This master thesis is based on own studies. A four-month study was performed at Hammarby Sjostad Research Station, which is located in Stockholm. One-stage deammonification process was evaluated in two different system configurations in pilot plant scale. The theoretical background for this thesis works is presented in the first part and where is presented negative impacts of nitrogen compounds in environment and requirements for purified wastewater in European Union (Sweden and Poland). In the next part of the thesis the nitrogen cycle is described and with focus on biological reactions for nitrogen removal. Especially, nitrification/denitrification and anammox processes are described with special focus on parameters affecting the anammox process performance and its advantages and disadvantages of using this process. Experimental results from the four-month study and evaluation of the microbial activity are described in the last part.

In this study NO2 and O3 gases which are the secondary pollutants in ambient air were sampled with designed passive tubes. The collected samples were extracted for UV-VIS spectrometric analysis. NO2 gases were converted to NO2- ions by extracting with water and then this solution was colored with Saltzman reagent (5:1 sulfanilamide and N-(1 naphthly)-ethlyene-diamine dihydrochloride). Then the absorbance of the solution was measured at 537 nm. O3 was extracted with 3-metyl-2-benzothiazolinone hydrozone hydrochloride solution which gives yellow color after extraction. The absorbance of the solution was measured at 430 nm. Before starting the experiment, the exposure time was optimized. For this purpose, 15 days in winter and 65 days in summer samplings were conducted. For both NO2 and O3 it was decided that 7 days of sampling period was required as an optimum sampling time in order to reach maximum collection efficiency values for both pollutants. To determine the uptake rates of the passive tubes, NO2 was sampled in fifteen weeks and O3 was sampled in thirteen weeks. During each sampling period passive tubes were placed nearby the active air sampling stations of Refik Saydam Hygiene Center, Air Quality Control and Research Laboratory in Ulus and Ke&ccedil / i&ouml / ren. Then uptake rates of NO2 and O3 passive tubes were determined by comparing passive and active sampler data. The uptake rate was calculated for NO2 and O3 as 0.91 x 103 cm3 h-1 and 1.71 x 103, respectively. Since sampling was done in ambient air the effect of meteorological parameters should be determined. Each meteorological parameters, wind speed, wind direction, relative humidity, temperature, pressure and solar ration were correlated with uptake rates. It was found out that none of these parameters had a significant effect on NO2 passive tube uptake rate. However, O3 passive tubes were affected from temperature, solar radiation and relative humidity. Temperature and solar radiation showed positive correlation with uptake rate, on the contrary, relative humidity was inversely correlated with uptake rate. Finally the pollution map of Ankara was created by taking samples from forty points in the city.

This thesis evaluates the performance of a deammonification system designed as a two-step tech-nology consisting of an initial partial nitritation followed by an Anammox process. Operation of a technical-scale pilot plant at the Himmerfjärden Wastewater Treatment Plant (Grödinge, Swe-den) has been assessed. Oxygen Uptake Rate (OUR) to evaluate the respiration activity of nitrifi-ers in the system and batch tests to assess reaction rates have also been applied in the study. It was found that the total inorganic nitrogen elimination strongly depended on the nitrite-to-ammonium ratio in the influent to the Anammox reactor, which was correlated with the per-formance of the partial nitritation phase. Therefore, a control strategy for oxidation of ammo-nium to nitrite has been proposed. Controlled oxygen supply to the partial nitritation reactor is obligatory to obtain a proper pH drop indicating oxidation of ammonia to nitrite at the adequate ratio. A very high nitrogen removal efficiency (an average of 84%) and stable operation of the system have been reached. Conductivity measurements were also used to monitor the system influent nitrogen load and the nitrogen removal in the Anammox reactor. The data gathered from the operation of the pilot plant enabled the use of multivariate data analysis to model the process behaviour and the assessment of the covariances between the process parameters. The options for full-scale implementation of the Anammox systems have been proposed as a result of the study. / QC 20101115

