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Wyoming INBRE Fall Research Network Retreat

September 12-14, 2019 Casper Mountain Trails Center

(9301 Casper Mountain Road, WY 82604)

Program

Casper Mountain Trails Center

Wyoming INBRE is supported by a grant from the National Institute of General Medical Sciences (2P20GM103432) from the National Institutes of Health. Any opinions expressed or results presented are solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Schedule of Events

The approximate driving time to the lodge from downtown Casper is about 20 minutes. All scientific presentations (oral and poster) are scheduled at the conference venue

(Casper Mountain Trails Center). Special note about meals: All breakfasts on conference days (September 13-14) are

on your own. Lunches on these days will be provided at the conference venue. The Thursday evening reception and Friday night dinner will be in the city of Casper (see details below for specific locations).

Thursday September 12, 2019

7:00-9:00pm Reception (3631 Monte Vista, Casper, WY 82601). Beverages and light snacks.

Friday September 13, 2019

(8:50-9:00am) (All morning speakers should load up their PowerPoint presentations on

the main computer in the Casper Mountain Trails Center). 9:00-9:30am Welcome and introductions. R. Scott Seville, Wyoming INBRE

Director/Principal Investigator 9:30-10:00am Northwest College Program Report

Bioinformatics and Basic Research in the Big Horn Basin: Opportunities for Project-Based Inquiry while Building Laboratory Skills and Theoretical Perspectives. Eric C. Atkinson Project Leader, Elise Kimble, Allan Childs, Uko Udodong, Michael F. Cuddy, and Monia Haselhorst.

10:00-10:30am Central Wyoming College Program Report

Fall 2019 Central Wyoming College INBRE Project Lead Report. William Finney Project Leader, Aaron Bender, Kirsten Kapp, Jacki Klancher, Steve McAllister, Tarissa Spoonhunter, and Tara Womack-Shultz.

10:30-11:00am Break 11:00-11:30am University of Wyoming and Casper College Program Report INBRE-Supported Undergraduate Research in Casper. Dagmara

Motriuk-Smith Project Leader 11:30am-12:00pm Laramie County Community College Program Report

Laramie County Community College Research Program Update. Ami Wangeline Project Leader, Courtney Springer, Marie Yearling, and Zachary Roehrs.

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12:00-1:30pm Lunch and discussion (on site)

(All afternoon speakers should load up their PowerPoint presentations on the main computer in the Nordic Ski Lodge before 1:25pm).

1:30-2:00pm Eastern Wyoming College Program Report Denitrification potential of soils with different land use and nitrogen inputs in semiarid ecosystems. Christopher Wenzel Project Leader.

2:00-2:30pm Western Wyoming Community College Program Report

INBRE Research at Western WY Community College. Bud Chew Project Leader, Dave Tanner and Josh Holmes.

2:30-2:50pm Student presentation: Preliminary Transcriptome Analysis of Adult

Culex tarsalis Mosquitoes Collected in Fremont County, Wyoming. Micah Conner and Aaron Bender (Central Wyoming College)

2:50-3:10pm Break 3:10-4:30pm Panel discussion: Planning for academic and professional

success in biomedical research/professional school. Panelists: Naomi Ward, Dagmara Motriuk-Smith, Aaron Bender, and Bud Chew. Moderator: R. Scott Seville

(4:15-4:30pm) (Students should set up their posters for tomorrow’s Poster

Session) 4:30pm Adjourn to travel down mountain 6:00-9:00pm Dinner and evening presentation (Location: Gruner Brothers

Brewery, 1301 Wilkins Cir, Casper, WY 82601). “Wyoming INBRE Phase 3- what’s in it for you.” R

Saturday September 14, 2019

(Casper Mountain Trails Center)

(8:50-9:00) (Morning speakers should load up their PowerPoint presentations on the main computer in the Casper Mountain Trails Center).

9:00-9:30am Northern Wyoming Community College District (Sheridan

College and Gillette College) Program Report Northern Wyoming Community College INBRE-Supported Research Activities. Ami Erickson Project Leader and Rob Milne.

