Nitrogen removal from municipal wastewater with the application of deammonification process offers an operational cost reduction, especially if it is combined with a maximal use of organic content of wastewater for biogas production. In this thesis, two approaches for integration of the deammonification process into the municipal wastewater treatment scheme were studied. The first approach is based on ammonium concentration from municipal wastewater by ion exchange followed by biological removal of ammonium from the concentrated stream by deammonification process. Experiments with synthetic and real municipal wastewater showed that strong acid cation resin is suitable for ammonium concentration due to its high exchange capacity and fast regeneration. Since NaCl was used for regeneration of ion exchange materials, spent regenerant had elevated salinity. The deammonification biomass was adapted to NaCl content of 10-15 g/L by step-wise salinity increase. The technology was tested in batch mode with 99.9 % of ammonium removal from wastewater with ion exchange and up to 95 % of nitrogen removal from spent regenerant by deammonification process. The second studied approach was to apply anammox process to low-concentrated municipal wastewater in a moving bed biofilm reactor (MBBR) and integrated fixed film activated sludge (IFAS) system without a pre-concentration step. After a 5 months period of transition to mainstream wastewater the pilot plant was operated during 22 months and stable performance of one-stage deammonification was proven. Clear advantage of IFAS system was shown. The highest stable nitrogen removal efficiency of 70 % and a nitrogen removal rate of 55 g N/(m3·d) was reached. Moreover, the influence of operation conditions on competition between ammonium oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) was studied by literature review, batch tests and continuous pilot plant operation. / <p>QC 20170309</p>

Aerobic activated sludge membrane bioreactors (AS-MBR) in municipal wastewater treatment are compact systems that can efficiently perform biological organic oxidation. However, aerobic processes require mechanical aeration accounting for over 40% of total expenditure of a wastewater facility. Additionally, a global urgency for nutrient (Nitrogen/Phosphorus) removal strategies due to surges of eutrophication events requires complex MBR configurations. An innovative and cost-effective process was developed with a dual income-stream: high-quality treated effluent and value-added microalgal biomass for several applications. The proposed process involved several integrated components; an ultrafiltration AS-MBR for organic oxidation followed by a microalgal membrane photobioreactor (MPBR) to remove nutrients (N/P) through assimilation while simultaneously photosynthetically generating dissolved oxygen effluent that was recirculated back into the AS-MBR, thereby reducing the need for mechanical aeration for oxidation. A lab-scale system was fed with a synthetic medium-strength municipal wastewater. The microalgal species C. vulgaris was initially tested in batch trials as a proof-of-concept study on its potential as a photosynthetic oxygenator for the AS-MBR and identify its nutrient utilization efficiencies. The MPBR and MBR were later constructed for continuous operation, with the aim to identify an optimal process configuration. The unit processes were subsequently isolated, where the AS-MBR was subjected to a modelled algal effluent to assesses the impact of varying influent characteristics and effluent recycle rates. A microbial community analysis was performed by high-throughput sequencing and a statistical data-driven modeling approach to assess treatment performances. The MPBR stage was then subjected to the effluent achieved by the AS-MBR stage under varying operating conditions to assess its treatment performance and the resulting algal biomass biochemical composition to identify its suitability for bioethanol, biodiesel, or animal feed production. The findings of this study ultimately confirmed the ability of C. vulgaris to support the AS-MBR for organic removal and fractional nutrient removal by supplying the oxygen demand, and further achieve an effluent polish stage for nutrient removal. The process configuration also demonstrated the ability to achieve a high microalgal biomass production with the potential of extracting valuable products as an added benefit of the wastewater treatment.