Understanding how nitrifiers react when exposed to low DO conditions could provide a greater understanding of low DO operations in full-scale biological wastewater treatment. Previous methods to observe nitrifier oxygen kinetics do exist in literature, however they are inefficient and labor intensive. Other more efficient methods require the use of selective inhibitors, which alter the characteristics of the biomass. This study developed a time and labor efficient respirometric method to distinctly measure oxygen half-saturation coefficients for both ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) without the use of selective inhibitors. By eliminating the use of inhibitory substances, representative biomass characteristics were maintained throughout the tests. The developed method, called the declining DO method, consisted of using a high-speed dissolved oxygen (DO) probe to measure relative oxygen uptake rates (OUR) within a batch reactor when varying substrates (ammonia and nitrite) were present in excess within the system. A forward model was developed based on Monod kinetics to simultaneously fit Monod curves to the experimental OUR data. These curves were fit by solving for optimum oxygen kinetic parameters representing endogenous respiration, NOB, and AOB. An inverse model using Markov chain Monte Carlo analysis was applied to the results found in the forward model to provide statistical validation of the proposed respirometric method. A separate method, called the substrate utilization rate test, was conducted in parallel with the declining DO tests to compare and verify oxygen half-saturation coefficient results. Parallel tests were conducted using biomass samples from three different Hampton Roads Sanitation District (HRSD) full-scale facilities. Operating conditions between the three HRSD facilities were considered when performing parallel testing, including averages for DO, solids retention time (SRT), and floc size. Average floc size was found to have a significant effect on the observed oxygen half-saturation values. Observed trends for the KO values estimated using the two methods remained consistent throughout all tests, where KO,NOB was always lower than KO,AOB. The comparison of the two methods highlighted some faults associated with the substrate utilization rate test, which is commonly used in literature to observe nitrifier oxygen kinetics. The declining DO method appeared to be more resistant to potential experimental error and required less than half the time compared to the substrate utilization rate test. The development of the declining DO method without the use of selective inhibitors provided a more time and labor efficient technique for estimating apparent KO values for NOB and AOB without sacrificing biomass characteristics representative of the full-scale treatment process. Biomass samples collected from variable treatment process conditions yielded consistent parallel test results, providing further evidence that the proposed declining DO method can be a robust and reliable technique for distinctly measuring apparent oxygen half-saturation values for NOB and AOB. / Master of Science / Wastewater treatment operations utilizing biological nitrogen removal (BNR) require a continuous supply of oxygen for aerobic processes. Energy costs associated with aeration generally accounts for at least 50% of the total energy consumption at conventional activated sludge wastewater treatment facilities. Operating aerobic zones at low average dissolved oxygen (DO) concentrations could be an effective way to significantly reduce aeration costs as well as material costs associated with BNR treatment processes. This study developed a method to measure oxygen kinetics for the two groups of autotrophic bacteria responsible for performing nitrogen removal. The method consisted of measuring relative oxygen uptake rates (OUR) within a batch reactor when varying substrates were available. This method is unique from previously developed techniques in that the use of selective inhibitors was not included, meaning the characteristics of the wastewater were largely unchanged and therefore better represent biomass conditions within the full-scale process. The results of the proposed method were verified using an alternate method for estimating oxygen kinetics. These two methods were conducted in parallel using biomass samples from several full-scale Hampton Roads Sanitation District wastewater treatment facilities utilizing a variety of process designs and operating conditions. Consistent results obtained between the two methods suggested the proposed method is an effective technique for distinctly measuring nitrifier oxygen kinetics.

The assessment of volatile organic compounds (VOCs) has become an area of particular interest in the field of atmospheric pollution due to their adverse health and environmental effects. This study is aimed to identify, quantify and characterize VOC in different urban areas and industrial areas in Bursa. The spatial distribution, seasonal variation as well as health risks assessment of VOC were discussed. Air samples were collected by means of sorbent passive sampling at over 50 sampling points in Bursa and analyzed by GC-thermal desorption. A total of seven weekly measurement periods were completed across the city center from 2005 to 2007. The source of VOC was apportioned with the commonly used receptor model, namely Factor Analysis. Motor vehicles and industrial solvent usage are the most abundant VOC sources to contribute to urban atmosphere in Bursa, 63% and 20%, respectively. The health risks of VOC were also evaluated. The questionnaire was filled out by selected people living in Bursa to obtain time-activity pattern for health risk assessment. Lifetime cancer risks were estimated with the measured VOC data. All the statistical parameters used to represent cancer risks for the selected compounds exceeded the stated level. Benzene had the highest adverse health effect among these compounds according to USEPA (2009) calculation with the mean cancer risk 3x10-4. In addition to intensive field sampling campaigns conducted in Bursa, the VOC measurement were completed for a period of six months to evaluate uptake rate of VOC in Ankara. Uptake rate equations depending on relative humidity and wind speed were developed for 25 VOCs.

