Branch Meeting Program

2020 Annual ASM Intermountain Branch Meeting ASM Intermountain Branch Meeting Hosted by Weber State University Saturday, December 5, 2020

Table of Contents Program ...... 2 Keynote Address ...... 3 Oral Sessions ...... 4 Oral Session A—Career Panel ...... 5 Oral Session B—AAR, CIV, and CPHM ...... 7 Oral Session C—AES and EEB ...... 9 Oral Session D—POM and HMB ...... 10 Oral Session E—MBP ...... 11 Employment Fair ...... 12 Poster Sessions ...... 15 Poster Session A—CIV and HMB ...... 16 Poster Session B—MBP ...... 17 Poster Session C—AAR, CPHM, and EEB ...... 18 Poster Session D—AES ...... 20 Oral Abstracts ...... 21 Poster Abstracts ...... 51 Author Index ...... 80

Program

9:00 – 10:00 Opening Session (https://weber.zoom.us/s/93785843584) Welcome Keynote Address—Dr. Dustin L. Williams, PhD, “Targeting Biofilms in Translational Research, Device Development, and Industrial Sectors”

10:00 – 11:30 Concurrent Oral Presentations Session A—Career Panel Session B—AAR, CIV, and CPHM Session C—AES and EEB Session D—POM and HMB Session E—MBP

11:30 – 12:15 Break for Lunch Employment Fair

12:15 – 1:00 Concurrent Poster Presentations Session A—CIV and HMB Session B—MBP Session C—AAR, CPHM, and EEB Session D—AES

1:00 – 1:30 Closing Session (https://weber.zoom.us/s/93785843584) Elections Memorials Awards

Keynote Address

Title

Targeting Biofilms in Translational Research, Device Development, and Industrial Sectors Abstract

Biofilm-related infections continue to complicate healthcare. For example, in cases of open fractures rates of infection have gone unchanged since their definition in the 1980s. This may be due to the inability of systemic antibiotics to achieve sufficient doses against biofilms that contaminate a wound site at the time of injury. All antibiotic therapies that are currently in clinical use have been developed, optimized and regulated for efficacy against planktonic bacteria. Novel strategies and technologies that target the biofilm phenotype may improve infection treatment and prevention. Work in our lab is multifaceted to address the biofilm problem including development of chemical compounds with potential to disperse and kill well-established biofilms, active release coatings that are optimized to eradicate biofilms, as well as a device to sustain local, high doses of antibiotic delivery. There is not likely any single method that will provide a solution, but as designs and devices are integrated into a polytherapy approach rates of biofilm- related infection may be reduced. Translational data from in vitro to in vivo testing indicates that the chemical and device approaches we are developing have promise to eradicate biofilms that contaminate and complicate sites of biofilm-related infection. The overall objective is to improve patient care and quality of life. Biography

Dustin L. Williams, PhD is an Associate Professor in the Department of Orthopaedics at the University of Utah and a Research Scientist with the Department of Veterans Affairs. He fuses his degrees in Microbiology (BS) and Bioengineering (PhD) to develop antimicrobial technologies that target the biofilm phenotype. He has launched two companies—CŪRZA and Purgo Scientific—out of the U of Utah. The combination of translational science and commercialization is aimed at improving patient care, reducing rates of biofilm-related infection, and mitigating bone pathologies (e.g., heterotopic ossification) that may be influenced by the presence of biofilms. Zoom

Link: https://weber.zoom.us/s/93785843584?pwd=d1hhMzVqdm1rRDQzdmpFdUJSOGJyUT09 Meeting ID: 937 8584 3584 Passcode: 769061

Oral Sessions

Oral Session A—Career Panel Purpose

This panel will give attendees the opportunity to interact with career microbiologists. Each panelist will present the trajectory of his or her unique career. Attendees will then have the opportunity to solicit advice about pursuing a career in microbiology. Biographies

Jeff Rogers is a Health Research Science Specialist with the U.S. Department of Veterans Affairs, within the Veterans Health Administration and University of Utah Division of Epidemiology. He currently performs epidemiological research within the Veterans healthcare system. He has experience in active-duty military service, hospital project management, microbiology research and data management roles. He is from South Ogden, UT and a husband and father of four children. He earned a BA degree from the University of Alaska Fairbanks and a BS degree from Weber State University in microbiology. He has also earned a master of health administration (MHA) degree from Capella University. He is currently finishing his master of public health (MPH) degree with a concentration in epidemiology from the University of Nebraska Medical Center. His interests are in infectious disease epidemiology, global health and management of infection control within hospital/clinical settings.

Jessi Dōne is a Nationally Registered, Certified Microbiologist at the baccalaureate level. She specializes in Food Safety, Quality, Pharmaceutical and Medical Device. Jessi holds a Master of Business Administration degree from Weber State University, Ogden, Utah; an AACSB accredited school. Her work and leadership positions include years of Food Safety, Quality and Regulatory Management as a Sr. Quality and Food Safety Manager, and SQF Practitioner, within an Extruded, Baked and Canned Products Plant, as well as, Senior Microbiologist and Director of Laboratory Operations within Dietary Supplements and Medical Device fields. Jessi has built, implemented and maintained Food Safety and Quality Plans, within a large manufacturing facility with multiple processes. Throughout her career Jessi has further developed her skills through attendance in various seminars and certifications like Better Process Control School (BPCS), Principles of Internal Auditing, FSPCA Preventive Controls for Human Food and FSPCA Preventive Controls for Animal Food, Thermal Processing Course for Low Acid Canned Foods (21 CFR 113), and is a contributing member of the American Society of Quality (ASQ) and American Society of Microbiology (ASM). Jessi is a Safe Quality Food Institute (SQFI) body of knowledge consultant for food processing, manufacturing and packaging plants. Jessi currently sits on the Weber State Microbiology Advisory Board as the Co-Chair Vice President and enjoys sharing her broad base of the Sciences, Risk Management and Mitigation, and passion for education and Food Science.

Nicholas Ashton received a B.S., M.S. and Ph.D. in bioengineering at The University of Utah completing his doctoral dissertation on The Adhesive Silk of the Aquatic Caddisfly Larva, a work conducted in search of design principles for better surgical glues, vascular embolizing agents, and tough hydrogels. He completed a Postdoctoral Fellowship in the Bone and Biofilm Research Laboratory at the University of Utah under the tutelage of Dustin Williams, PhD, an expert in anti-biofilm technologies. To continue his work towards better infection prevention and treatment technologies, he recently accepted a Research Instructor position in The Department of Orthopedics at The University of Utah. His personal research focuses on identifying antibiofilm agents which have been overlooked because of the assay failures used in current clinical and commercial R&D laboratories.

Chalice Gustavson has degrees in Chemistry, Criminal Justice, and Medical Laboratory Sciences from Weber State University and the University of Utah. She has had a variety of jobs—professional ice cream scooper, high school custodian, analytical chemist, quality control Manager, R&D chemist, project manager, and research assistant. She is currently a medical laboratory scientist at Lakeview Hospital in Bountiful. She lives in Clearfield with her husband, 11-year-old son and 17-year-old cat. Zoom

Link: https://weber.zoom.us/j/95131442556?pwd=aTN6bWhZWENZYXEwMDBWWXlCeHZ0UT09 Meeting ID: 951 3144 2556 Passcode: 328657

Oral Session B—AAR, CIV, and CPHM Tracks

Antimicrobial Agents and Resistance The Antimicrobial Agents and Resistance (AAR) track features the latest on the discovery and

development of new antimicrobial drugs in the pipeline, and their associated resistance mechanisms. The AAR track focuses on a broad range of topics, including surveillance and global epidemiology, mechanisms of resistance, antimicrobial discovery from pre-clinical investigations to post-marketing data, stewardship, novel approaches to infection diagnosis and treatment, and pharmacokinetic/pharmacodynamic principles to optimize antimicrobial design and use.

Clinical Infections and Vaccines The Clinical Infections and Vaccines (CIV) track encompasses clinical and epidemiological studies in adult and pediatric infectious diseases, global health issues, including emerging infectious diseases and tropical medicine, and recent developments and clinical studies in vaccines. This track is designed for infectious diseases physicians, clinical microbiologists, epidemiologists, public health officials and pharmacists to learn about the latest advances in clinical and basic science.

Clinical and Public Health Microbiology The Clinical and Public Health Microbiology (CPHM) track provides laboratory directors, medical technologists, clinicians, infection preventionists, and public health officials with the most recent developments and cutting-edge findings in the field. The CPHM track delves into multiple infectious disease sub-disciplines, including novel diagnostic technology, susceptibility testing, clinical laboratory biosafety and biosecurity, antimicrobial and diagnostic stewardship, surveillance and epidemiology, and “One health.”

Session Chair

Matthew Domek, Weber State University Zoom

Link: https://weber.zoom.us/j/96850815589?pwd=YU5vdTkvWk1xM3hETVZFaWExY3NiQT09 Meeting ID: 968 5081 5589 Passcode: 176970 Presentations

Tyler Smith Development of a Novel Anti-Biofilm Polyurethane Foam for use in Negative Pressure Wound Therapy

Jemi Ong Preliminary Results of an Antimicrobial Blue Light Device to Eradicate Bacteria at the Skin–Implant Interface of Osseointegrated Implants

Diana Calvopiña Convincing a Post-Translational Modification Machine to Make Novel antimicrobial peptides

Benjamin Ogilvie Alcohol-Free Hand Sanitizer Kills SARS-CoV-2

Tiana Scott A Signaling Pathway-Driven Bioinformatics Pipeline for Predicting Therapeutics Against Emerging Infectious Diseases

Justina Tavana Early Onset Alzheimer's Disease in Pacific Island Populations

Cierra Wheeler Identification of Staphylococcus aureus Genes of Interest for a Bovine Mastitis vaccine

Matthew Findlay Combating Vaccine-Hesitancy Using a Moral Foundations Approach

Oral Session C—AES and EEB Tracks

Applied and Environmental Science Applied Environmental Science (AES) fosters broad understanding and collaboration

between 17 sub-tracks, based on the fundamental shared interests in how microbes work in diverse environments. The AES track is designed for scientists with a variety of backgrounds in the basic sciences, engineering, public health and food production. Topics include basic and applied microbiology in environments from soil to food, lakes and oceans to drinking water systems, and deserts to urban population centers.

Ecology, Evolution and Biodiversity The Ecology, Evolution, and Biodiversity (EEB) track is defined by research that encompasses laboratory, clinical, engineered, and natural settings. The EEB track is aimed to advance ecological and evolutionary theory in microbes and other organisms in basic and applied research, and to promote microbial ecology, evolution, and biodiversity in biological education.

Session Chair

Lisa Wiltbank, Weber State University Zoom

Link: https://weber.zoom.us/j/92457185984?pwd=NXZaMnN4dlgzaDBDYlk5dG5vU3hBdz09 Meeting ID: 924 5718 5984 Passcode: 394489 Presentations

Sheri Motamedi Endemic Microbial Communities of Seafloor Mantle Rocks

Jyothsna Ganesh Microbial Community of the Rhizosphere of Ceanothus velutinus Improves the Plant’s Growth and Development under Greenhouse Conditions

Naveen Duhan deepNEC: A Novel Alignment-Free Tool for the Identification and Classification of Nitrification-Related Enzymes Using Deep Learning

Irene Knorr, Utah Agritourism: Operator Attitudes and Behaviors toward Zoonotic Disease Cody Zesiger

June Smith, Fermentation of Plant-Based Extracts by Dairy Lactic Acid Bacteria Niharika Mishra

Lizell Mejías Paucilactobacillus wasatchsensis WCD04 Biofilm Formation and Adherence to Stainless Steel

Dallin Leatham Antimicrobial Metabolites of Lactobacillus curvatus

Oral Session D—POM and HMB Tracks

Profession of Microbiology The Profession of Microbiology (POM) Track provides covers topics related to education, career choices, diversity and inclusion, professional development and science communication and outreach.

Host–Microbe Biology The Host–Microbe Biology (HMB) track explores research at the host–microbe interface. The HMB track addresses pathogenic and symbiotic relationships between microbes and the host, as well as the microbiome. Scientists studying genetics and physiology of microbes and viruses at the basic science level should expect to find topics covering the genetic, physiological and virulence mechanisms that shape host–microbe relationships.

Session Chair

Brian Poole, Brigham Young University—Provo Zoom

Link: https://byu.zoom.us/j/94155268218?pwd=K1pPZS9NdlpTNzU5MUpXQi9LcHdMQT09 Meeting ID: 941 5526 8218 Passcode: 757761 Presentations

Joel Griffitts Getting Students Close to Microbiology at a Distance

Israel Guerrero A Rag2−/−γc−/− Balb/c Mouse Model to Study Chikungunya Virus Pathogenesis

Kiara Whitley The Role of T Cell Co-Receptor CD5 in T Cell Metabolism, the Gut Microbiome, and Behavior

Yuko Sperry Role of CD5 in Oral Inflammation and Periodontal Disease

Brayden McGary Disruption of Mouse Respiratory Microbiome by Vaporized Nicotine

Madison Gray Chikungunya Virus Time Course Infection of Human Macrophages

Clayton Rawson Effects of Celiac Disease and a Gluten-Free Diet on the Human Gut Microbiome

Oral Session E—MBP Tracks

Molecular Biology and Physiology Molecular Biology and Physiology (MBP) covers a broad spectrum of microbiological phenomena. For scientists in the MBP track, the goal is to understand the basic mechanisms of microbial life at the molecular, cellular and multi-cellular level.

