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Microalgae have been in the center of research for several years due to their high production rates. The use of fresh-water algae in the production of biofuels coupled with wastewater treatment has become a topic of modern research. While most algal farming is performed in warm and sunny climate, this project focused on naturally occurring microalgae in Northern Sweden and their ability to reclaim wastewater and produce lipids. The four microalgae Chlorococcum sp. (MC1), Scotiellopsis reticulata (UFA-2), Coelastrella sp. (3-4) and Chlorella vulgaris (13-1) were grown under mixotrophic conditions in municipal wastewater (MWW), pulp and paper wastewater (PnP) and mixtures of both. Except of UFA-2, I found the growth of the tested species to be limited in pure PnP, however, mixtures of PnP and MWW were suitable for algal growth. The removal rates of total nitrogen achieved the goals regulated by the Swedish government for wastewater reclamation. Phosphorus, of which maximal levels according the Swedish regulations need to be below 0.5 mg/L, was efficiently removed by the strain 3-4 in PnP and in MWW + PnP (ratio 3+1) as well as by the strain 13-1 in MWW + PnP (ratio 3+1). The tested microalgae are therefore appropriate candidates to be used in sustainable wastewater treatment. The algal biomass composition was determined with the help of Fourier-transform infrared spectroscopy and an increase in the spectra for biomass grown in wastewaters within the fingerprint region (800 – 1800 cm-1) was observed. From this, I conclude that the lipid content was elevated in the biomass received after growth in PnP and all mixtures of MWW and PnP compared to the in BG11 grown biomass, which acts as a control group. A quantitative lipid analysis performed on the biomass of the strains UFA-2 and 3-4 confirmed higher lipid amounts after growth in PnP wastewater compared to growth in BG11. These findings show that the examined microalgae might have the potential to be used as a potential feedstock for biofuel productions after cultivation in local wastewaters.

Based on results of its Aquatic Species Program (1978-1996), which sought to develop algae-to-liquid fuel technology, the U.S. Department of Energy has suggested that algal wastewater treatment may be incorporated into biodiesel production schemes to reduce the operating costs of both processes. the purpose of the current research was to evaluate the triglycerides produced by wastewater-grown algae for their suitability as a fuel feedstock and to investigate the effectiveness of several solvent mixtures and extraction procedures at recovering lipids from fresh algae. The research involved two separate experiments. The first determined the quantity and quality of lipids produced over the lifetime of a batch culture of algae grown in a small outdoor high-rate pond. Transesterification of the algal triglycerides yielded mostly saturated and monounsaturated 16 and 18-carbon fatty acid methyl esters, together comprising approximately 8 to 30% of the biomass in the pond. The average triglyceride production rate during the grwoth phase of the culture was 0.97 grams per square meter of pond surface per day. The second experiment compared several industrially practicable extraction procedures to the Bligh and Dyer laboratory extraction method. The Bligh and Dyer laboratory extraction procedure provides excellent lipid recovery efficiency, but several factors limit its potential on an industrial scale. The Bligh and Dyer method requires a larger volume of solvents than other methods, uses the probable carcinogenic chemical chloroform, and involves a complex series of steps that are difficult to automate. A simple, low-energy extraction process using relatively non-toxic solvents was found to have an extraction efficiency comparable to that of the laboratory method.

The combustion dynamics and stability are dependent on the quality of mixing and vaporization of the liquid fuel in the pre-mixer. The vaporization characteristics of different blends of biofuel droplets injected into the air stream in the pre-mixer are modeled in this current study. The focus of this work is on the blended alternate fuels which are lately being considered for commercial use. Two major alternate fuels analyzed are ethanol and Rapeseed Methyl Esters (RME). Ethanol is being used as a substitute for gasoline, while RME is an alternative for diesel. In the current work, the vaporization characteristics of a single droplet in a simple pre-mixer has been studied for pure ethanol and RME in a hot air jet at a temperature of 800 K. In addition, the behavior of the fuels when they are mixed with conventional fuels like gasoline and diesel is also studied. Temperature gradients and vaporization efficiency for different blends of bio-conventional fuel mixture are compared with one another. The model was validated using an experiment involving convection heating of acoustically levitated fuel droplets and IR-thermography to visualize and quantify the vaporization characteristics of different biofuel blends downstream of the pre-mixer. Results show that the 20 µm droplets of ethanol-gasoline 50-50 blend is completely evaporated in 1.1 msec, while 400 µm droplets vaporized only 65% in 80 msec. In gasoline-ethanol blends, pure gasoline is more volatile than pure ethanol. In spite of having higher vapor pressure, ethanol vaporizes slowly compared to gasoline, due to the fact that latent heat of vaporization is higher for ethanol. For gasoline-ethanol blended fuels, ethanol component vaporizes faster. This is because in blended fuels gasoline and ethanol attain the same temperature and ethanol vapor pressure is higher than that for gasoline. In the case of RME-diesel blends, initially diesel vaporizes faster up to 550K, and above this temperature, vapor pressure of RME becomes dominant resulting in faster vaporization of RME. Current work also looks into the effect of non-volatile impurities present in biofuels. Depending on source and extraction process, fuels carry impurities which impact vaporization process. In this work these effects on ethanol blended fuel have been studied for different concentration of impurities. The presence of non-volatile impurities reduces the vaporization rate by reducing the mass fraction of the volatile component at the surface. However, impurities also increase the surface temperature of the droplet. Finally, the effects of hot and cold spots in the prevaporizer have been investigated. Due to inefficient design, prevaporizer may have local zones where the temperature of air increases or decreases very sharply. Droplets going through these abnormal temperature zones would vaporize at a different rate than others. Current study looks into these droplets to understand the vaporization pattern.