<p>This thesis evaluates the performance of a deammonification system designed as a two-step tech-nology consisting of an initial partial nitritation followed by an Anammox process. Operation of a technical-scale pilot plant at the Himmerfjärden Wastewater Treatment Plant (Grödinge, Swe-den) has been assessed. Oxygen Uptake Rate (OUR) to evaluate the respiration activity of nitrifi-ers in the system and batch tests to assess reaction rates have also been applied in the study. It was found that the total inorganic nitrogen elimination strongly depended on the nitrite-to-ammonium ratio in the influent to the Anammox reactor, which was correlated with the per-formance of the partial nitritation phase. Therefore, a control strategy for oxidation of ammo-nium to nitrite has been proposed. Controlled oxygen supply to the partial nitritation reactor is obligatory to obtain a proper pH drop indicating oxidation of ammonia to nitrite at the adequate ratio. A very high nitrogen removal efficiency (an average of 84%) and stable operation of the system have been reached. Conductivity measurements were also used to monitor the system influent nitrogen load and the nitrogen removal in the Anammox reactor. The data gathered from the operation of the pilot plant enabled the use of multivariate data analysis to model the process behaviour and the assessment of the covariances between the process parameters. The options for full-scale implementation of the Anammox systems have been proposed as a result of the study.</p>

<p>This master thesis in energy optimization was made during the autumn of 2006 at Käppala wastewater treatment plant in Lidingö, Stockholm. A preceding thesis, where all electricity consumption was mapped, showed that the aeration in the biological treatment is the single largest consumer in the plant, and it is therefore of interest to reduce this cost. The oxygen control strategy used at Käppala WWTP is working well from a nutrient removal point of view, but not from an economic one. The last aerobic zones have a very low oxygen consumption during low loading periods which give rise to enhanced dissolved oxygen concentrations with excessive costs and reduced denitrification as a result. But also during periods of normal loading unnecessary high oxygen concentration are sometimes given.</p><p>By modifying the aeration control strategy three full-scale experiments have been made, with the intention to reduce the air consumption. The experiments were carried out during week 37-50 in the autumn of 2006 and showed that savings could be made.</p><p>The regular oxygen control at Käppala WWTP controls the oxygen level in the aerobic compartment with two DO-setpoints; one in the first aerobic zone and one in the last. The zones in between are controlled by an airflow fractionation depending on the oxygen level in the first and last zone. In the first strategy to be evaluated, all four zones in the aerated part were individually controlled with its own setpoint. Two different setpoint combinations were tested. By using the fact that the efficiency in the oxygen transfer rate was higher at low airflows, savings of approximately 16 % were achieved. In the second strategy tested, an ammonia-feedback control combined with a DO-feedback controlled the DO-set point in the first aerobic zone. This strategy adjusted the DO- set points to the loading variations, and this gave a decreased airflow of approximately 9 %. Finally the two strategies were combined. All zones were then controlled individually with DO-set points set by an ammonium-feedback and a DO-feedback. The strategy gave savings in the airflow of approximately 18 %. In all three trials the aerated zones were more efficiently used, and the estimated savings are 550 000 SEK/year, and with a preserved nutrient removal efficiency.</p> / <p>Under hösten 2006 har ett examensarbete om energieffektivisering på Käppalaverket på Lidingö utförts. Ett föregående examensarbete där all elenergiförbrukning kartlades visade att blåsmaskinerna i biosteget står för den enskilt största förbrukningen i verket och det är därför av intresse att minska denna kostnad. Syrestyrningsstrategin som används på Käppalaverket fungerar mycket bra ur reningssynpunkt, men är inte optimal ur energisynpunkt. Dels luftas de första aeroba zonerna för mycket vid låg belastning vilket ger upphov till kraftigt förhöjda syrekoncentrationer i de sista aeroba zonerna med höga luftningskostnader och risk för försämrad denitrifikation, men även under normal belastning har det visat sig att onödigt höga syrekoncentrationer ibland ges.</p><p>Tre fullskaliga optimeringsförsök har utförts, med syfte att minska luftförbrukningen med bibehållen reningsgrad. Försöken pågick från vecka 37 till 50 hösten 2006, och visade att det finns möjlighet att spara energi genom att modifiera syrestyrningsstrategin.</p><p>Den reguljära syreregleringen i Käppalaverket styr syrehalten i den aeroba bassängen mot två syrebörvärden; ett i den första luftade zonen och ett i den sista. Luftflödet till de mellanliggande zonerna styrs av luftflödesandelar beroende på syrehalten i dessa två zoner. Den första strategin som utvärderades styrde istället samtliga zoner individuellt med egna börvärden, där två olika strukturer på de satta börvärdena användes. Genom att utnyttja en högre effektivitet i syreöverföringshastigheten vid låga luftflöden uppnåddes luftflödesbesparingar på ca 16 % i första försöket.</p><p>I den andra strategin styrdes syrebörvärdet i den första luftade zonen med hjälp av två återkopplingar, en från utgående ammoniumhalt och en från syrehalten i den sista luftade zonen.</p><p>Tack vare att strategin anpassade syrebörvärdena efter belastningen av syretärande ämnen erhölls luftflödesbesparingar på ca 9 %. Slutligen kombinerades de två strategierna; samtliga zoner styrdes individuellt med börvärden satta av en ammonium-återkoppling och en syre-återkoppling. Strategin medförde luftflödesbesparingar på ca 18 %. I samtliga försök utnyttjades de luftade zonerna bättre, och besparingspotentialen uträknad från 2005 års elpriser blev som mest 550 000 SEK/år, detta med en bibehållen reningsgrad.</p>