Session Chair

Michele Culumber, Weber State University Zoom

Link: https://weber.zoom.us/j/91079085651?pwd=SXdwZ1pNMEo0YW40YTVLZE41aWRxZz09 Meeting ID: 910 7908 5651 Passcode: 511355 Presentations

Erin Gaffney Investigating and Tuning Salt Tolerance of Purple Bacteria Using a Combined Bioinformatic and Electrochemical Approach

Colleen Newey Understanding ALS: PAS Kinase and Ataxin-2

Daniel Jackson Investigating the Impact of Host Environmental Factors on Pneumococcal UDP- Glucose Dehydrogenase Activity

Hannah Taylor Helicase-Mediated Mechanisms for Defense against Invasive Plasmids by the Type IV-A CRISPR–Cas Bacterial Immune System

Elise Overgaard Cellular Activity of ArtAB Toxin from Bovine Salmonella Typhimurium

Tyler Humpherys Immunotargeting of Membrane Associated Thymidine Kinase 1 in Colorectal Malignancies

Ed Ringger Exploring the Human Phageome through Sewage

Employment Fair

BioFire Diagnostics

With more than 25 years of molecular experience, BioFire Diagnostics, LLC sets the standard for molecular diagnostics through its pioneering advances in syndromic infectious disease testing. As bioMérieux’s Global Center of Excellence for Molecular Diagnostics, no other company has FDA-cleared and CE-IVD marked assays for more pathogens. BioFire Diagnostics has produced the easiest, fastest, and most comprehensive multiplex PCR testing available: the BioFire System. Zoom

Link: https://zoom.us/j/99977109508?pwd=SzRKZ0t1M3pWcVJIai8wUzR3RjJmdz09 Meeting ID: 999 7710 9508 Passcode: vyDY30 Contact Information

BioFire Recruiting Email: [email protected] Phone: 385-770-1132

Jacobs Technology

Jacobs is one of the world’s largest and most diverse providers of full-spectrum technical, professional and construction services for industrial, commercial and government organizations globally. The company employs over 54,000 people and operates in more than 25 countries around the world. Jacobs provides field, range, testing, chemical, and biological laboratory support to the U.S. Army at the West Desert Test Center (WDTC) located at Dugway Proving Ground (DPG), UT. As a major test site for the U.S. Army Test and Evaluation Command, the WDTC is responsible for the test and evaluation of chemical and biological (CB) defense systems, incendiary and smoke/obscuration munitions systems, and related operational concepts. In addition to these responsibilities, we also support non-CB testing at the WDTC for the DOD and other Federal agencies, including environmental assessment/impact evaluation and munitions demilitarization. Our Dugway Operation was established in 2004 with the award of the Field, Range, Chamber, and Laboratory Support Services (FRC&LSS) contract to Jacobs. Through the FRC&LSS contract, we manage an integrated team of contractors that includes Mellor Engineering with a composite staff size of more than 170 personnel. Zoom

Link: https://us02web.zoom.us/j/8262985892?pwd=Z0NUVHVNSWpvVlpGcklkL0h4OW81QT09 Meeting ID: 826 298 5892 Passcode: JDT Contact Information

Rick Pugh, Scientific Support Branch Manager Email: [email protected]

Poster Sessions

Poster Session A—CIV and HMB Tracks

Session Chair

Michaela Gazdik Stofer, Utah Valley University Zoom

Link: https://weber.zoom.us/j/98478910949?pwd=WE5rRG1haUNoS21aQzhFazB1eHlaUT09 Meeting ID: 984 7891 0949 Passcode: 255086 Posters

Brian Poole Short Variant of Turkey Hemmorhagic Enteritis Virus Vaccine Strain

Betsy Church The Behavior of Bacillus cereus in Human Breast Milk Pre- and Post- Pasteurization

Tallon Nielsen The Role of the Brain-Eating Amoeba Naegleria fowleri CD59-like Protein in Human Infections

Nicole Skalka CRISPR Deletion of Viral Receptor Genes in Human Cells

Kyle Reaveley The T Cell Co-receptor CD5 Alters Mouse Behavior and Gut Microbiome Composition

Isabella von Wallwitz Understanding the Effects of T Cell Co-Receptor CD5 on Metabolic Function and the Metabolome

Christopher Haynie The Role of CD5 in Regulating Levels of T Follicular Helper Cells

Poster Session B—MBP Tracks

Session Chair

Juliette Tinker, Boise State University Zoom

Link: https://boisestate.zoom.us/j/97405037441?pwd=RVFVVEIyS2s0eEF2cVNtc2xUZjlrZz09 Meeting ID: 974 0503 7441 Passcode: N7*xrr Posters

Carlos Moreno Measuring the Dragonfly (Tanypteryx hagen) Genome Size Using Flow Cytometry

Alex Benedict Optimizing Translation Efficiency of Recombinant Proteins in E. coli

Nolan Cole Epigenetic Profiling of Human Peripheral Blood Monocytes

Skyler Russell Chromatin Accessibility in C. elegans

Lucie Dearden The CCRL2 Atypical Chemokine Receptor Variant V180M Has a Higher Binding Affinity for Chemerin

Andrew Nelson The CCRL2 Atypical Chemokine Receptor Variant V180M Alters CCL4 Secretion in Leukocytes

Brigham Perry Determining the Role of PKMζ in Pancreatic Adenocarcinoma

Poster Session C—AAR, CPHM, and EEB Tracks

Antimicrobial Agents and Resistance The Antimicrobial Agents and Resistance (AAR) track features the latest on the discovery

and development of new antimicrobial drugs in the pipeline, and their associated resistance mechanisms. The AAR track focuses on a broad range of topics, including surveillance and global epidemiology, mechanisms of resistance, antimicrobial discovery from pre-clinical investigations to post-marketing data, stewardship, novel approaches to infection diagnosis and treatment, and pharmacokinetic/pharmacodynamic principles to optimize antimicrobial design and use.

Session Chair

Katrina Twing, Weber State University Zoom

Link: https://weber.zoom.us/j/97453200558?pwd=NjZXVVozeTdqRlhWVFRkRzVrR2JwQT09 Meeting ID: 974 5320 0558 Passcode: 198727 Posters

Katrina Twing Microbial Diversity of Rock-Hosted Serpentinite Subsurface Environment

Ashley Miller Gene Expression Changes in Plants Inoculated with Halophilic Bacteria and Grown in Salty Soil

Jaron Quirante Siderophore and Biofilm Properties of Halophilic Bacterial Strains That May Contribute to Growth Promotion of Inoculated Plants in Salty Soil

Kevin Vander Werff Comparative Genomic Analysis of M. foliorum Cluster EE Phages

Samuel Flor Discovery of the Human Phageome for Future Disease Diagnostics

Kawika Tupuola, The Effects of Ozone on Staphylococcus Xavier Stilson

Matthew Crook Survey of North Dakota Rhizobia to Improve Nodulation and Nitrogen Fixation in Chickpea

Poster Session D—AES Tracks

Applied and Environmental Science Applied Environmental Science (AES) fosters broad understanding and collaboration

Session Chair

Joel Griffitts, Brigham Young University—Provo Zoom

Link: https://byu.zoom.us/j/91717101035 Posters

Melinda Moss Microbial Production of Rare Sugars from Waste Lactose

Sterling Fuller Analysis of the Microbiome Composition from Low-Income Homes with Evaporative Coolers and Central Air Conditioners

Kate Sorensen Gluconate Metabolism by Paucilactobacillus wasatchensis WDC04 Can Be a Cause of Late Gas Defect in Aging Cheese

Niharika Mishra Effect of Inorganic Salts on the Growth of Probiotic Lactic Acid Bacteria

Chase Wahlstrom Selective Media for the Isolation of Paucilactobacillus wasatchensis

Josh Hansen Using a Virus Model System to Determine the Effect of Ozone in Locker Room Sanitation

Riley Nichols Can Automatic Hand Dryers Serve as a Microbial Reservoir for Contamination?

Oral Abstracts (alphabetical, by last name of presenter)

Convincing a Post-Translational Modification Machine to Make Novel Antimicrobial Peptides Calvopiña D, McPhie J, Wagstaff K, Price J, Griffitts JS Brigham Young University—Provo

Peptides are short chains of amino acids that are crucial in many biological pathways. Thiopeptides are a family of antimicrobial peptides that are characterized for having sulfur-containing heterocycles, and for being highly post-translationally modified. Thiopeptides exhibit intrinsic antimicrobial properties which make them promising candidates for the development of new antibiotics. We are studying the antimicrobial thiopeptide micrococcin, with the objective to understand how a specialized post- translational modification (PTM) machine interacts with the precursor peptide of micrococcin to rigidify its structure and confer the peptide with antimicrobial properties. We have discovered that the first 12 amino acids of the precursor peptide are not necessary for this post-translational modification. Once we have a deeper understanding of how the complex recognizes the native substrate peptide, we can use the PTM machine to make novel unnatural antimicrobial peptides.

deepNEC: A Novel Alignment-Free Tool for the Identification and Classification of Nitrification-Related Enzymes Using Deep Learning Duhan N, Norton JM, Kaundal R Utah State University

We propose deepNEC, a novel end-to-end feature selection and classification model training approach for nitrification-related enzymes prediction. The raw protein sequences encoding is used as an input, extracting sequential and convolutional features from raw encoded protein sequences based on classification rather than using traditional alignment-based methods for nitrification-related enzymes prediction. Two large datasets of protein sequences, enzymes and non-enzymes were used to train the models with protein sequence features like amino acid composition, dipeptide composition, conformation transition and distribution (CTD), NMBroto, conjoint, quasi order, etc. The k-fold cross-validation and independent testing were performed to validate our model training. deepNEC uses a three-tier approach for prediction; in the first phase, it will predict a query sequence as enzyme or non-enzyme; in second phase, it will predict whether the enzyme is an oxidoreductase or a non-oxidoreductase; in the third phase, for those enzymes predicted as oxidoreductases in phase-II, it will further predict and classify them into various nitrification-related enzyme classes. Among all, the DPC+NMBrot hybrid feature gave the best prediction performance (accuracy of 96.15% in k-fold training and 93.43% in independent testing) with a MCC (0.92 training and 0.87 independent testing) in phase I; in phase-II, the DPC feature gave the best prediction performance (accuracy of 99.82% in k-fold training and 94% independent testing) with a MCC (0.99 training and 0.88 independent testing); in phase-III, DPC feature gave the best prediction performance for 13 nitrification-related enzyme classes. We have also implemented homology-based method to remove false negatives. The tool can be accessed freely at http://bioinfo.usu.edu/deepNEC/ or standalone version can be downloaded from https://bitbucket.org/navduhan/deepnec/src/master/.

Combating Vaccine-Hesitancy Using a Moral Foundations Approach Cromar Z, Findlay M, Mills E, Turner E Utah Valley University

The World Health Organization included vaccine hesitancy (VH) in the top ten threats to global health in 2019. Currently amidst a global pandemic, multiple companies seek emergency FDA COVID-19 vaccine approval to fight the Coronavirus. With these vaccines nearing distribution, healthcare providers must reduce VH as the effectiveness of any vaccine is directly dependent upon the number of recipients. Further, the pandemic has exacerbated an already significant issue, as all age groups nationally are lagging in scheduled vaccinations. In Utah, this is especially concerning, as over the past decade the state has experienced the nation's third-largest decrease in vaccination compliance, with rates falling by almost 5%. Studies done in the U.S. have shown that the Moral Foundations Theory (MFT), a social psychological theory used to explain variation in human moral reasoning, can identify individuals with higher VH based on the core moral foundations with which they identify. This research suggests that messaging interventions (MI) that specifically appeal to the specific MFs of Liberty vs. Oppression, and Purity vs. Degradation could be more effective at reducing VH. This study explored those exact avenues, using MI in Utah. A survey containing one of three MI and a questionnaire designed to identify VH was presented to 800 Utah residents. Two of these video interventions delivered a pro-vaccine message, while the third operated as a control. In the two treatment videos, each message was delivered carefully, appealing to the MFs typically used in current interventions, and the MFs suggested by previous research. After preliminary analysis, this study found no significant difference in VH amongst all groups. This suggests that singularly using the MFT is not a viable method for effectively reducing VH. However, a follow-up survey is being sent to 500 of the respondents to evaluate for potential long-term changes in subjects' VH due to participation in this study.

Investigating and Tuning Salt Tolerance of Purple Bacteria Using a Combined Bioinformatic and Electrochemical Approach Gaffney EM, Grattieri M, Minteer SD University of Utah

Purple bacteria have been recently explored in environmental bioelectrochemical systems for their ability to generate energy from sunlight for applications in renewable energy generation, biosensing, and biosynthesis.1 Crucial to their deployment in such applications is the tolerance to a variety of environmental conditions.2 Among various environmental stresses, salinity is of interest due to fluctuating saline concentrations typical for industrial wastewaters.3 Specifically, the purple bacterium, Rhodobacter capsulatus, is a great candidate for microbial electrochemical systems owing to its well-studied photosynthetic electron transport chain allowing for wiring of the bacterium to an abiotic electrode.

In this presentation, the investigation of R. capsulatus adaptation to saline conditions will be discussed. We found that a slow increase of salinity values allowed for successful growth in up to 20 gl−1 NaCl, whereas immediate exposure to 20 gl−1 NaCl inhibited bacterial cell activity and growth. These results indicated a metabolic shift in the saline adapted strain, further analyzed through RNA sequencing to determine differential gene expression. The process of adaptation significantly impacted gene expression in nitrogen fixation mechanisms, the photosynthetic electron transport chain, osmoprotectant synthesis and intake, and flagellar systems of this bacterium. Insights obtained from the bioinformatic analysis allowed experimental triggering of rapid saline adaptation, resulting in an increase in bio-photocurrents when compared to the long-term saline adapted strain. Understanding the saline adaptation and bio- photocurrent generation in R. capsulatus allows for exciting opportunities in the field of environmental microbial electrochemistry.