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9:30-11:00am Student poster session

P1. Drug Resistant Bacteria Prevalence in Conventional vs. Organic Compost Amalgamated with Soil. Tawna R. Herrera, and Elise J. Kimble (Northwest College)

P2. CTE and the Accumulation of Tau Protein in the Brains of Bighorn Sheep. 1Tabitha Tyrrell, 1Eric Silk, 1Michael F. Cuddy, 1,2Eric C. Atkinson (1Northwest College, and 2University of Wyoming)

P3. Investigating the Link Between Melanism and Immune Function of Canids. Emilie Edwards and Eric C. Atkinson (Northwest College)

P4. The Ants Go Marching One-by-One: Metabarcoding the Hymenoptera Microbiome of the Big Horn Basin. Jacklyn A. Green, Richard A. Mangali, Tabitha N. Tyrrell, Tawna R. Herrera, Fantasia Critchfield, Camille L. Brandt, Sidney D. Brooks, Lara M. Chanthongthip, Emilie D. Edwards, Gareth G. Flowers, Drew Groll, Pascal Hagenimana, Abigail L. Hogan, Wyatt J. Horrocks, Brianna N. Kilpatrick, Chaz P. Krone, Carissa A. Schmidt, Laura Shoopman, Allan Childs, Elise Kimble, and Eric C. Atkinson (Northwest College).

P5. Spice Up Medicine. Fantasia Critchfield, Tayler Groll, Dylan Przygocki, and Uko Udodong (Northwest College).

P6. Extraction and Isolation of Medicinal Proteoglycans from Pearl and Blue Oyster Mushrooms. 1Thomas Haack and 2Eric Mechalke (1University of Wyoming at Casper, and 2Casper College)

P7. Molecular Categorization and Frequency of SHV-Like Antibiotic Resistance Genes in the Environment. 1Edwards Caitlyn, 1Burrough Hannah, and 2Chase John (1University of Wyoming at Casper, and 2Casper College)

P8. Understanding the Binding Specificity of PopZ Protein. 1Theressa Smith, 1Sarah Steinberg, 1Dagmara Motriuk-Smith, and 2Grant R. Bowman (1University of Wyoming at Casper, and 2University of Wyoming)

P9. Denitrification Gene Presence Relative to Land Use in Semiarid Environments. Brandon Phillips and Christopher Wenzel (Eastern Wyoming College)

P10. Preliminary Transcriptome Analysis of Adult Culex tarsalis Mosquitoes Collected in Fremont County, Wyoming. Kinyatta Kelvin and Bender Aaron (Central Wyoming College)

P11. Life History Factors Important in Incidence of White-nose Syndrome (Pseudogymnoascus destructans) among North American Bats. Candace Siirila and Eric C. Atkinson (Northwest College).

P12. Fishing for Water Quality.Gareth G.K. Flowers, Wyatt J. Horrocks, Michael F. Cuddy, and Eric C. Atkinson (Northwest College).

11:00am-1:00pm Lunch and INBRE Year 5 and Phase 4 program discussions-

students and faculty ~1:00pm- Wyoming INBRE Fall Network Retreat adjourns

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Presentations- Abstracts

Oral presentations are limited to 20 minutes (including questions) for all, except for Community College Program presentations which may last up to 30 minutes (including questions).

Student poster presentations (P) will be displayed in the Casper Mountain Trails Center (poster set-up is scheduled on FRIDAY September 13 from 4:15-4:30pm).

Oral Presentation Abstracts (All scientific abstracts are listed by order of presentation) 1. Bioinformatics and Basic Research in the Big Horn Basin: Opportunities for project-based inquiry while building laboratory skills and theoretical perspectives. Eric C. Atkinson, Elise Kimble, Allan Childs, Uko Udodong, Michael F. Cuddy, and Monia Haselhorst

Northwest College, 231 West 6th Street, Powell, WY 82435 Email: eric.atkinson@nwc.edu Abstract. At NWC, INBRE supported undergraduate research, exploration, and mentoring span topics from a) novel antibiotic research, b) characterization and modification of bactericidal compounds in spices, c) water quality within the Big Horn Basin, d) avian disease, e) characterization of antibiotic resistance in soils of varying agricultural use, to f) curriculum development. Our broadening trend continues in FY 2019/2020. We work to meet the following program goals: 1) enhance opportunities for WY community college undergraduates to better understand (and ultimately participate in) the field of biomedical research; and, 2) develop a pipeline of students with an interest in biomedical science who would then go on to complete their baccalaureate degrees and/or graduate degrees at UW. During 2018-2019, we implemented an applied bioinformatics investigation at the core of our 2-credit BIOL 2465-Research Problems in Biology course. Students employed a point-of-capture (POC) smart-phone based App (Anecdata.org), aseptically collecting native ant and wasp samples for microbial metabarcoding in collaboration with New Hampshire INBRE (UNH). Students learned bioinformatics/ genomics theory via weekly discussions on timely topics such as Neanderthal*Denisovan hybridization, Elizabeth Warren’s DNA characterization, and ethical controversies regarding CRISPR techniques applied to human embryos. Videos on lab protocols and Next Generation Sequencing (NGS) increased understanding while laboratory skills (pipetting, DNA extraction, PCR/amplification, and gel-electrophoresis) were taught via use of ant samples subsequently sent to Hubbard Center for Genome Studies. The QIIME2 pipeline and BASH were introduced as students attempted to correlate microbial diversity with hymenopteran taxa and command-line coding will be extended this year. Twenty-one students participated in our program; 14 students graduating with AS degrees (4 ASN), 2 of which transferred to UW with remaining students transferring to MSU, UM, SDSU, and UU. One transfer student applied for and was awarded an INBRE Transition Fellowship. Two students were accepted to summer REUs. Professional development for faculty was provided via MDIBL/UNH bioinformatics training, graduate course enrollment, and continued