Thesis advisor: Noah P. Snyder / Thesis advisor: Gail Kineke / Fluvial terraces form in both erosional and depositional landscapes and are important recorders of land-use, climate, and tectonic history. Terrace morphology consists of a flat surface bounded by valley walls and a steep-sloping scarp adjacent to the river channel. Combining these defining characteristics with high-resolution digital elevation models (DEMs) derived from airborne light detection and ranging (lidar) surveys, several methods have been developed to identify and map terraces. This research introduces a newly developed objective terrace mapping method and compares it with three existing DEM-based techniques to determine which is most applicable over entire watersheds. This work also tests multiple methods that use lidar DEMs to quantify the thickness and volume of fill terrace deposits identified upstream of dam sites. The preliminary application is to the Sheepscot River watershed, Maine, where strath and fill terraces are present and record Pleistocene deglaciation, Holocene eustatic forcing, and Anthropocene land-use change. Terraces were mapped at four former dam sites along the river using four separate methodologies and compared to manually delineated area. The methods tested were: (1) edge detection using MATLAB, (2) feature classification algorithms developed by Wood (1996), (3) spatial relationships between interpreted terraces and surrounding natural topography (Walter et al., 2007), and (4) the TerEx terrace mapping toolbox developed by Stout and Belmont (2013). Thickness and volume estimates of fill sediment were calculated at two of the study sites using three DEM-based models and compared to in situ data collected from soil pits, cut bank exposures, and ground penetrating radar surveys. The results from these comparisons served as the basis for selecting methods to map terraces throughout the watershed and quantify fill sediment upstream of current and historic dam sites. Along the main stem and West Branch of the Sheepscot River, terraces were identified along the longitudinal profile of the river using an algorithm developed by Finnegan and Balco (2013), which computes the elevation frequency distribution at regularly spaced cross-sections normal to the channel, and then mapped using the feature classification (Wood, 1996) method. For terraces upstream of current or historic dam sites, thickness and volume estimates were calculated using the two best performing datum surfaces. If all analyzed terraces are composed of impounded sediment, these DEM-based results suggest that terraces along the main stem and West Branch of the Sheepscot River potentially contain up to 1.5 x 106 m3 of fill. These findings suggest powerful new ways to quickly analyze landscape history over large regions using high-resolution, LiDAR DEMs while relying less heavily on the need for detailed and costly field data collection. / Thesis (MS) — Boston College, 2014. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Geology and Geophysics.

A dissertation submitted to the Faculty of Science, University of the Witwatersrand, Johannesburg, in fulfilment of the requirements for the degree of Masters of Science, 2017 / The tailoring of zeolites surface properties using organic functionalising agents displaying higher binding affinity for metal ions is a widely explored approach for water treatment. In this study, amine functionalised zeolites and phosphate functionalised zeolites were separately synthesised from similar natural zeolite precursors using reflux methods. The surface composition and morphological elucidations were achieved by characterising the adsorbents using Fourier Transform Infra-red spectroscopy (FTIR), thermogravimetric analysis (TGA), Zeta potential, Point of zero charge (pHPZC), and the Brunauer, Emmett and Teller analysis (BET). In case study 5.1, the sorption mechanisms of the uranyl ion onto amine functionalised zeolites (AMZ), activated carbon (AC) and natural zeolite (NZ) were studied as function of various environmental batch parameters. There was effective adsorption when uranium existed as uranyl ions: UO22+ and UO2OH+. The data fitted numerous kinetic and isotherm models suggesting that the equilibrium mechanisms were characteristic of a combination of chemisorption and physisorption for these three adsorbents. The Dubinin-Radushkevich (DR) model did not fit the data and therefore the energy values derived from it were not used to predict the mechanisms involved. However, the thermodynamic evaluations of parameters ∆H, ΔG and ∆S° showed that equilibrium mechanisms