References 1Grattieri M. 2020. Purple bacteria photo-bioelectrochemistry: Enthralling challenges and opportunities. Photochem. Photobio. Sci., 19:424–435. DOI: 10.1039/C9PP00470J 2Torres CI. 2014. On the importance of identifying, characterizing, and predicting fundamental phenomena towards microbial electrochemistry applications. Curr. Opin. Biotechnol., 27:107–114. DOI: 10.1016/j.copbio.2013.12.008 3Grattieri M, Minteer SD. 2018. Microbial fuel cells in saline and hypersaline environments: Advancements, challenges and future perspectives. Bioelectrochemistry, 120:127–137. DOI 10.1016/j.bioelechem.2017.12.004.

Microbial Community of the Rhizosphere of Ceanothus velutinus Improves the Plant’s Growth and Development under Greenhouse Conditions Ganesh J, Sun Y, Thomas A, Kaundal A Utah State University

Climatic change-induced environmental stresses affect crop production. Increase in world population increases the demand for food production. A need to develop sustainable agriculture under these stresses is required to develop resilient crops. One such approach is to explore natural resources like the rhizosphere microbiome of plants. The rhizosphere microbiome of a plant plays a significant role in the growth and development of plants and defense against various stresses. Here, we are investigating the microbiome of a native plant Ceanothus velutinus (snowbrush). Snowbrush is an evergreen native plant in western North America. It thrives well in harsh conditions and can act as ornamental plant in low water use landscaping but is difficult to propagate under landscape conditions. The microbial diversity analysis of bulk soil, rhizosphere, and endosphere of snowbrush revealed the presence of various Plant Growth Promoting Rhizobacteria (PGPR). The effect of native soil on cutting propagation showed increased rooting and callusing (13.33%) in treatment cuttings when compared to control (3.125%) under 50% native soil inoculation. The effect of native soil was observed in the growth of snowbrush plants from cuttings and seedlings in greenhouse conditions. The plants grown in 1-gallon pots inoculated with 200

ml of native soil showed an increase in growth, higher NO3 content (2–3 fold more), higher number of secondary shoots (6–9 times more), and a higher survival rate when compared to control. The microbial diversity analysis of the treated cuttings plants showed the presence of the same PGPR as in the native soil when compared to control cuttings. These results showed a significant role of rhizosphere microbes in the improvement of cutting propagation and growth of snowbrush plants in greenhouse conditions. Once the microbes are isolated and characterized for their role, we will test them to develop a propagation method from snowbrush cuttings for low water use landscaping.

Chikungunya Virus Time Course Infection of Human Macrophages Gray M Brigham Young University—Provo

Chikungunya virus (CHIKV) is an alphavirus spread by mosquitoes infecting 1.1 million people per year around the world, including the Western Hemisphere in 2015. CHIKV is an arbovirus spread by Aedes albopictus mosquitoes as vectors. Since then CHIKV has evolved and found a new mosquito host, Aedes aegypti. For someone who is infected with CHIKV their life will be forever altered, with arthralgia and myalgia causing debilitating pain for weeks, months, and even years, affecting the livelihood of the patient. As of today, there is no cure, treatment, or vaccine for most alphaviruses. In current research of the CHIKV human macrophages have been identified as an important cell to the success of the virus. After the virus is transferred to a human via mosquito, an immune response is triggered including macrophages. It has been discovered that after a macrophage begins the phagocytosis of CHIKV, the virus causes the macrophage to then undergo induced apoptosis, catalyzing the virus spread in the body. The question that still remains is when does the macrophage get infected to the point of apoptosis? We evaluated the transcriptional response of human macrophage cells infected with CHIKV at three different time points. Samples of RNA were collected from either mock-infected or infected monocyte-derived macrophages at 24 or 48 hours after infection. We then generated cDNA and performed RNA-sequencing on the material from the duplicate samples taken at each of three time points (i.e., six samples total). We received 30–60 million sequencing reads per sample on an Illumina NovaSeq instrument. We then analyzed fastq data including differential gene expression, enriched functional annotations, and modulated intracellular signaling pathways. Our analysis on potential therapeutic drugs continues as we get closer to our goal of furthering the information and development of creating a prophylactic, therapeutic, or vaccine.

Getting Students Close to Microbiology at a Distance Griffitts JS, Benedict AB, Calvopiña D Brigham Young University—Provo

The past year has compelled educators to provide students with remote experiences that approximate the real thing. For the MMBIO 360 (Microbial Genetics) course at BYU, a sequence of laboratory activities were filmed and made available to students, in conjunction with short in-person experiences tailored to give them a feel for the most critical techniques. In this presentation, a logical series of lab activities in bacterial genetics will be outlined, with a description of student assessment strategies that aim to train real-world skills in design, analysis, and communication.

A Rag2−/−γc−/− Balb/c Mouse Model to Study Chikungunya Virus Pathogenesis Guerrero Arguero I, Villalva C, Valdoz J, Viazzo R, Van Ry PM, Berges BK, Robison RA Brigham Young University—Provo

Chikungunya virus (CHIKV) is an emerging alphavirus that causes a febrile disease that is typically manifested by myalgia, maculopapular rash, and severe polyarthritis, but reports of neurological disease in patients have become increasingly common. In this study, we assessed the role of Rag2 and γc immune functions and how their absence plays a role in CHIKV pathogenesis . We evaluated CHIKV infection of Rag2−/−γc−/− Balb/c and wild-type mice to determine the role of host adaptive and innate immune systems in CHIKV pathogenesis. CHIKV-inoculated Rag2−/−γc−/− mice developed local inflammation in the infected paw and joint damage at 8 days post-infection (dpi). We also detected abundant viral RNA in serum, liver, spleen, and brain tissue of Rag2−/−γc−/− mice. Additionally, we detected an increase in IL-12p70, IL-6, and IFN-γ (pro-inflammatory cytokines) and MCP-1 (a monocyte chemokine) in serum at 8 dpi. This study provides a foundation for studying CHIKV-induced arthralgia and neuropathy in a Rag2−/−γc−/− model that closely resembles many aspects of CHIKV-associated human pathology.

Immunotargeting of Membrane-Associated Thymidine Kinase 1 in Colorectal Malignancies Velázquez EJ, Humpherys TB, Cress JD, Bellini DM, Smith KR, O’Neill KL Brigham Young University—Provo

The objective of this project is to provide evidence that membrane-associated Thymidine Kinase 1 (TK1) is a suitable target for the immunotargeting of colorectal cancer. TK1 is a nucleotide salvage enzyme that is upregulated on various types of tumors including gastrointestinal and colorectal malignancies. Recent evidence has shown that TK1 is uniquely associated with the cell membrane in colorectal cancer cell lines and colon tumors from patients. This suggests TK1 could possibly be an immunotherapeutic target for the treatment of colorectal tumors. To target membrane-associated TK1, we developed novel anti TK1 antibodies. The antibodies specificity to bind to TK1 were validated through siRNA TK1 knockdown and flow cytometry. A screening of HT-29 and SW620 colon adenocarcinoma cell lines for TK1 revealed a 34% and 16% TK1 positivity on the cell membrane of each cell respectively. When we used our custom anti- TK1 antibody (10E8) it outperformed commercial options showing higher sensitivity to detect membrane- associated TK1 (>50% TK1-positive cells). Both malignant colon tumors and normal colon tissue from patients were dissociated and analyzed by flow cytometry. No significant levels of TK1 on the cell membrane of normal colon tissue were found. In contrast, up to 42% of the dissociated malignant colon cells were TK1-positive. The potential of the novel anti-TK1 antibody 10E8 to generate an immune response was then explored through antibody-dependent cell-mediated cytotoxicity (ADCC) experiments. The ADCC response by mononuclear cells (MNC) against TK1-overexpressing colon cells (HT-29) was measured in the presence of anti-TK1 antibodies (10E8) using ImageXpress pico cell imaging. After 48 hours, a 50% decrease in the number of HT-29 cells was measured, which was significantly different from the isotype control (p = 0.0216). We conclude that the immunotargeting of membrane-associated TK1 in colorectal malignancy is possible with the use of anti-TK1 antibodies.

Investigating the Impact of Host Environmental Factors on Pneumococcal UDP-Glucose Dehydrogenase Activity Jackson D, Martin JE Idaho State University

Capsular polysaccharide (CPS) is a major virulence determinant for human pathogenic bacteria. Although the functional roles of CPS in bacterial virulence have been established, knowledge of how CPS production is regulated remains limited. Streptococcus pneumoniae (pneumococcus) CPS expression levels and overall thickness change in response to available environmental factors, such as oxygen, carbohydrate, and metal ions. These factors can vary significantly between host environmental niches and infection stage. Since thicker CPS levels are unfavorable for pneumococcal colonization but necessary for evasion of the host defense systems, we postulated that CPS expression must be precisely regulated. In this study, we investigate the impact of oxygen and manganese (Mn) on the activity of the pneumococcal UDP- glucose dehydrogenase CpsK, a key enzyme required for CPS biosynthesis. CpsK harbors a single conserved cysteine (C259) in its active site that we hypothesize is prone to oxidation. We show that that CpsK activity is inhibited by hydrogen peroxide (H2O2) above 100 µM in a time dependent manner via possible oxidation of C259. Incubation with Mn prior to H2O2 treatment does not reduce the inhibition of CpsK activity by H2O2. Future experiments planned will assess the oxidation state of C259 by SDS–PAGE using an AMS–trypsin digestion assay. In toto, the data collected will expand our understanding of how available host environmental factors affect pneumococcal CPS expression, possibly providing new avenues for therapeutic treatments of pneumococcal disease.

Utah Agritourism: Operator Attitudes and Behaviors toward Zoonotic Disease Knorr I, Rood K, Wilson D, Walker-Bravo A, Miner D, Hepworth K, Peterson D, Davis K, Zesiger C, Stock M Utah State University

The risk of contracting a zoonotic disease increases at agritourism events that feature animals. Utah experienced a rise in human infections of Shiga toxin-producing Escherichia coli that was linked with petting zoos in 2018. Utah State University Extension, in collaboration with the Utah Department of Agriculture and Foods and Utah Public Health, conducted an online survey to assess factors influencing transmission, biosecurity interventions, and whether education and outreach were needed amongst agritourism operators. Respondents (n=31) represented 14 counties across Utah and Idaho. Agritourism visitation was the greatest October–December. 74% of operations allowed direct animal contact and 35% allowed eating in animal areas. Most (68%) gave verbal instructions about disease and hand washing, but only 35% had written instruction (e.g. signage). 74% had washing stations and/or sanitizer present. Respondents preferred online and printed materials over other educational experiences (in-person workshops and others). Agritourism is a new and critical demographic for USU Extension to reach.

Antimicrobial Metabolites of Lactobacillus curvatus Leatham D , Oberg T, Oberg C Weber State University

Using the genome of Lactobacillus curvatus WSU1, a bioinformatics analysis revealed the organism contained the genes for a potential metabolomic pathway which utilized lactic acid to produce propanol and propionic acid which are two potential antimicrobials. No specific gene was found for the conversion of the intermediate 1,2-propanediol to lactaldehyde but a likely candidate was proposed. Lactic acid bacteria (LAB) can often produce 1,2-propanediol from acetate and hydroxyacetate. It is hypothesized that like Lactobacillus reuteri, L. curvatus can utilize glycerol to produce 3-hydroxypropionic acid and 3- hydroxypropionaldehyde (reuterin) both of which are known antimicrobials. All of these processes are strictly anaerobic thus all cultures were grown in medicine bottles containing septa where the headspace was purged with 95% N2 and 5% CO2. The cultures were grown in carbohydrate restricted MRS containing either lactate, 1,2-propanediol, hydroxyacetate, acetate, or glycerol at a 80 mM concentration. The proposed metabolomic pathway may need B12 in order to function thus the cultures were grown in duplicates with half containing B12. A GC analysis will be done to test for the presence of 3- hydroxypropionaldehyde, propionic acid, propanol, and 3-hydroxypropionic acid at day 0, 1 and 7 of growth. The presence of certain products could prove the functionality of the proposed metabolic pathway and/or if L. curvatus can utilize glycerol similar to L. reuteri. If L. curvatus is able to produce antimicrobial compounds it would be of vital interest to the food industry. As a common non-starter lactic acid bacteria (NSLAB), it could be added to products to help inhibit the growth of pathogens or spoilage organisms.

Disruption of Mouse Respiratory Microbiome by Vaporized Nicotine McGary BC, Packard JF, Clark DN Weber State University

Vaping is the use of electronic cigarettes (e-cig) that contain nicotine, but not smoke. Vaping is seen as a safer alternative to traditional cigarettes and has increased substantially over the past ten years, with 3 to 5% of Americans reporting regular e-cig use. Despite potential advantages compared to smoking, the CDC reported over 2,800 hospitalizations and/or deaths this year due to electronic vaping associated lung injury (EVALI) in the United States. There is much debate over the nature of the danger caused by vaping; importantly, little is known about the effects of nicotine vapor on the respiratory microbiome. To this end, we examined the microbiome of the upper and lower respiratory tract of four mice—two control mice and two that received vaporized nicotine (10.0 mg/ml once daily for 28 days). Post-mortem respiratory tissue lavage was processed to extract DNA and PCR-amplify 16S ribosomal sequences, which were used to prepare an Illumina DNA sequence library. Operational taxonomic units were mapped using BLASTn to compile counts of species-specific total reads. This sequencing analysis revealed a 50-fold increase in Staphylococcus saprophyticus in both the upper and lower respiratory tract of each of the nicotine-treated mice (p < 0.005), and several other bacteria only infected the nicotine-treated lungs. On the other hand, several species representing the normal microbiome were reduced by greater than two-fold in the nicotine-treated isolates. These findings represent a disruption of the microbiome of the mouse respiratory tract due to vaping, which could represent potential changes and infections in humans.

