The Lake Washington, and Sammamish basin contains a complex mix of life history strategies of Oncorhynchus nerka. These life history strategies include 1) Anadromous sockeye that remain in the lake for one or two years before migrating to the ocean and returning to fresh water as mature adults; 2) “Residual” sockeye salmon that breed with and are genetically a part of the anadromous population but do not migrate to the ocean, and 3) “kokanee” salmon that are genetically distinct from anadromous and residual sockeye. Kokanee salmon are native to the basin and are thought to have evolved when glaciers or other barriers restricted access to the ocean. Although once extremely abundant throughout the basin, native kokanee are now thought to be found only in Lake Sammamish. Large numbers of kokanee-like fish continue to occasionally migrate from Lake Washington into the Sammamish river and its tributaries. Intriguingly, these “mystery nerka” migrate and spawn later than the sockeye/residual population and may represent a fourth distinct O. nerka population (e.g. a remnant of native Lake Washington kokanee, or a newly evolved kokanee population derived from sockeye ancestors). Sockeye, residuals, and kokanee use gill rakers, bony extensions in the throat, to capture prey. In other cases where kokanee have evolved from sockeye ancestors, the number and size of gill rakers differs – a reflection of the of the different types environments they mature in and the different types of prey available in freshwater vs. saltwater. In this research we document the variation in gill raker number and length in the Lake Washington/Sammamish populations of O. nerka in an attempt to 1) investigate trophic adaptations within the basin associated with life history strategy and location of maturation, and 2) to assess the relationship of “mystery nerka” to the other populations known to occur in the basin.

Freshwater is one of the most valuable resources in Washington State. In recent decades, water supply has been affected due to climate change and population growth. Understanding changes in water supply and demand is crucial for ensuring an abundance of water for residential, economic, and industrial needs. The proposed research analyzes changes in the streamflow regime of the Cedar and Tolt Rivers which provide drinking water for the greater Seattle area. The main goal is to calculate the water budgets for the Cedar and Tolt watersheds and estimate how the inputs and outputs to these budgets change over the 21st century. An existing ensemble of streamflow projections for the Cedar and Tolt Rivers are used to analyze changes in water supply. The mean streamflow for each month is compared between a 30-year control period (water years 1971-2000) and a 30-year future period (water years 2031-2060). For each of these periods, I determine “optimistic” and “pessimistic” scenarios for the streamflow. For the “drought” month the highest streamflow value is considered as “optimistic”, and the lowest as “pessimistic” since the goal is to assess potential shortages. I use existing monthly demand values provided by Seattle Public Utilities and create different future scenarios, based on the predictions of population and employment growth. Supply and demand values are compared to evaluate (1) the potential for water shortage and (2) water management and conservation methods to satisfy the unmet demand. One potential water management method is the construction of a new reservoir. The results of the research are aimed at helping to inform society and water managers about the potential changes in the water system. Based on this information, they might be able to introduce changes in their future plans to accommodate the predicted needs.

Climate change has increased the prevalence and intensity of wildfire events in the Pacific Northwest. Exposure to wildfire smoke has been linked to several adverse health outcomes, most notably respiratory-related morbidity and mortality. Local public health response to reducing a community’s exposure has primarily focused on communicating risks to the public and provide suggestions to reduce exposure risk. Effectively communicating the health effects of wildfire smoke and the interventions possible to reducing exposure is imperative for reducing adverse health outcomes during smoke events. In order to discern the messages the public received, we conducted a content analysis of wildfire smoke risk information communicated by local and state government organizations and by the mainstream media during the August 2018, statewide wildfire smoke event in Washington state. The Extended Parallel Process efficacy and risk framework was used to assess the presence and message content of wildfire smoke risk and efficacy information. Summary statistics were calculated to identify common messages about efficacy recommendations, vulnerable populations, and trusted sources of public health information. With these results, we hope public health jurisdictions will better understand the extent of their wildfire smoke risk messages and improve their standing as trusted sources of information in the community. Through the use of the extended parallel process framework-- a “recipe” for future wildfire season communication can be created.