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Plants are exposed to various environmental contaminants through irrigation with reclamation water and land application of municipal biosolids. Plants have been shown to take up contaminants from soil and groundwater, and to some extent, metabolize them in their tissues. These mechanisms have potential important implications for the environment and human health. First, as plants constitute the basis of the terrestrial food chain, accumulation of toxic chemicals or their metabolites inside plant tissues may lead to contamination of animals and humans. Second, the recognition of the capability of plants to take up and metabolize contaminants has led to the development of a plant-based remediation technology, referred to as phytoremediation. Phytoremediation is defined as the use of higher plants for the removal of environmental contaminants from soil and groundwater. Although phytoremediation is conceptually attractive as a green, environmental-friendly technology, the metabolism of xenobiotic compounds by plants is often slow and incomplete, possibly resulting in the accumulation of toxic pollutants and/or their metabolites inside plant tissues. Without further detoxification, phytoremediation may result in pollution transfer, potentially threatening the food chain, and eventually humans. Gaining further knowledge about the fate of environmental contaminants inside plant tissues is therefore of paramount importance for conducting environmental risk assessment and enhancing the efficiency of phytoremediation applications. It's an attractive concept today to cultivate plants on contaminated lands, in order to combine the benefits of phytoremediation with plant-based biofuel production. Unlike conventional plant bioenergy production, plant biomass grown on marginal contaminated soil will not compete with land for food production. However, the effect of contaminants on the plant biomass and bioenergy feedstock yield have received little attention. Molecular biology techniques, such as high-throughput gene expression analysis, constitute powerful tools to understand the molecular bases of the plant metabolism and response to environmental contaminants. The objective of this thesis is to understand the physiological and transcriptional responses of two model plants, thale cress (Arabidopsis thaliana) and soybean (Glycine max), exposed to various classes of contaminants, including silver nanoparticles (AgNPs), pharmaceuticals (zanamivir - ZAN and oseltamivir phosphate - OSP), explosives (2,4,6-trinitrotoluene - TNT), and polychlorinated biphenyls (PCBs). Detection of the contaminants inside plants tissues was performed using advance analytical methods, including inductively-coupled plasma - mass spectrometry (ICP-MS), gas-chromatography - mass spectrometry (GC-MS), and liquid-chromatography (LC-MS). The effects of contaminants on plants were assessed by recording various plant metrics, including biomass, root and shoot length, and soybean production. The transcriptional response of plants to exposure to selected contaminants (AgNPs, OSP, and ZAN) was investigated using whole-genome expression microarrays and reverse-transcription real-time (quantitative) PCR (RT-qPCR). In the first experimental phase of this research, the effects of AgNPs and soluble silver (Ag+) on A. thaliana plants were investigated. AgNPs are widely used nanomaterials, which have raised environmental concerns because of their toxicity to most living organisms, including plants. Exposure of hydroponic A. thaliana plants for 14 days to 20-nm AgNPs resulted in a slight increase of the biomass at low concentrations (1.0 and 2.5 mg / L) and a significant decrease of the biomass at higher concentrations (5.0 to 100 mg / L). Exposure to Ag+ for 14 days resulted in a significant reduction of the biomass after 14 days at concentration at and above 5.0 mg / L. Genome-wide expression microarrays revealed that exposure of A. thaliana to AgNPs and Ag+ at the concentration of 5 mg / L for 14 days resulted in differential expression of many genes involved in the plant response to stress and to biotic and abiotic stimuli. Although distinct gene expression patterns developed upon exposure to AgNPs and Ag+, a significant overlap of differentially expressed genes was observed between the two treatments, suggesting that AgNP-induced stress originated partly from silver toxicity and partly from nanoparticle-specific effects. In the second experimental phase of this research, the effects of the antiviral drugs, OSP and ZAN, on A. thaliana were investigated using an approach similar as the one described above. OSP and ZAN are pharmaceutical drugs that currently constitute the last line of defense against influenza infection. These drugs have been widely detected in