Paucilactobacillus wasatchsensis WCD04 Biofilm Formation and Adherence to Stainless Steel Mejías LA, Culumber MC, Oberg CJ Weber State University

Paucilactobacillus wasatchsensis WCD04 is a gram-negative non-starter lactic acid bacterium (NSLAB). Its presence is characterized by gas production during the cheese aging process, which causes splits and cracks the cheese. This NSLAB is thought to be an environmental contaminant, but its reservoir is unknown. Plb. wasatchsensis does not survive heat exchange processing, so its persistence in cheese manufacturing could be caused by post-pasteurization contamination and biofilm formation in cheese vats. We investigated different physiological conditions in which Plb. wasatchsensis reaches optimal growth for biofilm formation on stainless steel. Biofilms were grown on cleaned, sterilized, stainless steel washers (5.92 cm2) in the wells of sterile, 24-well, polystyrene culture plates. Washers were added to 2 ml of MRS+R broth amended independently for each experiment and wells were inoculated with either 10 or 100 µl of a four-day culture of Plb. wasatchensis WDC04. The variables tested were pH (4, 5, or 6), galactose concentration (0.5%, 1%, or 2% v/v), lactose concentration (1% and 2%), and glucose (1% and 2%). Biofilms were allowed to develop for one week at 30 °C. The washers were aseptically removed and rinsed with sterile distilled water to remove planktonic cells. Attached biofilm was removed by vortexing in sterile saline for 1 min. The solution was serially diluted and plated in duplicate on MRS+R agar and incubated at 30 °C for five days. Biofilm growth was below detection at pH 4 and 7. Biofilm formation was greatest at pH 6 with 1% or 2% galactose (3.4 × 104 cfu/cm2 and 6.8 × 104 cfu/cm2). Similar growth was observed with 1% lactose (5.2 × 104 cfu/cm2). Understanding the optimal growth conditions will provide us with opportunities to test methods for the prevention or removal of WDC04 biofilms that may occur in dairy processing facilities.

Endemic Microbial Communities of Seafloor Mantle Rocks Motamedi S, Twing KI, Pendleton HL, Brazelton WJ University of Utah

Ultramafic rocks in Earth’s mantle represent a tremendous reservoir of carbon and reducing power. Mixing of these rocks with overlying seawater due to tectonic uplift causes an exothermic reaction known as ‘serpentinization’ that also releases hydrogen gas (H2), methane (CH4), and small organic molecules.

During the International Ocean Discovery Program Expedition 357 to the Atlantis Massif, rocks were collected from a subseafloor site of active serpentinization for the first time. The problem of contamination of subsurface samples was a primary concern during all stages of this project.

Many precautions were taken to keep potential contaminants away from the endemic microbial communities of the rock samples. As soon as the rock samples were brought up to the ship, the outer surfaces of these rocks were flamed to remove any contaminants from the seawater. Frozen rock samples were sent to the Kochi Institute for Core Sample Research in Japan, where the rock samples were sectioned with a custom-designed rock saw in a microbiologically-clean facility and crushed into fine grains in preparation for DNA extraction. In the Brazelton lab at the University of Utah, DNA extraction was conducted in an air-purified room with low levels of dust particles. Environmental DNA was purified with an Aurora DNA purification system to remove any remaining mineral impurities from the rocks.

This research project has produced the first census of the microbial diversity within the rocky serpentinite subsurface that underlies the Lost City Hydrothermal Field through cultivation-independent environmental DNA sequencing methods and have implications for life detection experiments, including the design of sampling strategies, the handling of samples in situ and in the lab, and the analysis of large datasets.

Understanding ALS: PAS kinase and Ataxin-2 Newey C, Pape JA, Grose JH Brigham Young University—Provo

When Lou Gehrig was diagnosed with Amyotrophic lateral sclerosis (ALS) in 1939 there was no cure. He died 2 years later in 1941. Eighty years later there is still no cure and the average survival time for people diagnosed with ALS is 3–5 years. As we search for a cure, the protein Ataxin-2 has been shown to extend life of ALS model mice (TDP-43 mice) when therapeutically reduced (Becker et al., 2017). While mutations in Ataxin-2 have been shown to increase likelihood of developing ALS, there is little understood about the molecular pathway behind this phenotype. Some evidence for how this is occurring is that Ataxin-2 has been shown to sequester proteins to stress granules, which is normally a protective cell mechanism however it may become pathological in the development of a variety of diseases including ALS. One protein of note that has previously been discovered to be sequestered to the stress granules by Ataxin-2 is TORC1, a protein complex involved in nutrient, energy and redox sensing as well as protein synthesis. Using the power of yeast genetics, we recently discovered that a nutrient sensing protein kinase PAS kinase is directly phosphorylating Ataxin-2 and is positively regulating the sequestration of Ataxin-2 and TORC1 to stress granules. We have also discovered that Ataxin-2 is involved in the sequestration of PTC6 to stress granules. PTC6 is a protein involved in mitophagy, a cellular processes which clears out damaged mitochondria. We have characterized the positive regulation of Ataxin-2 by PAS kinase and we are characterizing how Ataxin-2 is involved in the sequestration of both TORC1 and PTC6 to the stress granules, how this affects mitochondrial health, and what this implies about the development of ALS and other neurodegenerative diseases.

Alcohol-Free Hand Sanitizer Kills SARS-CoV-2 Ogilvie BH, Solis-Leal A, López JB, Poole BD, Robison RA, Berges BK Brigham Young University—Provo

SARS-CoV-2 is the virus responsible for the current global pandemic, COVID-19. Because this virus is novel, little is known about its sensitivity to disinfection. In this study, we performed suspension tests against SARS-CoV-2 using three commercially available quaternary ammonium compound (Quat) disinfectants and one laboratory-made 0.2% benzalkonium chloride solution. Three of the four formulations completely inactivated the virus within 15 seconds of contact, even in the presence of a soil load or when diluted in hard water. We conclude that Quats rapidly inactivate SARS-CoV-2, making them potentially useful for controlling SARS-CoV-2 spread in hospitals and the community.

Preliminary Results of an Antimicrobial Blue Light Device to Eradicate Bacteria at the Skin–Implant Interface of Osseointegrated Implants Ong J, Godfrey R, Peterson T, Tam J, Drake L, Farinelli B, Isaacson B, Pasquina P, Epperson RT, Williams DL University of Utah

Percutaneous osseointegrated prosthetics (POPs) are becoming available for amputees as an alternative to prosthetic socket technology. POPs make it easier for amputees to attach and detach prostheses, and improve mechanical force feedback through the implant. Unfortunately, the nature of percutaneous implants makes them highly susceptible to infection. Antimicrobial blue light (aBL) is hypothesized to manage bacterial burden and biofilm formation at the skin–implant interface. Additionally, we are testing the efficacy of oregano oil and CZ (anti-biofilm compound). Once tested independently, we will observe any synergistic effects between aBL and the oregano or CZ to improve our treatment’s effectiveness for managing bioburden.

First, we developed an ex vivo system to simulate the POP environment. Biofilms of Staphylococcus aureus are grown on titanium coupons in a CDC biofilm reactor for 72 hours, scraped off the surface, and inoculated at the site of the simulated skin–implant interface. During the aBL experiments, the inoculated sheep skin is exposed to aBL for 1 hour two times, 6 h apart, which constituted a 1-day treatment. For a 1-day treatment of oregano and CZ, each is formulated into a gel, after which 1.5 mL of gel is aseptically applied to each skin sample. Each sample is then gauze wrapped and placed in the incubator for 24 h. The three methods are currently being extended to a 7-day treatment plan.

Preliminary data indicates a 1-day treatment of any of the three methods (aBL, CZ, oregano), reduces

bioburden by 1–2 log10 units. While not yet completed, our 7-day plan data demonstrates significant reduction in CFU counts each additional day treatment is applied.

Overall, an aBL device, CZ gel, or oregano gel may help kill biofilm at the skin–implant interface in POPs, but all need more than a single application to be effective. We will focus on finishing the 7-day treatment experiments for each method after which we will test for synergistic effects.

Cellular Activity of ArtAB Toxin from Bovine Salmonella Typhimurium Overgaard EM, Tinker JK Boise State University

Salmonellosis is the most common bacterial foodborne illness in the United States. Non-typhoidal strains of Salmonella can infect both humans and animals; resulting in acute gastroenteritis. Bovines can harbor sub-clinical infections and are a main reservoir of disease for humans. Despite the availability of Salmonella vaccines for bovines, the incidence of the disease has not declined, and a more effective vaccine is needed. Bacterial AB5 toxins are key virulence factors and important antigens in some licensed vaccines. S. Typhimurium phage-type DT014 harbors an AB5-type toxin, ArtAB, that is not well- characterized. The goal of this study was to determine the in vitro activity of ArtAB on different cell types. S. Typhimurium artAB was previously cloned into E. coli and purified using D-galactose and metal ion affinity chromatography. Confocal microscopy revealed internalization of E. coli purified ArtAB-HIS into Vero cells. Cytotoxicity assays with purified ArtAB on Vero and CHO epithelial cells indicated a slow cytotoxic response at high concentrations after more than 18 hours of incubation. During short incubation times, ArtAB stimulated cell growth and metabolic activity, even at high concentrations. Analysis of cellular structure indicated that ArtAB caused a clustering phenotype, similar to that of pertussis toxin (PT) and distinct from the cellular elongation caused by cholera toxin (CT) on CHO cells. On Vero cells, ArtAB uniquely stimulated dendrite formation at lower concentrations. Findings indicate that purified ArtAB can bind to and enter tissue culture cells and that this toxin has a reproducible in vitro cellular activity similar to that of PT. In future studies, we will assess the activity of purified ArtAB in vivo. Findings will determine the contributions of ArtAB to toxicity and virulence in bovines and will inform the construction of a novel mucosal Salmonella bovine vaccine that will utilize the antigen and potential adjuvant activity of ArtAB.

Impact of Celiac Disease and a Gluten-Free Diet on the Microbiome of the Human G.I. Tract Rawson C, Hooper VA, Hansen RJ, Gazdik Stofer MA Utah Valley University

The human microbiome is the community of microbes associated with the human body. Imbalances in the microbiome is associated with a variety of symptoms. Celiac Disease (CD) is characterized by an immune response in the small intestine that is triggered by the protein gluten, found in wheat. Previous studies have identified microbial dysbiosis in patients with CD. To better understand changes in the microbiome of celiac patients we examined the microbiome diversity and composition in CD patients compared to healthy individuals on gluten-free diets. This will help identify what microbiome changes are associated with CD compared to those that are caused by the CD patients lack gluten in their diet. We analyzed: those diagnosed with CD and on a gluten-free diet, those who do not have CD and are on a gluten-free diet, and those who do not have CD and are not on a specific diet. The dietary and disease influences on the microbiome of the gastrointestinal tract was analyzed via a single stool sample collected from each study subject. Microbial DNA was extracted using the QIAamp PowerFecal DNA kit following manufacturer's specifications (Qiagen); the V3/V4 region of the 16s rRNA was sequenced using the 600 cycle v3 MiSeq kit (Illumina). Sequenced data was statistically analyzed using a Kruskal–Wallis analysis. At the phylum level data between the groups was not statistically significant. However, at the genus level, there was statistical significant with two genera. The first was the archaeon Methanobrevibacter, which was found only in the control group (p = 0.212). The second was the bacteria Dialister, which two-fold higher in then CD group compared to the control and was nine-fold higher in the control group compared to the GF group (p = 0.0204). Further analysis is needed to determine if these microbiome changes could have physiological relevance.

Exploring the Human Phageome through Sewage Ayers H, Bustos Y, Correa E, Ramsey J, Reed J, Ringger E, Soule C, Soule S, Telford M, Flor S, Breakwell DP, Grose JH Brigham Young University—Provo

The microbiome plays a pivotal role in maintaining an organism’s health. Many human diseases, including cancer and obesity are linked to specific changes in the microbiome. Bacteriophages, which often infect a very specific range of bacterial strains, may provide the ability to diagnose or predict gut-related diseases earlier than current methods. Current assays of the human microbiome do not allow for specificity between pathogenic and nonpathogenic strains of bacteria, and cannot distinguish between bacterial strains of the same species. Studying the human phageome can provide a more accurate view of any changes in an organism’s microbiome. So far, only the CrAss phage family has been found to be common in all human gut flora; however, we hypothesize that other common phage families exist. Previous human phageome studies attempted to find common human phages by filtration of fecal samples followed by sequencing of any found viral DNA. Instead, we performed PCR amplification on sewage samples directly, with primers specific to Serratia and Klebsiella phage families. Specifically, we tested whether phages belonging to these families were found abundantly in sewage samples collected in Utah County. We found that several of these phages were indeed detectable by running PCR directly on sewage samples, which is a promising initial result, as it demonstrates that PCR is a viable way to test raw samples for the presence of a phage. Future work in this direction will include performing similar tests directly on individual sewage samples. Our findings set a precedent for an expanded view of the human phageome and more specific observations of any given individual’s or population’s collective microbiome. If more phage families are found to be common to large groups of individuals, this would open the door for further research in disease diagnostics.

A Signaling Pathway-Driven Bioinformatics Pipeline for Predicting Therapeutics against Emerging Infectious Diseases Scott TM, Jensen S, Pickett BE Brigham Young University—Provo

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel Betacoronavirus that was first reported in Wuhan, China in December of 2019. Its associated disease has been named coronavirus disease-2019 (COVID-19), which has now become associated with a worldwide pandemic. The devastating effects of this pandemic have highlighted the need to quickly identify potential prophylactic or therapeutic treatments that can reduce the signs, symptoms, and/or spread of disease when dealing with a novel infectious agent. We constructed a novel computational pipeline that predicts drugs and biologics that could be repurposed to combat a novel infectious disease. Specifically, this workflow analyzes RNA sequencing data to determine differentially expressed genes, enriched Gene Ontology (GO) terms, and dysregulated pathways in infected cells, which can then be used to identify FDA-approved drugs that target proteins within these pathways. We used this pipeline to perform a meta-analysis of RNA sequencing data from cells infected with three Betacoronaviruses including severe acute respiratory syndrome coronavirus (SARS-CoV; SARS), Middle East respiratory syndrome coronavirus (MERS-CoV; MERS), and SARS-CoV-2, as well as respiratory syncytial virus (RSV) and influenza A virus (IAV) to identify therapeutics that could be used to treat COVID-19. This analysis identified twenty-seven existing drugs, most of which already have FDA-approval, that are predicted to counter the effects of SARS-CoV-2 infection. These results are validated by separate studies involving canakinumab, anakinra, tocilizumab, sarilumab, baricitinib, and others. While the results reported here are specific to Betacoronaviruses, such as SARS-CoV-2, our bioinformatics pipeline can be used to quickly identify candidate drugs for treating future emerging infectious diseases.

