Abstracts (sorted by last name of primary presenter)
Jennifer Abshire, Southwestern Oklahoma State University
1:15-3:15 in LSR 4
Pseudomonas syringae motility appendages: roles in surface colonization and survival
Jennifer Abshire and Regina McGrane
Pseudomonas syringae is a bacterial phytopathogen that causes disease in economically important crops. P. syringae is found in many different environments, each with varying levels of moisture. Therefore, the appendage it relies on for motility varies. The removal of genes encoding for these motility factors significantly impairs pathogenicity. The objective of this project was to evaluate the impact of P. syringae motility related appendages on colonization of varying environments. To characterize the role of flagella and pili, movement of deletion mutants was observed in saturated, liquid, and natural surface environments. Flagella mutants had significantly reduced motility in both saturated and liquid conditions, whereas pili mutants had significantly reduced motility in saturated conditions only. This indicates that flagella are the most relied upon appendage for motility and that pili have minor impacts. Similarly, on simulated natural surfaces flagella were the primary movement appendage. To characterize the impact of flagella and pili on colonization of plants, attachment to leaves and seeds as well as growth on leaves were evaluated. Mutants lacking flagella or both flagella and pili were impaired in leaf colonization but showed seed attachment similar to the wild type. However, mutants lacking both the flagella and pili had greater levels of attachment to leaves. To characterize the impact of flagella and pili on antibiotic sensitivity, mutants and the wild type were evaluated using a Kirby-Bauer sensitivity assay. Mutants were more sensitive to ciprofloxacin than the wild type, which is opposite to observations made in a different P. syringae strain. These results indicate that removal of the genes encoding for motility factors such as pili and flagella, directly impacts pathogenicity. Limiting the motility of P. syringae could halt or severely lessen the symptoms associated with this disease-causing pathogen and increase crop yields of host plants.
Neha Amberkar, Texas A&M University - Texarkana
1:15-3:15 in LSR 3
Microplastic Pollution in East Texas Freshwaters and Water Treatment Plants
Neha Amberkar and Vaishnavi Ramachar
Usage of plastic in modern society generates many benefits, yet there is the issue of emerging concern regarding the accumulation and pollution of plastics in aquatic environments as well as its effects in the ecosystem. Microplastics are plastic fragments smaller than 5 mm. They are especially harmful as they can easily be ingested by wildlife, affecting the food web. As this waste is discarded, the breakdown of larger plastics results in excessive accumulation in the environment as microscopic particles. This study focuses on identifying microplastic pollution in the inflow and outflow of freshwaters around water treatment plants in East Texas. To analyze the amount of microplastic fragments as well as corresponding size, water samples were collected from the surface, middle, and sediment layers. To understand microplastic pollution in raw unfiltered water as well as the effect a standard treatment plant has on eliminating these wastes before re-release into the ecosystem, samples were collected from the water that passes in and out of the sewage treatment plant. Once collected, the water samples were then divided into portions, filtered, and remaining microplastics were then observed under a microscope. Size, composition, and overall density of the microplastic fragments were analyzed. Densities were also observed corresponding to the surface, middle, or sediment layer of the waters to determine where these microplastics aggregate and have the strongest presence. Conclusions were then hypothesized for the implications of this pollution in the local ecosystem and on the larger environmental scale.
Nana Amponsah, Angelo State University
1:15-3:15 in TXH 2
Predator Avoidance Behavior in Physa acuta
Nana Amponsah and Ben Skipper
The senses of prey animals have been finely tuned by natural selection to detect potential predators before a predation attempt is begun. For aquatic invertebrates, chemosensation is the primary means by which predators are detected. However, in polluted waters, the chemosensory abilities of prey animals may be negatively impacted through interference. In such cases, prey animals may not be able to sense predators before a predation attempt. This study investigates predator avoidance behaviors of Physa acuta, a freshwater snail, exposed to two potential predators and scent of a distressed conspecific in both clean and polluted waters. Snails were observed in isolation for a period of thirty minutes. The first ten minutes allowed snails to acclimate to the new tank. In the second ten minutes we recorded the snail’s movement in the absence of predator information. Prior to the third ten minutes, we added scent of a predaceous beetle, crayfish, or water (control) and then continued recording snail behavior. We observed no significant difference in movement and behavior after the addition of beetle scent and water. However, there was significant decrease in movement after addition of crayfish scent, with some snails escaping to dry areas in the test tank. Tests in polluted water, where Physa acuta may be unable to use chemosensation (effectively), are forthcoming.
Kia Baeza, St. Edward's University
1:15-3:15 in TXH 2
Commonly Used Agricultural Pesticides and their Effect on the Immunological Function of Honeybee Populations
Kia Baeza
Colony collapse disorder (CCD) is a current phenomenon which is causing rapid declines in honeybee populations. While researchers have yet to find a direct cause of CCD, it is hypothesized that it may be the result of a combination of multiple factors such as global warming, parasitic Varroa mites, and the use of pesticides. The goal of this study was to identify the effect of four commonly used pesticides in agriculture (imidacloprid, glyphosate, thiamethoxam, and clothianidin) on the immune function of honeybees. For each pesticide, 150 honeybees were collected randomly from a set of hives in the Austin area and starved for four to six hours. A pesticide sugar water solution with biological relevant concentrations was then fed to each bee with a micropipette. Bees were then left for zero (control), two, or four hours after feeding before hemolymph was extracted. Hemolymph was added to buffer, lysed with glass beads, and placed at a -80oC to stop protein activity. Samples are currently undergoing several colorimetric assays utilizing a spectrophotometer to determine immunological protein activity including a RED 660 assay for overall protein concentrations, prophenoloxidase, catalase, and peroxidase. Anecdotally, bees subjected to the pesticide glyphosate appear to have higher mortality over time compared to the other pesticides tested. Bees that ingested glyphosate had significantly lower protein concentrations than all other pesticides tested. Prophenoloxidase activity appears highest when bees are exposed to clothianidin, while imidacloprid significantly decreases activity four hours after exposure. Results from this study will help determine what effects, if any, pesticides contribute to the onset of CCD.
Anna Bailey, Oral Roberts University
11:15-11:30 in DBU
Gene Annotation of the Hypothetical Protein-Coding Genes of Chlamydia caviae
Anna Bailey
Genetic information for a multitude of microorganisms has become readily available due to technological innovations in the field of genome sequencing. However, several of these novel genome sequences remain unexplored and require manual annotation to uncover their hypothetical protein-coding genes. Through the use of publicly available online bioinformatics programs developed by the United States government as well as researchers across the globe, primary amino acid sequences derived from hypothetical gene sequences can be used to predict their function. Examples of properties identified by these programs include protein families, conserved domains, signal peptides, transmembrane regions, open reading frames, and protein localization. The purpose of this research project is to predict the functions of five unannotated hypothetical protein-coding genes in the bacterium Chlamydia caviae, which infects the mucosal epithelium of rodents. The genes CCA_RS04585, CCA_RS04590, CCA_RS04595, CCA_RS04600, and CCA_RS04605, were analyzed and are predicted to code for the following: glycerol-3-phosphate dehydrogenase (NAD(P) (+)), a membrane protein, two type-III secretion system family proteins, and phosphoenolpyruvate carboxykinase, respectively. These predictions provide some insight into the proteome of Chlamydia caviae. Ultimately, the proposed gene annotations must be validated through molecular cloning and biochemical methods to determine whether Chlamydia caviae expresses these proteins and whether they perform their bioinformatically predicted functions.
Kaylee Ballard, The University of Tulsa
1:15-3:15 in LSR 3
Next Generation Sequencing of Staphylococcus Species Isolated from Oklahoma Retail Mushrooms
Kaylee Ballard, Madison Bickle, Leena Neyaz, Anand B. Karki, and Mohamed K. Fakhr
Staphylococcus species are important pathogens and are responsible for various infections associated with serious community and hospital acquired diseases. Most studies characterizing Staphylococcus spp. are biased toward clinical strains vs. those isolated from food. Extended study on the genomic structure and virulence potential of various species of Staphylococcus from retail mushroom is lacking in the literature. In a preliminary study conducted in our laboratory, the prevalence of Staphylococcus spp. isolated from Oklahoma retail mushrooms was significant, and several strains were phenotypically resistant to a wide range of antibiotics. Most of these strains harbor antibiotic resistance genes like mecA gene in addition to some enterotoxin genes. The objective of the current study was to determine the genomic sequence using next generation sequencing (NGS) technology of 12 various Staphylococcus species previously isolated from Oklahoma retail mushrooms. Twelve strains of Staphylococcus species (3 S. fleurettii, 2 S. sciuri, 1 S. saprophyticus, 1 S. vitulinus, 1 S. xylosus, 1 S. succinus, 1 S. pasteuri, 1 S. warneri and 1 S. haemolyticus) were streaked on TSA plates from stock cultures (-70oC) and then grown as liquid cultures in TSB media overnight. Genomic DNA was extracted from the overnight cultures using DNeasy blood and tissue kit (Qiagen) according to the manufacturer instructions. The twelve strains were also screened for the presence of plasmids using the Qiagen MediPrep plasmid isolation kit. Plasmids with various sizes appeared to be prevalent among the tested strains. Next generation sequencing library preparation are currently underway using the Illumina Nextera XT DNA library preparation kit and it will be run on the Illumina MiSeq Sequencer. Genome assembly will be performed using the CLC Genomics Workbench and the microbial genome finishing modules. Comparative genomics among the twelve sequenced Staphylococcus strains will be explored to detect genes responsible for virulence and antimicrobial resistance.
Kaylee Ballard, The University of Tulsa
9:30-9:45 in DBU
Influence of Staphylococcus aureus Co-Contamination On Survival of Campylobacter Strains At Low Temperature, Biofild Formation, and Aerotolerance
Kaylee Ballard, Anand B. Karki, Claudia Harper, and Mohamed K. Fakhr
Campylobacter, which causes the diarrheal illness Campylobacteriosis, is one of the leading bacterial foodborne pathogens in the United States. High prevalence of the bacteria has been found in retail meat and liver products, most of which are also contaminated with Staphylococcus aureus. In adverse conditions, the presence of Staphylococcus aureus has shown to have an influence on the survival and biofilm formation of Campylobacter. This study investigates the effect of Staphylococcus aureus on the survival of Campylobacter at low temperatures commonly used in retail meat storage, its biofilm formation, and aerotolerance. Six strains of Campylobacter [C. jejuni (T1-21, NCTC11168, OD2-67) and C. coli (HC2-48, WAS-33, ZV1-224)] were incubated in MHB media as well as filter sterilized S. aureus grown MHB media from two strains of S. aureus (B4-59C and B6-55A). Assays for survival with viable count, biofilm formation and aertolerance assays were performed. Results showed there was a higher survival rate for prolonged time at low temperatures in the Campylobacter strains grown in MHB media containing S. aureus metabolites versus the MHB media control. Aertolerance was also shown to be enhanced with the presence of S. aureus metabolites. This indicates that the extracellular metabolites of S. aureus influence the survival of Campylobacter. There were no consistent results on the influence of S. aureus grown media on Campylobacter biofilm formation. Similar experiments including S. aureus cells instead of the filtrate media were also conducted. Presence of S. aureus cells enhanced survival of strains of Campylobacter at low temperature as well as biofilm formation.
Emily Bedea, Southwestern Oklahoma State University
1:15-3:15 in TXH 1
In-silico analysis and homology modeling of β-carbonic anhydrase from Streptococcus sanguinis, an opportunistic pathogen involved in subacute infective endocarditis
Emily Bedea and Vijayakumar Somalinga
Streptococcus sanguinis is one of the leading causes of infective endocarditis (IE) in a susceptible population. Although several virulence factors involved in IE have been identified, the exact mechanism by which S. sanguinis colonizes endocardium remains unclear. In addition to some well-characterized virulence factors, S. sanguinis also contains virulence-associated proteins that are not well characterized. One such protein that is associated with virulence is carbonic anhydrase (CA) that catalyzes the reversible hydration of CO2 to bicarbonate. The goal of this study is to identify and characterize carbonic anhydrase from S. sanguinis. We have identified a β-CA from S. sanguinis (SsaCanB) using BLAST analysis that shows high homology to other well-studied β-carbonic anhydrases. SsCanB is an 18.2 kDa protein with no N-terminal signal peptide indicating its cytoplasmic localization. Sequence alignment of S. sanguinis with closely related homologs shows conserved domains typical of β-CA including residues important for metal coordination and protein-protein interactions. Homology modeling and subsequent structure analysis of SsCanB reveals that both proteins have α/β fold typical of β-CA’s. Analysis of the active site of SsaCanB shows conserved residues involved in zinc ion coordination similar to other well-studied β-CA’s. Furthermore, the catalytic dyad (Asp/Arg) required for the activation of water molecule coordinated with zinc ion is also conserved in SsaCanB.