were exothermically, randomly and spontaneously favoured for all adsorbents at temperatures ranging between 22 and 40oC. The adsorption capacity of 0.452 mg g-1 was achieved at pH 3 by 500 mg AC dosage using 20 mL volume of 10 mg L-1 uranyl ion solution after equilibrating for 6 h within the temperature ranges of 22 to 30oC. Under the same conditions of sorbent dosage of 500 mg, uranyl solution volume of 20 mL and 10 mg L-1 U(VI) solution concentration, the maximum adsorption capacity of 0.506 mg g-1 for NZ and 0.480 mg g-1 for AMZ were both achieved at pH 4 after equilibration time of 21 h and 6 h with the optimum temperature range of 22 to 30oC, respectively. The model results predict that intraparticle diffusion thorough pores decreased in the order AC ˃ NZ ˃ AMZ while estimating that chemisorption occurred in a reverse order. On the basis of the modelled data, it was deduced that amine functionalisation of natural zeolites improves their chemisorption capability for uranyl ion and can therefore be used as a cost efficient adsorbent for small-scale remediation of contaminated aquatic systems. In another case study 5.2, the surface properties of successfully prepared aminomethyl phosphonic acid functionalised natural zeolite (APZ) were compared to those of commercial silica polyamine composites (SPC) for uranium uptake in batch aqueous solutions. The FTIR spectrum revealed that (3-aminotrimethyl) phosphonic acid functional groups were successfully grafted onto natural zeolite. The TGA analysis showed that the APZ had higher thermal stability and fewer active sites compared to SPC. The optimum adsorption capacity (qe) of 49 mg g-1 and 44 mg g-1 uranium was achieved using 25 mg SPC and 100 mg APZ, respectively at pH 4, 25oC after 1 and 6 h equilibrating time. The data best fitted the pseudo second-order kinetic model and Freundlich isotherm model. The thermodynamic studies showed that adsorption occurred chemically and exothermically for both APZ and SPC. The overall selectivity order for APZ was; Na ˃ Mn ≥ U ˃ Ca ˃ Fe and for SPC was; Fe ˃ Mn ≥ Ca ˃ U˃ Na. The findings showed that phosphate- and amine-functionalised zeolite bind strongly to uranium compared to the unmodified natural zeolite and other conventional adsorbents such as activated carbon. Their selectivity for this element was commendable. With further improvements in the synthetic protocols e.g. by using microwave-based methods, it should be possible to obtain functionalised zeolite that has superior properties to SPC. / XL2017

A interação entre fungos e metais, utilizada pela biorremediação, pode resultar na remoção do metal de locais ou resíduos contaminados. Os objetivos desse trabalho foram investigar a tolerância e o comportamento de nove espécies de fungos isolados de local contaminado com Cu2+ e avaliar a capacidade de remoção de Cu2+ daquela com resistência diferenciada. A partir da inoculação em meio de cultura batata dextrose ágar acrescido de diferentes concentrações de Cu2+, o crescimento e a macromorfologia foram analisados. P. ochrochloron demonstrou tolerância distinta e foi selecionada para os testes de remoção, com variação da concentração de Cu2+ e do tempo de contato. Tanto a biomassa morta, seca e pulverizada quanto a biomassa em crescimento foram capazes de interagir com o Cu2+, embora a capacidade de remoção e a quantidade de Cu2+ removido tenham sido superiores pela biomassa morta. Ficou evidente que os fungos apresentam potencial a ser explorado, sendo importante o avanço de pesquisas nesse campo para o desenvolvimento de eficientes processos de biorremediação. / The interaction between fungi and metals, used by bioremediation, can result in metal removal from contaminated sites or waste. The aim of this work was to investigate the tolerance and the behaviour of nine fungal species isolated from a Cu2+ contaminated site and to evaluate the Cu2+ removal capacity by species with differentiated resistance. From the inoculation in potato dextrose agar culture medium added with different Cu2+ concentrations, the growth and the macromorphology were analyzed. P. ochrochloron demonstrated distinct tolerance and was selected for the removal tests, with variation of Cu2+ concentration and contact time. The dead, dry and pulverized biomass as well as the growing biomass were able to interact with Cu2+, although the removal capacity and the amount of removed Cu2+ have been higher by the dead biomass. It was evident that the fungi have potential to be explored, being the advancement of researches in this field important for the development of efficient bioremediation processes.