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analysis and sample development via avian disease research in which students gained other skills important to field-based biomedical research.

2. Fall 2019 Central Wyoming College INBRE Project Lead Report. 1William Finney, 1Aaron Bender, 2Kirsten Kapp, 3Jacki Klancher, 1Steve McAllister, 1Tarissa Spoonhunter, and 1Tara Womack-Shultz

1Central Wyoming College, 2660 Peck Ave., Riverton, WY 82501 2Central Wyoming College, 240 S. Glenwood St #124, P.O. Box, Jackson, WY 83001 3Central Wyoming College, 120 Enterprise Blvd., Lander, WY 82520

Email: bill@cwc.edu Abstract. INBRE supported faculty at Central Wyoming College continue engage undergraduate students in research and develop broadly applicable skills in wide-ranging projects. Dr. Aaron Bender works with students on sequencing the genome of C. tarsalis as part of their continuing work studying the West Nile Virus. Professor Kirsten Kapp continues her work investigating the sources of microplastics in natural waters. Professor Jacki Klancher has recently returned from her most recent Interdiciplinary Climate Change Expidition (ICCE). Dr. Tarissa Spoonhunter continues her work in science education on the Wind River Reservation. Professor Tara Womack-Shultz is beginning a new collaborative project involving circadian rhythms and binge eating. 3. INBRE-Supported Undergraduate Research in Casper. Dagmara Motriuk-Smith

University of Wyoming at Casper, 125 College Dr., Casper, WY 82601 (reporting also for Casper College) Email: motriuk@uwyo.edu Abstract. The goal of the INBRE supported research is to train undergraduate students, attending Casper College and University of Wyoming at Casper, in conducting biomedical research. The INBRE internship was awarded to seven students in the spring, four in the summer, and seven are expected to apply this fall. The research projects concentrated on studying antibiotic resistance genes in the environment, expression of recombinant spider silk-like proteins, intrinsically disordered proteins, morphology and phylogeny of eimerian parasites, properties of medicinal glycoproteins. More recently, a collaborative grant awarded to Dr. Teulé-Finley and Dr. Cherrington will support research investigating effects of citrullination on tubulin function. Two Casper College faculty members Dr. Chase and Dr. Mechalke-were awarded The School of Science Outstanding Faculty of the Month Award. Lastly, three Casper students participated in a fluorescent microscopy workshop. 4. Laramie County Community College Research Program Update. Ami Wangeline, Courtney Springer, Marie Yearling, and Zachary Roehrs

Laramie County Community College, 1400 E. College Dr., Cheyenne, WY 82007

Email: awangeli@lccc.wy.edu

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Abstract. The mission of the Laramie County Community College (LCCC) IDeA Network for Biomedical Research Excellence (INBRE) research group is to improve the access of students to authentic research experiences. LCCC as an institution is in its third year of the American Association of Community Colleges, Pathways Project redevelopment and much of our curriculum is being redesigned at the program level. Specifically, for our INBRE program, these changes will actually expand access to our undergraduate research courses across all of our science degrees. As for the research itself, our main foci this year have been winding down several ongoing projects including sequencing of the Alternaria genome and transcriptome in collaboration with the University of Wyoming (UW) Bioinformatics core and the Yellowstone Fire Study in collaboration with UW-Casper and University of Oklahoma. Last, we have acquired a variety of new instrumentation from various Wyoming State laboratories and are working with the LCCC foundation on new endowed funds earmarked for undergraduate research, both of which will open new avenues for student research and opportunities. 5. Denitrification potential of soils with different land use and nitrogen inputs in semiarid ecosystems. Christopher Wenzel