Alexa Benavides, Houston Baptist University
1:15-3:15 in LSR 4
The Very Hungry Superworm: Superworms' ability to digest Polystyrene
Alexa Benavides, Julia Ta, Daisy Lopez and Agnieszka Czopik
Polystyrene (PS) is a synthetic polymer made from styrene and is commonly used in commercial packaging due to its ability to repel water and resist corrosive substances. However, the same chemical stability that makes PS so useful also poses a threat to the environment, as PS is non-biodegradable and consequently, amasses in landfills. Research published by Stanford University in 2015 provides a possible solution to this environmental crisis, since it showed that the larvae of mealworms (Tenebrio molitor) host a specific strain of gut bacteria (Exiguobacterium sp. strain YT2) that are able to break down PS into biodegradable compounds, such as water and carbon dioxide. To further expand on this idea, we fed mealworms that were maintained on only polystyrene, for seven weeks, to one group of superworms, and another group of superworms were fed mealworms that were maintained on only oatmeal for the same amount of time. We then fed PS to both groups of superworms and calculated the amount of PS eaten per worm in order to analyze if changing the worm microbiota will have an impact on their ability to digest PS. We plan to further our studies by analyzing crumbs from the PS that was fed to the superworms to determine whether there are enzymes remaining on it that will help it degrade.
Jillian Blake, Oral Roberts University
9:00-9:15 in TXH
Attraction of swarming paper wasp sexuals (Polistes spp.) to the chemical attractant N-3-methylbutyl acetamide
Jillian Blake
Before the beginning of the experiment, common passageways used by the paper wasps to enter inside the ORU dormitories were sealed in hopes of decreasing the number of wasps entering the buildings. This study investigated the numbers of paper wasps (Polistes spp.) swarming atop dormitories after the limitation of dormitory access in comparison to data from previous years. The objective was to make a weekly observation counts of swarming paper wasps on the roofs of the Francis and Michael Cardone dormitories at Oral Roberts University. With the new changes in dorm accessibility, it was predicted that the overall wasp count would be lower than that of previous years. These counts found that two distinct peaks of wasp activity occurred in early and late October due to variable weather temperature. As expected, the overall wasp count was lower than total wasp counts in years dating back to 2011. However, this reduction may be due to the second study, trapping wasps atop the dorms. Three sets of wasp traps were placed: two sets atop the Francis dormitory and one set along the fence lining the west side of Fred Creek. The traps collected a total of 680 wasps, but the effectiveness of the N-3-methylbutyl acetamide attractant compared to the control was not significant. Thus, it was concluded that further experimentation would be necessary to gather more significant data.
Hannah Butterfield, Austin College
11:30-11:45 in TXH
Investigating Synergy Between PA28γ and IKKε
Hannah Butterfield and Lance Barton
Cancer is characterized by excessive cellular proliferation and by dysregulation of signaling pathways related to growth. PA28γ is a proteasome activator frequently increased in cancer and involved in regulation of multiple cellular pathways that affect proliferation, including the Nuclear Factor κB (NFκB) pathway. Inhibitor of Nuclear Factor κB Kinase ε (IKKε) is a breast cancer oncogene that affects multiple pathways that implicate PA28γ, including the activation of NFκB. This study aims to elucidate whether PA28γ is required for normal IKKε activity. I hypothesize that PA28γ enhances the overall expression and nuclear export of IKKε, and aids IKKε-mediated cell signaling. I am assessing this through evaluating differences in IKKε nuclear, cytosolic, and overall concentration in PA28γ+/+ and PA28γ-/- murine embryonic fibroblast (MEF) cells. I am also evaluating differences in IKKε-implicated cellular processes including transcriptional activation and animal weight gain. The data does not support a role for PA28γ in regulation of IKKε.
Constance Byers, Texas A&M University - Texarkana
11:00-11:15 in DBU
Microplastic Pollution in East Texas Freshwaters and Water Treatment Plants
Constance Byers and Roya Morakabian
Usage of plastic in modern society generates many benefits, yet there is the issue of emerging concern regarding the accumulation and pollution of plastics in aquatic environments as well as its effects in the ecosystem. Microplastics are plastic fragments smaller than 5 mm. They are especially harmful as they can easily be ingested by wildlife, affecting the food web. As this waste is discarded, the breakdown of larger plastics results in excessive accumulation in the environment as microscopic particles. This study focuses on identifying microplastic pollution in the inflow and outflow of freshwaters around water treatment plants in East Texas. To analyze the amount of microplastic fragments as well as corresponding size, water samples were collected from the surface, middle, and sediment layers. To understand microplastic pollution in raw unfiltered water as well as the effect a standard treatment plant has on eliminating these wastes before re-release into the ecosystem, samples were collected from the water that passes in and out of the sewage treatment plant. Once collected, the water samples were then divided into portions, filtered, and remaining microplastics were then observed under a microscope. Size, composition, and overall density of the microplastic fragments were analyzed. Densities were also observed corresponding to the surface, middle, or sediment layer of the waters to determine where these microplastics aggregate and have the strongest presence. Conclusions were then hypothesized for the implications of this pollution in the local ecosystem and on the larger environmental scale.
Jailene Canales, University of Central Oklahoma
11:15-11:30 in TXH
Characterizing Early Developmental Defects in an Avian Model of Maternal PKU
Jailene Canales, Austin McDonough, Morgan Massey, and Nikki J Seagraves
Maternal phenylketonuria [MPKU] is a syndrome of multiple congenital anomalies including cardiovascular malformations [CVMs], brain and growth restriction when a mother with Phenylketonuria [PKU] does not control her dietary intake of Phenylalanine [Phe]. In this study, we aim to establish and characterize an avian model of MPKU. We focused on early developmental defects. We investigated the effect of 2500μM Phe exposure by in-ovo yolk injection. Following the injection, the embryos underwent further development for 48 hours until dissection was performed. At HH14-17, India ink was injected into the yolk as a contrast dye. Images were taken of embryos and they were scored based on Drake et. al (2006.). Embryos exposed to high Phe displayed gross morphological changes including developmental and growth delays, anterior and posterior abnormalities, and torsion defects. Histological analysis is underway to determine changes in heart development. Currently there is no data interrogating the mechanism by which Phe causes heart defects. We plan to utilize this model to define the mechanism of Phe cardiac teratogenicity which is critical for improving MPKU treatments and outcomes.
John Cantley, University of Texas at Tyler
1:15-3:15 in LSR 3
Pseudomonas aeruginosa Antibiotic Resistance Gene Profile in East Texas Clinical Isolates
John Cantley, Shandis Fancher, and Ali Azghani
Pseudomonas aeruginosa is an opportunistic, Gram-negative bacterium and causative agent of acute nosocomial, community-acquired pneumonia, or chronic infections. Clinical isolates of this organism harbor intrinsic and acquired antibiotic resistant genes. Improper antibiotic prescription or use may select for mutations in genes coded for ß lactamase, porins, or efflux pumps. These mutations confer resistance across multiple drugs, make the treatment of infections more challenging, and increase the rate of infection occurrence and mortality. The goal of this project is to identify antibiotic resistant genes carried by clinical isolates of P. aeruginosa in East Texas. We utilized quantitative real-time PCR on recently collected clinical isolates of P. aeruginosa in order to determine the presence of targeted antibiotic resistant genes. Bacterial subcultures were adjusted to an optical density of 0.8 (OD600), and total DNA was isolated using the boiling method for 10 min at 100 ºC. Cold ethyl alcohol was used to enhance precipitation of a salt free DNA pellet. Samples were checked for concentration and purity using a NanoDrop spectrophotometer. Quantitative real-time PCR was performed using targeted primers on the DNA extracts using 40 thermal cycles of 20 sec at 95ºC, 20 sec of 60ºC, and 30 sec of 72ºC. We are in the process of applying the primers for aforementioned genes on each of these isolates. Our far-reaching goal is to compare the rate of antibiotic resistant strains of P. aeruginosa in East Texas to global database in the context of clinicians standard care and patients compliance.
Angela Chen, Angelo State University
9:00-9:15 in LSR
Atypical Expression of Cell Cycle Genes in a Drosophila Model of Alzheimer’s Disease
Angela Chen
Understanding the genetics of the cell cycle is important for studying normal development and physiology as it guides development, function, and maintenance in our bodies. Atypical gene expression in cell cycle genes can lead to pathology such as developmental disorders and neurological diseases. Alzheimer’s disease is the most common cause of dementia and is a significant financial burden on the United States economy. Atypical expression of Cdk4 has been studied in the brain of Alzheimer’s disease patients. Asp and Mcph1 are genes that are related to the dysregulation of the cell cycle, and mutations in these genes can result in microcephaly, a human neurodevelopmental disease. However, there have been no studies so far to determine the gene expression of Mcph1, Asp, and Cdk4 in the Drosophila model of Alzheimer’s disease. The goal of this research was to develop primers to allow neurodevelopmental gene expression to be studied both qualitatively and quantitatively for neurological diseases such as microcephaly and Alzheimer’s disease. Functional primer sets were developed and expression levels of Asp, Mcph1, and Cdk4 were tested using qualitative and quantitative RT-PCR. Aberrant expression of these genes was observed in the disease models compared to the wild type strain. Characterizing the role of these genes may reveal potential use in inducing adult neural stem cells to proliferate for use in future in novel therapeutics and regenerative medicine for Alzheimer’s disease.
Clay Clarkson, Texas A&M University - Corpus Christi
1:15-3:15 in TXH 1
Quantifying the relationship between methane concentration and mcrA genes from a cultured Methanolobus spp. isolated from surfaces sediments of Tranquitas Creek, Kingsville Texas
Clay Clarkson and Brandi Kiel Reese
Methanogens are a diverse taxonomic group within the archaeal domain that are capable of methanogenesis. Methanogenesis was once thought to be unique to the phylum Euryarchaeota, however; the essential gene for methanogenesis, methyl-coenzyme M reductase (mcrA), has been discovered amongst other phyla including the renamed miscellaneous Crenarchaeota group: Bathyarchaeota. Members of Bathyarchaeota are widespread amongst environments and contain variants of mcrA, suggesting a potential ability to use other methylated alkanes other than those known today. Though, little is known about this phyla’s contribution to global methane fluxes and the exact substrates for methanogenesis. In attempt to identify and quantify methane fluxes of potential methanogenic members outside the Euryarchaeota phyla, an enrichment approach followed by subsequent transfers was employed to obtain a methanogen isolate in culture. Surface sediments of a freshwater stream system was used for the enrichment phase. Taxonomic identification of the pure cultures was done by sequencing on the 16S rRNA gene. Methane flux per individual cell was calculated by using the relationship between methane concentration and mcrA gene abundances during exponential growth phases. Quantification of methane and mcrA genes in the pure cultures was determined by gas chromatography and droplet-digital PCR. This study aims to demonstrate a potential use of mcrA gene counts as a proxy for quantifying methane fluxes.
Katherine Collins, Austin College
9:15-9:30 in LSR
Barriers to plant establishment from seed in tallgrass prairie restoration
Katherine P. Collins
Conversion to agriculture during the late 19th and early 20th Centuries severely reduced the extent of the tallgrass prairies once prominent throughout the central United States. In an on-going tallgrass prairie restoration project at Austin College’s Sneed Prairie, students track restoration progress on nine fields managed three different ways. Success of grass establishment from planted seeds has varied dramatically among fields, and even among experimental replicates subjected to the same restoration techniques. Three fields have had dramatic success of grass establishment from planted seeds. Six fields have had less, and in some cases much less, success. The objective of this project is to test hypotheses with potential to explain the lack of widespread establishment from planted seeds in the latter six fields.
Cristin Connor, University of the Ozarks
11:45-12:00 in LSR
Temperature and the Variability of Vertebrae in Dasypus novemcinctus
Cristin Connor and Ashley Saunders
The nine-banded armadillo is a fascinating creature and exceptionally useful for scientific research. This armadillo always gives birth to four genetically identical offspring which minimizes the variables that must be accounted for in an experiment. Another interesting fact, and the focus of this study, is that the number of vertebrae in this species is variable. With only 40% of a studied population having the median number of vertebrae (Asher, et. al.) This is believed to be due to variation in the body temperature of the mother during gestation. 77% of the armadillos studied had the median number of vertebrae but three litters of offspring had abnormal counts. Two of these litters were part of the control group and all three were in a normal range of variation, only about 2-3 degrees between day and night. This brings up an issue with the temperature hypothesis because it did not appear to be the causative agent in this experiment. Perhaps a next step would be food stress, as would be experienced in the wild, which could cause more natural occurrence of vertebral variation.