Short-term exposure to diesel exhaust particulate matter can cause headaches, dizziness, and irritation of the nose, throat, and eyes. Prolonged exposure has been shown to increase the risk of developing cardiovascular disease, respiratory disease and lung cancer. Because heavy machinery is often fueled by diesel, occupations such as coal miners, truck drivers, railroad workers, and construction workers are at high risk of exposure. Non-occupational exposures are also of concern, especially in locations impacted by high volumes of vehicle traffic. A potential way to determine diesel exhaust exposure is by measuring the amount of nitrated polycyclic aromatic hydrocarbons (NPAHs) in household dust. There is no current method for measuring NPAHs in dust. We conducted a literature review of methods for measuring related chemicals in household dust and measuring NPAHs in other matrices. We tested three different methods before establishing the optimized sample preparation and cleanup process using silica gel solid phase extraction followed by analysis using high performance liquid chromatohraphy with tandem mass spectrometry detection (HPLC/MS/MS). To evaluate method performance we analyzed replicates of spiked and unspiked household dust and silica gel. The method was used to analyze 42 household dust samples collected in two communities – one with expected high and one with expected low traffic-related air pollution. Future research should include comparisons between dust NPAH measurements and other measures of diesel exhaust exposure, including NP (nitropyrene) metabolites in urine, air filter NP, or a priori predicted exposure based on home location.

In the era of new technologies, the functions of mobile apps cover all aspects of our lives. Social networking apps expand our social and business groups, as well as increase job and entertainment opportunities dramatically. Other types of apps, such as travel and buying & selling apps, enable people to get both tangible and intangible products without leaving home. The fast development of mobile apps, however, made it difficult for elders over 65 to understand and learn, and the physical condition of elders presents barriers to operating new technologies. Overall, the lack of accessible design on mobile apps has caused elders to not have the equal opportunity to obtain information and enjoy the same conveniences as other age groups; and new technologies have gradually made the older generation feel abandoned. In the past six months, I conducted a competitive analysis of the current situation of using mobile apps by the elderly to gain a broad view on the user experience of mobile apps for elders. I designed and distributed a questionnaire survey among members of the Yakima Serious Table Tennis Club to understand the barriers they encountered when using various types of mobile apps. I interviewed two retirees from different fields of employment and cultural backgrounds about their views on the usability of different kinds of mobile apps. Ultimately, referring to my collected data and the Web Content Accessibility Guidelines version 2.1, I analyzed possible changes that could be made by some popular apps to improve the accessibility for elders. I created interface templates of an accessible mobile social media for elders using Adobe Experience Design; then I edited it to a final version while doing usability tests on the templates. The conclusions from this research could be used to help design applications that are suitable for more age groups, and to ultimately make society more inclusive by letting elders have equal opportunity to enjoy new technologies.

 Antibiotic resistance is a growing threat to public health. This resistance, coupled with a dearth of new antibiotics, makes the study and development of antibiotics of critical importance. Many antibiotics derive from microbial pathways that synthesize complex natural products, and engineering these pathways to produce new antibiotics is an exciting prospect. I aim to use such a strategy to produce variants of the klebsidin lasso peptide, known for its distinct lariat knot structure and its antibiotic activity. Klebsidin is produced by three biosynthetic enzymes that modify and export a ribosomally-synthesized precursor peptide. We have developed an assay that relies on cellular growth to (1) investigate how mutation influences the antibiotic activity and biosynthesis of klebsidin, and to (2) identify variants of klebsidin that overcome a known resistance mutation. When expressed in E. coli, klebsidin inhibits growth of its host cell. I expressed a library of approximately 10,000 klebsidin variants, each within a single cell, and used next-generation DNA sequencing to count the frequency of each variant before and after a growth selection. Functional klebsidin variants should decrease in frequency after selection, whereas null variants should increase in frequency. Using these sequencing data, I generated functional scores for a majority of single amino acid mutations within klebsidin, identifying positions that tolerate mutation and positions for which mutation abolishes bioactivity. To better understand these trends, I am performing mass spectrometry experiments to study the biosynthesis of key variants. In addition, we are interested in how mutagenesis of klebsidin can combat known resistance to lasso peptides. By performing the same growth selection in an E. coli strain resistant to wild type klebsidin, we hope to identify variants that overcome this resistance. Together, these studies will provide a framework for understanding lasso peptide engineering and identifying novel peptide-based antibiotics.