Eastern Wyoming College, 3200 West C Street Torrington, WY 82240 Email: chris.wenzel@ewc.wy.edu Abstract. The INBRE program at Eastern Wyoming College (EWC) dates back to 2009. Since that time 14 undergraduate students have participated, 13 of which have gone on to pursue or are planning to pursue higher degrees in Biology or Biomedical related fields. Research at the EWC-INBRE Laboratory has focused on two major areas including 1) microbial and plant DNA quantification; 2) polymerase chain reaction (PCR) to determine the presence or absence of microbial genes involved in denitrification in soils and nitrate uptake in plants. In addition, UW laboratory facilities are being used for A) soil microbial gas production as influenced by variation in environmental conditions; and B) RT-qPCR to quantify denitrifying genes. Research has been supported by equipment purchases including, a) Nano-Drop UV-vis for DNA quantification; b) Bio-Rad T100 Thermal Cycler for gene detection; and c) So Low -40 freezer for nucleic acid preservation. The INBRE program has been an invaluable addition to the Pre-professional and Science based programs on our campus. It has provided hands-on biomedical research experiences and scholarship opportunities for students, and enhanced the caliber of our science faculty thereby attracting better-prepared students to our campus. Anthropogenic activities have altered the global cycle of nitrogen substantially. Land use and nitrogen-based fertilizer inputs are known to influence groundwater nitrate levels and nitrous oxide gas fluxes from soil ecosystems. However, little is known about the relationship between these gas fluxes, the denitrification potential of the microbial community (i.e. presence of denitrification genes), and nitrogen inputs in relation to land use in semiarid ecosystems. Thus, the objective of this study is to determine denitrification potential at five sites with different land uses (i.e. grazed-grassland, cropland, urban, wetland, sub-irrigated), and varying nitrate and water inputs. Soil collected from the different sites was incubated at varying water potentials (dry, field

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capacity, and saturated), and potassium nitrate was added to all treatments, except the dry treatment group. During the incubation, nitrous oxide gas fluxes were measured and at the end the soil was analyzed for presence of microbial genes involved in denitrification (nirS/K, norB, and nosZ genes). Gene presence differed for all land uses, with the grazed-grassland site showing the lowest percent frequency for all genes. Preliminary analyses indicate the addition of nitrate and water conditions result in variations in nitrous oxide gas fluxes, but there were no common patterns across land uses. Further data analysis will provide a better understanding of the dynamics between biotic and abiotic variables influencing denitrification in semiarid ecosystems. 6. INBRE Research at Western WY Community College. Bud Chew, Dave Tanner, and Josh Holmes

Western Wyoming Community College, 2500 College Drive, Rock Springs, WY 82901

Email: bchew@westernwyoming.edu Abstract. INBRE research at WWCC is designed to expose promising freshman and sophomore students to undergraduate research, to encourage students to consider a career that includes research, to enhance students’ opportunities in a competitive world, and to further scientific knowledge through presentation and publication of research work. This year, 8 students are participating in INBRE funded research; topics include evolutionary biology/ecology, and cardiovascular physiology. Three faculty members have established INBRE research programs. Research at WWCC would not be possible without the WY INBRE grant. The opportunities presented by INBRE have allowed the college to recruit and retain better faculty members; students involved in INBRE research at WWCC become better, more confident students, while not only developing enhanced lab/field skills, but also responsibility, perseverance, and leadership skills. Student presentation: Preliminary Transcriptome Analysis of Adult Culex tarsalis Mosquitoes Collected in Fremont County, Wyoming. Micah Conner and Aaron Bender

Central Wyoming College, 2660 Peck Avenue, Riverton, WY 82501 Email: mrc0616@cwc.edu Abstract. Fremont County is a hotspot for West Nile virus (WNV) infection in Wyoming. It is known that the mosquito Culex tarsalis (C. tarsalis) serves as the primary insect vector for WNV in western states such as Wyoming. Although a great deal is known about the transmission, replication, and genetics of WNV, very little is known about the genome structure and gene expression profile of C. tarsalis. Our research team captured mixed populations of wild mosquitos in Fremont County. These were sorted and adult C. tarsalis were positively identified. Total RNA from this pool was isolated, column-purified and quantified. Two parallel samples of high quality total RNA were submitted for Illumina next-generation RNA sequencing. Our extremely comprehensive dataset consists of nearly 350 million paired-end reads. The average data yield for the 2 samples is approximately 50,000 megabases with a Q score of >/= 30 for over 91% of called bases and has a mean quality score of nearly 38. In collaboration with the University of Wyoming INBRE bioinformatics core, our research team intends to