Fermentation of Plant-Based Extracts by Dairy Lactic Acid Bacteria Smith J, Mishra N, Oberg CJ, Culumber MC Weber State University

Plant-based alternatives to fermented dairy products are growing in popularity and economic importance. However, the qualities of fermented dairy products are difficult to recreate with plant-based extracts and little is known about the metabolism of lactic acid bacteria (LAB) in plant extracts. This study evaluated the ability of LAB to cause a pH change in almond, coconut, or oat-derived beverages through fermentation. Cultures used included single strains of Streptococcus thermophilus, Lactobacillus rhamnosus, Bifidobacterium animalis, and Lactobacillus casei, and mixtures of Lactobacillus sp., S. thermophilus, and Bifidobacterium sp. that are commercially used for fermentation or probiotic supplementation. Carbohydrate use by individual LAB cultures was first evaluated using API 50 CH panels. All cultures were able to use galactose, glucose, fructose, mannose, esculin, and tagatose. The ability of these LAB to ferment carbohydrates present in the plant extracts supports our hypothesis that these organisms can actively ferment substrates present in plant extracts. Three commercial plant-based beverages were incubated with commercial LAB cultures at three inoculum levels, 0.5, 1.0, and 2.0% w/v, and pH monitored during incubation at 37 °C over 7 hours. Acid production in coconut and almond extract was only observed at the highest inoculum level. However, three of the LAB strains, S. thermophilus YFLO1, Lb. casei 431, Lb. rhamnosus LGG produced acid in the oat extract at all inoculum levels. The pH dropped from 7.5 to 5.75 within 240 minutes. When inoculated into oat extract, the Lactobacillus sp. survived 5 hours of incubation with little change in the CFU/ml. We also tested the ability of B. animalis subsp. lactis BB12, to survive or grow in plant-based beverages. We observed a lower pH (5.5) improved survival of BB12 in oat extract by 155% over 14 days. Results showed fermentation is LAB strain dependent based upon the type of plant extract being fermented.

Development of a Novel Anti-Biofilm Polyurethane Foam for use in Negative Pressure Wound Therapy Smith TB, Rawson K, Neuberger T, Bell I, Hooper N, Brown S, Parkin M, Williams DL University of Utah

Background: Negative pressure wound therapy (NPWT) is an increasingly prevalent treatment for high energy, traumatic battlefield-related injuries, which are typically caused by a blast from an improvised explosive device or a rocket propelled grenade. Pathogenic biofilm can contaminate these wounds and lead to chronic infections because the structure of polyurethane (PU) foam used in NPWT may harbor and encourage increased biofilm growth. To date, Granufoam Silver is the only variation of PU foam for NPWT that possesses any degree of antimicrobial efficacy but is minimally effective against biofilms.

Methods: Since Granufoam Silver is a 10.0% w/w of silver nanoparticles on a PU foam matrix, we produced prototypes of 2.5%, 5.0%, and 10.0% w/w of active CZ-01179 to PU foam. Minimum inhibitory concentration (MIC) values and antibiofilm efficacy of CZ-01179 were determined using in vitro broth dilution assays. Ex vivo testing was performed by inoculating pig skin explants with MRSA and A. baumannii biofilms, followed by a two-day incubation period. The skins were treated with Granufoam Silver and 10% w/w CZ-01179 + PU in the presence of NPWT for 1, 3, and 7 days. In vivo antibiofilm efficacy is currently being analyzed by surgically creating 4×4 cm porcine excision wounds, followed by a 3-day inoculation period. The wounds are then treated with Granufoam Silver or 10.0% w/w CZ-01179-PU under NPWT for 7 days.

Results: Broth dilution testing showed an 8 log10 and 5 log10 reduction of MRSA and a 9 log10 and 6 log10 reduction of A. baumannii against 10.0% and 5.0% w/w CZ-01179-PU over 24 h, respectively. Ex vivo testing resulted in a 4 log10 (7 days) reduction against MRSA and 9 log10 (7 day) reduction against A. baumannii for 10% w/w CZ-01179-PU. For these same time periods Granufoam Silver expressed a <1 log10 reduction against both MRSA and A. baumannii. In-vivo efficacy testing through the 7-day end point has thus resulted in a 4 log10 reduction against A. baumannii and a 3 log10 reduction against MRSA. In vivo testing continues.

Conclusion: Data indicates that the novel antibiofilm compound can be loaded into PU foam repeatably and display antimicrobial activity. Furthermore, in vitro, ex vivo, and in vivo data indicates that CZ-01179 possess greater antibiofilm efficacy against MRSA and A. baumannii in comparison to Granufoam Silver.

Role of CD5 in Oral Inflammation and Periodontal Disease Sperry YK, Townsend J, Mahler K, Cardon D, Téllez Freitas CM, Weber KS Brigham Young University—Provo

In 2016 the World Health Organization reported that oral diseases affected half of the global population. Oral diseases, such as periodontal disease, can be caused by poor oral hygiene and/or tobacco use. Prolonged exposure to bacteria leads to chronic inflammation. Lymphocytes invade the site and initiate inflammation which can cause tooth loss and is a risk factor for heart and lung disease. Previous studies have shown that patients with severe periodontitis have increased autoreactive B lymphocytes expressing high frequencies of CD5. CD5 is a T cell co-receptor that regulates T cell development and function. We propose to investigate the role of CD5 in helper T cell activation and its effect in oral inflammation. We will use mice genetically altered to have T cells with or without CD5. We hypothesize that CD5 enhances T cell activation in periodontal disease. We will test this hypothesis by co-culturing T cells expressing or lacking CD5 with supernatant from oral mucosal or gingival epithelial cells that have been exposed to LPS (lipopolysaccharide, a major component of the outer membrane of Gram-negative bacteria). We hope to characterize the T cell response based on cell surface marker expression, cytokine production, proliferation, and migration.

Early Onset Alzheimer's Disease in Pacific Island Populations Tavana JP, Weber KS, Jensen JL, Hansen DV, Kauwe JSK Brigham Young University—Provo

Alzheimer’s disease (AD) is the most common form of dementia and affects nearly 50 million people worldwide. Early onset AD (EOAD) is responsible for less than 5% of AD cases with an estimated prevalence of 41.2 per 100,000 persons at risk.1 According to the 2010 census, Asian Americans and Pacific Islanders (AAPI) are the fastest growing minority group in the United States.2 In Samoa, Alzheimer’s disease ranked sixth and in Tonga ranked tenth among causes of mortality in 2018.3 The prevalence and incidence of Alzheimer’s disease and related dementias within AAPI communities and in indigenous PI’s is not well understood. Data on AAPI subpopulations in the US and in the Pacific Islands is lacking. We have initiated a collaborative research project to correct this deficit by establishing a Pacific Islander (PI) dementia cohort in Utah, Samoa, and Tonga. To do this, we are developing and validating dementia screening tools in their native languages that are optimized for these populations. Second, we are enrolling a PI cohort in Utah. Third, we will enroll indigenous Samoan and Tongan dementia cohorts. And finally, we will conduct whole genome sequencing and variant prioritization on DNA samples collected from our dementia subjects. In summary, this study provides opportunities to observe very rare and population-specific genetic variation in PI populations with early onset Alzheimer’s Disesase. The gathered information will not only benefit those involved but also contribute to our understanding of Alzheimer’s disease progression by identifying and investigating genetic factors for Alzheimer’s disease. Continued genetic research in diverse populations will maximize genetic discovery and reduce health disparities in these populations.4

Helicase-Mediated Mechanisms for Defense against Invasive Plasmids by the Type IV-A CRISPR–Cas Bacterial Immune System Taylor HN, Domgaard H, Metcalf J, Jackson RN Utah State University

Clustered regularly interspaced short palindromic repeats–CRISPR associated (CRISPR–Cas) systems are used by bacteria and archaea to defend against invasive mobile genetic elements (MGEs), such as bacteriophage and plasmids. CRISPR–Cas systems are highly diverse and work through various mechanisms to identify and eliminate invasive MGEs. Type IV CRISPR–Cas systems are the least studied CRISPR–Cas type. Type IV-A systems, a subtype of type IV CRISPR–Cas, have recently been shown to defend against invasive plasmids but the mechanisms through which this defense occurs are unknown. To determine the mechanisms employed by the type IV-A system, we recombinantly expressed and purified a type IV-A Csf ribonucleoprotein (RNP) complex and CasDinG helicase from Pseuodomonas aeruginosa. We used a series of in vitro biochemical experiments, including EMSAs and helicase assays, to characterize the helicase activity of CasDinG, show binding of the Csf RNP complex to a nucleic acid substrate through complementarity to a CRISPR-derived RNA, and the apparent recruitment of CasDinG to a substrate by the Csf RNP complex. We proposed a model of type IV-A CRISPR–Cas defense. This work fills a major gap in the CRISPR–Cas field and uncovers a unique mechanism of prokaryotic defense.

Identification of Staphylococcus aureus Genes of Interest for a Bovine Mastitis Vaccine Wheeler C, Scarbrough D, Tinker JK Boise State University

Bovine mastitis, painful inflammation of the udder, caused by an infection from Staphylococcus aureus, remains to be a formidable opponent within the US Dairy industry. Mastitis infection costs the dairy industry billions in losses, but there has yet to be a vaccine developed to effectively protect against S. aureus infection. Our lab has previously constructed an S. aureus vaccine that includes the targeted conserved S. aureus genes, isdA and clfA. While the results of vaccine trials revealed an antigen-specific immune response among inoculated cows, it was not successful in preventing infection in a challenge study (Misra et al. 2018). Through proteomics, genomics and the use of bioinformatic tools such as PSortB, Vaxign, and the NCBI database, we have identified six top genes of interest: cvfC, fib, metN2, ssaA, isaA, and lysM. Currently, we are investigating homology among these genes within different S. aureus genomes and confirming presence of these genes through PCR methods. Successful identification of the conservation of these genes will promote cross-protection against different strains of S. aureus. Future use of reverse vaccinology methods will enable epitope predictions, antigenicity and immunogenicity specific to S. aureus. Results will support the construction of an effective multi-valent vaccine to prevent bovine mastitis.

The Role of T Cell Co-Receptor CD5 in T Cell Metabolism, the Gut Microbiome, and Behavior Whitley KV, Téllez Freitas CM, Cox T, Magoffin W, Reaveley K, von Wallwitz I, Mahler K, Haynie C, Tall A, Teasdale J, Weber KS Brigham Young University—Provo

Helper T cells are well known as the coordinators of the adaptive immune system and are critical to an effective immune system response to infection or cancer. Helper T cell activation is regulated by three primary interactions: The T cell receptor (TCR) and peptide presented by MHC II, followed by co-receptor simulation or inhibition and cytokine regulation. CD5 is a T cell co-receptor that regulates thymocyte selection and peripheral T cell activation. Removal of CD5 has been shown to not only affect T cell functions such as metabolism and calcium signaling, but also systemically affect gut microbiome composition and mouse behavior. To better understand the systematic roles of CD5 and the microbiome, we bred wild type and CD5 knockout mice to generate offspring that have different genotypes but have the same microbiome. Our preliminary results suggest CD5-driven and gut microbiome mechanisms that regulate mouse behavior, indicating that balance between the immune system, the gut microbiome, and the brain must be considered in the development of immunotherapies to increase effectiveness and success.

Poster Abstracts (alphabetical, by last name of presenter)

Optimizing Translation Efficiency of Recombinant Proteins in E. coli Benedict A, Chamberlain J, Griffitts JS Weber State University

The most common source of loss at Donor Human Milk banks is milk that tests positive for Bacillus cereus after pasteurization. This bacterium is a problem because of its ability to form endospores which survive the pasteurization process. Bacillus cereus can cause serious complications in compromised infants. My objective is to see if pre-pasteurization B. cereus concentrations can predict post-pasteurization contamination. If we could use pre-pasteurization counts for Bacillus, it would prevent loss of pasteurized human donor milk, and decrease loss of revenue for milk banks. Breast milk samples were inoculated with B. cereus, which was quantified on nutrient agar with 2% agar before and after pasteurization. Milk was pasteurized at 62.5 °C for 30 minutes. Initially, pasteurization reduced the number of B. cereus colonies by more than half compared to the pre-pasteurized samples. (One test went from 16 CFUs/ml to 2 CFUs/ml). However, if the milk was inoculated with Bacillus and frozen for storage before pasteurization, the number of CFUs/ml of B. cereus post-pasteurization increased or was similar to pre-pasteurization. These preliminary results may indicate that freezing the B. cereus activates endospore formation and could have a significant impact on how donor milk is stored prior to pasteurization.

The Behavior of Bacillus cereus in Human Breast Milk Pre- and Post-Pasteurization Church B, Culumber MC Weber State University

Epigenetic Profiling of Human Peripheral Blood Monocytes Cole N, Adams K, Johnson SM Brigham Young University—Provo

Identifying the epigenetic mechanisms at play within cell-types is one of the early steps in understanding crucial differences between healthy and diseased cells. Peripheral blood monocytes play a crucial role in both the innate and adaptive immune system. We explore differences that exists within the epigenetic profiles of human monocytes based on sex. To determine if chromatin accessibility is sexually dimorphic, we collected blood samples from 6 volunteer adults (3 men and 3 women). Monocytes were then isolated from each volunteer. The ATAC-seq protocol was followed, and data analysis was performed. The results of this analysis will determine to what extent chromatin accessibility of human monocytes is impacted by sex. These differences could have implications for therapeutics targeting the immune system.