wastewater effluents, especially during the influenza season, and they have the potential to contaminate agricultural plants through irrigation and land application of biosolids. Exposure of A. thaliana to OSP showed a significant decrease in the plants biomass at the concentrations of 20 and 100 mg / L, although no significant effect on the biomass was recorded upon exposure to ZAN (up to 100 mg / L), suggesting low acute toxicity of these compounds on plants. On the other hand, Arabidopsis exposure to OSP and ZAN at 20 mg / L resulted in significant transcriptional changes, including up- and down-regulation of many genes involved in the plant response to oxidative stresses and response to stimuli. Comparison with an Arabidopsis gene expression database (Genevestigator), revealed that many genes significantly up- and down-regulated by exposure to OSP and/or ZAN were similarly affected by exposure to biotic and abiotic stresses, toxic chemicals, and hormonal stimuli, suggesting that OSP and ZAN have negative chronic effects on plant health. The third experimental phase of this thesis focuses on the effects of two important persistent pollutants, TNT and PCBs, on the growth of soybean plants, with the objective of assessing the potential of using energy crops for the combined benefit of land remediation and biofuel (biodiesel) production. Explosives, such as TNT, are common toxic contaminants frequently observed at explosive manufacturing sites and military training ranges. PCBs are ubiquitous and toxic contaminants that are found in virtually every compartment of the environment. Short-term growth inhibition tests conducted with TNT and selected PCBs (e.g., 2,4'-dichlorobiphenyl - 2,4'-DCB) showed that these compounds exerted no or mild observable effects on plant growth even when applied at very high concentrations (i.e., 100 to 250 mg / kg soil, respectively). Analysis of TNT and 2,4'-DCB in exposed plant tissues showed average concentrations of 30 - 40 ng/g of TNT and 9,000 to 17,000 ng/g of 2,4'-DCB, which is consistent with biotransformation of TNT inside plant tissues. On the other hand, long-term exposure experiments show that exposure to TNT significantly affected soybean growth and production of bean in TNT-exposed plants (25 - 50 mg / kg soil). Exposure to TNT resulted in a significant decrease of the biomass of harvested beans after 120 days, which may have important consequences on the yield of biodiesel obtained from plants grown on contaminated land. Soybean were then exposed to 2,4'-DCB and its major transformation products, 4-OH-2,4'-DCB). Although high concentrations of the parent PCB (100 and 200 mg / kg) resulted in significant decrease of the biomass, high concentrations of the OH-metabolite resulted in increase of the plant biomass. Future research work will include the determination of the molecular bases of the effects - both positive and negative - of TNT, PCBs, and OH-PCBs on soybean plants and beans. / Civil Engineering

<p> In this thesis, the rate-limiting step is determined in a biofuel cell with a bio-anode, a Nation membrane and a conventional, platinum-based cathode using reference electrode method. It was discovered by surprise that the cathode overpotential dominated the cell overpotential. Na + in the membrane was found to hinder the W transport. The cathode overpotential increased due to the presence of Na + in the membrane and at the cathode. The limited H+ transport causes the increase of the cathode overpotential. H+ transport is the rate-limiting step in our biofuel cell, rather than commonly believed electron transport. Moreover, the cell power output degradation is not due to the conventionally believed depletion of the fuel substrate, inter-penetration of the fuel and oxidizer and the degradation of the biocatalysts, but the limited W transport in our biofuel cell. </p> <p> The existing oxygen reduction mechanism at the cathode was questioned and revised. When Na+ occupies all sulfonate groups in the membrane, only the Na+ from the buffer can pass through the membrane. The oxygen reacts with the water transported with Na+ and electrons to produce OH", which balances with the transported Na+ to keep electroneutrality at the cathode. </p> <p> Tris buffer without Na + was utilized as alternative anolyte in the biofuel cell. It was found that the cell with Tris buffer had a poorer performance in comparison with sodium phosphate buffer due to the increases of the anode and cathode overpotentials. Tris buffer does not constitute a solution to the problem. </p> <p> This work represents a step toward a more complete understanding of the properties of biofuel cells. To improve biofuel cell output, the herein identified H+ transport limitation in Na + contained Nation needs to be overcome. </p> / Thesis / Master of Applied Science (MASc)