Estudou-se nesta pesquisa o processo de lodos ativados para pós-tratamento do efluente proveniente de um reator anaeróbio compartimentado, que recebe esgoto sanitário. Isso se deu em vários ensaios em sistema descontínuo (\"batch\'\') para avaliar a biodegradabilidade dessa água residuária e em um ensaio em sistema contínuo com finalidade de melhorar a qualidade do efluente final e determinar os coeficientes cinéticos, os parâmetros a\' e b \', a taxa de utilização de oxigênio e o coeficiente &#945. Também realizaram-se ensaios hidrodinâmicos para verificação do tipo de reator utilizado. A pesquisa desenvolveu-se com base em uma célula de aeração, em escala de laboratório, composta por quatro câmaras (reatores) que possuíam tanque de aeração e decantador secundário justapostos, separados por uma cortina fixa. Não houve controle de temperatura nos dois sistemas utilizados. No ensaio em sistema contínuo quatro reatores foram operados simultaneamente com as respectivas idades do lodo de 5, 10, 15 e 20 dias durante 35 dias. O tempo de detenção hidráulico foi mantido próximo de 8 horas. Nesse ensaio os reatores quando se encontravam estabilizados dinamicamente, apresentaram remoção superior a 95% para DBO bruta, 90% para DBO filtrada, 87% para DQO bruta, 80% para DQO filtrada e 95% para SST. Mesmo não sendo realizado processo adicional para remoção de nutrientes, os reatores, após a estabilização dinâmica, apresentaram remoção de fosfato de até 28% e de nitrogênio amoniacal até níveis de não serem detectados pelos métodos de análises utilizados. Os ensaios hidrodinâmicos mostraram que os reatores são do tipo mistura completa. / In this work the activated sludge process for post-treatment of effluent anaerobic baffled reactor which receives sanitary sewage was studied. This was established during various experiments in batch system to evaluate the biodegradable of this wastewater and also in an experiment of continuous system aiming to improve the quality of the final effluent and to determine the kinetic coefficients, the a\' and b\' parameters, the oxygen uptake rate and the &#945 coefficient. Hydrodynamic tests to verify the type of reactor used in this research were also perfonned. The work was developed and based on an aeration cell bench-scale, composed of four chambers (reactors) which had an aeration tank and a secondary settling tank in the same chamber, separated by a fixed baffle. Temperature control was not used in both systems. In the experiment of continuous system, four reactors were simultaneously operated with sludge age of 5, 10, 15 and 20 days during 35 days. The hidraulic retention time was maintained at around 8 hours. When the reactors were dynamically stabilized, they showed an average removal superior to 95% for BOD (biochemical oxygen demand), 90% for filtrated BOD, 87% for COD (chemical oxygen demand), 80% for filtrated COD and 95% for TSS (total suspended solids). Without additional process of nutrient removal, after the dynamic stabilization the reactors presented up to 28% of total phosphate removal and ammonia nitrogen up to levels of not being detected by the analyses methods used. The hydrodynamic tests showed that the reactors are of the complete-mix type.