A presente pesquisa teve como objetivo a otimização de processos de precipitação química na remoção de fósforo de esgotos sanitários mediante a utilização de sais de ferro como coagulante. Analisou-se a remoção de matéria orgânica e produção de lodo obtidas. O trabalho foi desenvolvido com efluentes provenientes de duas estações de tratamento da SABESP, sendo elas a ETE Barueri lodos ativados convencional e ETE Ribeirão Pires tratamento anaeróbio com reatores UASB. Os ensaios de Jar-Test foram conduzidos em laboratório e o coagulante utilizado foi o cloreto férrico. Os resultados obtidos demonstram que a dosagem do coagulante, em ambos os efluentes estudados, resulta na remoção de fósforo total a valores inferiores a 1 mg/L. As dosagens de coagulante necessárias foram de 80 mg/L para a ETE Barueri e 60 mg/L para a ETE Ribeirão Pires. A remoção de carga orgânica também foi alcançada e valores de DBO5,20 inferiores a 10 mg/L são obtidos com dosagens inferiores às necessárias para a remoção de fósforo. A dosagem de coagulante necessária para a remoção de DBO5,20 a valores inferiores a 10 mg/L foi de 40 mg/L. A produção de lodo é o principal problema encontrado quando se utiliza precipitação química, a quantidade de lodo produzida é significativa e deve ser levada em consideração no dimensionamento do sistema de tratamento e disposição final. Incrementos na produção de lodo da ordem de 113% foram obtidos para a ETE Barueri e 51% para a ETE Ribeirão Pires quando foram consideradas as dosagens de coagulante necessárias a remoção de fósforo totais a valores inferiores a 1 mg/L. / The aim of the present study was the chemical precipitation optimization of phosphorus from domestic wastewater with a ferric salt as coagulant. The organic matter removal and the sludge production obtained were analyzed. The study was developed with effluents from two wastewater plants from SABESP, such as ETE Barueri conventional activated sludge and ETE Ribeirão Pires anaerobic treatment with UASB reactors. Jar-Test tests were done in laboratory and the coagulant used was the ferric chloride. The results show that the coagulant dosage, in both effluents studied, results in a total phosphorus removal to values lower than 1 mg/L. The necessary coagulant dosages were 80 mg/L to ETE Barueri and 60 mg/L to ETE Ribeirão Pires. The organic matter removal also was obtained and values of BOD lower than 10 mg/L are obtained to lower dosages than the necessary dosage to phosphorus removal. The necessary coagulant dosage to BOD removal to values lower than 10 mg/L was 40 mg/L. The sludge production is the main problem when chemical precipitation is used, the amount of sludge produced is significant and must be taken into consideration in the system project of treatment and final disposal. Increases in the sludge production were around 113% to ETE Barueri and 51% to ETE Ribeirão Pires when the necessary coagulants dosages were considered to total phosphorus removal to values lower than 1 mg/L.

Shadows are ubiquitous in image and video, and their removal is of interest in both Computer Vision and Graphics. In this thesis, four methods for interactive shadow removal from single images are presented. Their improvements are made in user interaction, quality and robustness of shadow removal. We also show our state-of-the-art ground truth data set with variable scene categories for shadow removal and applications for shadow editing and its extension to video data processing.

O presente trabalho avaliou a remoção do nitrogênio amoniacal de liquido percolado de aterro sanitário através do processo de nitritação/desnitritação (via curta) utilizando a própria matéria orgânica presente no lixiviado como fonte de carbono para o processo de desnitritação. Foi também avaliada a remoção de nitrogênio amoniacal do lixiviado através do processo de stripping de amônia. Para o processo de stripping foram utilizados dois sistemas, o primeiro era constituído por um reator (R-2) com volume útil de 20L mantido sobre agitação mecânica (palheta) e o segundo era composto de uma coluna cilíndrica, com volume de lixiviado de 10L, mantida sob aeração. Além da remoção de nitrogênio houve também uma diminuição na concentração de DQO, DBO e COT durante os ciclos de stripping. A taxa volumétrica de remoção de amônia durante os ciclos de stripping variou entre 78 e 86,95 mg N-NH3/L.dia. Para a avaliação da remoção de nitrogênio através da via curta (via nitrito), foi utilizado um reator operado em bateladas seqüenciais (SBR- 1) com volume útil de 20 L. O reator foi operado por um período de aproximadamente 1 ano. A avaliação do processo de nitritação/desnitritação se deu em 4 