Bailee Cummings, Southern Nazarene University
1:15-3:15 in LSR 3
Mosquito Sampling and Arbovirus Surveillance in Oklahoma
Bailee Cummings, Randall Welles, Helen Arango, Nicholas Back, and Caio Martinelle França
Vector-borne diseases are a major burden for public health. In the U.S., more than 640,000 cases of vector-borne diseases were reported from 2004 to 2016. In the past 13 years, mosquito-borne disease cases have tripled. Culex mosquitos are responsible for establishing endemic West Nile virus across the U.S. The current extended range of invasive Aedes mosquitos dramatically changes the landscape for arboviral risk in the U.S. because they can transmit diseases like Zika, Dengue, and Chikungunya. According to the Oklahoma State Department of Health, West Nile virus (WNV) was introduced to Oklahoma in 2002. Mosquitoes carrying WNV can infect persons of any age; however, those over the age of 50 are at greater risk of developing serious illness affecting the nervous system. From 2002-2018, the State of Oklahoma registered over 779 cases of WNV and 58 deaths. Three WNV outbreaks have occurred in Oklahoma in 2003, 2007, and 2012. This accelerating spread of arboviruses highlights the need for a proactive mosquito surveillance. Mosquitoes were collected from multiple counties in Oklahoma using CDC miniature light traps resulting in 7929 mosquitoes captured. Mosquitoes were identified to 15 different specifies from four main genera: Aedes, Culex, Anopheles, and Psorophora. Nucleic acid was extracted from field-collected mosquitos using DNA/RNA ShieldTM from 50 mosquito pools. Surveillance of viruses in mosquito pools was performed with RT-qPCR using universal primers for detection of flavivirus-specific viral nucleic acid. We identified three RT-PCR positive pools for the presence of the genus Flavivirus. We are currently working on whole viral genome sequencing for these positive samples in order to obtain phylogeographic information of the viruses.
Caitlyn Cutrer, Northwestern State University
9:45-10:00 in LSR
Identifying Leptospira interrogens in feral hog populations using the MALDI-TOF
Caitlyn Cutrer, Jordan Bringedahl, and William Simpson
Leptospira interrogens is a zoonotic pathogen that causes leptospirosis. Feral hogs (Sus scrofa) are known carriers of this bacterium and due to the hogs’ reproductive prowess their population is beginning to encroach into city water reservoirs creating a human health concern. Traditionally, identification of Leptospira sp. is done by using the microscopic agglutination test; however, this can be expensive and time consuming. Alternatively, by using MALDI-TOF we can identify multiple species of Leptospira using mass spectrum profiles. Blood was opportunistically collected from 23 feral hogs across Natchitoches Parish. The serum was separated via centrifugation. 0.5 mls of a respective serum was then pipetted into Fletcher’s medium modified with 0.05% hemin. Once growth was observed (~5-7 days), 1 ml of medium was aseptically pipetted out into a 1.5 ml tube and centrifuged, the supernatant was decanted, and the pellet spotted on a MSP 96 target polished steel plate and overlaid with 1 µl of HCCA. Samples were analyzed using the MALDI-TOF Biotyper software (v4.1.70). The results have yielded mixed results ranging from no growth, to the enrichment of Aeromonas hydrophila, Stenotrophomonas maltophilia, and multiple Pseudomonas sp.; however, some samples yielded mass spectra that could not be readily identified, due to a lack of Leptospira sp. profiles in the current version of the Biotyper database. Thus, we are working on constructing our own library of Leptospira sp. using pure cultures obtained from ATCC and will retroactively assess the previous non-identified spectra obtained from the various serum enrichments.
Erin Dempsey, East Central University
10:45-11:00 in TXH
Plant Pathogenic Fungal Species on Melons from Guatemala
Erin Dempsey, Patrick McDowell, Matt Broge, C. Biles, and A. Howard
Melons (Cantaloupe; Cucumis melo var. cantaloupensis) were purchased from local grocers in 2016 through 2019 and observed for post-harvest diseases. Of the 80 melons purchased in 2019, 75% developed fruit rot symptoms caused by fungi. The diseased tissue indicated that the majority of lesions were caused by Diaporthe spp. (syn; Phomopsis), and to a lesser extent lesions were caused by Alternaria and Fusarium spp. Plant pathogens such as Diaporthe spp. enter the surface of the melon fruit early in development and remain latent until fruit maturity. While ripe fruit is harvested and imported with no external evidence of disease, internal fruit rot becomes evident as the fruit matures. The objective of this study was to characterize Diaporthe spp. imported in Guatemalan melons. Fungal isolates were characterized based on culture growth characteristics, spore morphology, and DNA analysis. Guatemalan isolates were morphologically similar to D. sojae and D. curcurbitae. Deoxyribonucleic acid (DNA) was extracted from fungal hyphae and purified polymerase chain reactions (PCR) products were Eurofin, Inc. for sequencing. Sequencing analysis demonstrated that some of the isolates were a match for D. pterocarpi and species within the D. arecae complex Our finding of pathogenic Diaporthe spp. suggest that plant pathogens are carried across international borders and imported into the United States. Further analysis is being conducted on the melons collected in 2018 and 2019.
Cuishan Deng, East Central University
1:15-3:15 in LSR 4
The difference in leaf-tying caterpillar communities between forest edge and interior habitats
Cuishan Deng, Norbu Gurung, and H. George Wang
Edge effect is an important factor in shaping community patterns, particularly in forest habitats. We conducted an experiment to investigate the potential edge effect on forest arthropod communities at the Nature Conservancy’s Pontotoc Ridge Preserve. We set up artificial leaf ties on red oak trees in both edge and interior habitats, and conducted two censuses in summer 2018. We recorded the morphospecies and the number of all arthropods in the leaf ties during each census. We calculated the Simpson Diversity Index for all arthropods on each tree and compared them between the two habitats using mixed-effects modeling. We also compared the number of individuals between the two habitats using Poisson regression. There was no significant difference between the two habitats for either metrics. We then compared the densities of caterpillars (Lepidoptera) between the two habitats, and found them to occur more on trees in the interior habitat. Similarly, the number of aphids (Aphidae) was significantly higher on trees in the interior habitat.
Kade Ezell, Southwestern Oklahoma State University
1:15-3:15 in TXH 2
Antimicrobial activity of phytopathogen biosurfactants against human pathogens
Kade Ezell and Regina McGrane
Antimicrobials play a major role in the fight against bacterial growth in many areas of the modern world. Without the benefits of antimicrobials, the medical field, food industry, and the overall well-being of all life would be significantly impacted. The gram-negative, bacterial, phytopathogen Pseudomonas syringae produces the biosurfactants syringafactin and rhamnolipid while colonizing plant tissue. These biosurfactants lower surface tension, act as lubricants for bacterial swarming motility, and increase diffusion of plant nutrients. Our laboratory has observed repulsion of P. syringae deletion mutants lacking biosurfactant production and leaf colonizing bacteria when inoculated on swarming agar media in close proximity to wild type P. syringae. We hypothesize that biosurfactants may be important for P. syringae competition in the environment through repulsion of nearby bacteria. The effects of syringafactin and rhamnolipid on bacterial human pathogens have not been studied. The objective of this work is to evaluate the motility of human pathogens in the presence of P. syringae and to determine if biosurfactants are responsible for repulsion or growth inhibition. To evaluate the impact of P. syringae on the behavior of human pathogens, strains were inoculated in close proximity to P. syringae and swarming motility was observed. Results show that P. syringae repelled all but two gram-negative human pathogens tested but did not impact gram-positive human pathogens. Repelled gram-negative strains were also challenged with purified syringafactin and rhamnolipid. Results demonstrate that syringafactin plays a role in observed repulsion. Preliminary studies suggest syringafactin may also inhibit the survival of gram-negative human pathogens. Collectively, this work supports the proposal that P. syringae utilizes biosurfactants to outcompete closely related bacteria. Due to the potential antimicrobial properties possessed by syringafactin, P. syringae could be used to make antibiotics that attack gram-negative bacteria that live in competition with P. syringae.
Emily Falcon, University of Central Oklahoma
1:15-3:15 in LSR 4
Documenting the Expansion of an Invasion of Mediterranean Geckos (Hemidactylus turcicus) at the University of Central Oklahoma and its Surrounding Area
Emily C. Falcon, Samah Houmam, Gary D. Thomas, Laura B. Kimmel, and Allyson M. Fenwick
The Mediterranean gecko (Hemidactylus turcicus) is an exotic, nocturnal species characterized by slow dispersal which makes it a good model for studying invasions. These geckos were intentionally, repeatedly introduced to the University of Central Oklahoma (UCO) during 1963-1965 and 1985-1997. Surveys 2005–2010 and 2014–2018 documented the spread of geckos from seven to 30 buildings on campus, and six buildings off-campus in the surrounding community. Seven additional buildings on campus and one building off-campus were inspected but were uninhabited. We collected 213 tail tissue samples with a goal of having 20 samples from each building. We will go back to surveyed buildings where geckos were not observed during the fall. We will also survey new buildings. Based on genotyping of 16 previously published microsatellite loci, we found two subpopulations on and off campus. Using analyses with STRUCTURE and ARLEQUIN, we expect the buildings farther from the introduction site to cluster together, and to have more genetic differentiation compared to the source population. This project continues to monitor and document the geographic and genetic progress of a population of exotic species as it slowly expands. Data generated will help answer questions about other exotic and possibly harmful species and their adaptations to urban areas.
Shandis Fancher, University of Texas at Tyler
1:15-3:15 in LSR 4
Determining the Mechanism of Pseudomonas Elastase Induced Activation of EGFR
Shandis Fancher, Dustin DeLong, Dustin Patterson, and Ali Azghani
Pseudomonas aeruginosa is a Gram-negative bacterium which functions in an opportunistic manner in immunocompromised patients. Several toxins and proteases play a significant role in the pathogenesis of P. aeruginosa acute infections as well as in Cystic Fibrosis associated chronic lung inflammation. We have reported that elastase isolated from P. aeruginosa (PE) is an activator of inflammatory pathways in vitro. This study examines the mechanisms of PE-induced inflammatory responses of lung epithelial cells in an effort to determine a more effective treatment modality. Our working hypothesis for this project is that PE activates epithelial growth factor receptors (EGFR) indirectly through activation of a secondary soluble ligand. Indirect interactions between PE and EGFR expressed on human pulmonary epithelial cell line A549 were analyzed using Virus Like Particle (VLP) encapsulated elastase. Cells were maintained in DMEM/F-12 with 10% FBS and 1% antibiotic-antimycotic solution. For experiments, cells were grown to confluence in 6-well plates, semi-starved in 2% FBS overnight, and starved in HBSS for 2 hours before treatment. Target treatments included commercial PE, recombinant PE (rPE), and rPE encapsulated in Virus Like Particles (VLP/rPE) constructed in the lab. Cells were treated with HBSS and empty VLP as negative controls and EGF as a positive control. Cells were lysed post-treatment for protein extraction and concentration determination, and SDS-PAGE analysis. Western blots were performed using primary antibodies targeting phosphorylated EGFR. We found that EGF, free PE, and rPE activate EGFR while encapsulated PE failed to do so. Our preliminary data indicates that PE activates EGFR by direct ligand/receptor interactions, the nature of which will be explored in successive studies.
Yesica Garcia, Southwestern Oklahoma State University
1:15-3:15 in LSR 3
Isolating Rhizobacteria for Promoting Growth of Winter Wheat
Yesica Garcia and Regina McGrane
Increasing crop yields is of critical importance for feeding an ever-growing population. Plant growth promoting rhizobacteria (PGPR) have been found to increase crop yield in various ways including inducing systemic resistance, regulating plant-growth promoting hormones, and increasing nutrient intake by assimilating nutrients into useable forms. PGPR are also appealing because of the smaller amounts needed to induce benefits compared to synthetic fertilizers. Winter wheat is a major crop in Oklahoma; more than five million acres were planted across the state in 2011. It also serves as pasture for the production of millions of pounds of beef. Currently, approximately two pounds of nitrogen are used to produce one bushel of profitable winter wheat yields. In an attempt to positively impact wheat yields, we sought to isolate PGPR from soil in the Weatherford, Oklahoma area. The objective of the experiment was to identify PGPR that most evidently increased winter wheat yield. Soil samples were collected, and bacteria were isolated via serial dilution plating. The isolated strains were then introduced onto the winter wheat seeds, plants were grown for two weeks, and yield was quantified by measuring total fresh weight, stem and shoot length, and stem and shoot weight. Plants with potential PGPR were compared to control plants that were not inoculated. In this work, eighteen strains were tested; none significantly increased the measured indicators of yield. However, we observed that some plants exhibited greener pigmentation and greater leaf number, suggesting the rhizobacteria may induce alternative health benefits. In future experiments, these phenotypes will also be quantified, and rhizobacteria will be isolated from a broader variety of soil types. Collectively, this work aims to identify rhizobacteria for use in agriculture with the purpose of benefiting crop yields.