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assemble the transcriptome and search for gene-expression profiles that may lead to a better understanding of the WNV transmission process. Finally, using a combination of qRT-PCR and bioinformatic analysis of our sequencing dataset, we will ask whether other members of the flaviviridae are carried by C. tarsalis and whether WNV can integrate into its host’s genome. 7. Northern Wyoming Community College INBRE-Supported Research Activities. Ami Erickson and Rob Milne

NWCCD-Sheridan College, 3059 Coffeen Avenue, Sheridan WY 82801 Email: aerickson@sheridan.edu Abstract. Northern Wyoming Community College has participated in Wyoming INBRE since its origin. During this time, many community college professors and undergraduate students have had the chance to conduct research and attend conferences, accumulating a variety of science skills and experiences. Six Sheridan College students were actively involved in molecular biology, tissue culture and chemistry-related research during the past academic year. Over the past two years, two students have been awarded the UW INBRE Transition Fellow. This summer two students had the opportunity to present research posters at the ASHS annual conference. Their research included the use of tissue culture to propagate native plants and the evaluation of the health and productivity of the Sheridan R&E trial vineyard. Over the 2018-2019 academic year our activities have included completing the second year of an INBRE DRPP Collaborative grant in partnership with Dr. Nic Blouin and Dr. Andrew Parsekian, while obtaining a Wyoming INBRE Research Network equipment grant, which allowed for the purchase of a Raman microscope system with analysis software. Raman Spectroscopy is the only method for uniquely confirming the presence of graphene. The research purposes for the RAMAN microscope system are to test vapor deposition synthesis conditions using various feedstocks, temperatures and metal substrates to optimize graphene synthesis for the ultimate purpose of developing novel biomedical sensors. Our goal for the next year are to continue developing student research opportunities in molecular biology, tissue culture, bioinformatics and chemistry, while encouraging collaboration opportunities between UW and NWCCD faculty.

Poster Abstracts P1. Drug Resistant Bacteria Prevalence in Conventional vs. Organic Compost Amalgamated with Soil. Tawna R. Herrera, and Elise J. Kimble

Northwest College, 231 West 6th Street, Powell, WY 82435 Email: Tawna.herrera@nwc.edu Abstract. Humans use compost for agriculture, horticulture, and soil sciences. Could drug resistant bacteria be more prevalent in organic compost mixed soil or conventional compost mixed soil? Environmental bacteria in conventional compost are exposed to antibiotics that could select for increased resistance to antibiotic treatments. To test this hypothesis, we have taken samples of compost from five farms located in Park County,

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Wyoming; three organic and two conventional. Compost samples are initially spread on TSA agar medium plates followed by isolation of bacteria. With complete isolation of bacteria, a suite of antibiotic discs is then placed on lawns of bacteria using the Kirby-Bauer method. Preliminary results of organic compost have yielded fifteen disparate drug resistant bacterial types out of 50 isolates. Conventional compost bacteria isolates however, had fourteen drug resistant bacterium out of 44 isolates thus far. Tentative identification of the drug resistant bacteria will provide an indication whether agricultural practices lead to greater exposure to pathogenic drug resistant bacteria. P2. CTE and the Accumulation of Tau Protein in the Brains of Bighorn Sheep. 1Tabitha Tyrrell, 1Eric Silk, 1Michael F. Cuddy, 1,2Eric C. Atkinson

1Northwest College, 231 West 6th Street, Powell, WY 82435 2University of Wyoming, 1000 E. University Avenue, Laramie, WY 82071

Email: tabitha.tyrrell@nwc.edu Abstract. Bighorn sheep (Ovis canadensis) have long-been assumed to be immune to the consequences that arise following repetitive blunt trauma to the head, such as a traumatic brain injury. However, little to no research has been conducted to make any conclusive arguments supporting this hypothesis. Chronic traumatic encephalopathy (CTE) is a neurodegenerative disease in which tau proteins detach from the microtubules they are normally associated with and become hyperphosphorylated to form aggregations that gradually spread throughout the brain. Although tau is related to many different neurodegenerative diseases (tauopathies), this specific type of protein aggregation is unique to CTE. To determine whether or not bighorn sheep present with this specific tauopathy, we collected the brains of three sheep that were donated by a local taxidermist for our research. The brains were initially stored in the refrigerator for approximately three weeks and were then fixed in formaldehyde. The brains were then separated into the left and right hemispheres with a dissection blade. The halves were dehydrated with 90% ethanol and embedded in paraffin. The brains were then sectioned into sections of 5-10 microns using a manual microtome. We tested for tau protein aggregations by using a Gallyas silver stain. This stain was selected due to its high sensitivity to neurofibrillary tangles and protein aggregations within brain tissue. Further examination of the specimens will determine whether or not the brains present with the aforementioned tauopathy, chronic traumatic encephalopathy. P3. Investigating the Link Between Melanism and Immune Function of Canids. Emilie Edwards and Eric C. Atkinson