Poly- and perfluoroalkyl substances (PFAS) are ubiquitous contaminants across the globe, can bioaccumulate in aquatic taxa, and potentially biomagnify in food webs. Consequently, research examining the influence of PFAS on wildlife is warranted. Amphibians are sensitive to contaminants such as PFAS because of their porous skin and associations with aquatic habitats where contaminants accumulate. Because PFAS tend to bioaccumulate and can adversely affect the endocrine system, there is a need to examine uptake rates to inform ecotoxicology studies, as well as a need to examine sublethal effects. To address these knowledge gaps I conducted two experiments. First, I exposed larval northern leopard frogs (<i>Rana pipiens</i>), American toads (<i>Anaxyrus americanus</i>), and eastern tiger salamanders (<i>Ambystoma tigrinum</i>) to PFAS chemicals perfluorooctanoic acid (PFOA) or perfluorooctane sulfonate (PFOS) at concentrations of 10 or 1000 ppb for 10 days and sampled them every 48 hours during the exposure period. In the next experiment, I examined the effects of PFAS exposure via contaminated substrate on the survival and growth of post metamorphic amphibians of the same species. I found that, for all species, body burdens often reached steady state within 48 to 96 h of exposure. Steady-state body burdens of PFOA ranged from 3,819–16,481 ng/g dry weight among treatments and species (corresponding BCFs of 0.5 to 2.5), while PFOS body burdens ranged from 6,955–489,958 ng/g dry weight (corresponding BCFs of 47–259) among treatments and species. These data suggest that steady state is rapidly reached in larval amphibians exposed to PFAS, particularly regarding PFOS. This reflects a high potential for trophic transfer of PFAS within food webs because amphibians are often low in trophic position and are important prey for many aquatic and terrestrial species. In post-metamorphic amphibians, there was no influence of PFAS on survival or mass. However, significant effects on snout-vent length were observed in all species, and body condition differences were observed for two of my species. I found that all leopard frogs increased in scaled mass index (SMI) when exposed to a PFAS treatment, indicating an increased body condition. Toads exhibited a more variable SMI pattern across treatments, with no outstanding trends, and tiger salamanders did not differ significantly across treatments. These data suggest that sublethal effects vary greatly depending on the species, possibly due to life history traits. Future research examining biomagnification potential is warranted to determine the influence of PFAS on food webs. Additionally, there is a need to determine the physiological mechanisms underlying the observed effects of PFAS exposure.<br>

Escherichia coli (E. coli) is an attractive host for production of recombinant proteins, since it generally provides a rapid and economical means to achieve high product quantities. In this thesis, the impact of the glucose uptake rate on the production of recombinant proteins was studied, aiming at improving and optimising production of recombinant proteins in E. coli. E. coli can be cultivated to high cell densities in bioreactors by applying the fed-batch technique, which offers a means to control the glucose uptake rate. One objective of this study was to find a method for control of the glucose uptake rate in small-scale cultivation, such as microtitre plates and shake flasks. Strains with mutations in the phosphotransferase system (PTS) where used for this purpose. The mutants had lower uptake rates of glucose, resulting in lower growth rates and lower accumulation of acetic acid in comparison to the wild type. By using the mutants in batch cultivations, the formation of acetic acid to levels detrimental to cell growth could be avoided, and ten times higher cell density was reached. Thus, the use of the mutant strains represent a novel, simple alternative to fed-batch cultures.   The PTS mutants were applied for production of integral membrane proteins in order to investigate if the reduced glucose uptake rate of the mutants was beneficial for their production. The mutants were able to produce three out of five integral membrane proteins that were not possible to produce by the wild-type strain. The expression level of one selected membrane protein was increased when using the mutants and the expression level appeared to be a function of strain, glucose uptake rate and acetic acid accumulation. For production purposes, it is not uncommon that the recombinant proteins are secreted to the E. coli periplasm. However, one drawback with secretion is the undesired leakage of periplasmic products to the medium. The leakage of the product to the medium was studied as a function of the feed rate of glucose in fed-batch cultivations and they were found to correlate. It was also shown that the amount of outer membrane proteins was affected by the feed rate of glucose and by secretion of a recombinant product to the periplasm. The cell surface is another compartment where recombinant proteins can be expressed. Surface display of proteins is a potentially attractive production strategy since it offers a simple purification scheme and possibilities for on-cell protein characterisation, and may in some cases also be the only viable option. The AIDA-autotransporter was applied for surface display of the Z domain of staphylococcal protein A under control of the aidA promoter. Z was expressed in an active form and was accessible to the medium. Expression was favoured by growth in minimal medium and it seemed likely that expression was higher at higher feed rates of glucose during fed-batch cultivation. A repetitive batch process was developed, where relatively high cell densities were achieved whilst maintaining a high expression level of Z. / QC 20110608