Selena Gomez, Oral Roberts University
10:00-10:15 in LSR
Canine diets in the digital age: an examination of dog owner’s feeding practices and external signs of health in dogs
Selena Gomez
Dogs have been known as man’s best friend for years, they serve practical roles in society such as police dogs and service animals but more commonly serve as family members. As dogs transitioned from pets to family members, the feeding practices of owners changed from table scraps to commercial dog food. The popularity of commercial dog food increased because dog nutrition was considered a science best left for the experts, marketing strategies used by dog food companies, and convenient packaging and storage of dog food. Today most dogs eat commercial dog foods which contain by-products and have been recalled due to chemical contamination and toxic nutrient amounts which have led to debate about their nutritional quality and increased the prevalence of alternative diets. A survey was administered to dog owners in a convenience sample to explore the dog’s role in the household, owners’ feeding practices, factors influencing owners’ choice in dog food, and external signs of health (coat, skin, teeth, energy, stool, and body condition) in dogs. This study found that the majority of owners considered their dog family. Most owners fed commercial dog food and many fed diets had aspects of alternative dog foods including real meat as the first ingredient, no by-products, no grains, and all natural ingredients. There was no significant difference (P > 0.05) between dog food manufacturers and their effects on the external signs of health. Also, there was no significant difference (P > 0.05) between the dog owners’ blind estimation of body condition score and open body condition score estimation.
Constance Green, East Central University
1:15-3:15 in TXH 1
Finding Novel Antibiotic Producing Bacteria
Constance Green and April Nesbit
The demand for new antibiotics is extremely high due to an increase in antibiotic-resistant bacteria. This is causing common infections to become untreatable. Antibiotics are naturally occurring chemicals secreted from bacteria or fungi that kill other microbes. Thus, antibiotic producing bacteria are typically found in locations with high levels of competition with other microbes, such as the soil. Our goal is to use soil samples to find bacteria that produce previously unknown antibiotics in the hopes of postponing a post-antibiotic era. After testing sixteen sites for potential antibiotic-producing bacteria, we have found four isolates, which show inhibition of Enterococcus faecium with some also inhibiting Escherichia coli. Identification of these isolates using staining, biochemical testing, and 16S sequencing indicates that they are all Gram positive, endospore-forming bacilli in the Genus Bacillus. Future studies are to determine the species and isolate the secreted chemical causing inhibition of E. faecium and E. coli.
Virginia Green, University of Dallas
1:15-3:15 in LSR 4
Mortality during Pseudomonas aeruginosa chronic infection of Drosophila melanogaster dependent on mating experience of host
Virginia Green, Eli Wallace, Deanna M. Soper, and William L. Cody
Evolutionary trade-offs are extremely significant in understanding why certain phenotypes evolve and persist in different species. Such a trade-off may exist between reproduction and bacterial resistance in Drosophila melanogaster. Recent studies suggest that reproduction may negatively affect immunity, which in turn influences the coevolution of virulence in pathogens such as Pseudomonas aeruginosa. Drosophila melanogaster males are known to produce seminal proteins that reduce female life span, and studies suggest that these proteins may reduce females’ ability to resist acute infection. Characterization of this trade- off will further illuminate the immunological cost of reproduction in fruit flies, leading to a more complete understanding of the coevolutionary dynamics of bacterial virulence and host resistance. While decreased immune function following mating has been noted previously, acute models of infection using thoracic puncture have traditionally been used rather than the chronic oral route. Given the influence of route of infection on host immune response, a thorough analysis of the effects of sexual experience must examine whether induced immunodeficiency is also observed against chronic infection. In this study, we orally infected groups of virgin and experienced flies with P. aeruginosa and compared mortality rates to determine whether either group displayed increased resistance to infection. Experienced female flies showed increased mortality after chronic infection compared to virgin females. These observations suggest that the act of mating reduces female flies’ ability to resist oral bacterial infection, possibly as a result of exposure to seminal proteins. These findings may shed light on the coevolutionary past of host-pathogen pairs and suggest that coevolution infection models may need to be expanded to account for such factors as host experience.
Joselyn Gutierrez, West Texas A&M University
10:00-10:15 in TXH
The Effects of Summer and Winter Burning on Herpetofauna in a Sand Sage Prairie Ecosystem
Joselyn M. Gutierrez, Richard T. Kazmaier, and Matthew W. Poole
Fire is a natural component in prairie ecosystems, and controlled burning is one of the most common tools used to rehabilitate prairie habitat. However, most research addressing impacts of prescribed fire have focused on short-term responses, and most studies that have addressed the impact of fire on herpetofauna have focused on burning that is conducted during the dormant season. From 2004– 2009, we explored short-term responses to seasonality of fire on small vertebrates in a sandsage prairie ecosystem. In 2018, we reinitiated research in the same study plots to begin evaluating longer-term effects in the system. Drift fence arrays with pitfall traps were used to sample herpetofauna in a randomized complete block design with 5 blocks and 3 treatments (summer-burned, winter-burned, and unburned) during early and late season sampling periods. We regressed year of sampling against abundances and community metrics to evaluate linear responses to the different burning regimes over this time period. In the early season, green toads tended to increase over time with winter-burning (P = 0.095), but six-lined racerunners, plains narrowmouths, and most community metrics tended to increase over time with summer-burning (P < 0.092). Both six-lined racerunners and plains blind snakes increased over time in both burning treatments (P < 0.004), while both species diversity and species richness trended to slightly increase over time with winter-burning (P < 0.101) in the late season. Trends detected were generally weak, perhaps because of a long-term severe drought that has persisted in the region and could be masking treatment effects. Continued monitoring of these populations should elucidate treatment effects. However, our results suggest that summer-burning may be more beneficial to herpetofaunal communities than other treatments.
Brendan Harrison, Southwestern Oklahoma State University
1:15-3:15 in LSR 4
How sex ratios & population density affect sexual conflict over pairing duration in Hyalella amphipods
Brendan Harrison and Rickey Cothran
Research on sexual conflict, i.e. different evolutionary interests between the sexes, has exploded over the past few decades, but we know relatively little about how demographics affect sexual conflict in nature. Sex ratio is a demographic parameter that can affect sexual conflict through male-female encounter rates, which in-turn affects harassment experienced by females. Population density can have similar effects on female-male encounter rates. Under such scenarios the energy necessary to resist male harassment may alter the outcome of sexual conflict over mating. We tested the effects of sex ratio and density on pairing duration in a freshwater amphipod in the genus Hyalella. Males prefer longer pairing durations than females because pairing allows males to monopolize access to receptive females. While pairing for long periods is costly for both sexes (e.g., decreased foraging efficiency and increased predation risk), the net cost is higher for females because they do not reap the benefits of increased mating success. We hypothesized that females would experience longer pairing durations in male-biased and denser populations. To test these hypotheses, we set up populations that varied in sex ratio (female-biased, male-biased, or equal sex ratios) and population density (high or low). We discovered that guarding duration increased significantly from female-biased treatments to equal sex ratio treatments and again from equal sex ratio treatments to male-biased treatments, while no significant difference was found between low and high density treatments. These results indicate that density has no effect on guarding duration whereas sex ratio does. In nature Hyalella populations are usually female-biased because males are more likely to be eaten by predators. Our results suggest that changes in predator populations could upset the natural female-bias of these populations and increase the intensity of conflict, which would have cascading effects on population health by lowering female fitness.
Emily Hernandez and Elizabeth Gwartney, Oklahoma City University
1:15-3:15 in TXH 1
The Enemy of my Enemy is my Friend: Mycobacteriophage and the Treatment of Drug-Resistant Tuberculosis
Emily Hernandez, Loren Matrone, Elizabeth Gwartney, and Greg Mullen
Drug Resistant Tuberculosis (TB) is a worldwide health crisis being faced today. According to the World Health Organization, roughly 240,000 people died from drug resistant TB in 2016 alone. In response to this global emergency, new methods of treatment are being developed and tested in the lab setting, one of which being by Mycobacteriophage. Mycobacteriophage are viruses that infect mycobacteria and spread through internal reproduction. This form of therapy will include upwards of four or five different phages to avoid issues with resistance, which calls for identification of unique types of phage that enter the bacteria through different pathways. The goal of our study is to characterize OCU1 and OCU2 phage by cloning portions of their genomes for sequencing and by also identifying possible mechanisms of resistance by isolating phage-resistant mycobacteria. We have high hopes that the research being done within the OCU Biology department will leave a mark on the journey to identifying a solution to drug resistant TB.
Fiorella Hernandez, St. Edward's University
9:15-9:30 in DBU
Examining unc-33 expression using GFP transcriptional reporter
Fiorella Hernandez, Hailey Trombley, and Andrea Holgado
An emerging avenue of research aimed towards the development of therapies for Alzheimer’s Disease (AD) includes the investigation of a Microtubule Associated Protein (MAP) called CRMP2. Recent studies demonstrate that animal models for Alzheimer’s disease have increased levels of inactivated CRMP2. Besides its role in Alzheimer’s Disease, CRMP2 and its C. elegans orthologue UNC-33 were shown to control axonal elongation and microtubule assembly. Additionally, current research suggests that CRMP2/UNC-33 functionality can be regulated by autophagy, the engulfing of cytoplasmic elements into the autophagolysosome for degradation. To study the latter, we began the characterization of a transgenic C. elegans strain that expresses GFP under the regulation of an unc-33 promoter. The first steps of this characterization consisted in the sequencing of the transgene integrated in these nematodes. Secondly, we determined the expression pattern of GFP in intact nematodes throughout their life cycle. Lastly, we investigated colocalization of the GFP reporter with known cellular markers. Findings from these studies include: the sequencing of a 2,700 base pair promoter that maps to the 5’ UTR of the unc-33 gene; a neuronal localization of the GFP reporter at all stages of the C. elegans life cycle; and a distinctive cellular expression pattern in nematodes with enhanced autophagy. These findings corroborate previous research and provide further evidence for the autophagy-based regulation of the expression profile of CRMP2/unc-33.
Fernanda Hernandez Sanchez, University of the Ozarks
10:45-11:00 in DBU
Novel Approaches for Investigating Cancer Growth Inhibition and Male Contraception by DEC-TEC and PROTAC
Fernanda Hernández-Sánchez, Zhuang Jin, and Martin M. Matzuk
The bromodomain protein 4 (BRD4) and bromodomain testis-specific protein (BRDT) are attractive therapeutic targets for cancer and male contraception, respectively. In this work, a series of dual inhibitors for BRD4 and BRDT were designed and synthesized based on DNA-Encoded Chemistry Technology (DEC-TEC) and proteolysis targeting chimera (PROTAC). In vitro activities of the inhibitors on the bromodomain 1 (BD1) and the bromodomain 2 (BD2) of the two protein targets (BRD4 and BRDT) were determined by the alphascreen assay. Cell growth inhibitory activities of the inhibitors on cell MV4;11 were also evaluated.
Minh-Nguyet Hoang, University of Dallas
10:30-10:45 in DBU
Novel Glial Cells Affect Sensitivity to Nociception in Third Instar Larvae of Drosophila melanogaster
Minh-Nguyet Hoang, Justin Patawaran, Julia Krause, Catherine Hand, Deanna M. Soper, and Drew Stenesen
Pain is a sensory stimulus with a variety of therapeutic applications. Several genes involved in nociception within Drosophila melanogaster have been identified. Beta-alanine transporter (BalaT) is known to function in glial cells supporting the adult fly visual system; however, its role in larvae has remains unexplored. Here we identify novel BalaT expressing cells at tracheal branches points of wandering third instar larvae. Sensory neurons extend processes along tracheal branches and affect the organism’s response to pain. To examine the role of these new bilateral tracheal (BLT) cells in nociception, we performed temperature-response assays with BalaT loss-of-function mutants. Our data demonstrates an increased sensitivity to pain in BalaT mutants relative to wild type controls. From our current findings, we propose that BLT cells are glial and function in support of sensory neurons transmitting information related to painful stimuli.