fases distintas; na primeira fase o sistema foi alimentado com 2L de lixiviado bruto, na segunda fase o sistema R-1 foi alimentado com lixiviado pré tratado (NNH3lixiviado @ 1200 mg/L), na fase três o sistema foi também alimentado com 2 L de percolado pré-tratado (N-NH3lixiviado @ 900 mg/L) e por fim, na fase 4 o sistema foi alimentado com 4L de lixiviado pré-tratado (N-NH3lixiviado @ 900 mg/L ). Nas duas primeiras fases da pesquisa a remoção de nitrogênio foi relativamente eficiente, variando entre 80 e 90%, porém mesmo com a concentração de amônia livre variando entre 0,18 e 20,7 mg/L não houve a inibição da nitratação durante a etapa aerada dos ciclos de tratamento. Durante as fases 1 e 2, a fase anóxica foi relativamente longa sendo que, as taxas específicas de desnitritação variaram entre 0,0100 e 0,0148 Kg NO3 -, NO2 -/ Kg SSV.dia. Na fase 3, após a mudança na alimentação do sistema, o reator R-1 entrou em regime de equilíbrio e a inibição da nitratação foi praticamente total (acima de 95%). Mesmo com a completa inibição da nitratação, a etapa anóxica durante os ciclos de tratamento da fase 3 continuou sendo relativamente longa (72 h em média) remetendo assim, a uma falta de matéria orgânica para a redução do nitrito durante a fase anóxica. Na fase 4, apesar da não inibição da nitratação durante os primeiros ciclos, nos ciclos que se seguiram a inibição foi quase total, demonstrando claramente a adaptação do sistema às condições necessárias ao acúmulo de nitrito. A manutenção do pH da massa líquida próximo a 8,3 foi determinante para o acúmulo de nitrito durante todas as fases da pesquisa. Nas fases 1 e 2 houve a necessidade de controle do pH com adição de alcalinizante, já nas fases 3 e 4 esse controle não foi necessário. / The present work evaluates the biological ammonia removal of a landfill leachate, through nitritation/denitritation as well as the utilization of the organic compounds present in the landfill leachate, as carbon source to denitritation process. It also evaluates the ammonia removal of landfill leachate by the ammonia stripping process. It has been used two systems for the stripping process, the first one was constituted of a reactor (R-2) with 20L useful volume kept on mechanical shaking (vane) and the second one was composed of a cylindrical column, with a 10L of landfill leachate volume, kept on aeration. Beyond the biological ammonia removal it also had a reduction in the COD, BOD and TOC concentration during the stripping cycles. The ammonia removal volumetric rate during the stripping cycles have varied between 78 e 86,95 mg N-NH3/L.day. In order to evaluate the ammonia removal through the short cut (via nitrite) one has used a sequencing batch reactor (SBR-1) with a 20L useful volume. The reactor was been operated by a period of approximately 1 year. The assessment of nitritation/denitritation process has happened in 4 distinct phases: in the first one the system was fed on 2L of heavy landfill leachate; in the second the system R-1 was fed on pre-treated landfill leachate (N-NH3leachate @ 1200 mg/L); in the third phase the system was also fed on 2L of pre-treated landfill leachate (N-NH3leachate @ 900 mg/L); and finally, in the fourth phase it was fed on 4L of pre-treated landfill leachate (N-NH3leachate @ 900 mg/L). In the two first phases of this research the biological ammonia removal was been relatively efficient, varying between 80 and 90%, however, even with the free ammonia varying between 0,18 and 20,70 mg/L, it has not had nitratation inhibition during the aerated stage of the treatment cycles. During phases 1 and 2 the anoxic phase was relatively long and specific denitritation rates have varied between 0,0100 and 0,0148 Kg NO3 -, NO2 -/ Kg VSS.day. In phase 3, after the change of system feeding, the R-1 reactor has entered in balance regime and the nitratation inhibition has been practically total (above 95%). Even with the complete nitratation inhibition, the anoxic phase during the phase 3 treatment cycles have continued being relatively long (72h on average), thus sending to a lack of organic compounds for the nitrite reduction in this phase. In phase 4, although the not inhibition of nitratation during the first cycles, in the followed cycles the inhibition has been almost total, demonstrating clearly the system adaptation to the necessary conditions to the nitrite accumulation. The maintenance of pH of liquid mass next to 8,3 has been determinative for the accumulation of nitrite during all phases of this research. In phases 1 and 2 it have been necessary to control the pH alkalinizing the system, already in phases 3 and 4 this control has not been necessary.