Northwest College, 231 West 6th Street, Powell, WY 82435 Email: emilie.edwards@nwc.edu Abstract. The documented cause of melanism in most species of mammals, fish, and birds is caused by a mutation in the Melanocortin 1 receptor (Mc1r) gene. However, in wolves (Canis lupus; n=8) and coyotes (Canis latrans; n=10), it is caused by a mutation in the K locus, which encodes the beta-defensin protein that acts as an alternative ligand for the Mc1r gene. These β-defensin proteins are important microbiomial factors that help to link the innate and adaptive immune responses. The antibodies that are

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generated to fight a disease are left behind even after the disease has run its course, and DNA signatures of those pathogens may be present as well. We used data from the Yellowstone National Park service to compare the coat types and health of wolves during one calendar year to begin investigating this link. P4. The Ants Go Marching One-by-One: Metabarcoding the Hymenoptera Microbiome of the Big Horn Basin. Jacklyn A. Green, Richard A. Mangali, Tabitha N. Tyrrell, Tawna R. Herrera, Fantasia Critchfield, Camille L. Brandt, Sidney D. Brooks, Lara M. Chanthongthip, Emilie D. Edwards, Gareth G. Flowers, Drew Groll, Pascal Hagenimana, Abigail L. Hogan, Wyatt J. Horrocks, Brianna N. Kilpatrick, Chaz P. Krone, Carissa A. Schmidt, Laura Shoopman, Allan Childs, Elise Kimble, and Eric C. Atkinson

Northwest College, 231 West 6th Street, Powell, WY 82435 Email: jacklyn.green@nwc.edu Abstract. Ants comprise a ubiquitous family (Formicidae) of the Hymenoptera contributing hugely to the functioning of ecosystems. To investigate the microbiome of ants and wasps (Vespidae) inhabiting the Big Horn Basin, we aseptically collected over 50 samples using point-of-collection software (Anecdata.org) to document spatial distribution from areas east and south of Lovell, WY to Belfry, MT. Bioinformatics theory and application became the central theme of Biology 2465 “Research Problems in Biology”, introducing undergraduates to genomic analyses. We extracted DNA from whole-body homogenates, amplified 16S ribosomal DNA (V4 region) following Earth Microbiome Project protocols, and sequenced on Illumina platform. Metabarcode data were characterized via the QIIME2 pipeline to compare patterns of microbial diversity across space and ant/wasp taxa. We will be investigating bacterial diversity further this year assessing spatial and taxonomic patterns potentially with genomic structure of the ants (Pogonomyrmex occidentalis) themselves. P5. Spice Up Medicine. Fantasia Critchfield, Tayler Groll, Dylan Przygocki, and Uko Udodong

Northwest College, 231 West 6th Street, Powell, WY 82435 Email: Fantasia.critchfield@nwc.edu Abstract. Spices have a long history in medicine treating a broad range of ailments through natural resources. However, the active compounds in these spices have very low solubility in water. Eugenol makes up over half of the chemicals found in cloves (spice), a compound with only 2.46 mg/ml aqueous solubility at 25°C. Cloves can be used in the treatment of acne, yeast infections, as well as food poisoning, septic shock, and typhoid fever. Increasing the solubility and bioavailability of drug products are usually created through formulation or the use of prodrugs. In this research, we will be using carbohydrate derivatives to mediate solubility. With the use of glucose, mannose, and galactose as glycosyl donors, we will prepare glycosides using the active ingredients of spices as glycosyl acceptors. A change in solubility could result in a change in activity profile. If the activity profile were to increase, these altered spices

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could reveal more potent remedies, leading to a greater prevalence in modern medicine. P6. Extraction and Isolation of Medicinal Proteoglycans from Pearl and Blue Oyster Mushrooms. 1Thomas Haack and 2Eric Mechalke