Joshua Holsing, Oral Roberts University
11:30-11:45 in DBU
Antimicrobial Effects of Electrically Modified Lyophilic Silver Hydrosol in vitro and in vivo
Joshua Holsing
Silver has been known for many medicinal uses in the forms of colloidal silver and silver nanoparticles, both of which pose possible health risks when improperly applied. Silver in the form of an electrically modified hydrosol bypasses the adverse effects of prior forms, yet still maintains potent antimicrobial properties. This study focuses on the efficacy of this electrically modified silver hydrosol against A. faecalis, E. coli, MRSA, P. aeruginosa and S. epidermidis by utilizing a modified Kirby-Bauer disk diffusion method to measure the zone of inhibition (ZOI) against the concentration of the hydrosol; and against S. aureus, S. epidermidis, and P. aeruginosa by measuring dosage contingent percent decrease on live epithelial tissue to determine if efficacy is the result of concentration or dosage based treatment. The concentration dependent test results concluded statistically significant ZOI (all p < 0.01), yet all but S. epidermidis treatments failed post hoc test significance. The dosage dependent test concluded a near total eradication of cultured bacteria up to a 100% decrease in bacterial growth (p < 0.0463) with no change between treated and untreated epithelial cells. These results indicate safe usage of the silver hydrosol in vivo and support the hypothesis of high efficacy dose dependent treatment on multiple nosocomial gram negative and gram positive bacterial strains. Possible future research could fixate on the silver hydrosol’s interaction within a living system with exhaustive testing on the hepatocytes for presence of silver.
Aubrey Howard, West Texas A&M University
1:15-3:15 in TXH 1
Microscopic Characterization of the Aeroallergen and Reduction in Reduction Index on Using Air Purifiers With AHPCO and Plasma Nanotechnology
Aubrey Howard, Nabarun Ghosh, Jon Bennert, and Jeff Bennert
Allergies are a hypersensitivity of the immune system to normally benign substances. The Panhandle of Texas sees diagnosis rates typically double that of the state average. Seasonal allergies can have a strong impact on the quality of everyday life due to the presence of aeroallergens in the air such as pollen. In order to aid diagnosis of seasonal allergies and characterize the aeroallergens in the Texas Panhandle, research must be conducted to establish a foundation for determining the trends of data and provide a reference for identification of allergenic weed species. Similar studies have been conducted, however they have not included native angiosperms specific to the Texas Panhandle, making it more difficult for individuals to identify plant origins of pollen. In order to address this concern, a set of observations must be made analyzing the morphological characterization of pollen structure to aid in diagnosis and establish a reference for researchers for identification of pollen grains to the plant they have originated. We have recorded the GPS location, collected the whole plant where possible for making herbaria and the flowers for the pollen analysis. Flowering plants have been collected and their pollen analyzed and characterized. Micrographs were taken using an Olympus DP74 digital camera attached to a Olympus BX40 microscope and analyzed using cellSense digital software. The most significant aeroallergens recorded were the pollens like grass pollen (Poaceae), Short Ragweed (Ambrosia artemisiifolia), Common Sunflower (Helianthus annuus), Hairy Sunflower (Helianthus hirsutus), Silverleaf Nightshade (Solanum elaeagnifolium) and Lamb’s Quarters (Chenopodium album) as well as fungal spores like Alternaria, ascospores from Pezizales, Drechslera, Stachybotrys, Cladosporium and Curvularia. Further analyses of the gathered data are needed for prediction on the distribution of pollen and pollen seasons. We aim to incorporate the AHPCO and Plasma Nanotechnology for developing an advanced air purification system to reduce the indoor particulate matters including all forms of aeroallergen. This data collecting stage of research will help establish a foundation for future research and is the first crucial step for the hypothetico-deductive approach.
Nancy Huynh, Texas Wesleyan University
1:15-3:15 in LSR 4
The effect of active ingredients in mouthwash on Streptococcus sobrinus biofilm formation
Nancy Huynh and Christopher Parker
A collection of microorganisms in the human body can either be healthy or cause diseases depending on the bacterial organism. Microbes that adhere to surfaces in the mouth that form biofilms can negatively affect overall health. The microbes, Streptococcus mutans and Streptococcus sobrinus, may be associated with cavities and lead to “rampant” tooth decay. We tested the effect of ethanol in mouthwash on the formation of S. sobrinus biofilm. This research can provide insight into whether alcohol in mouthwash is effective on inhibiting S. sobrinus biofilm formation. In this study, S. sobrinus biofilm formation capabilities were investigated under varying levels of ethanol concentration in Listerine® Zero mouthwash. The results of the One-Way Analysis of Variance (ANOVA) statistical test showed that there was no difference between the varying levels of ethanol concentrations. Thus, ethanol levels in mouthwash may not inhibit S. sobrinus biofilm formation, and additional research is required to test the active ingredients from essential oils and antibacterial activities of mouthwashes.
Mary Kadatska, Oral Roberts University
9:15-9:30 in TXH
Effects of trinitrobenzene induced colitis on vasoactive intestinal peptide, calcitonin gene-related peptide and morphology of colon in rats
Mary Kadatska
The primary pain medications used today are opioid based. However, they have caused a decrease in life expectancy in years as recent as 2015-2017 leading to the opioid crisis because they are widely known to cause development of tolerance and to promote substance abuse disorders with opioids as well as other drugs, (Koenig 2018, Dai et al. 2018). These circumstances call for a search for a replacement for opioid medications. In this study, we looked primarily at vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP), and at basic morphology of the epithelial lining of the intestinal mucosa. VIP is known to have certain motile, secretory, and anti-inflammatory functions in the colon (Del Valle-Pinero et al. 2015, Abad et al. 2005). CGRP is known to have vasodilatory, nociceptive, secretory, and motor functions which can be linked to pain and inflammation (Cottrell et al. 2012). The experimental rat was treated with 2,4,6-trinitrobenzene sulfonic acid (TNBS), and its CGRP/VIP colon levels were measured by measuring mean grey intensity (MGI) of antibody stains particular to CGRP and VIP and compared with naïve and control (ethanol treated) colons. Our results showed a significant increase in VIP MGI per nerve fiber (p0.05).
Jack Koszykowski, Oral Roberts University
10:30-10:45 in LSR
The Effects of Azadirachta indica (neem) Extract on RP62A Staphylococcus epidermidis Biofilm Formation
Jack Koszykowski
Biofilms are dense microbial communities of bacteria. In biofilms, bacteria possess a higher defense than their planktonic forms. Within these close-knit communities, bacteria transformation is abundant due to a communication process referred to as quorum sensing. Because of this, bacteria within biofilms often possess antibiotic resistance traits. Due to the increased pathogenicity of the bacteria, biofilms form a severe threat to humans with indwelling medical devices on which biofilms can form. Thus, the search for novel compounds that can inhibit or deter biofilm growth has been a frequented area of study. In this study, varying concentrations of leaf extracts from Azadirachta indica, or neem, were tested on RP62A, a biofilm forming strain of Staphylococcus epidermidis. Neem was chosen due to its wide variety of medicinal properties. The goal was to inhibit biofilm formation. The Crystal Violet Assay used to determine biofilm levels showed that as the concentrations of neem increased the absorption reading increased significantly. The dead/alive assay used to determine bacterial viability showed that as the concentration of neem increased, bacteria viability decreased, with the exception of neem that wasn’t dissolved in DMSO. The results were largely inconclusive. While the neem showed to possess antibacterial activity, the assay used to determine biofilm levels did not yield any definitive data due to the characters of the neem within the wells of the 96-well microtiter plate. The assay to determine bacteria viability showed that 1/2 concentration of neem had the highest effectiveness in preventing both biofilm and planktonic growth.
BreAnna Le, Southwestern Oklahoma State University
1:15-3:15 in TXH 1
Investigating effects of innate immune agonists on co-receptor B7-H4 in B lymphocytes
BreAnna Le and Christopher G. Horton
The immune system depends on communication from innate immune cells, which act quickly upon pathogenic microbes through stimulation of their pattern recognition receptors, to adaptive immune cells, which respond accordingly to signals such as cytokines and cell surface proteins (co-receptors). Co-receptors can help activate or inhibit the functions of the adaptive immune response and can also lead to specific adaptive immune cell differentiation. B7-H4 is predominantly viewed as an inhibitory co-receptor expressed in non-lymphoid tissues, especially among tumors where it functions to impair anti-tumor immunity. We have observed a mild, agonist-specific increase in B7-H4 expression in mouse primary B cells in previous studies. From these results, we hypothesized that exposing B lymphocytes to different degrees of pathogen-like stimuli will alter the expression of B7-H4. In our experiments, while maintaining actively growing cultures of an A20 cell line, we stimulated B lymphocytes with varying concentrations of imiquimod, LPS, DMXAA, and polyIC for 6-72 hours, and observed their expression of B7-H4 by flow cytometry. Our results demonstrated that stimulation with these agonists, particularly imiquimod, LPS, and DMXAA after a 72 hour incubation, lead to significant alterations in B7-H4’s expression, while cells stimulated with polyIC showed no significant expression when compared to unstimulated cells. Although all three concentrations used for imiquimod, LPS, and DMXAA on average showed significance, the most dramatic effects were observed using imiquimod. Future studies such as using qRT-PCR to insure the presence of the gene will be needed for more in-depth characterization, but our results thus far begin to shed light on the possible mechanism of how B7-H4 expression is initiated. Most importantly, B7-H4 studies such as these can lead to possible cancer/tumor treatments and vaccinations, in which inhibiting B7-H4’s expression will hinder its interference with anti-tumor immunity.
Sidney Lyons, East Central University
1:15-3:15 in TXH 2
The effect of Iron Oxide Nano particles on development, and fluctuating asymmetry of Zophobas morio larvae
Sidney Lyons, Randall Maples, and George Wang
Nanoparticles are used in a variety of ways to introduce large materials on the molecular level. We examined the potential effect of nano particles on darkling beetle (Zophobas morio) larvae development. We hypothesized that the iron oxide nano particles would stunt the growth of the beetle larvae, and cause fluctuating asymmetry in the antenna and limbs of adults. We believe this because fluctuating asymmetry is an indicator of stress on an organism. We reared a total of 40 Z. morio larvae divided into two groups. We fed the larvae in the treatment group carrots coated with a small amount of iron oxide nano particles, while larvae in the control group were fed organic carrots. The larvae were allowed to pupate and enclose in the lab. We recorded the mass and length of the pupae, and measured the antenna length, and leg length of adults.
Monica A. Martinez, Austin College
10:00-10:15 in DBU
Estradiol Effects on Hibernating Physiology of Ground Squirrels
Monica A. Vania Martinez and Jessica Healy
Estradiol is a sex hormone with anorexigenic effects. Previous literature reports that exogenous estradiol reduces food intake and body fat in non-hibernating rodents, yet few studies confirm whether this pattern is applicable to the Thirteen-Lined Ground Squirrel (Ictidomys tridecemlineatus). In 2018, 5 individuals were implanted with capsules that contained either vehicle (control) or exogenous estradiol (Est+) and food intake, body composition, and metabolic rate were measured during the summer active period. Preliminary analysis suggests that Est+ animals may have increased metabolic rate compared with controls during the implant period, but it is unknown if estradiol implantation has long term effects on torpid metabolic rate. Individuals were aroused from an initial torpor at 18, 15, and 5°C and their oxygen consumption was recorded as a proxy for metabolic rate. Observing the effects of exogenous estradiol on torpid metabolic rate will give more insight into the physiological mechanisms underlying energy balance and allocation within Ictidomys tridecemlineatus.
Susan McGuire, Texas A&M University - Corpus Christi
1:15-3:15 in TXH 2
Investigating Nutrient and Organic Carbon Distribution in Wetland Ecosystems
Susan McGuire, Rachel Weisend, and Brandi Kiel Reese
Wetlands provide many ecosystem services for both animals and humans. Wetland communities in the Gulf Coast are changing as mangroves are expanding throughout the region and into previous northern limits. This project will investigate the carbon and nutrient distribution within the sediment of the three vegetation types (mangrove-dominated, Spartina-dominated, and seagrass-dominated) that can be found within coastal wetlands. We measured nitrate, nitrite, ammonium and carbon in order to understand the changes in geochemical signals and sediment properties with respect to depth, seasonality, and vegetation in a wetland system. This will help understand ecosystem shifts across a spatial and temporal scale. We found that there were no significant differences between different seasons within the same vegetation type. However, mangrove-dominated sediment was significantly different from Spartina-dominated sediment (p value < 0.05) and seagrass-dominated sediment (p value < 0.05), but when comparing Spartina-dominated sediment to seagrass-dominated sediment there was no significant difference. Also, all November samples were significantly different than all August samples (p value < 0.05), but there was no significant difference when comparing November samples to June sample, or June to August. This information will help us asses changes in macrofauna, meiofauna, and the microbial community within a wetland ecosystem.
