi inferior ao método fornecido pelo fabricante e com um LD de 574,2 e 127,1 μg L-1 para sulfato e nitrato, respectivamente. As análises espectrofotométricas foram eficientes para medir sulfato e nitrato entre o intervalo de 40 e 200 mg L-1. A associação da cinética microbiana de crescimento e consumo de sulfato foi realizada em cultura pura de BRS e em cultura mista de amostras ambientais. Para testes com cultura pura foi utilizada uma cepa de BRS considerada como modelo experimental para o grupo, a bactéria Desulfovibrio vulgaris. Para testes com cultura mista foram utilizados consórcios microbianos coletados de amostras de água produzida de petróleo e do Rio Camarajipe, Salvador-BA. Foi observado que a D. vulgaris apresentou uma taxa de crescimento de 0,67 h-1 em meio rico contendo sulfato, mas é capaz de crescer na presença de nitrato com taxas próximas de 0,34h-1 e um consumo de nitrato de ≅22mg L-1h-1. Esse resultado ilustra a ineficiência do uso isolado de nitrato para controlar BRS em poços de petróleo. Os testes relacionando a variação da densidade de D. vulgaris em meio de cultura com fonte de carbono e aceptor de eletron (SO4 2-) em concentrações padronizadas demostrou alta correlação linear (R2 ≥ 99%). Esse resultado comprova viabilidade de usar o Modelo de Monod para quantificar biomassa dessa cultura em apenas 12 horas de teste, ao invés de 28 dias como é o caso da técnica de NMP. A sensibilidade do método esta entre as densidades de 102-108 cel/mL. Os resultados foram menos significativos com culturas mistas de BRS provinda de amostras ambientais, mas, os resultados sugerem que ajustes na técnica podem melhorar seu desempenho. / Sulphate reducing bacteria (SRB) are of significant environmental and industrial importance. SRB produces significant amounts of sulphite that can cause bio-corrosion on industrial installations and be toxic to humans. Monitoring the presence of such bacteria is an activity frequently required in order to determine treatment and prevent sulphite production during petrol exploration. Injection of biocides is commonly used for this purpose and should be proportional to SRB present in the environment in order to avoid economic and environmental drawbacks. The objective of this work was to develop an indirect method for monitoring SRB population in situ based on the consumption ratios of electron acceptors such as sulphate. First, it was necessary to adapt a chromatographic method for detecting ions such as sulphate and nitrate, but the approach has also showed to be successful for separating 15 other anions such as tungstate, sulphite, phosphate, molibdate, oxalate, nitrite, chlorate, pyruvate, butyrate, propionate, acetate, fluorite, formiate, succinate and citrate. The chromatographic approach is robust to pass the “INMETRO” and “ANVISA”, the Brazilian regulation agencies. Additionally, the chromatographic approach was also tested against a spectrophotometric method for determine sulphate. The chromatographic run achieved was of 45 min only, compared to a longer period suggested by the manufacturer and with a detection limit of 574.2 and 127,1mg l-1 for sulphate and nitrate, respectively. The spectrophotometric approach was efficient for detecting sulfphate and nitrate at the range of 40 to 200mg l-1. Sulphate consumption versus the biomass activity of SRB was first tested using pure culture and later with environmental mixed cultures. Desulfovibrio vulgaris was used as a biological model representing SRB for the tests with pure culture. Two different consortiums were used for testing environmental SRB mixed cultures, one collected at the Camarajipe River, Salvador-BA, and cells grown from industrial produced water obtained from oil wells. It was observed that D. vulgaris showed a growth rate of about 0,67 h-1 in rich media with sulphate as sole electron acceptor, but the culture was also able to grow on nitrate with an average consumption of ≅22mg L-1h-1 and a growth rate of 0.34 h-1. This result suggest the inefficiency of using nitrate as means to control SRB in oil wells once some SRB can utilize it as electron acceptors. Tests correlating the variation of D. vulgaris initial biomass with sulphate consumption rate at standard conditions of carbon source and electron acceptor concentrations showed to be of statistical significance (R2 ≥ 99%). This result strongly support the tested method, which is based on the Monod model, for measuring bacterial biomass within only 12 hours instead of 28 days such as required by the most probable number technique (MPN). The sensitivity of the method was set between 102-108 cel/ml, but the results were less significant when tested with mixed environmental cultures. Nonetheless, the result showed that some adjustment in the technique may improve its performance.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-03-24T10:41:02Z No. of bitstreams: 3 Dissertação - Jaquelinne Pires Vital da Costa - 2017 - parte 1.pdf: 4379999 bytes, checksum: b3803a48e0c37eb318b174d9582a33e1 (MD5) Dissertação - Jaquelinne Pires Vital da Costa - 2017 - parte 2.pdf: 16358112 bytes, checksum: 7c49d76a1bb2c8ca62b7fe392e7e0bb1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-03-24T11:04:59Z (GMT) No. of bitstreams: 3 Dissertação - Jaquelinne Pires Vital da Costa - 2017 - parte 1.pdf: 4379999 bytes, checksum: b3803a48e0c37eb318b174d9582a33e1 (MD5) Dissertação - Jaquelinne Pires Vital da Costa - 2017 - parte 2.pdf: 16358112 bytes, checksum: 7c49d76a1bb2c8ca62b7fe392e7e0bb1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-03-24T11:04:59Z (GMT). No. of bitstreams: 3 Dissertação - Jaquelinne Pires Vital da Costa - 2017 - parte 1.pdf: 4379999 bytes, checksum: b3803a48e0c37eb318b174d9582a33e1 (MD5) Dissertação - Jaquelinne Pires Vital da Costa - 2017 - parte 2.pdf: 16358112 bytes, checksum: 7c49d76a1bb2c8ca62b7fe392e7e0bb1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Research into new sources of natural pigments has grown, mainly due to the toxic effects caused by synthetic dyes used in food, pharmaceuticals, textiles and cosmetics. In this context, the microbial pigments are presented as an alternative to supply this demand. There are several microorganisms capable of producing pigments, among them the filamentous fungus, Monascus. This is capable of producing yellow, orange and red pigments, and its production depends on the conditions under which it is grown. Thus, agroindustrial wastes have emerged as a cheap and profitable option for the production of biopigments. The aim of the present study was to evaluate the use of cheese whey as a substrate for the production of pigments by Monascus ruber CCT 3802. For this purpose, we first sought to evaluate the radial growth of the fungus in solid medium under serum concentrations of cheese powder and different pH values. The results demonstrated the potential of whey as a substrate for fungus growth, since there was a significant difference between the growth rates obtained for each medium when compared to the control medium (without cheese whey). As for pH, growth rates of the fungus were higher at pH 6.0. At extreme pH values, such as pH 2.0 and 8.0, the growth of the fungus presented inhibition and the visual aspect differed, showing yellow and red coloration, respectively. This demonstrates the clear influence of pH on the production of pigments produced by M. ruber. Culture media containing glucose and cheese whey at different concentrations were studied and the results showed higher pigment production was obtained in the media containing only glucose. For this reason, the third part of the study sought to use lactose, after the enzymatic hydrolysis process, as a carbon source for the production of pigments by Monascus ruber. Culture media containing glucose, lactose and hydrolyzed lactose were incubated at 30°C in shaker for 7 days. Although it contained less than 43% of glucose, the medium containing hydrolyzed lactose presented maximum pigment production of 7.58 UA510, surpassing the culture containing glucose. The pigment produced in hydrolyzed lactose appeared lighter, but redder and more yellow than in glucose. The consumption of reducing sugars and biomass production for the culture containing lactose only were inhibited, with a decline in pigment production after 72 hours of cultivation. The results presented in this research contribute to the fact that future studies optimize the hydrolysis of lactose and, consequently, cheese whey, using it as a substrate for the production of biopigments by Monascus ruber. / A pesquisa por novas fontes de pigmentos naturais tem crescido, principalmente devido aos efeitos tóxicos causados por corantes sintéticos usados em alimentos, produtos farmacêuticos, têxteis e cosméticos. Nesse contexto, os pigmentos microbianos apresentam-se como alternativa para suprir essa demanda. Vários são os micro-organismos capazes de produzir pigmentos e, dentre eles, destaca-se o fungo filamentoso, Monascus. Este é capaz de produzir pigmentos amarelo, laranja e vermelho, e sua produção depende das condições em que é cultivado. Assim, os resíduos agroindustriais têm surgido como opção barata e rentável para a produção de biopigmentos. O presente estudo teve como objetivo avaliar o aproveitamento do soro de queijo como substrato para a produção de pigmentos por Monascus ruber CCT 3802. Para isso, primeiramente, buscou-se avaliar o crescimento radial do fungo em meio sólido sob concentrações de soro de queijo em pó e diferentes valores de pH. Os resultados demonstraram a potencialidade do soro de queijo como substrato para o crescimento do fungo, visto que houve diferença significativa entre as velocidades de crescimento obtidas para cada meio quando comparadas ao meio controle (sem soro de queijo). Quanto ao pH, as velocidades de crescimento do fungo foram maiores em pH 6,0. Em valores de pH extremos, como pH 2,0 e 8,0, o crescimento do fungo apresentou inibição e o aspecto visual diferiu-se, apresentando coloração amarela e vermelha, respectivamente. Isso demonstra a clara influência do pH na produção de pigmentos produzidos por M. ruber. Em estudos preliminares, buscou-se avaliar a produção de pigmentos alimentares por Monascus ruber em cultivo submerso, usando o soro de queijo como substrato. Meios de cultivo contendo glicose e soro de queijo em diferentes concentrações foram estudados e os resultados demonstraram maior produção de pigmentos foi obtida nos meios contendo apenas glicose. Por esse motivo, buscou-se utilizar a lactose, após o processo de hidrólise enzimática, como fonte de carbono para a produção de pigmentos por Monascus ruber. Meios de cultivo contendo glicose, lactose e lactose hidrolisada foram incubados a 30ºC em shaker, durante 7 dias. Apesar de conter menos de 43% de glicose, o meio contendo lactose hidrolisada apresentou produção máxima de pigmentos de 7,58 UA510, superando o cultivo contendo glicose. O pigmento produzido em lactose hidrolisada apresentou-se mais claro, porém mais vermelho e mais amarelo que em glicose. O consumo de açúcares redutores e produção de biomassa para o cultivo contendo apenas lactose foram inibidos, havendo declínio na produção de pigmentos após 72h de cultivo. Os resultados apresentados nesta pesquisa contribuem para que estudos futuros otimizem a hidrólise da lactose e, consequentemente, do soro de queijo, aproveitando-o como substrato para produção de biopigmentos por Monascus ruber.