1University of Wyoming at Casper, 125 College Dr., Casper, WY 82601 2Casper College, 125 College Dr., Casper, WY 82601

Email: thomashaackcollege@yahoo.com Abstract. Many common edible mushrooms have been documented to display medicinal properties. Proteoglycans found in oyster mushrooms were correlated with anti-tumor and immunomodulatory responses with Sarcoma-180 cancer cells. The purpose of this study is to isolate the best growth and extraction method to maximize the amount of proteoglycans separated from mycelia and sporocarp. Pearl and blue oyster mushrooms were grown in different chemical and physical environments in order to determine the mycelia growth for each mushroom. Several centrifuge, dialysis, and ion/size exclusion chromatography methods provide proteoglycans from samples to find the greatest percent yield of medicinal protein from each mycelia and sporocarp. P7. Molecular Categorization and Frequency of SHV-Like Antibiotic Resistance Genes in the Environment. 1Caitlyn Edwards, 1Hannah Burrough, and 2John Chase

1University of Wyoming at Casper, 125 College Dr., Casper, WY 82601 2Casper College, 125 College Dr., Casper, WY 82601

Email: cmille77@uwyo.edu Abstract. The emergence and spread of antimicrobial resistant genes poses a threat to healthcare systems worldwide. Although this issue is significant, little research has assessed the environmental factors contributing to the rise of antibiotic resistance. Environmental microbiota represent one of the most diverse reservoirs of antimicrobial resistance, known as the resistome. The resistome includes not only the genes that confer resistance to clinical pathogens, but also to the nonpathogenic species that dominate the environment. While the resistome is ancient, modern day selective pressures are undoubtedly shaping its current form, degree of mobility, and allele frequency. OBJECTIVE: The primary focus of this project is on eukaryote, prokaryote, and viral populations in the environmental resistome. Over 100 water samples have been taken from aqueous systems, including freshwater, mineral hot springs, and wastewater, throughout Wyoming. Metagenomic DNA for beta-lactamase genes are being isolated in order to investigate the molecular categorization and frequency of these genes. The changes between the microbiome and resistome of these water sources are being analyzed to gain a better understanding of where beta-lactamase genes occur in the environment. The current goal of this project is to continue the ongoing analysis of any SHV-like antibiotic resistant genes found in the samples through the creation of metagenomic

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DNA libraries. After library creation, DNA will be sequenced followed by bioinformatics analysis. The long-term goal is to present a more accurate picture of the molecular categorization and frequency of SHV-like genes in the environment. P8. Understanding the Binding Specificity of PopZ Protein. 1Theressa Smith, 1Sarah Steinberg, 1Dagmara Motriuk-Smith, and 2Grant R. Bowman

1University of Wyoming at Casper, 125 College Dr., Casper, WY 82601 2University of Wyoming, 1000 E. University Avenue, Laramie, WY 82071

Email: theressa.smith.mn@gmail.com Abstract. Caulobacter crescentus is a gram negative bacteria in the class of Alphaproteobacteria. It is known to express an intrinsically disordered protein known as Polar Organizing Protein Z (PopZ). PopZ is responsible for cell organization and polar localization during asymmetric cell division of C. crescentus. The interactions between PopZ and a specific binding partner can be observed by expression of recombinant DNA in Escherichia coli. Induction with Isopropyl β-D-1-thiogalactopyranoside (IPTG) induces expression of mCherry fused with PopZ. Induction with arabinose results in expression of green fluorescent protein (GFP) fused with any of the binding partners. Therefore, the interactions between binding partners and PopZ can be observed by using fluorescent microscopy. Previous studies have optimized experimental conditions of positive control expression, using ChpT and ParB, which co-localize with wild type PopZ. Current research includes screening interactions of PopZ mutants that have either been altered at the N-terminus or the PED regions, with proteins involved in cell division. P9. Denitrification Gene Presence Relative to Land Use in Semiarid Environments. Brandon Phillips and Christopher Wenzel