Shayla Miller, Southwestern Oklahoma State University
1:15-3:15 in TXH 2
The Battle of the Sexes: How female condition affects conflict over mate guarding duration in Hyalella amphipods
Shayla Miller and Rickey Cothran
Sexual conflict, different evolutionary interests between females and males, is common in nature and is expected to shape the evolution of mating traits and affect the productivity of populations. Most research has focused on male “offense” traits with relatively less research on female “defense” traits. We explored how female condition affects conflict over mate guarding duration in a freshwater amphipod species in the genus Hyalella. We hypothesized that females in poor condition would experience longer mate guarding periods because they are unable to resist male pairing attempts. We tested this hypothesis by varying the amount of spirulina, a high nutrient protein powder, in female’s diets. Females received either a high-quality diet or a low quality-diet. Females from each diet treatment were then exposed to either one or three males representing a low and high harassment treatment, respectively. We found a significant effect of harassment treatment but no significant effect of female diet on pairing duration. It is possible that our diet treatment did not significantly affect female condition or other female traits, e.g., morphological traits are important in resolving conflict over pairing duration. Our results show that harassment levels do have an effect on pairing duration. Longer pairing durations could affect female amphipod fecundity by reducing foraging efficiency, which could have a negative impact on the overall health of an amphipod population.
Taylor Miller, Oral Roberts University
11:45-12:00 in DBU
Anti-cancer effects of compounds secreted by Flammulina velutipes
Taylor Miller
Compounds collected from various mushroom species, including Flammulina velutipes, have been shown to have immunomodulatory and antitumor effects (Seklic et al. 2016). This experiment was performed to investigate the cytotoxic activity of F. velutipes secretions on MCF10A breast epithelial cells and MDA MB 468 breast adenocarcinoma cells. It was hypothesized that there would be a significant difference in cell viability between the two cell lines when treated with the mushroom secretions. The first preliminary cytotoxicity assay revealed a decrease in cell viability in both the MCF10A cells and in the MDA MB 468 cells. A student’s t-test also revealed a significant difference in cell viability between the two populations, the tumorigenic cells having the higher average percent viability. The next cytotoxicity assay measured the average percent viability of the cell lines at varying concentrations. A significant difference was found at 1X and 0.5X concentrations, the highest difference being at the 0.5X concentration. The third and final cytotoxicity assay tested only the 0.5X concentration of compound on the cell lines. This showed that the non-tumorigenic MCF10A cells had a significantly higher average cell viability percentage than the tumorigenic MDA MB 468 cells. The results support the hypothesis that F. velutipes secretions exhibit cytotoxic activity in cancer cells. There is evidence to support the idea of selective cytotoxicity between normal epithelial tissue and cancerous tissue.
Julio Molina-Pineda, University of the Ozarks
1:15-3:15 in TXH 1
Antioxidant Carbon Nanoparticle Results in Novel Auditory Response in Mice
Julio A. Molina-Pineda and Fred A. Pereira
Deafness is a common side effect in people undergoing chemotherapeutic treatment with Cisplatin, a chemical proven to be ototoxic due to its oxidative damage that results in disturbance of the endocochlear potential. An antioxidant carbon nanoparticle has been found to protect the ear from such damage and to enhance auditory response as well. Two experiments were performed: one in which the nanoparticle dosage and its effects were investigated by studying the effect of three continuous doses. And a second experiment in which the inner ear and brain’s potassium channels mRNA expression was compared between nanoparticle treated and untreated mice. Preliminary findings suggest that the nanoparticle increases and delays auditory response, and that higher dosage helps in maintaining the enhancement rather than increasing it. Furthermore, the mRNA analysis suggests a possible mechanism for the enhancement caused by the nanoparticle.
Julio Molina-Pineda, University of the Ozarks
11:15-11:30 in LSR
Maternal Commensal Bacteria Inhibits Intrauterine ZIKV Infection and Fetal Growth Restriction in a Mouse Model
Julio Molina-Pineda, Maxim Seferovic, Gregory Valentine, and Kjersti Aagaard
ZIKV has been associated with congenital infection, microcephaly, and pregnancy loss due to viral mutations that facilitate maternal to fetal viral spread. Commensal bacterial microbes have been demonstrated to modulate host immunology and viral infection. We therefore hypothesized that commensal bacteria would influence maternal to fetal transmission of ZIKA virus. Employing a mouse model of Zika infection, we compared ZIKV transmission rate and pup infectivity between normal conventional (C) and germ free (GF) gnotobiotic mice. Pregnant Swiss-Webster mice were given four doses of 1x104 PFU of a contemporaneous ZIKA strain (ZIKV) or a mock injection (M) on embryonic days 4 through 7. Conventionalized (GF exposed to bacteria at weaning) or immune inhibiting anti-IFNI1 antibody were negative and positive controls, respectively. Tissues were recovered at e18.5 just prior to delivery and TRIzol extraction performed for genomic viral RNA. TaqMan based qPCR was performed from cDNA and quantified against ZIKV standard curves. ZIKA infected placentas were positive in 82% of demised fetus compared to 52% of live pups (p=0.04). No virus was found in either maternal or fetal brains, however, virus was recovered from maternal spleen and uterus, and was highest by an order of magnitude in fetal placental tissue overall.
Nicole Mountz, Oral Roberts University
9:30-9:45 in TXH
Cytotoxic Effects of Inonotus obliquus Secretions on Non-Tumor and Tumor Cells
Nicole Mountz
The recent development of targeted cancer therapies allows for treatment that is selective of molecular expressions unique to cancer cells. Extractions of Inonotus obliquus have shown antineoplastic properties that could provide naturally-derived targeted therapies with reduced harm to normal cells. This study aimed to investigate the cytotoxic activity of I. obliquus secretions against the proliferation of human breast carcinoma cells and compared to normal human mammary epithelial cells. I. obliquus mycelia were grown and passaged on a potato/dextrose/yeast agar supplemented with woodchips (20 ºC). Secretions were harvested from mycelia with Opti-MEM cell medium (48 hours; 37 ºC; 0% CO2), purified and stored at 4 °C. Purified secretions were tested on MDA-MB-468 cells and MCF 10A cells with an MTT cytotoxicity assay (24 hours; 5% CO2; 37 ºC). Absorbance values were measured at a ratio of OD570 to OD605 and used to calculate cell viability percentage. MTT cytotoxicity assay results showed a significantly greater cell viability of normal mammary epithelial cells than of breast carcinoma cells after 24 hours of exposure (22.1% vs. 2.26% respectively; P < 0.01). Results indicate the presence of cancer-specific cytotoxic activity within I. obliquus secretions. Furthermore, these results suggest the potential use of I. obliquus secretions as a cancer-targeted treatment.
McKayla Muse, University of Central Oklahoma
11:30-11:45 in LSR
Analysis of Proliferation and Migration in Phenylalanine, Retinoic Acid, and 4-diethylaminobenzaldehyde Treated Cells
McKayla Muse, Kayley Pate, Gabriel Rucci, and Nikki J Seagraves
Maternal phenylketonuria [MPKU] is a syndrome of multiple congenital anomalies including cardiovascular malformations [CVMs], and brain and growth restriction when a mother with Phenylketonuria [PKU] does not control her dietary intake of Phenylalanine [Phe]. However, the mechanisms responsible for Phe-induced CVMs are poorly understood. Our lab has preliminary evidence that high levels of Phe could inhibit Retinoic Acid [RA] signaling, which typically promotes the expression of genes such as proliferation, migration, and differentiation. Proliferation and migration of the neural crest cells are important in formation of the outflow tract (OFT) and aortic arch arteries (AAA). We hypothesize that Phe inhibits migration and proliferation, which may contribute to the defects seen in MPKU. We also looked at the effects of exposure to RA and 4-diethylaminobenzaldehyde [DEAB], a known RA inhibitor. We conducted in-vitro proliferation and migration assays on several cell types to determine if proliferation and migration was affected by Phe, RA, and DEAB exposure. Images were analyzed with ImageJ and GraphPad Prism. Present research suggests that Phe exposure causes a significant decrease in proliferation of cells. It is also shown that RA increases or does not affect proliferation, and that DEAB decreases cell proliferation. In this way, Phe is similar to DEAB, which suggests that it also acts as an RA inhibitor. This could contribute to the CVMs observed in MPKU. The results of migration studies showed that Phe acted similar to DEAB, where the rate of migration was slower than that of RA, which is known to increase migration. This suggests that Phe may act as an inhibitor of migration. This is significant because it eludes to a potential mechanism that Phe could affect RA signaling, thus resulting in the types of defects observed in human MPKU.
Lois Nguapa, Oral Roberts University
10:45-11:00 in LSR
Bioinformatic Prediction of Gene Function in Streptobacillus moniliformis
Lois Nguapa
Since the completion of the Human Genome Project, DNA sequencing technologies have made vast advancements in the sequencing of organismal genomes. Determining function, on the other hand, still proves challenging, as this requires the time-consuming investigation of a single gene and its protein product in vitro and in vivo. In the last couple of decades, the amount of genomic and proteomic data, filled with thousands of hypothetical genes, has increased to a large degree. This information, deposited in publicly available databases, can be used to predict gene function from a DNA sequence alone using online bioinformatics programs developed by the United States government and researchers around the globe. The purpose of this research is to predict the functions of hypothetical bacterial protein-coding genes that have been identified in the organism Streptobacillus moniliformis. Five hypothetical genes were analyzed from the Streptobacillus moniliformis DSM12112 genome: SMON_RS07405, SMON_RS07640, SMON_RS07645, SMON_RS07655, and SMON_RS07660. Various bioinformatic methods were applied to both the raw nucleotide sequence and amino acid sequences of these genes and compared against those of other well-characterized genes from other organisms. SMON_RS07405 was predicted to a methyltransferase, SMON_RS07640 and SMON_RS07645 were predicted to be ABC permeases, SMON_RS07655 was predicted to be an alpha/beta hydrolase, and SMON_RS07660 was predicted to be a tetratricopeptide (TPR) repeat. These findings cannot be confirmed until the proteins have been expressed in model organisms and tested in the laboratory biochemically.
Toi Nguyen, Texas Wesleyan University
1:15-3:15 in TXH 1
Identification and Distribution of Wolbachia in Natural Culex quinquefasciatus Populations in Tarrant County, TX.
Toi Nguyen
Wolbachia, the most prevalent genus of endosymbiont in invertebrates, has been utilized successfully as a biological control agent against mosquito-borne viruses. Therefore, knowing the distribution of Wolbachia in the natural mosquito populations is essential to implement the control programs. Culex quinquefasciatus is the primary vector transmitting West Nile Virus in Tarrant County, TX. In this project, polymerase chain reactions (PCRs) using general Wolbachia surface protein (wsp) primers were performed to detect Wolbachia in one hundred mosquito samples from the natural Cx. quinquefasciatus populations in six zip code areas of Tarrant County, TX. Overall, sixty-four mosquito samples were found infected with Wolbachia. Further contingency analysis using the estimated odds ratios and their 95% CI’s revealed that the natural Cx. quinquefasciatus population at one zip code (76051) was more likely to be detected with Wolbachia than at the other five zip codes. In addition, we will present the results from PCRs using group- and strain-specific wsp primers as well as phylogenetic analysis.
Sophia Ostertag, University of Dallas
1:15-3:15 in TXH 1
Coevolution of host-pathogen interactions in a Pseudomonas aeruginosa chronic infection model of Drosophila melanogaster
Sophia Ostertag, Sarah Tripplehorn, Natalie Sandlin, Ana Henriquez, Deanna Soper, and William Cody
Pseudomonas aeruginosa is an antibiotic resistant, opportunistic pathogen that causes both acute and chronic infections in a variety of hosts. P. aeruginosa is a major cause of acute human infections like UTIs, ventilator-associated pneumonia infections, and surgical site infections. Notably, chronic P. aeruginosa infections are the leading cause of morbidity and mortality in cystic fibrosis patients. The innate immune system in Drosophila melanogaster makes it a suitable host for examining these acute and chronic infections. This study will provide insight into the evolutionary response of antibiotic resistant bacterial strains, with the intent of understanding the implications of host-pathogen interactions. With repeated coexposure of these species, virulence, immunity, and reproductive traits alter in a dynamic coevolutionary relationship. The Red Queen hypothesis postulates that pathogenic species serve to increase the level of genetic diversity within a population. With pathogenic selection pressures, the host population may develop random mutations that allow for its survival under that environmental circumstance. The pathogenic population may similarly develop adaptive mutations. Three experimentally enforced evolutionary treatment groups will be used: host evolution, pathogen evolution, and coevolution. In the host evolution treatment group, successive generations of D. melanogaster are exposed to stock samples of PAO1. In the pathogen evolution treatment group, successive generations of lethal PAO1 are extracted from deceased flies and used to infect a stock population of D. melanogaster. In the coevolution treatment group, successive generations of offspring are infected with the lethal strain of PAO1, which is extracted from deceased flies of the previous generation. The variation in mortality among the host species will be characterized, with the goal to provide insight on the coevolutionary host-pathogen interaction, virulence factors, and the survivability of the host.