Survey of North Dakota Rhizobia to Improve Nodulation and Nitrogen Fixation in Chickpea Laughter C, Paryzek J, Fonseka D, Crook MB, Wiley-Kalil A Weber State University

Chickpea is becoming an increasingly important crop in North Dakota, particularly in drier regions of the state where growers are looking for high value crops that can tolerate semi-arid growing conditions to break up disease cycles and balance risk in crop rotations. Fertilizer input costs are much lower for chickpea than for non-legume crops due to the ability of these plants to benefit from nitrogen fixation, where rhizobia induce the formation of nodules on the plant roots and fix atmospheric nitrogen for the plant host. This benefits not only the original chickpea crop, but crops grown in the subsequent growing seasons. However, good nodulation of the chickpea crop is not guaranteed. Nodulation requires either inoculation or that the correct species of rhizobia already be present in the soil. Researchers in western North Dakota have observed poor or no nodulation in chickpea crops. We hypothesized that commonly used chickpea inoculants are not well-adapted to North Dakota soils, which impairs their ability to effectively nodulate chickpea. Thus we undertook a survey of chickpea-nodulating rhizobia that are native to North Dakota soils in order to determine whether their nodulation and nitrogen fixation rates with chickpea are superior to commonly used inoculants. In 2018, soil was collected prior to planting from 31 fields in six counties in North Dakota (Divide, Williams, Golden Valley, Burke, Ward, and Bottineau) and Sheridan County, Montana, and analyzed for nitrate content. Frontier and Sierra chickpea varieties were planted in the different soil samples and grown in the greenhouse for 3–4 weeks. Five plants were also collected from each field during the growing season to assess their nodulation rate. Rhizobia were isolated from 2 nodules per plant (both greenhouse plants and field plants) and identified by 16S sequencing; for some strains three additional genes (recA, gyrB, and atpD) were sequenced for phylogenetic analysis. We plan to test these strains for their ability to nodulate and fix nitrogen both in laboratory and field settings with an eye toward developing a commercial chickpea inoculant with high performance in North Dakota soils.

The CCRL2 Atypical Chemokine Receptor Variant V180M Has a Higher Binding Affinity for Chemerin Dearden L, Nelson A, González Murcia JD, Weinert A, Téllez Freitas CM, Ferrel MN, Grose JH, Ridge PG, Wilson E, Kauwe JSK, Weber KS Brigham Young University—Provo

A recent genome-wide association study of 59 cerebrospinal fluid proteins with connection to Alzheimer’s disease (AD) demonstrated an association between decreased levels of chemokine ligand 4 (CCL4) and an atypical chemokine receptor, C-C chemokine receptor-like 2 variant V180M (CCRL2-V180M). Low levels of CCL4 are associated with a decreased risk of AD development as well as other inflammatory diseases. In this study, we characterized the biological function of the CCRL2-V180M allele compared to the wild-type allele in both CCRL2 receptor isoforms (CRAM-A and CRAM-B). We characterized the CCRL2-V180M variant by analyzing predicted folding protein structure and hydrophilicity interactions. We transfected CHO-k1 cells with plasmids carrying the wild-type (WT) or V180M variant of the CCRL2 gene and analyzed binding affinity with known receptor ligand chemerin in both WT and V180M cells. Protein model analysis did not predict CCRL2-V180M causing a structural change compared to the CCRL2-WT protein, but it did predict changes of hydrophilicity interactions between CCRL2-V180M and CCRL2-WT in both CRAM-A and CRAM-B isoforms. Cell based binding assays revealed that both CRAM-A-V180M and CRAM-B-V180M have higher binding affinity for chemerin compared to CCRL2-WT. These findings provide molecular and biological mechanistic insights into the GWAS association between CCRL2-V180M and decreased levels of CCL4 in CSF, providing a potential biological mechanism for a previously reported association of CCRL2 variants and decreased CCL4 levels with reduced risk of AD.

Discovery of the Human Phageome for Future Disease Diagnostics Ayers H, Bustos Y, Correa E, Ramsey J, Reed H, Ringger E, Soule C, Soule S, Telford M, Flor S, Breakwell DP, Grose JH Brigham Young University—Provo

Analysis of the Microbiome Composition from Low-Income Homes with Evaporative Coolers and Central Air Conditioners Fuller S, Lambson L, Johnston JD, Weber KS Brigham Young University—Provo

Recent work suggests that evaporative coolers increase the level and diversity of bioaerosols. We conducted a cross-sectional study of metropolitan, low-income homes in Utah with evaporative coolers and central air conditioners. Dust samples were collected from four locations in each home and analyzed for dust-mite allergens, endotoxins, and β-(1→3)-D-glucans. We found that dust mite antigens, endotoxins, and β-(1→3)-D-glucans were all significantly higher in evaporative cooler versus central air conditioning homes. The composition of microbiome populations in homes plays a key role in developmental and daily health, with a critical role in asthma and allergy development. Using high through-put sequencing we will analyze the bacterial, fungal, and archaeal community in homes using evaporative coolers and central air conditioners. Analysis of the V3 & V4 region of the 16S rRNA gene will be performed to identify and categorize the taxa in dust from these homes. Microbial DNA will be isolated, and bar codes will be added to enable multiplexed sequencing. High through-put sequencing will be performed in an ISeq100 and identification of the microbial community will then be performed and microbial populations from homes with evaporative coolers and central air conditioners will be compared.

Using a Virus Model System to Determine the Effect of Ozone in Locker Room Sanitation Hansen J, Coles J, Madsen C, Steiner C, Nyone C, Scharmann B, Scharmann S, Mishra N, Oberg CJ McKay Dee Family Medicine Residency, Intermountain Healthcare

Viral infections can spread rapidly through a team with surface contamination a common method of transmission. Ozone has been shown in several laboratory studies to be effective at inactivating viruses. We examined the effectiveness of ozone for viral inactivation on solid surfaces in an active Division I collegiate locker room. Sterile petri plates were inoculated with E. coli bacteriophage T7, used as the viral model system, and placed in triplicate 3 to 27 feet from an ozonator (Extreme Ozone Co.) in a collegiate locker room. Plates were exposed to ozone for either 2 or 3 h. Virus was harvested from the plates with sterile saline, diluted, and enumerated using the soft agar overlay procedure with E. coli B. After incubating for 24 h at 37 °C, viral plaques were counted and compared to controls not exposed to ozone. A peak ozone level of 3.6 ppm with an overall average of 1.6 ppm was measured during the 3 h exposure. Results showed a significant decrease (50–97%) in viral load for 8/20 test plates. After the 2 h trial, no viral inactivation was observed. For 3 h of exposure, ozone reduced viral counts by up to two logs on some exposed plates, but the effect was variable in plates laid next to each and did not correlate to the distance from the ozonator. While duration of ozone exposure appears to play a role in viral inactivation (2 h vs 3 h), other real-world factors such as room ventilation and humidity could also play a role. Results are consistent with laboratory studies which found a 1–2 log decrease in viral load after ozone exposure. In comparison, use of a similar methodology with Staphylococcus aureus and E. coli showed consistent kill rates of <90%. Our results indicate that typical ozonation treatment of locker rooms may have more variable real-world results compared to experiments done in controlled laboratory settings and ozone at this concentration should not solely be relied upon for locker room virus control.

The Role of CD5 in Regulating Levels of T Follicular Helper Cells Haynie CJ, Whitley KV, Téllez Freitas CM, Weber KS Brigham Young University—Provo

Helper T cells (TH) play a critical role in the adaptive immune response. Naïve T cells differentiate upon activation based upon multiple factors including cytokine signaling, transcription factor regulation and co- receptor presence. Among the various TH subsets, T follicular helper cells (TFH) are responsible for aiding activation of B cells for antibody production. It is well established that T cell co-receptors influence T cell activation and function and a recent report has shown that changes in tonic signaling play a key role in TFH cell-lineage decisions. CD5 is an inhibitory co-receptor expressed on the surface of T cells and is known to regulate thymocyte selection and T cell receptor (TCR) signaling. We compared wild-type C57/B6 and knockout CD5−/− mice and found that the knockout mice have a distinctly different gut microbiome and free IgA levels in serum was lower in CD5 knockout mice. Because of these observations, we are interested −/− in determining how TFH cells in wild-type C57/B6 and CD5 knockout mice are involved in regulating the immune–gut interface.

Gene Expression Changes in Plants Inoculated with Halophilic Bacteria and Grown in Salty Soil Miller A, Knowles A, Hill J, Pickett BE, Nielsen BL Brigham Young University—Provo

We have isolated several halophilic bacteria from native halophytes growing near Goshen, Utah at the south end of Utah Lake. These isolates have been screened as plant inoculants for stimulation of alfalfa growth in salty soil. One Halomonas strain supports consistent growth stimulation under these conditions. This raises the question as to what is the molecular plant response to bacterial inoculation? We have isolated RNA from control and inoculated plants grown in the presence and absence of 1% NaCl. Gene expression levels and relative changes in each treatment were analyzed by RNAseq and quantitative reverse-transcriptase PCR (qRT-PCR). Several genes were significantly up- and down-regulated in inoculated plants grown in the presence of salt, including some genes such as catalase that may directly relate to the plant growth stimulation levels observed.

Effect of Inorganic Salts on the Growth of Probiotic Lactic Acid Bacteria Mishra N, Oberg CJ, Culumber MC Weber State University

Dairy-based products are often used as delivery systems for probiotics. Unfortunately, many people miss the benefits of probiotic-enhanced products because they cannot consume dairy foods due to lactose intolerance or allergies. Our research explores using plant-based products as a delivery system for probiotics. Previous research has shown promising results, but many commercial products contain phosphate salts which are known to inhibit some probiotic lactobacilli. Our research examined the growth of lactic acid bacteria (LAB) in a plant-based product, “oat milk,” and the impact of phosphate salt preservatives on LAB growth. Oat-based beverages with phosphate-containing salts were inoculated with 24 h cultures of commercial probiotic strains: Lactobacillus acidophilus LA-5, Bifidobacterium BB12, Lb. casei 431, Lb. casei F-19, Lb. rhamnosis BLC-48, and Lb. rhamnosis LGG. LAB survival was enumerated using MRS agar for Lactobacillus and MRS+cystine for Bifidobacterium strains. Plates were incubated anaerobically at 35 °C with counts done at days 0, 7, and 14.

Results showed some strains of LAB grew in the oat-beverages with or without added salts and that all cultures survived, regardless of the composition, however, most strains preferred growth in specific brands. Simple Truth “oat milk” brand gave the best growth results for most strains. We expect differences in probiotic strain growth was due to the composition of each brand. The protocol was repeated using only Chobani oat-beverage products that were either treated with phosphate salts or left untreated. All cultures survived independent of the phosphate concentration, but most did not grow over the 14 days of refrigerated incubation. Four of five strains survived approximately 10-fold better in the treated product. These results show the addition of phosphate-containing salts as stabilizers in “oat milk” does not inhibit most probiotic strains so it can be used as a probiotic delivery system.

Measuring the Dragonfly (Tanypteryx hagen) Genome Size Using Flow Cytometry Moreno C, Frandsen P, Weber KS Brigham Young University—Provo

Flow cytometry has been a reliable method for estimating the genome size of various organisms including alfalfa, tomato, beetles, fruit flies, cattle, mice etc. In fact, Pflug et al. found flow cytometry was more reliable and accurate at estimating the genome sizes of beetles than next-generation sequencing methods. Here we are using flow cytometry to estimate the size of the dragonfly genome (Tanypteryx hagen) which will be valuable to comparing it to its closest relatives and in generating a phylogenetic tree. To accomplish this, dragonfly tissue and tissue from organisms with a known genome size (Drosophila melanogaster, C57BL/6 mouse, and Lycopersicum solanum) are homogenized to release intact nuclei and stained with propidium iodide (PI). The standards used for flow cytometry genome size estimation are organisms with a relatively close size to the predicted dragonfly genome size. The samples are analyzed using flow cytometry and the mean intensities of the G1 peaks of the standards are used to estimate the dragonfly genome size. The initial genome size estimate for T. hagen is ~2.6 Gbp.

Microbial Production of Rare Sugars from Waste Lactose Moss M, Griffitts JS, Taylor B Brigham Young University—Provo

Conversion of waste lactose into low-calorie, low-glycemic index rare sugars is an attractive technological goal because 1) it offers a long-term source of high-demand, nutritionally beneficial carbohydrates and 2) it reduces the environmental impact associated with lactose and upstream dairy effluents. Recombinantly expressing β -galactosidase, glucose isomerase, D-allulose epimerase and L-arabinose isomerase enzymes in a single system should make it possible to convert both lactose monomers to rare sugars; glucose to allulose and galactose to tagatose. Enzyme fusions may increase the overall efficacy of the multi-enzyme system or at the very least reduce the burden of isolating and purifying the enzymes separately. Increasing host cell permeability to lactose by over-expressing lacY could be one way to generate a whole-cell system in E. coli wherein the sugar conversions can take place inside the cell where the enzymes are produced without the need for enzyme purification. Utilizing a secretion tag in a S. cerevisiae vector is another feasible method to produce a whole-cell system where enzymes accumulate extracellularly during the cell proliferation step so that the sugar conversion can happen in tandem with cell growth.