The Methow Valley is a community located in Okanogan County, WA that has experienced wildfires in 4 of the past 5 years. The Methow Valley Clean Air Project (MVCAP) is a local volunteer group that promotes air quality awareness through outreach and programming. MVCAP’s Purple Air Network was designed to provide access to spatial air quality information to help community members make decisions about protective actions, to identify relatively clean air spaces, and to serve as a public health invervention for wildfire smoke. In collaboration with MVCAP, we worked to install a network of 20 purple air monitors in the summer of 2018. The monitors were placed in homes of “Clean Air Ambassadors” who committed to maintain and promote the monitor. For calibration, each monitor was placed according to federal guidelines and two were collocated with nephelometers maintained by Washington Department of Ecology. During the summer of 2018, two wildfires burned nearby which allowed the monitors to be utilized in a wildfire smoke event. The data from the monitors was downloaded and compared to the nephelometer data using linear regression. I worked to establish a correction factor by analyzing the data and found that the Purple Air monitor over reported the PM2.5 concentration by a factor of 0.53 and even more at high concentrations. The network proved the importance of high spatial concentration monitoring by capturing the air quality variation. In some areas, the air quality was good while in others it was hazardous. Having the network is a tool for individuals to know what the air quality is near their homes and other places in their community. It allowed individuals to access local air quality data and make choices about poor air quality. The next step for MVCAP is designing an online interface that applies the correction factor directly to accurately communicate risk.

Campylobacter jejuni is a Gram-negative spirochete bacteria that colonizes the guts of many bird species. They are pathogenic to humans and typically consumed with undercooked poultry or contaminated water. In this study, two species of crow, separated by 7,000 miles, have been examined for their ability to carry C. jejuni. Crow fecal samples were collected from Bothell, Washington (WA) and Kolkata, India (KK). All of the crow Campylobacter isolates were confirmed by qPCR and PCR methods to be C. jejuni. No other species are isolated. We compared these isolates by method of MLST (Multilocus Sequence Typing) and searched for known allelic profiles on the PubMLST database.Thirteen  isolates were analyzed for seven housekeeping genes : aspA, glnA, glyA, gltA, pgm, tkt, uncA. Using primers specified in the PubMLST database portions of the genes were amplified by PCR, verified by gel-electrophoresis and sent to Eurofin Genomics for sequencing. Once the sequence files were returned, they were aligned and assembled using Mega7 software. An allele number was obtained for each sequence for each sequence from PubMLST database. The allelic profile of the 7 genes for each isolate was used to obtain the Sequence type (ST) of the isolate. MLST of 15 crow isolates that represented different phylogenetic clusters based on fla-SVR sequencing, showed a majority of the WA isolates to belong to sequence types ST2678, ST9271, ST3322, ST5472. The Kolkata isolates were ST8288 and ST5472. All types were shown to occur in wild birds. One crow’s ST3174 from Kolkata has been shown to occur in human stool, while 4 isolates including Kolkata and Washington were of unknown ST’s. Although Kolkata and WA isolates belonged to different ST’s, majority were from wild birds that have rarely been isolated from humans.

Skin plays an important function as a highly innervated sensory organ. This sensory function is critical to organism survival, but the skin, and its axons, are easily damaged and need to be repaired to maintain homeostasis. Zebrafish are an excellent model to study tissue repair because they regenerate tissue efficiently and share conserved skin architecture with other vertebrates. A cell type shown in previous lab experiments to contribute to skin repair is a macrophage like cell, the Langerhans Cell. I hypothesize that if Langerhans Cells are not present within zebrafish epidermis then tissue repair will be incomplete or delayed. I investigated these cells using two different methods. First, I analyzed mutations that cause a loss of function in genes required for Langerhans Cell development. Second, I used the transgenic ablation technique to inducibly kill Langerhans Cells in the skin by addition of the antibiotic metronidazole. I imaged fluorescent Langerhans Cells in the zebrafish epidermis in both of these genetic scenarios to quantify the reduction in Langerhans Cells. Following the completion of these experiments, we expect to see that a lack of Langerhans Cells will induce slower or halted recovery from damage. In the loss of function method, I expect that various mutants will not have as many Langerhans Cells and that following induced ablation, Langerhans Cells will die out when the drug is introduced and slowly recover following metronidazole removal. The key implication of this is finding more about our own biology that can lead to faster healing. In theory, if Langerhans Cells have a major effect on this process then knowing more about our own skin can help us translate this understanding to our own healing process.