Amanda Pang, East Central University
1:15-3:15 in LSR 3
Growth Inhibition of Chaetomium globosum After Exposure to NSAIDs
Amanda Min Xing Pang, Leah Dudley, Hannah Taff, Tyler Rowsey, and Charles Biles.
Molds (fungi) are primary allergens in homes and businesses that cause a range of respiratory problems. Chaetomium globosum is one of the most common fungal species that causes water-damage in buildings. This research was conducted to determine the inhibitory activity of NSAIDs on C. globosum. Previous experiments indicated that low levels of NSAIDs inhibited growth of C. globosum when applied to sheetrock. Recent experiments have tested the hypothesis that NSAIDs enter the fungal cells and shut down respiratory channels of the mitochondria. This research was conducted in order to understand whether NSAID solutions must be present on the substrate before inoculation, or are able to penetrate the cell wall of existing spores and prevent germination. Subcultures of C. globosum ascospores were quantified using a hematocytometer with a light microscope, then placed in different concentrations of various NSAID solutions for 24 hours. The ascospores were washed twice by centrifugation at 15,000 rpm in sterile water. After washing, the ascospores were pipetted onto 9cm Petri media. In the initial experiment after the first centrifugation, 900 μL of the supernatant was removed and also placed on PDA media in 9cm Petri dishes. This was to determine if the ascospores were centrifuged to the bottom 100 μL of the solution. Data from the supernatant experiment showed only 1-3 ascospores were present in the extracted supernatant. Fungi undergo vacuolation under stress, which results in enlarged vacuoles in the hyphal cells. Fungi on control and treatment plates were observed under light microscopy. Acetylsalicylic acid (Aspirin) completely inhibit growth at 10 mM concentration after washing the ascospores. In contrast, Naproxen (Aleve™) displayed no inhibitory effects on C. globosum growth.
Kayle Patatanian, University of Central Oklahoma
1:15-3:15 in TXH 1
Analysis of cardiac teratogenicity of phenylalanine using the avian model: the role of the focal adhesion pathway Analysis of cardiac teratogenicity of phenylalanine using the avian model: the role of the focal adhesion pathway
Kayle Patatanian, Jamie Watson, and Nikki Seagraves
Maternal phenylketonuria (MPKU) is characterized by the teratogenic effects of phenylalanine (Phe). High levels of intrauterine Phe cause cardiovascular malformations, intellectual impairment, microcephaly, and low gestation mass. Mothers with PKU must maintain a strict diet to insure serum Phe concentrations remain below 360 µM to avoid these congenital defects. Previous functional genomic studies done by our lab have determined that the focal adhesion pathway (FAP) is significantly enriched under Phe treatment. Six genes associated with this pathway are variably expressed. Our project aims to visualize their expression during stages HH10, 12, and14 in chicken embryo heart and out flow tract tissue using in-situ hybridization. Hepatocyte growth factor is one of the six genes under investigation and has been shown to promote angiogenesis through its effects on cell survival, differentiation, and motility.
Blake Pate, Dallas Baptist University
9:30-9:45 in LSR
Concordance of HPV genotyping between histopathology and trigger cytology
C. Blake Pate
The human papillomavirus is the most sexually transmitted virus in the United States with over 14 million individuals being infected by the virus each year. The most oncogenic HPV genotypes, 16 and 18, give rise to over 70% of all cervical cancers, while other genotypes can cause genital warts and other problematic conditions. It is for this reason that research be conducted to improve the effectiveness the HPV vaccine and the current methods of genotyping in order to aid the public in the prevention of such a debilitating virus. In this study, HPV genotyping results acquired from clinical routine pap smear (cytological) results from patients enrolled in the HPV Vaccine Effectiveness Study at Yale University were compared to the biopsy (histopathological) results in order to determine the sensitivity, specificity, and accuracy of the cytological reports. The concordant analysis confirmed our hypothesis that the HPV genotyping from the trigger cytology report would be greater than 90% concordant with that obtained from the biopsy when collected within 12 months apart. These findings open the door for increasing interest in developing more accurate and dependable HPV genotyping techniques in cytological specimens to allow for greater ease in acquiring HPV test results. In addition, these results are currently allowing researchers in the HPV Vaccine Effectiveness Study to enroll patients into the study more rapidly using cytological genotyping results rather than the more time-intensive histopathological specimens. A prospective four year study that aims to confirm these findings with a larger number of specimens is now underway to establish HPV surveillance tools that are most cost and resource efficient.
Kayley Pate, University of Central Oklahoma
1:15-3:15 in TXH 2
Analysis of Proliferation and Migration in Phenylalanine, Retinoic Acid, and 4-diethylaminobenzaldehyde Treated Cells
Kayley Pate and Mckayla Muse
Maternal phenylketonuria [MPKU] is a syndrome of multiple congenital anomalies including cardiovascular malformations [CVMs], and brain and growth restriction when a mother with Phenylketonuria [PKU] does not control her dietary intake of Phenylalanine [Phe]. However, the mechanisms responsible for Phe-induced CVMs are poorly understood. Our lab has preliminary evidence that high levels of Phe could inhibit Retinoic Acid [RA] signaling, which typically promotes the expression of genes such as proliferation, migration, and differentiation. Proliferation and migration of the neural crest cells are important in formation of the outflow tract (OFT) and aortic arch arteries (AAA). We hypothesize that Phe inhibits migration and proliferation, which may contribute to the defects seen in MPKU. We also looked at the effects of exposure to RA and 4-diethylaminobenzaldehyde [DEAB], a known RA inhibitor. We conducted in-vitro proliferation and migration assays on several cell types to determine if proliferation and migration was affected by Phe, RA, and DEAB exposure. Images were analyzed with ImageJ and GraphPad Prism. Present research suggests that Phe exposure causes a significant decrease in proliferation of cells. It is also shown that RA increases or does not affect proliferation, and that DEAB decreases cell proliferation. In this way, Phe is similar to DEAB, which suggests that it also acts as an RA inhibitor. This could contribute to the CVMs observed in MPKU. The results of migration studies showed that Phe acted similar to DEAB, where the rate of migration was slower than that of RA, which is known to increase migration. This suggests that Phe may act as an inhibitor of migration. This is significant because it eludes to a potential mechanism that Phe could affect RA signaling, thus resulting in the types of defects observed in human MPKU.
Vincent Phan, University of Dallas
1:15-3:15 in TXH 2
Novel Therapeutics for Head and Neck Cancer
Vincent Q.D. Phan, Caroline R. Delma, and Mohan Natarajan
Head and Neck Cancer is one of the most potent cancers today, and treatment for it hasn’t improved over the last 2 decades. Head and Neck Squamous Cell Carcinoma (HNSCC) has a 5-year survival rate of 50%. This means that people may not even make it past 5 years after they put their cancer into remission. This is motivation for us to try to find better alternatives that can preserve life. We experimented with EC359, which is a pipeline drug that was made to treat mainly pancreatic cancer but can inhibit the group of Head and Neck Cancer. We also used the cancer cell lines HSC-3 (tongue cancer) and CAL-27 (salivary gland cancer) because they had the most metastatic potential against the body. We did several different in-vitro tests with EC359 against the cancer cells. We would grow cancer cells in T-flasks until we had enough cells to start experiments with. We did Fold Growth and MTT assays to see the cytotoxicity of EC359 against the cancer cell lines. We followed that with Fluorescence Microscopy to see whether apoptosis or necrosis was occurring within the cells. We followed this with western blotting to try to understand which apoptotic pathway EC359 took. Then we finished with a Scratch Assay to see if EC359 had any effect on the metastatic capabilities of CAL-27 and HSC-3. EC359 showed cytotoxicity in CAL-27 and HSC-3 at low Nano-molar range. EC359 showed the ability to suppress the viability of CAL-27 and HSC-3 cells and inhibited their proliferation. EC359 induced apoptosis in CAL-27 and HSC-3 cells by initiating the intrinsic apoptotic pathway. EC359 exhibited slight inhibition on the migration of CAL-27 but seemed ineffective against HSC-3 cells.
Jennifer Prophet, Southwestern Oklahoma State University
1:15-3:15 in LSR 3
Timing of Avian Migration Onset Through the Oklahoma City Area 1995-2017 Using NOAA Weather Data
Jennifer Prophet, Amy Hofeld, and Zach Jones
Avian migration timing and duration may be linked to large-scale climate patterns and reflect shifts in average global temperatures. The National Centers for Environmental Information (NCEI) provides an archive of NOAA weather radar data scans occurring every ten minutes dating back to 1995. Our primary data of interest are 1) reflectivity and 2) radial velocity. Aerial density and flight direction were collected for available years and compared to global temperatures to examine seasonal migration effects. We used KTLX (OKC) weather radar data to determine time, duration, intensity and direction of seasonal avian migrations. Fall migrations began in September and spring migrations in March between 1995-2017, while the global average temperature steadily increased by 2.358 ̊C. Fall migration trended toward later onset dates, while Spring migration trended toward earlier. Fall migration onset exhibits a stronger positive relationship with temperature than spring, potentially because inter-year March temperature patterns vary more than those of September. Ground truthing of radar data using eBird sightings revealed large-scale migratory events in Oklahoma during the second week of the fall migratory period. Comparatively, in spring migration there was a continuous migratory event occurring throughout the first 3 weeks of migration. This continuous rate over time may also contribute to the weaker relationship between migratory onset and temperature in the spring.
Rebecca Romero, West Texas A&M University
1:15-3:15 in TXH 2
Current aspects of PM 2.5 and analyzing the aeroallergen of the Texas panhandle
Rebecca Romero and Nabarun Ghosh
PM 2.5 stands for particulate matter 2.5, also called particulate pollution and is air pollutant particles not visible with the naked eye due to their microscopic size of 2.5 microns. Hazardous Aeroallergen particles of this size are dangerous because they pass the body’s defenses into the lungs and bloodstream. Hazardous PM 2.5 have the potential to lead to death. Data shows out of 6 people who die from air pollution, one person dies from PM 2.5. In this study, I’ll analyze PM 2.5 and Aeroallergens in the Texas panhandle and conduct studies on how these factors affect this area. One concern the Texas panhandle encounters is the air pollution coming from the feedlots here. Also, humans spend most of their time indoors; and using air purifiers inside can help maintain cleaner air indoors. I will use the BX Olympus Digital microscope to analyze the PM 2.5 and the aeroallergen that are prevalent in the Texas panhandle. I will follow the standard procedure to collect dust particles floating in the air as designed by AAAAI (American Academy of Allergy, Asthma, and Immunology). I will use clean glass slides on which I shall place double sticky tape and save them in a clean slide box. I will put double sided sticky tape on a clean glass slide and place the slide on the roof. I will allow the slides to be exposed to the air for 24 hours, 48 hours, and 72 hours. The prepared slides will be viewed using a BX 40 Digital Olympus Microscope. Being able to go cleaner indoors using the Advanced Hydrated Photo Catalytic Oxidation (AHPCO) Nanotechnology to purify the air and going greener outdoors, will assist the world in decreasing the steady incline of harmful air pollution and create a better environment for our health.
Ruqea Saheb, Texas Wesleyan University
1:15-3:15 in TXH 2
The Change in Oral Microflora in Response to Coffee Intake
Ruqea M. Saheb
The human microbiome presents a diverse array of microorganisms. Specifically, the human oral microflora is composed of 600 taxa at the species level. The distinct habitats of mouth influence the colonization of microorganisms due to varying environmental conditions such as pH levels, oxygen exposure, temperature, and nutrients. Therefore, the disruption of a specific colonized habitat may lead to disruption in the oral microflora. Coffee exerts antibacterial activity due to its rich composition of phenolic compounds, which has been studied to inhibit the growth of the common decay-causing bacteria, Streptococcus mutans. In this study, genomic DNA was collected from students at Texas Wesleyan University and represented coffee consumers and non-coffee consumers. The genomic DNA was sequenced in order to characterize the oral microflora based on the 16s rRNA sequence. We hypothesized that the oral microflora decreases in species richness and relative abundance in response to coffee intake. In addition, we hypothesized that there will be significant differences in metabolic pathways present in these two groups. Our preliminary findings support these hypotheses in that the two groups differ in species richness, species abundance, and metabolic pathways.