The CCRL2 Atypical Chemokine Receptor Variant V180M Alters CCL4 Secretion in Leukocytes Nelson A, González Murcia JD, Dearden L, Weinert A, Téllez Freitas CM, Ferrel MN, Grose JH, Ridge PG, Wilson E, Kauwe JSK, Weber KS Brigham Young University—Provo

A genome-wide association study of cerebrospinal fluid proteins with connection to Alzheimer’s disease (AD) demonstrated an association between decreased levels of chemokine ligand 4 (CCL4) and an atypical chemokine receptor, C-C chemokine receptor-like 2 variant V180M (CCRL2-V180M). Low levels of CCL4 are associated with a decreased risk of AD development as well as other inflammatory diseases. Using a novel co-culture method, in which chemokine receptor transfectants are cultured with chemokine and peripheral blood mononuclear cells (PBMCs), we analyzed CCRL2-V180M and WT interactions with CCL19 in the presence of PBMCs to determine CCL4 secretion. We found that CRAM-A-V180M interactions with CCL19 in the presence of PBMCs results in increased CCL4 secretion when compared to CCRL2-WT transfected cells, whereas CRAM-B-V180M interactions with CCL19 in the presence of PBMCs results in decreased CCL4 secretion when compared to CCRL2-WT transfected cells. Understanding how this atypical chemokine receptor allele decreases CCL4 and possibly inflammation could lead to novel therapeutic solutions for AD and other diseases that that are exacerbated by the inflammatory response.

Can Automatic Hand Dryers Serve as a Microbial Reservoir for Contamination? Packard H, Nichols R, Domek MJ, Oberg CJ Weber State University

Automatic electric high-speed hand dryers are considered an environmentally friendly alternative as they reduce paper waste and are considered more sanitary than the paper towel dispensers since they eliminate direct contact with the dispenser and towels. Increasingly, these hand dryers are the only option in public restrooms. Our purpose for doing this research was to determine if high-speed automatic electric hand dryers in public restrooms are antiseptic or if they can serve as a source of contamination to hands during drying. Initially, university restrooms were selected for sampling that have variable degrees of foot traffic. Sampling was then done in men’s and women’s restrooms in three buildings with four bathrooms tested in each building. Testing was conducted by swabbing a 5 cm2 area of the top, middle, and bottom of each hand dryer using 3M Quickswabs. Pour plates containing TSA were used to enumerate swab samples with plates counted at 48 hours after incubation at 37 °C. Results showed the bottom of dryers in both the men’s (average of 311 CFU/5 cm2) and women’s (average of 299 CFU/5 cm2) restrooms had the most contamination. The middle section was the second most contaminated for both men’s and women’s restrooms averaging 144 CFU/5 cm2 and 145 CFU/5 cm2, respectively. The top was the least contaminated for both men’s (average 107 CFU/5 cm2) and women’s (average 51 CFU/5 cm2) restrooms. Presumptive plating on mannitol salt agar (MSA) and violet red bile agar (VRBA) of bottom dryer sections showed Staphylococcus and coliforms were present. Overall there was no difference between the two brands of dryers (Dyson Airblade vs Mediclinics Dualflow Plus) and no statistical difference between men’s and women’s restrooms. Differences were observed based on sampling location inside the dryer and for restrooms in higher traffic areas which had a higher level of microbial contamination, probably based on increased use. Results showed that high-speed automatic electric hand dryers can serve as a source of contamination during hand drying. Since the use of swabs for sampling only recovers between 1 and 10% of the total organisms present, the actual level of microbial contamination in restrooms was at least a log greater than actual plate counts. As a preventative measure, the inside of these dryers should be cleaned on a daily basis to prevent people from contaminating their hands immediately after washing them.

The Role of the Brain-Eating Amoeba Naegleria fowleri CD59-Like Protein in Human Infections Gee J, Nielsen TJ, Clark DN Weber State University

The brain-eating amoeba Naegleria fowleri is a free-living amoeba capable of an extremely rare and fatal infection of the human central nervous system known as primary amoebic meningoencephalitis. Several dozen of these rapidly progressing infections have occurred in the US, which is unfortunately most often diagnosed at death. Experimental drugs, such as amphotericin, azithromycin, and miltefosine have cured relatively few. Important in the infection process, it is believed that N. fowleri CD59-like complement regulatory protein plays a protective role in resistance to lytic cell death caused by complement. Complement proteins are found in blood serum and can bind to and promote the killing of invaders, but this reaction can be blocked by CD59. To evaluate the ability of CD59-like protein to camouflage N. fowleri against complement, anti-CD59 antibodies and serum complement were added to cultured N. fowleri. In addition, N. fowleri was treated with combinations of the experimental drugs amphotericin, azithromycin, and miltefosine. Using an infection model using cultured human HeLa cells, levels of cell death were measured in both N. fowleri (necrosis levels) and the HeLa cells they infect (apoptosis levels). Neutralization of CD59 with antibody did not consistently increase Naegleria death by complement, but combinations of drugs killed Naegleria and protected HeLa cells. These results provide a greater understanding of the pathogenesis and treatment of this devastating infection.

Determining the Role of PKMζ in Pancreatic Adenocarcinoma Teasdale J, Perry B, Ghosh S, Perry A, Weber KS Brigham Young University—Provo

Identifying novel proteins, or locating regions of abnormal protein level expression, serves a critical role in emerging therapeutic methods for genetic based diseases. Proteins that fulfill either of these requirements in cancerous growths attract particular attention, as they serve as a backbone for immunotherapies, and other emerging methods of treatment. Our project focuses on the protein PKMζ, which was thought to be highly expressed exclusively within neural tissue. Recent work has found a discrepancy in this protein's concentration within pancreatic adenocarcinoma (PAAD). PAAD comprises 85% of all pancreatic cancers and has a five-year survival rate of only 9%. Understanding the role of PKMζ in PAAD may open avenues to treatments that are less invasive and more effective. We are utilizing qPCR to test the relative abundance of PKMζ within PAAD tissues. We are also measuring the relative abundance of PKMζ in other tissue samples to validate whether or not it may serve a therapeutic role in the pancreas.

Short Variant of Turkey Hemmorhagic Enteritis Virus Vaccine Strain Quaye A, Ord B, VanDerWerff KJ, Poole BD Brigham Young University—Provo

Background Hemorrhagic enteritis (HE) is a disease affecting turkeys characterized by weight loss, bloody diarrhea, immunosuppression (IS), splenomegaly, and up to 60% mortality. This disease is caused by turkey hemorrhagic enteritis virus (THEV), belonging to the genus Siadenovirus. The THEV-A Virginia Avirulent Strain (VAS) has been used as a live vaccine for decades but still manifests immunosuppression. Our initial goal was to clone the whole virus genome into a plasmid for further studies.

Methods A commercial THEV vaccine was purchased from Hygieia Biological Labs and the entire genome (~26 kb) was cloned into a fragment of plasmid pPG012 to form a circular plasmid (~28 kb) using the NEBuilder HiFi kit. PCR was used to screen for successfully transformed colonies. Clones were examined by restriction enzyme fingerprinting, PCR, and Sanger sequencing.

Results Whole-genome plasmids were made using NEBuilder. PmeI and EcoRI digestion showed identical bands to the predicted pattern. However, one BamHI cut site near the 3′ end was missing. PCR and sequence analysis showed that the cloned 3′ end was 288 bp shorter than expected. PCR using the vaccine DNA as template revealed two bands, one corresponding to the amplicon size with a 288bp deletion and the other to the expected size with no deletion. Sequencing found that the longer band had the published sequence, while the shorter band had the same 288bp deletion as found in the plasmid.

Conclusions The 288bp deletion at the 3′ end of the virus genome is not an artefact from our cloning procedure but is a variant of the genome derived from the vaccine. The vaccine contains two variants of THEV, one being the published VAS strain without any sequence deletions, while the other variant is a novel variant, hitherto undocumented. Further studies will investigate the replicative fitness and contribution of this shorter variant to infectivity and virulence.

Siderophore and Biofilm Properties of Halophilic Bacterial Strains that May Contribute to Growth Promotion of Inoculated Plants in Salty Soil Quirante J, Porter C, Quirante T, Nielsen BL Brigham Young University—Provo

Several halophilic (salt-tolerant) bacterial strains were isolated from native halophytes (salt-tolerant plants) near Utah Lake. The strains were tested for maximum salt tolerance, with some capable of growing in 4 M NaCl. These strains were then tested as inoculants of alfalfa and some grass varieties to determine if they are able to stimulate growth in salty soil. Three isolates showed significant growth stimulation activity. To determine what properties the bacteria may have that would contribute to plant growth stimulation, each strain was tested for siderophore production, biofilm formation, and other biochemical properties. We found that our growth-promoting strains have some but not all of these properties. We are now testing more strains to determine which properties are most common in plant growth-promoting isolates.

The T Cell Co-Receptor CD5 Alters Mouse Behavior and Gut Microbiome Composition Reaveley K, Whitley KV, Téllez Freitas CM, Cox T, Magoffin W, von Wallwitz I, Mahler K, Haynie C, Tall A, Teasdale J, Weber KS Brigham Young University—Provo

Behavior is affected by both psychological and biological factors. The central nervous system, immune system, and gut microbiota work together to affect our mental health and cognitive behaviors. Immune cell development is also influenced by and influences the gut microbiota composition. T cells help regulate immune responses to foreign and commensal microbes. T cell activation is regulated by co-receptor proteins which can dramatically affect T cell function and immune responses. CD5, a T cell co-receptor, modulates T cell activation by inhibiting T cell activation by self-proteins and helps prevent potential autoimmune effects. Our project studies examined the CD5 protein co-receptor and its effects on the microbiota, immune response, and central nervous system by testing mice that have the protein (CD5WT), mice that don't have the protein (CD5KO) and their progeny. We hypothesize that CD5 alters the gut microbiota which, in turn, alters the cognitive behavior of these mice. To investigate this hypothesis, we have conducted a series of behavioral tests based on both genotype and microbiome composition and found significant differences in both behavior and microbiome composition due to CD5. This project improves our understanding of the intersection of the immune system, gut microbiota, and the central nervous system.

Chromatin Accessibility in C. elegans Russell S, Carter J, Johnson SM Brigham Young University—Provo

The dauer state generated through distinct temperatures, starvation, and other variables can initiate this stress-resistant state in the maturation of C. elegans. Utilizing starvation to induce dauer state, the goal of this experiment is to observe the varying elements of chromatin accessibility between C. elegans who underwent the dauer state and those that did not. These differences are to be observed by following an ATAC-seq protocol presented and developed in “Chromatin accessibility dynamics reveal novel functional enhancers in C. elegans”. We expect to find evidence to further support the presently observed variations and further clarify elements enhancing chromatin accessibility.

CRISPR Deletion of Viral Receptor Genes in Human Cells Skalka N, Conroy M, Spencer J, Aranda J, Demler T, Ludlow R, DeSanti C, Clark DN Weber State University

Enterovirus 71 (EV71) and herpes simplex 1 (HSV-1) are viruses that cause skin lesions in humans. EV71 is a virus that causes hand foot and mouth disease (HFMD) and primarily affects young children. In recent outbreaks in Asia, however, there have been reports of more serious and life-threatening brain infections. HSV-1 is a lifelong infection, causing genital herpes and cold sores, which affects 50 to 80 percent of US adults. In this experiment, we use CRISPR to edit the human genome in cultured cells (HEK293 and HeLa) to decrease the infectivity of these two viruses by deleting their receptors. To delete these receptor genes, a guide RNA (gRNA) was designed for each receptor using the Broad Institute gRNA design tool (ANXA2, SCARB2, and SELPLG for EV71, and Nectin-1 and HVEM for HSV-1). Plasmids that express each gRNA and the CRISPR cutting enzyme, Cas9, were transfected into human cells using the base plasmid All_in_one_CRISPR. This plasmid contains a dsRed fluorescent protein and a hygromycin selectable marker for selection of edited cells. Plaque assays then compare infectivity in parent cells versus receptor- deleted cells. Because viruses use combinations of receptors, the end goal is to determine which receptors are most critical for attachment and entry into cells in order to target those receptors for virus inhibition.

Gluconate Metabolism by Paucilactobacillus wasatchensis WDC04 Can Be a Cause of Late Gas Defect in Aging Cheese Sorensen K, Oberg CJ, Domek MJ, Culumber MC, Young S, Oberg T, McMahon D Weber State University

Paucilactobacillus wasatchensis, a nonstarter lactic acid bacteria, can cause late gas production, and splits and cracks in aging cheese when it metabolizes 6-carbon sugars in cheese, particularly galactose, to a 5- carbon sugar, resulting in the release of CO2. Previous studies have not explained late gas production in aging cheese when no galactose is present. Based on the genome sequence of Plb. wasatchensis WDC04, genes for potential metabolic pathways were mapped using Knowledgebase Predictive Biology software. This metabolic modeling predicted Plb. wasatchensis WDC04 could use gluconate. Gluconate contains 6 carbons and Plb. wasatchensis WDC04 contains genes to convert it to ribose-5-P using phosphogluconate dehydrogenase by a decarboxylating step, producing CO2 during its metabolism. The goal of this study was to determine if sodium gluconate, often added to cheese to reduce calcium lactate crystal formation, could be metabolized by Plb. wasatchensis WDC04 resulting in gas production. Carbohydrate restricted MRS (CR-MRS) was mixed with varying ratios of ribose, sodium gluconate or D-galactose (total sugar content of 1%). Oxyrase® (1.8% v/v) was also used to create an aging cheese environment in selected tubes. Tubes were inoculated with a 4 d culture of Plb. wasatchensis WDCO4 and results recorded over 8 d. When inoculated into CR-MRS containing only sodium gluconate, Plb. wasatchensis WDC04 grew, confirming gluconate utilization. Of the ten ratios used, Plb. wasatchensis WDC04 produced gas in six with the most gas production resulting from the ratio of 100% sodium gluconate with no added ribose or galactose. Results showed that Plb. wasatchensis WDC04 can utilize sodium gluconate to produce CO2 gas, which could cause late gas formation in aging cheese. The presence of sodium gluconate in cheese thus becomes another risk factor for unwanted gas production, and formation of splits and cracks.