by Li Sze Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1990. / Bibliography: leaves 234-259. / ACKNOWLEDGEMENT --- p.i / ABSTRACT --- p.ii / CONTENTS : / Chapter CHAPTER 1: --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Our Environment Is Polluted --- p.1 / Chapter 1.2 --- Heavy Metal Contamination --- p.3 / Chapter 1.3 --- The Effect of Cadmium and Some Related Metals on Environment --- p.5 / Chapter 1.4 --- The Uses of Microorganisms in Cleaning Up Environment --- p.9 / Chapter 1.5 --- Mechanisms of Cadmium Uptake in Cadmium Accumulating Strains --- p.10 / Chapter 1.6 --- Techniques for Cell Immobilization --- p.13 / Chapter 1.7 --- Prospect --- p.20 / Chapter CHAPTER 2: --- ISOLATION OF CADMUIM ACCUMULATNIG MICROORGANISMS --- p.22 / Chapter 2.1 --- Introduction --- p.22 / Chapter 2.2 --- Materials and Methods --- p.25 / Chapter 2.2.1 --- Recipes Used for Growing Various Organisms --- p.25 / Chapter 2.2.2 --- Methods Used for Collecting Organisms to be Tested --- p.27 / Chapter 2.2.3 --- Observation of Samples by Microscope --- p.28 / Chapter 2.2.4 --- Enrichment of Cadmium Resistant Microorganisms --- p.28 / Chapter 2.2.5 --- Selection and Isolation of Cadmium Resistant Microorganisms --- p.29 / Chapter 2.2.6 --- Purification of Microbial Colonies --- p.30 / Chapter 2.2.7 --- Preliminary Classification of Selected Microorganisms --- p.30 / Chapter 2.2.8 --- Screening of Cadmium Accumulating Strains --- p.30 / Chapter 2.2.9 --- Cadmium Analysis --- p.31 / Chapter 2.3 --- Result --- p.32 / Chapter 2.3.1 --- Selection of Cadmium Resistant --- p.32 / Chapter 2.3.2 --- Cadmium Resistance of Isolates --- p.36 / Chapter 2.3.3 --- Screening of Cadmium Accumulating Microorganisms --- p.38 / Chapter 2.4 --- Discussion --- p.39 / Chapter CHAPTER 3: --- GENERAL CHARACTERIZATION OF STRAIN 1000A --- p.43 / Chapter 3.1 --- Introduction --- p.43 / Chapter 3.1.1 --- Various Factors Affecting the Accumulation of Cadmium of Strain 1000A --- p.43 / Chapter 3.1.2 --- Identification --- p.44 / Chapter 3.2 --- Materials and Methods --- p.45 / Chapter 3.2.1 --- "Preparation of Solutions, Antibiotics and Reagents" --- p.45 / Chapter 3.2.2 --- Culture Media Used --- p.47 / Chapter 3.2.3 --- Growth Kenetics Determination --- p.48 / Chapter 3.2.4 --- Determination of the Effect of Cadmium Concentration on Cd-uptake in Free Cells --- p.49 / Chapter 3.2.5 --- Determination of the Effect of Phosphate Concentration on Cd-uptake in Free Cell --- p.49 / Chapter 3.2.6 --- Determination of the Cd-uptake in Free Cells in Continuous Cultures --- p.50 / Chapter 3.2.7 --- Determination of Antibiotic Resistance of Strain 1000A --- p.51 / Chapter 3.2.8 --- Dstermination of Relationship between Chloramphenicol Resistance and Cd-uptake --- p.52 / Chapter 3.2.9 --- Cadmium Analysis --- p.52 / Chapter 3.2.10 --- Determination of Inorganic Precipitation of Cadmium --- p.53 / Chapter 3.2.11 --- Assimilation Tests --- p.54 / Chapter 3.2.12 --- Identification of Strain 1000A --- p.55 / Chapter 3.3 --- Result --- p.55 / Chapter 3.3.1 --- Growth Kinetics of Strain 1000A in Cadmium Supplemented Peptone Medium --- p.55 / Chapter 3.3.2 --- Cd-uptake of Strain 1000A at Various Cadiuin Concentration --- p.65 / Chapter 3.3.3 --- Effect of Phosphate concentration on Cd-uptake of Strain 1000A --- p.65 / Chapter 3.3.4 --- Cd-uptake of Strain 1000A in Continuous Cultures --- p.70 / Chapter 3.3.5 --- Inorganic Precipitation of Cadmium Phosphate --- p.75 / Chapter 3.3.6 --- Determination of Antibiotic-Resistance of Strain 1000A --- p.78 / Chapter 3.3.7 --- Effect of Chloramphenicol on Cd-uptake and Cadmium Resistance of Strain 1000A --- p.82 / Chapter 3.3.8 --- Determination of the Effect of Tetracyclin --- p.85 / Chapter 3.3.9 --- Assimilation Tests --- p.94 / Chapter 3.3.10 --- Identification of Strain 1000A --- p.94 / Chapter 3.4 --- Discussion --- p.97 / Chapter CHAPTER 4: --- DETERMINATION OF CADMIUM UPTAKE MECHANISM IN P. PICKETTI 1000A --- p.102 / Chapter 4.1 --- Introduction --- p.102 / Chapter 4.2 --- Materials and Methods --- p.105 / Chapter 4.2.1 --- Preparation of Solutions and Reagents --- p.105 / Chapter 4.2.2 --- Preparation of Reagents for SDS-PAGE --- p.105 / Chapter 4.2.3 --- Recipes for Growing Cells --- p.107 / Chapter 4.2.4 --- Protein Determination --- p.108 / Chapter 4.2.5 --- Examination of Cadmium Accommodation in P. picketti 1000A by Transmission Electron Microscope --- p.108 / Chapter 4.2.6 --- SDS-polyacrylamide Gel Electrophoretic Determination of Protein Profiles --- p.109 / Chapter 4.2.7 --- Phosphate Assay --- p.111 / Chapter 4.2.8 --- Orthophosphate Estimation --- p.112 / Chapter 4.2.9 --- Sulphide Analysis --- p.112 / Chapter 4.2.10 --- Cadmium Analysis --- p.113 / Chapter 4.2.11 --- Cd-binding Determination through Column Separation --- p.113 / Chapter 4.2.12 --- Cd-binding Determinate ion through SDS Electrophoresis --- p.114 / Chapter 4.2.13 --- Determination of Cadmium Distribution of Cells --- p.115 / Chapter 4.3 --- Result --- p.116 / Chapter 4.3.1 --- SDS-PAGE Determination of Protein Profiles of P. picketti 1000A --- p.116 / Chapter 4.3.2 --- Determination of Cd-binding Protein of P. picketti 1000A --- p.121 / Chapter 4.3.3 --- "Determination of the Relationship of Cellular Cadmium, Sulphide and Phosphate" --- p.131 / Chapter 4.3.4 --- Examination of Cadmium Accumulation of P. picketti 1000A by Transmission Electron Microscope --- p.142 / Chapter 4.3.5 --- Cadmium Distribution of Cadmium-Accommodated Cells --- p.148 / Chapter 4.4 --- Discussion --- p.152 / Chapter CHAPTER 5: --- CORRELATION AMONG METALS IN HEAVY METAL UPTAKE --- p.158 / Chapter 5.1 --- Introduction --- p.158 / Chapter 5.2 --- Materials and Methods --- p.158 / Chapter 5.2.1 --- Preparation of Solutions --- p.159 / Chapter 5.2.2 --- "Determination of Effect of Zn+2 ," --- p.160 / Chapter 5.2.3 --- Determination of Effect of Cu+2 . --- p.161 / Chapter 5.2.4 --- "Correlation among Cd+2, Cu+2 and Zn+2" --- p.161 / Chapter 5.2.5 --- Growth Kenetics Determination --- p.162 / Chapter 5.2.6 --- Cell Sample Preparation --- p.162 / Chapter 5.2.7 --- Orthophosphate Estimation --- p.162 / Chapter 5.2.8 --- Metal Analysis --- p.163 / Chapter 5.3 --- Result --- p.163 / Chapter 5.3.1 --- Effect of Zn+2 --- p.163 / Chapter 5.3.2 --- Effect of Cu+2 --- p.173 / Chapter 5.3.3 --- "Correlation among Cd+2, Cu+2 and Zn+2" --- p.178 / Chapter 5.4 --- Discussion --- p.195 / Chapter CHAPTER 6: --- HEAVY METAL UPTAKE OF IMMOBILIZED CELL --- p.197 / Chapter 6.1 --- Introduction --- p.197 / Chapter 6.2 --- Materials and Methods --- p.199 / Chapter 6.2.1 --- Preparation of Solutions and Medium --- p.199 / Chapter 6.2.2 --- Harvesting of Cells --- p.199 / Chapter 6.2.3 --- Immobilization of Cells --- p.199 / Chapter 6.2.4 --- Determination of the Effect of Temperature --- p.200 / Chapter 6.2.5 --- Determination of Optimum Cell Concentration in Polyacrylamide Gel --- p.201 / Chapter 6.2.6 --- Determination of pH Effect on Cd-uptake --- p.201 / Chapter 6.2.7 --- Pretreatment with 70% Methanol --- p.202 / Chapter 6.2.8 --- Combined Pretreatment with Methanol and NaOH --- p.202 / Chapter 6.2.9 --- Effect of Phosphate on Cd-uptake of Immobilized Cell --- p.202 / Chapter 6.2.10 --- Comparison of Cadmium- and Copper-uptakes in Cells Immobilized in K-carrageenan and in Polyacrylamide --- p.203 / Chapter 6.3 --- Result --- p.204 / Chapter 6.3.1 --- Effect of Temperature on Cd-uptake --- p.204 / Chapter 6.3.2 --- Determination of Optimum Cell Concentration in Polyacrylamide Gel --- p.204 / Chapter 6.3.3 --- Effect of pH on Cd-uptake of Immobilized Cells --- p.207 / Chapter 6.3.4 --- Effect of Methanol on Cd-uptake --- p.210 / Chapter 6.3.5 --- Combined Effect of pH and Methanol on Cd-uptake --- p.213 / Chapter 6.3.6 --- Effect of Phosphate on Cd-uptake of Immobilized Cells --- p.213 / Chapter 6.3.7 --- Comparison between Cadmium- and Copper-uptake of Cells Immobilized in K-carrageenan and in Polyacrylamide --- p.220 / Chapter 6.4 --- Discussion --- p.228 / Chapter CHAPTER 7: --- CONCLUSION --- p.232 / REFERENCES --- p.234