Eastern Wyoming College, 3200 West C Street Torrington, WY 82240 Email: phillipsb4046@ewcmail.wy.edu Abstract. Denitrification is a process whereby nitrate or nitrite is reduced to a gaseous form of nitrogen (N2 or N2O), most commonly by microbes. Biological denitrifying activities, which play an important role in the global nitrogen cycle, are characteristic of both prokaryotes and several fungi. Denitrification potential can be used as an indicator of both soil health and environmental quality. A synopsis of the genes involved in denitrification (nirS/K, norB, nosZ) along with their relative presence in soil under various land uses (i.e. grazed-grassland, cropland, urban, wetland, sub-irrigated) is presented. An enhanced understanding of the role these genes play in soil relative to various land uses will improve our understanding of nitrogen cycling in soil and the potential to prevent nitrates from contaminating groundwater and nitrous oxide from contributing to the greenhouse effect and global climate change. P10. Preliminary Transcriptome Analysis of Adult Culex tarsalis Mosquitoes Collected in Fremont County, Wyoming. Kelvin Kinyatta and Aaron Bender

Central Wyoming College, 2660 Peck. Avenue, Riverton WY 82501

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Email: kmk0609@cwc.edu Abstract. Fremont County is a hotspot for West Nile virus (WNV) infection in Wyoming. It is known that the mosquito Culex tarsalis (C. tarsalis) serves as the primary insect vector for WNV in western states such as Wyoming. Although a great deal is known about the transmission, replication, and genetics of WNV, very little is known about the genome structure and gene expression profile of C. tarsalis. Our research team captured mixed populations of wild mosquitos in Fremont County. These were sorted and adult C. tarsalis were positively identified. Total RNA from this pool was isolated, column-purified and quantified. Two parallel samples of high quality total RNA were submitted for Illumina next-generation RNA sequencing. Our extremely comprehensive dataset consists of nearly 350 million paired-end reads. The average data yield for the 2 samples is approximately 50,000 megabases with a Q score of >/= 30 for over 91% of called bases and has a mean quality score of nearly 38. In collaboration with the University of Wyoming INBRE bioinformatics core, our research team intends to assemble the transcriptome and search for gene-expression profiles that may lead to a better understanding of the WNV transmission process. Finally, using a combination of qRT-PCR and bioinformatic analysis of our sequencing dataset, we will ask whether other members of the flaviviridae are carried by C. tarsalis and whether WNV can integrate into its host’s genome. P11. Life History Factors Important in Incidence of White-nose Syndrome (Pseudogymnoascus destructans) among North American Bats. Candace Siirila and Eric C. Atkinson

Northwest College, 231 West 6th Street, Powell, WY 82435 Email: candace.siirila@nwc.edu Abstract. Life and History. How do the two of these go together? It seems that bats are affected by not one, but both of these things together. It is important to learn how the life history of bats can teach us about deadly fungal pathogens such as Pseudogymnoascus destructans (Pd), the causative agent of White-nose Syndrome, and how they might spread throughout the United States. I investigated the life history characteristics of North American bats to elucidate patterns that may predispose certain species to infection by Pd. I classified bats by migratory pattern (fully migratory, partially migratory, or resident), roosting behavior (colonial versus singly), roosting habitat (cave, tree, or dispersed locations), and whether the species enters torpor. I reduced this matrix via Principal Components Analysis yielding a model in which the first two factors accounted for over 63% of the variability in Pd infection status. Hence, by studying the life history characteristics of bats, we can predict not only which species are susceptible to this disease but, with environmental information, also the geographical susceptibility each species may exhibit. P12. Fishing for Water Quality Gareth G.K. Flowers, Wyatt J. Horrocks, Michael F. Cuddy, and Eric C. Atkinson

Northwest College, 231 West 6th Street, Powell, WY 82435 Email: wyatt.horrocks@nwc.edu

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Abstract. Water quality is important, especially in the Big Horn Basin, where both lotic and lentic water systems play a major role in agriculture as well as fish habitat. Water quality indicators such as pH, salinity, TDS, conductivity, and turbidity are all useful in determining water quality and can be tested either in the field or lab. We collected geospatial information and water samples from several sources representative of the Shoshone River and the Clark’s Fork of the Yellowstone River watersheds in the Big Horn Basin. Our data indicate that, regardless of the source, the pH remained close to 8.5, while the TDS and salinity varied by several hundred parts-per-million. This suggests that the solid and salt compounds entering these waters are not noticeably affecting acidity/basicity. We extracted DNA (eDNA) from water samples to seek insight to how microorganisms are affected by the water quality. Ultimately, this project aims to create a GIS map of the Big Horn Basin highlighting how water quality indicators change throughout the Basin.

Acknowledgements Wyoming INBRE is supported in part by a grant from the National Institute of General Medical Sciences (2P20GM103432) from the National Institutes of Health. Any opinions expressed or results presented are solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Special thanks to the administration and staff of the University of Wyoming/ National Park Service Research Station for their assistance in organizing and running the conference.

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