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Microbubbles are small gas-filled bubbles with diameters ranging from 50 to 1 μm, and less than 200 nm are called nanobubbles. Their small sizes and large specific surface area result in a high gas dissolution rate and long lifetime in liquid. Ozone is a strong oxidant that destroys microorganisms and only produces oxygen as the final by-product in fresh water. However, due to the poor stability of aqueous ozone, critical gas waste happens during treatments which leads to a high economic loss. Microbubbles have shown promising enhancement of ozone treatment. In previous studies, ozone microbubbles exhibited excellent efficacy in the removal of organic contaminants and inactivation of microorganisms including bacteria, spores, and fungi, but few articles discuss the virus inactivation of ozone microbubbles treatment. Human noroviruses (NoVs) are the primary cause of foodborne illnesses in the US, and the development of effective inactivation methods is crucial. Because of the absence of suitable in vitro cultivation methods for NoVs and the constraints of the available infectivity models for these viruses, most of the studies about inactivation use surrogate viruses that are similar to NoVs in genetics and structure. Tulane virus is a NoV surrogate that can identify the same putative co-factor. This study focuses on the influence of treatment time, disinfectant air exposure, and the presence of organic contaminants on the inactivation efficacy of ozone microbubbles or millimeter bubbles. The results demonstrate that more than one log10 reduction was produced when the Tulane virus was exposed to ozone millimeter bubbles and ozone microbubbles for a short period of time, even in the presence of high organic load (FBS), and the protective effect of the organic load was shown when the disinfectant induced volume increased. The findings indicate that conducting further research on ozone microbubbles in aqueous applications in food-related applications is useful.

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Currently, human norovirus is the leading cause of acute gastroenteritis and accounts for most cases of foodborne illnesses in the United States each year. Due to its tissue culture inefficiency, studies of human norovirus have been crippled for more than forty years.Tulane virus (TV) stands out as a suitable surrogate of human norovirus given its high amino acid identity with human norovirus and its well-established cell culture system. It was first isolated from rhesus macaques (Macaca mulatta) in 2008 and identified as a novel Calicivirusrepresenting a new genus, Recovirus genus (Farkas et al., 2008). However, there are still unanswered questions about its infectious cycle and the essential factors for its infection. In this study, we have obtained a TV variant (the 9-6-17 strain) that has lost the binding ability to the B-type histo-blood group antigen (HBGA), which was proposed to be the receptor of both TV and human norovirus. In the first chapter, we outline how the sequence analysis,structural biology studies, and mutagenesis studies of the 9-6-17 TV strain have shed light on the interaction with its host cell receptor. To investigate the key residues for HBGA binding, we established the full-length infectious clone of the 9-6-17 TV strain. We present a highly selective transformation of serine 367, located in the predicted HBGA binding site, into a lysine residu e. Our results advance the understanding of genetic changes in TV required for adaptation to cell culture environments. Cryo-EM is an awarding winning technique that has been the greatest scientific breakthrough in recent years. It was awarded the Nobel Prize in Chemistry in 2017. Despite the technological advances of the direct electron detector and image processing software, several major roadblocks remain for high-resolution structure determination with cryo-EM. In the later chapters, we explored the most efficient way of using VPP to enhance image contrast, how to tackle the airwater interface problem by encapsulating target protein, how to reach a higher resolution by refining high order parameters, and the helical indexing problem in real space. These technical advances would benefit the whole cryo-EM community by providing convenient tools or insights for future directions.

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