Wastewater treatment has been a major issue in undeveloped and developing countries due to the lack of access to water filtration systems to treat wastewater. Without clean water resources, it has affected agriculture, water storage, and daily activities. According to Global Affairs Canada, eighty percent of illnesses in developing countries are linked to a lack of proper wastewater treatment. In this project, an eco-friendly and lower cost of chemical, ferrate, has been generated and analyzed to combat global wastewater treatment.To generate more stable and user friendly ferrate. The precursors of ferrate are synthesized in solutions. These solutions are freeze dried to form stable precursors in solid form. These precursors can be mixed to give an easy route to make ferrate on demand. More importantly, they can be added directly to wastewater to generate ferrate for disinfection. In the second part of this project we have come up with an analytical technique to quantify the concentration of ferrate in solutions using UV instrument.The ferrate solution can be easily transferred to developing countries to use in wastewater treatment without generating harmful chemicals to humans or the environment. In future research, the research will focus on the mass production of ferrate as well as creating a more stable form.

Separation of proteins using metal ligand complexes is a well-established practice in the field of bioengineering and biochemistry.  electrospray ionization mass spectrometry (ESI-MS) can be used to identify bio and organic molecules. Previous studies have detected chelated metal ions using ESI-MS, this project focuses on the identification of a metal ligand complex comprised of a tridentate chelating agent Iminodiacetic acid (IDA) and a bidentate ligand, Histidine. By coordinating copper with IDA a binary complex is formed, this allows for the detection of copper by proxy of the IDA. By selecting a unique fragment related to IDA we can target in tandem mass spectrometry (MS/MS) for greater sensitivity; the complex can be selected for analysis out of solution. We will use this novel approach to build a parent-ion scanning technique to monitor metal-ligand complexes extracted from environmental matrices. first the ideal solution parameters are determined to maximize the complex formation and detection of the Cu-IDA complex. So far, a high ratio of copper to IDA coupled with a basic buffer have yielded the best data. Creating a ternary complex comprised of copper IDA and an imidazole ring containing compound, histidine. Selectively tuning to the peaks associated with the copper IDA complex, the ternary complexes can be selected for in depth analysis of its structure and bonding properties. Future work could focus on identification of metal ligand complexes from soil samples with other compounds containing imidazole rings such as the neonicotinoid imidacloprid which has been indicated in bee colony collapse. By chelating solid with IDA complex formed from pentacoordinate copper ions could be detected despite the low relative concentration.

The CRISPR-Cas system is an adaptive immune system that provides protection to bacteria against bacteriophages. This system offers a clear advantage for bacteria as they need to defend themselves against the threat of bacteriophages. However, bacteria that have CRISPR systems can experience autoimmune damage, if they take up a spacer targeting a lysogen incorporated into their genome. This threat gives bacteria an incentive to encode CRISPR inhibitors. Through a metagenomics screen to identify novel inhibitors of the CRISPR-Cas9 system, it was found that a bacterial nickel binding chaperone protein, HypA, inhibits Cas9. This unexpected discovery may offer insight into how bacteria regulate their CRISPR systems and prevent autoimmunity. Through my work, I have determined that a diverse set of homologs can inhibit Cas9. I have also determined that HypA may act to destabilize Cas9’s structure. In order to investigate whether HypA can prevent autoimmune damage, I co-expressed HypA and a self-targeting Cas9, and found that HypA can protect the bacterial genome from Cas9. This work may provide insight into the evolution and regulation of the CRISPR-Cas system in bacteria, by helping explain how bacteria navigate the challenges caused by having an immune system. This work may also illuminate why some bacteria retain and others lose their CRISPR systems when they encode self-targeting CRISPR spacers. 