Clallam County has recently been experiencing concerning air quality conditions due to smoke drift from wildfire events in nearby areas. The objective of this study was to assess the wildfire smoke health-risk communication needs of organizations that serve the public in Clallam County. Communication needs were assessed by surveying 10 organizations that serve sensitive populations. Surveys were conducted in person or over the phone, and summary statistics were calculated. In addition, a low-cost air monitor was installed to track summer time air quality. The low-cost air monitor was installed adjacent to a nephelometer administered by the Olympic Region Clean Air Agency (ORCAA). This was done to determine the accuracy of the low-cost air monitor by comparing air quality values registered from both devices. The majority (60%) of organizations responding to the survey reported that they had not received information about the health risks of wildfire smoke. Nearly all organizations (90%) reported that they have the capacity and are willing to communicate the health-risks of wildfire smoke to the people they serve in Clallam County. Analysis of air quality data shows the low-cost monitor may be a useful device in determining air quality conditions. The correlation coefficient for the daily averages (from July 12 to August 24) between the ORCAA nephelometer and the low-cost air monitor was determined to be 0.98, but closer examination of data for hourly averages show a correlation coefficient as low as 0.82. Future wildfire smoke events in Clallam County require public health interventions to address health-risk communication needs of sensitive populations. Implementation of a low-cost air monitor network accessible by the public is a promising prospect to protect sensitive populations in Clallam County.

In this study we looked at the presence of streptomycin resistance genes, strA, strB, and aadA and the insertion sequence IS133 for the bacteria Escherichia coli obtained from the wetlands located within the University of Washington Bothell. The wetlands serve as a roost for more than 15000 crows all through the autumn and winter months. The hypothesis of this study was that the presence of antibiotic resistance may be able to spread through the vector of American crows. The genes tested in this study encode for factors that play a role in streptomycin resistance (Sugiyama, Masanori, and Osamu Nimi). The IS was tested because it would indicate presence of transposon TN5393 which in turn would suggest the presence of strAB genes on conjugative plasmids (Zhao, J., and H. Dang) and thus capable of horizontal gene transfer among the bacteria. The samples were collected from crow feces and water runoff within the wetlands. 23 water samples and 37 fecal samples, were testedby TaqMan™ quantitative PCR for the presence of the three streptomycin resistance genes, strA, strB, and aadA. Once the streptomycin genes were detected, the samples were tested to see whether the genes were located on a plasmid by testing for IS133 insertion and the TN5393 transposon. Our results show that 53% of the isolates tested were positive for strA, 57% were positive for strB, 5% were positive for aadA, 22% were positive for IS133, and 22% were positive for both strA and strB. Plasmids isolated from one water runoff sample and a fecal sample showed the presence of aadA and strB. The plasmids were successfully transformed into DH5 Alpha E. coli and were ampicillin resistant but not streptomycin resistant. These results indicate that there may have been other plasmids in the cell that rendered the samples resistant to ampicillin.

Beside its academia, University of Washington Bothell (UWB) is also known for its phenomenal nature where more than 15,000 crows roost every evening, especially during the autumn and winter months, in the wetlands. Since these crows have infested these wetlands, and because free living birds can be significant contributors of antibiotic resistant (AR) bacteria to the environment they were chosen for this study. E. coli, which lives as a harmless commensal in the gut of all animal and birds, has proved to be not only an indicator of fecal coliform but also of antibiotic resistance present in the environment. This study seeks to find out the different sequence types(STs) of E. coli found in UWB wetlands using the Multi Locus Sequence Typing (MLST) technique, to find out about the lineage, and potentially how these isolates have ended up in the UWB wetlands. Antibiotic resistance possessed by the isolates was performed against 13 different antibiotics by other students in the laboratory. MLST was performed according to methods specified in the EntroBase database; Specifically 7 housekeeping genes, were amplified and sequenced. Extended spectrum beta lactamase (ESBL) and non-ESBL containing isolates were found. Two non-ESBL isolates, one from water and one from the fecal samples collected on the same date, with the same antibiotic resistant pattern, turned out to be from the human isolated clone ST 58, suggesting a link between crow and water. ESBL E.coli isolate ST 131, a highly virulent, and multi drug resistant isolate was found in 6 fecal and 1 water isolate. Therefore it is concluded that crows are potential vectors of spreading multiple drug resistant strains in wetlands, which poses health risk since these strains may be carried further during the winter months.