The Effects of Ozone on Staphylococcus Stilson X, Scharmann B, Tupuola K, Scharmann B, Madsen C, Oberg CJ Weber State University

Staphylococcus aureus is a common human-borne bacterium found in 40% of the population, primarily in their nasal passages. This bacterium, a common cause of in-hospital postoperative infections, can also be found in the athletic community, residing on uniforms, training table, playing fields, practice mats and even team mates. Outbreaks of S. aureus within athletic teams is not uncommon. In sports venues, ozone has been used to sterilize swimming pools, athletic equipment, and other surfaces prone to bacterial contamination. This research was conducted to determine if a commercial ozone generator was effective is killing S. aureus in a locker room. Petri dishes (TSA) inoculated with a nonpathogenic strain of S. aureus were put in strategic locations (in triplicate) in WSU training and locker rooms. Lids were removed from the inoculated petri plates and three ozone generators, one for the training room and one for each of the two locker rooms were turned on for a set time. In trial one, they were on for 120 minutes for the two football locker rooms and 90 minutes for the training room, while in trial two they were all on for 180 minutes. Additionally, uninoculated control plates were placed in each room as a test for air borne contaminants that might fall on the plates during the experimental protocol. Three hours after ozone infusion, petri dishes were carefully collected, then incubated for 48 h at 37 °C. Following incubation, the number of S. aureus colonies on each plate were recorded. Two hours into the run cycle, ozone measurements were made using an Ozone Monitor InDevR Model 205. Results for trial one showed an overall S. aureus reduction of 78.7 ± 8.3%, while trial two showed an increase in the overall reduction to 93 ± 1.8%. There was a modest correlation between S. aureus survival and distance from the ozone generator in trial one (R2 = 63). Average ozone readings increased from a background of 17 ppb to 1042 ppb at ground level.

Microbial Diversity of Rock-Hosted Serpentinite Subsurface Environment Twing KI, Brazelton WJ, Schrenk MO Weber State University

Serpentinization is a widespread geochemical process involving the aqueous alteration of ultramafic rocks, which results in high pH (>10) and highly reducing pore-fluids. These fluids contain an abundance of hydrogen and abiogenic organic molecules, which can potentially fuel microbial communities in this system. Previous studies have looked at the microbial composition of serpentinite fluids and found two main taxa, Clostridia and Betaproteobacteria. However, the serpentinite subsurface is a rock-hosted environment and to date no research has investigated the microbes living associated with the rocks in the serpentinite environment.

In 2011 the Coast Range Ophiolite Microbial Observatory (CROMO) was established in northern California, allowing for the ongoing monitoring of serpentinite fluids, as well as characterization of the minerals and mineral-associated communities within the serpentinite subsurface environment. The drilling of two wells at CROMO yielded 30- and 45-meter cores. Microbial community composition of these cores, as assessed by 16S rRNA gene sequencing preformed as part of the Census of Deep Life, indicate that different microbes are associated with the rocks than are found in the fluids. These data, combined with geological and geochemical data from the cores, will help elucidate what environmental factors drive community composition within serpentinite rocks.

Comparative Genomic Analysis of Microbacterium foliorum Cluster EE Phages Downey JL, Griffin BT, Jensen AR, Ludwig A, Spencer JB, Scott TM, Vander Werff KS, Pickett BE, Grose JH, Breakwell DP Brigham Young University—Provo

Bacteriophages are grouped into different clusters based on genome synteny and percent nucleotide identity (PNI). PNI matrix was generated using Clustal2.1 on MAFFT. Analysis using Phamerator and NCBI to compare Microbacterium foliorum cluster EE phage genomes revealed that though largely similar, cluster EE phages differ in several ways. Phamerator groups protein coding genes into different “phamilies” or “phams” based on protein similarity. A large portion of all cluster EE genomes are composed of the same phams at their responding loci, suggesting high genome similarity. While this is the case for most genes, there are certain loci that contain considerable pham variability. Due to these differing loci, it is proposed that four subclusters of phages are created within the entire EE cluster. These subclusters were also identified by a phylogenetic tree using all cluster EE phages. Phage genomes are highly variable, and this is true of cluster EE phages. Some cluster EE phages have unique coding sequences that create their own pham. The phylogenetic tree supports the novelty of several phages, for example phages Efeko, BonaeVitae and Naby.

Understanding the Effects of T Cell Co-Receptor CD5 on Metabolic Function and the Metabolome von Wallwitz I, Whitley KV, Téllez Freitas CM, Cox T, Magoffin W, Reaveley K, Mahler K, Haynie C, Tall A, Teasdale J, Weber KS Brigham Young University—Provo

Metabolic function in immune cells is critical for effective activation and destruction of foreign pathogens. As T cells are activated, their metabolic profile is significantly changed due to signaling cascades mediated by the T cell receptor (TCR) and co-receptors found on their surface. Because T cell activation and proliferation require large amounts of biomass and energy indicative of anabolic growth, the metabolic profile of activated T cells also changes, resulting in a greater utilization of aerobic glycolysis. CD5 is a T cell co-receptor that regulates thymocyte selection and peripheral T cell activation. Removal of CD5 has been shown to affect metabolic T cell function as well as systemically affect the metabolome as a whole. It is also well understood that the microbiome can also influence the metabolome. So, to better understand how CD5 specifically affects systemic metabolic function, we bred wild type and CD5 knockout mice to generate offspring that have different genotypes but have the same microbiome and measured their serum metabolomics. Our preliminary results suggest CD5-driven and gut microbiome mechanisms that regulate the metabolome, indicating that balance between the immune system and the gut microbiome must be considered in the development of immunotherapies to increase effectiveness and success.

Selective Media for the Isolation of Paucilactobacillus wasatchensis Wahlstrom RC, Domek MJ, Oberg CJ Weber State University

The obligate heterofermentative bacteria Paucilactobacillus wasatchensis has been shown to cause late gas blowing in aged cheeses, which results in defects such as splitting and crumbling of the cheese block. The ability to quickly and accurately isolate Plb. wasatchensis, especially when it is present at low concentrations compared to other bacteria in a cheese sample, could be very beneficial to the dairy industry. However, the current protocol for isolating Plb. wasatchensis is time intensive and imprecise. The goal of this study is to accurately detect Plb. wasatchensis when as few as 103 CFU’s per gram are present within 72 hours, as well as be able to inhibit competing SLAB and NSLAB with one media. Testing was conducted using 24 well plates in a Tecan infinite 2000 plate reader, in which 7 SLAB and NSLAB strains were tested in triplicate along with the Plb. wasatchensis type strain, WDC04. Each well was filled with carbohydrate restricted MRS (CR-MRS) broth containing 1% ribose, 2% Oxyrase and .01% 2- deoxyglucose, a glucose analog. Results showed that under these conditions WDC04 could complete its logarithmic growth phase in 28 hours while the glycolysis inhibitor, 2-deoxyglucose, limited the growth of the 7 other SLAB and NSLAB strains. Lacticaseibacillus casei and Lacticaseibacillus paracasei, two of the most common NSLAB strains, showed the greatest level of inhibition between MRS broth (OD600 = 1.28 at 28 h) and CR-MRS+2-deoxyglucose (OD600 = 0.60 and 0.54, respectively) after 28 h. Preliminary results for the incorporation of 2-deoxyglucose into CR-MRS agar as a selective plating media for Plb. wasatchensis show promise. This method could be used to determine the presence of Plb. wasatchensis in cheese when it is low concentrations (103 CFUs/g) versus the high concentration of SLAB (108 CFUs/gram) that obscure its detection with current isolation techniques.

Author Index

Adams K ...... 54 Frandsen P ...... 63 Aranda J ...... 73 Fuller S ...... 58 Ashton N ...... 6 Gaffney EM ...... 25 Ayers H ...... 42, 57 Ganesh J ...... 26 Bell I ...... 45 Gazdik Stofer MA ...... 41 Bellini DM ...... 30 Gee J ...... 67 Benedict A ...... 52 Ghosh S ...... 68 Benedict AB ...... 28 Godfrey R ...... 39 Berges BK ...... 29, 38 González Murcia JD...... 56, 65 Brazelton WJ ...... 36, 76 Grattieri M ...... 25 Breakwell DP ...... 42, 57, 77 Gray M ...... 27 Brown S ...... 45 Griffin BT ...... 77 Bustos Y ...... 42, 57 Griffitts JS ...... 22, 28, 52, 64, 70 Calvopiña D ...... 22, 28 Grose JH ...... 37, 42, 56, 57, 60, 65, 77 Cardon D ...... 46 Guerrero Arguero I ...... 29 Carter J ...... 72 Gustavson C ...... 6 Chamberlain J ...... 52 Hansen DV ...... 47 Church B ...... 53 Hansen J ...... 59 Clark DN ...... 34, 67, 73 Hansen RJ ...... 41 Cole J ...... 59 Haynie C ...... 50, 71, 78 Cole N ...... 54 Hepworth K ...... 32 Conroy M...... 73 Hill J ...... 61 Correa E ...... 42, 57 Hooper N...... 45 Cox T ...... 50, 71, 78 Hooper VA...... 41 Cress JD ...... 30 Humpherys TB ...... 30 Cromar Z ...... 24 Isaacson B ...... 39 Crook MB ...... 55 Jackson D ...... 31 Culumber MC ...... 35, 44, 53, 62, 74 Jackson RN ...... 48 Davis K ...... 32 Jensen AR ...... 77 Dearden L ...... 56, 65 Jensen JL ...... 47 Demler T ...... 73 Jensen S ...... 43 DeSanti C ...... 73 Johnson SM ...... 54, 72 Domek MJ ...... 66, 74, 79 Johnston JD ...... 58 Domgaard H ...... 48 Kaundal A ...... 26 Dōne J ...... 5 Kaundal R ...... 23 Downey JL ...... 77 Kauwe JSK ...... 47, 56, 65 Drake L ...... 39 Knorr I ...... 32 Duhan N ...... 23 Knowles A ...... 61 Epperson RT ...... 39 Lambson L ...... 58 Farinelli B ...... 39 Laughter C ...... 55 Ferrel MN ...... 56, 65 Leatham D ...... 33 Findlay M ...... 24 López JB ...... 38 Flor S ...... 42, 57 Ludlow R ...... 73 Fonseka D ...... 55 Ludwig A ...... 77

Madsen C ...... 59, 75 Rawson C ...... 41 Magoffin W ...... 50, 71, 78 Rawson K ...... 45 Mahler K ...... 46, 50, 71, 78 Reaveley K ...... 50, 71, 78 Martin JE ...... 31 Reed H...... 57 McGary BC...... 34 Reed J ...... 42 McMahon D ...... 74 Ridge PG ...... 56, 65 McPhie J ...... 22 Ringger E ...... 42, 57 Mejías LA ...... 35 Robison RA ...... 29, 38 Metcalf J ...... 48 Rogers J ...... 5 Miller A ...... 61 Rood K ...... 32 Mills E ...... 24 Russell S ...... 72 Miner D ...... 32 Scarbrough D ...... 49 Minteer SD ...... 25 Scharmann B ...... 59, 75 Mishra N ...... 44, 59, 62 Scharmann S ...... 59 Moreno C ...... 63 Schrenk MO ...... 76 Moss M ...... 64 Scott TM ...... 43, 77 Motamedi S ...... 36 Skalka N ...... 73 Nelson A ...... 56, 65 Smith J ...... 44 Neuberger T ...... 45 Smith KR ...... 30 Newey C ...... 37 Smith TB ...... 45 Nichols R...... 66 Solis-Leal A ...... 38 Nielsen BL ...... 61 Sorensen K ...... 74 Nielsen TJ ...... 67 Soule C ...... 42, 57 Norton JM ...... 23 Soule S ...... 42, 57 Nyone C ...... 59 Spencer J (WSU) ...... 73 O’Neill KL ...... 30 Spencer JB (BYU) ...... 77 Oberg CJ ...... 33, 35, 44, 59, 62, 66, 74, 75, 79 Sperry YK ...... 46 Oberg T ...... 33, 74 Steiner C ...... 59 Ogilvie BH ...... 38 Stilson X ...... 75 Ong J ...... 39 Stock M ...... 32 Ord B ...... 69 Sun Y ...... 26 Overgaard EM ...... 40 Tall A ...... 50, 71, 78 Packard H ...... 66 Tam J ...... 39 Packard JF ...... 34 Tavana JP ...... 47 Pape JA ...... 37 Taylor B ...... 64 Parkin M ...... 45 Taylor HN ...... 48 Paryzek J ...... 55 Teasdale J ...... 50, 68, 71, 78 Pasquina P ...... 39 Telford M ...... 42, 57 Pendleton HL ...... 36 Téllez Freitas CM ...... 46, 50, 56, 65, 71, 78 Perry A ...... 68 Thomas A ...... 26 Perry B ...... 68 Tinker JK ...... 40, 49 Peterson D ...... 32 Townsend J ...... 46 Peterson T ...... 39 Tupuola K ...... 75 Pickett BE ...... 43, 61, 77 Turner E ...... 24 Poole BD ...... 38, 69 Twing KI ...... 36, 76 Price J ...... 22 Valdoz J ...... 29 Quaye A ...... 69 Van Ry PM ...... 29 Ramsey J ...... 42, 57 Vander Werff KS ...... 77

VanDerWerff KJ ...... 69 Weinert A ...... 56, 65 Velázquez EJ ...... 30 Wheeler C ...... 49 Viazzo R ...... 29 Whitley KV ...... 50, 71, 78 Villalva C ...... 29 Wiley-Kalil A ...... 55 von Wallwitz I ...... 50, 71, 78 Williams DL ...... 3, 39, 45 Wagstaff K ...... 22 Wilson D ...... 32 Wahlstrom RC ...... 79 Wilson E ...... 56, 65 Walker-Bravo A ...... 32 Young S ...... 74 Weber KS .... 46, 47, 50, 56, 58, 63, 65, 68, 71, 78 Zesiger C...... 32