Probiotics are living organisms that when ingested have been linked with health benefits to the gut and improvement of conditions such as irritable bowel syndrome (IBS). The gut contains a plethora of microorganism populations that make up the microbiota. To understand how these populations communicate with each other, and the tissues surrounding them, it is imperative to identify and characterize the method of communication. Extracellular vesicles are one such possible method. Extracellular vesicles (EVs) are lipid-bilayer delineated sacks of material secreted from cells. It has been established that EVs are used as a waste disposal system. However, new research revealed that EVs can be used by the cell for methods of communication. Furthermore, EVs are now being linked to cell-to-cell and cell-to-organism communication. If EVs have been linked to communication, then characterizing them is one step closer to understanding how probiotic bacteria function. Previous studies have mainly characterized EVs by their size and divided them into 3 main groups: exosomes (40-150 nm), microvesicles (100-1000 nm), and apoptic bodies (>2000 nm). However, an analysis of proteins found in these EVs has not been performed yet. Here we compare EV proteins to proteins in the cell, to determine which protein fractions are secreted by cells through vesiculation for signaling purposes. To separate the cellular fraction from the EVs fraction, cell suspensions were centrifuged. First, the cells were pelleted and collected at 300x g. The leftover supernatant was spun at 16,000x g to pellet the EVs. Then, proteins from cells and EVs were solubilized and digested with trypsin. The tryptic peptides will be analyzed using liquid chromatography-mass spectrometry. A comparison of proteins in cells and EVs, and their relative concentrations can help us learn more about how probiotic EVs function in the gut.

Baylisascaris procyonis is a predominant parasitic infection of raccoons (Procyon lotor) in the Pacific Northwest, commonly referred to as “raccoon roundworm”. Raccoons serve as definitive hosts of the parasite, harboring adult worms in their intestine and shedding eggs in their feces. Infection can be spread to humans, dogs, birds and rodents through incidental consumption of eggs or other infected animals. Maturation of eggs occurs in the gut before larvae travel to other tissues including the liver, heart, lungs, brain and eyes. Infection causes encephalitis, liver damage, blindness, seizures, coma and death. Understanding the regional prevalence of B. procyonis infection is important for targeting resources for effective treatment. This research seeks to determine the prevalence of B. procyonis infection between geographically distinct raccoon populations of the Pacific Northwest. We expect a greater prevalence of B. procyonis infection in urban groups due to higher population densities. Fecal samples were collected between 2013 and 2020 from three categories of geographical regions: urban, rural and island. Samples were taken from the greater Seattle area (urban), surrounding regions of the Puget Sound (rural) and Blakely Island (island). Wet mounts were prepared from flotations using 1 g of fecal sample in aqueous ZnSO₄ or sugar solution. Samples were examined using light microscopy to identify the presence of B. procyonis eggs and nematode larvae. Current data shows a greater prevalence (p = 0.018) of B. procyonis eggs in urban populations compared to rural and island populations. There appears to be no difference in nematode larvae prevalence between geographical locations (p = 0.586) suggesting nematode infections in rural and island populations are likely not B. procyonis. This data provides valuable information to educate the public about the risk of B. procyonis infection and take preventative measures to protect humans and domestic animals. 

The search for renewable energy, electricity-generating wind turbines were first introduced by Charles F. Brush in 1888. Wind turbines use the principles of turning the mechanical energy of the wind into useful electrical energy that is able to produce work while also having no direct emissions towards the atmosphere. Using Betz’s law derived from the principles of conservation of mass and momentum of the air stream flowing, we construct and test a model wind turbine maximizing the power generated due to the varying angular velocities, from which testing data are used to iteratively design blade aerodynamics and assignation angle. For the particular model used thus far, the data indicate that with angular velocity lower than or equal to 4.124 rad/s and 13.359 rad/s a maximum efficiency of 22-23% is achieved. The blade designs are flat and angled blades, which are 3D printed and tested its efficiency on the model with pitches of 15°,30° and 45° to accommodate the motor to generate the optimal use of power input, therefore maximum power output. The result shows that 30° pitch is the most optimal angle for both blades design, with the flat blade generating 29000% more power than the angled blade. Blade pitch of 15° is the least efficient, resulting in no power generated with angled blade design, and significantly lower power in flat blade design compared to the other pitch. This discovery is essential to the future development of renewable energy especially in revolutionizing the wind turbine to be more efficient.