Enhanced Biological Phosphorus Removal (EBPR) is an established process in wastewater treatment that uses bacteria to reduce phosphorus levels below regulatory discharge limits. Recently, in light of growing political concern over phosphorus sustainability, EBPR has also been recognised as a platform from which phosphorus may be recovered and recycled onto land as fertiliser. Operating EBPR to optimise performance and efficiency is therefore extremely important, but remains a challenge due to poor understanding of these bacterial ecosystems. Raman spectroscopy is a non-invasive, label-free, culture-independent technique capable of analysing live, single cells. Despite its advantages, Raman spectroscopy has been applied to study EBPR bacteria in just a handful of studies and thus has a low profile in this field of research. More work is required to investigate potential areas of application for Raman spectroscopy in EBPR research. The principal thesis presented here is that Raman spectroscopy could be used as a tool to improve EBPR. The Raman spectra used for this investigation were acquired from individual EBPR bacteria dried onto a calcium fluoride substrate. The bacterial samples were collected from three different sources, namely lab-scale sequencing batch reactors located in Edinburgh (University of Edinburgh, UK) and Boston (Northeastern University, USA), and a full-scale EBPR plant in Slough (Thames Water, UK). Using these spectra, some potential applications and limitations of Raman spectroscopy for improving EBPR were explored. In this foundation work, a particular emphasis on spectral analysis methods was kept in light of the benefits of automating analysis as well as the need for standardisation to be able to compare results between different studies and groups. Nine methods were compared for baselining Raman spectra of individual EBPR bacteria. From these, the “small-window moving average” (SWiMA) method was determined to be the best baselining technique for our purposes at the current time. In agreement with earlier studies, the Raman spectroscopic signatures of three key EBPR metabolites – polyphosphate, polyhydroxyalkanoate (PHA) and glycogen – were shown to be clearly identifiable in individual EBPR bacteria when present. The Raman shifts of characteristic spectral bands arising from polyphosphate were shown to vary significantly between samples and the implications of this were discussed. Examples of how the Raman spectra of individual bacteria can be modelled with multivariate tools to open up new areas for research were given. MCR modelling was demonstrated to offer a novel way to normalise the Raman spectra of individual EBPR bacteria prior to quantitative analysis. With the instrumental set-up in this work, the limit of detection (LOD) of aqueous polyphosphate samples was estimated to be approximately 0.08 M and 0.02 M for 10 second and 200 second acquisitions respectively. Future work is required to research ways in which a more comparable form of polyphosphate ‘standard’ might be prepared so that direct correlation can be drawn between measurements made on such a standard and measurements made in bacterial cells. Overall, several applications and challenges of Raman spectroscopy for the investigation of EBPR bacteria are presented in this work together with recommendation for how to process the spectral data. The conclusions drawn from this work indicate that Raman spectroscopy could be used as a tool to improve EBPR but further work is required to refine and apply these methods.