Anthony Sanchez, St. Edward's University
11:00-11:15 in TXH
UNC-33, a microtubule associated protein, plays a key role in neuronal autophagy
Anthony Sanchez and Andrea Holgado
Autophagy is a self-degradative mechanism used within cells to maintain homeostasis in recycling cellular waste and protein aggregates through an autophagosomal-lysosomal system. The regulation of autophagy has been implicated in axonal development via CRMP-2/UNC-33, a microtubule associated protein (MAP) that serves a key role in neuronal development. To better evaluate the relationship between CRMP-2/UNC-33 and autophagy, we studied autophagy in unc-33 mutants by monitoring autophagosome maturation and pathway flux. Autophagy was induced via starvation and monitored using double fluorescently tagged autophagosomal proteins (dFP). The cleavage of the dFP into monomeric fluorescent protein (mFP) upon arrival at the lysosome was used as a representation of flux through the pathway. Our findings show that unc-33 mutants have defects in autophagic flux under starved conditions. Further analysis of autophagosome trafficking is currently underway to elucidate a potential mechanism for the link between MAPs, axonal development and homeostatic mechanisms such as autophagy.
Faith Sawyer, Dallas Baptist University
9:00-9:15 in DBU
MRSA: The Missing Link
Faith Sawyer
The emergence of antibiotic-resistant bacteria has multiplied the danger of Staphylococcus aureus infections, which are the most common cause of skin and soft tissue infections (Alam, et al., 2015). When the infecting S. aureus bacteria acquires antibiotic resistance factors, complications increase and difficulty in patient treatment rises. In an attempt to understand the bacteria’s process of evolving from non-antibiotic resistant staphylococci bacteria, scientists have examined the DNA of various Methicillin-resistant Staphylococcus aureus strains, focusing mainly on the mobile genetic elements (MGEs) contained within the genome. While MRSA’s evolution from non-resistant strains through MGEs may reveal answers, examination of horizontally transferred genetic material through the lens of absence—an area that has not been subjected to much in-depth research—may provide substantial clues. When compared to antibiotic-susceptible strains, are there genes missing from the MRSA genome? To investigate this possibility, the NCBI database was used to obtain the complete genomes of an antibiotic-susceptible S. aureus strain (MSSA), and two different isolates of MRSA from the USA300 strain. Using the bioinformatics software UGENE, the MRSA isolates were individually compared to the MSSA strain by creating dot plots of their genomes. Any notable interruptions in nucleotide base pair matches on the dot plots—indicating a genetic sequence that was found in the MSSA strain but not found in the MRSA isolate—was analyzed with NCBI nucleotide BLAST to predict its function. Findings showed that specific enzymes and bacteriophage proteins were absent from a prophage region in the MRSA isolates.
Abigail Sequeira, University of Dallas
9:45-10:00 in TXH
Parasitic infection and exposure alters mate choice in Potamopyrgus antipodarum
Abigail Sequeira and Deanna M. Soper
Parasites can manipulate host traits in a variety of ways, such as alterations to host behavior, morphology, and physiology. Most research in this field examines parasitic alterations on post-infected hosts, not under parasite exposure. Potamopyrgus antipodarum, a freshwater snail from New Zealand, is known to coevolve with a sterilizing trematode parasite in glacial lakes. In one previous study, Potamopyrgus antipodarum increased mating rates and number of partners upon parasitic exposure alone. In this study, we exposed two different field-collected populations of P. antipodarum to the parasite Microphallus and observed the number of mating and rejecting events when placed with males from sympatric and allopatric populations. Use of field-collected snails meant that a proportion of female snails were already infected, and post-test dissections revealed this to be the case. We found that females altered mating behavior, but only when exposed and infected. This data indicates that exposure to parasites alone can alter female behavior, but only when females are infected. These results highlight the importance of examining host behavioral alterations upon exposure to parasites, but that future tests should include both infected and uninfected hosts.
Max Shaw, West Texas A&M University
1:15-3:15 in LSR 3
Analysis of the amino acid content for beef, chicken and turkey bone broths
Max Shaw and Nick Flynn
Amino acids provide essential building blocks for the proteins that provide structural support and help control signals within our bodies thus allowing us to live and function normally on a daily basis. Quantifying key amino acids in various health products may help us better understand their contribution to general health and support or repudiate claims made by supplement companies. For this project we quantified 19 individual amino acids in bone broth from beef, chicken and turkey. Amino acids from commercial samples of each broth were analyzed using an HPLC method with ninhydrin post column derivatization for quantification. Norleucine was used as the internal standard for analysis. The comparison of the broth concentrations to meat and PAP concentrations allowed us to see that broths may have an ability to contribute vital amino acids to one’s diet, but do not appear to be a better source.
Theresa Stein, Southwestern Oklahoma State University
1:15-3:15 in LSR 4
The effects of stream order, season, and drainage on the abundance of two caddisfly families
Theresa Stein, Amy Hofeld, and Skyler Mills
Rivers are constantly flowing and vary in different locations. Therefore, one area of a river or stream may differ from another in abundance of organisms. However, streams remain connected, as changes upstream may affect areas downstream. We hypothesized that stream order, season, and drainage would affect the abundance of two caddisfly families (Leptoceridae and Hydroptilidae). Species within each of these families have been identified as species of conservation concern by the Oklahoma Department of Wildlife and Conservation. We collected adult caddisflies in the Kiamichi River, Little River, and Spring Creek drainages using manned and unmanned light traps. Samples were collected during both spring and summer months. For both families, we found no difference in abundance between seasons or among drainages. We found that the abundance of leptocerids was significantly higher in third order streams than first or second order streams. However, stream order did not have a significant effect on hydroptilid abundance. We conclude that stream order affects the abundance of Leptoceridae in eastern Oklahoma streams. This information can be used to optimize sampling effort to detect species of conservation concern in this family of caddisflies.
Zoey Stormes, Angelo State University
1:15-3:15 in LSR 3
The Effects of HEPA Air Filters on Airborne Mold Spores in College Dormitories
Bethany Chapman, Lily Ellzey, Zoey Stormes, and Nicholas Negovetich
Every year thousands of students across the nation are required to live in university campus dorms, residence buildings maintained by the university that may have been in operation for years. After an article discussing the death of a University of Maryland immunocompromised student had mold implicated in her cause of death and fellow students at our university who are mold sensitive complained of their symptoms bothering them since arriving on campus, we became intrigued on methods universities or students could employ in individual dorm rooms to decrease overall mold in their dorm. One easy fix is to replace the standard air filter with a better one. This study investigates the effects of changing air filters to a higher MERV quality in dorm rooms on airborne mold spores. Through exposing SDA agar plates to the ambient air in dorm rooms, the mold in two different buildings of the same residence hall was assessed. A second set of agar plates was exposed after changing the air filters in each room, allowing for measurement of the effect new filters have on mold spore counts. Assessment of the before (x=2.05 spores/plate) and after plates (x=1.78 spores/plate) revealed no significant difference between the two, indicating that changing air filters does not reduce the number of mold spores in the tested dorm. However, there appears to be significant differences in the quantity of mold spores between the individual dorms.
Emma Thornburg, Austin College
11:45-12:00 in TXH
Investigating the Positive and Negative Regulators of PA28γ in Cancer
Emma Thornburg and Lance Barton
PA28γ is an ATP and ubiquitin-independent proteasome activator implicated in the development of cancer. Many of the downstream targets of PA28γ can benefit cancer development, and multiple cancers demonstrate increased expression of PA28γ. While the benefits of elevated PA28γ activity have been studied, the mechanisms by which PA28γ expression is regulated have not been fully elucidated. Two key regulators of PA28γ expression, miRNA-7 and p38, may be involved in the increased expression of PA28γ in cancer cells. I hypothesize that in cell lines with increasing amounts of PA28γ, p38 activity will be increased concomitantly, while miRNA-7 expression will be decreased. I also hypothesize that the decrease in miRNA-7 could have a synergistic effect on p38 signaling, leading to an overall larger increase in PA28γ in cancer cell lines. Results confirm a role for both p38 and miRNA-7 signaling in the increased expression of PA28γ in a tumorigenic cell line and a metastasized cancer cell line. Interestingly, p38 appears to also play a role in the regulation of miRNA-7 in PA28γ +/+ cells. Finally, results show that p38 inhibition decreases cellular proliferation while miRNA-7 inhibition increases cellular proliferation. Overall, these results demonstrate that the regulation of PA28γ by p38 and miRNA-7 may play a role in cancer development through increased proliferation of cells.
Keith Tittle, Texas Wesleyan University
11:00-11:15 in LSR
Species-area Relationships across Conserved lands in Texas: Does iNaturalist Work?
Keith Allen Tittle
Species-area relationships provide insights into ecological communities that in turn can be used to determine where conservation efforts are needed. Observations on iNaturalist provide a basis for estimating species richness using citizen science. iNaturalist has been used to collect species richness data from more than 60 parks and conserved areas near 3 major urban areas of North and Central Texas. According to the theory of island biogeography, area size predicts species richness. Species-area relationships are being assessed using linear regression following log transformation of area size and species number. The models for plant species reveal that richness increases as area size increases. However, much of species richness remains unexplained by area size alone. Human disturbance can accelerate extinction rates in conserved lands across Texas. iNaturalist can lessen the impact of this disturbance.
Rachel Vargas, St. Edward's University
1:15-3:15 in LSR 3
Hemocyte Protein Analysis as an Indicator of Immunological Response in Tigrosa helluo Following Leg Autotomy
Rachel Vargas
Autotomy is a survival mechanism utilized by various organism, in which an individual voluntarily loses a body part in response to an external stressor, often in the form of a predator attack. While this capability is a benefit towards immediate survival, there are notable impediments towards the overall survival of an autotomized individual. Behavioral and physiological alterations are commonly present following the autotomy event. The molecular components aiding in the recuperation of an organism after the loss of a leg have been of particular interest as they are indicative of the organism’s overall immunological activity. In invertebrates, the hemocytes incorporated within hemolymph are important factors in cellular immunity. Within reponses to leg autotomy, hemocyte interactions with foreign matter entering through an open wound can be assessed through the various proteins involved with the hemocyte activity. By analyzing the proteins present in the hemolymph of the wolf spider Tigrosa helluo after autotomy, we were able to identify the overall presence and function of the proteins associated with this stress response. The cost of this survival technique was indirectly inferred through the quantification of different proteins released during autotomy as a function of the immunological response. Protein concentration and prophenoloxidase activity, an precursor for the organisms encapsulation response, was measured in both the controlled and autotomized female Tigrosa helluo and the control sample of the males. While there was no significant differences between either of the parameters across any of the sample groups, the observed patterns of this preliminary data suggests a possible difference between protein activity in females and males, and a difference in the prophenoloxidase activity in autotomize individuals compared to non-autotomized individuals.
Brent Wesley, Angelo State University
9:45-10:00 in DBU
Genetic Barcoding Analysis of Agave
Brent Wesley, Loren Ammerman, and Bonnie Amos
Century plants, or Agave, grow in the southwestern United States and three species are known from the Big Bend National Park region. At least one of these species, Agave havardiana, provides a food source for and is pollinated by Mexican long-nosed bats (Leptonycteris nivalis). It can be difficult to determine which species of plant a bat has visited from morphological analysis of the pollen that clings to the fur. Our goal was to take a molecular approach and identify a barcoding gene that would be useful to be able to distinguish species of Agave plants from their pollen. Another goal was to determine how genetically different two species that grow in Big Bend National Park are from each other (A. havardiana and A. lechuguilla). We collected leaves from 36 plants (16 A. havardiana and 13 A. lechuguilla) and extracted DNA. PCR was performed with forward and reverse primers for 5 genes (4 chloroplast genes and 1 nuclear gene region) that have been useful for distinguishing other species of plants. PCR products were purified and sequenced. We analyzed the similarities and differences between the genes from each species, calculated divergent values, and determined the best primer set for pollen barcoding viability. The best primer set, or gene, will be the one with consistent, species-specific DNA differences to clearly distinguish between species of Agave in the region. We found that 3 of the genes were identical among the samples of the two species. One nuclear gene, ITS2, showed a low level of sequence divergence with a pairwise divergence value of 1.38% using a Tamura 3 parameter with gamma distribution. We conclude that the best gene for species level barcoding of agave is ITS2 because of the evidence of sequence divergence.
Dani Whiting, Southwestern Oklahoma State University
1:15-3:15 in TXH 1
Raptor Perch Locations and Types in Southwestern Oklahoma
Dani Whiting and Zach Jones
Migrating and overwintering raptor species use a variety of hunting methods and perching types to search for and initiate prey captures from rural and agricultural landscapes surrounding Weatherford, OK. The primary objective of this study was to identify species preferences for perch types (primarily a range of power transmission line pole sizes and materials, as well as pole density) and landscape context (amount of tilled, non-tilled pasture, woodland, man-made structures other than power poles, etc.) near chosen hunting locations. Two 48-km routes were sampled beginning in fall 2018, one north and one south of Weatherford, OK. Preliminary data show the following trends: American Kestrels (Falco sparverius) primarily used wires between poles; Red-Tailed Hawks (Buteo jamaicensis) indicated a preference for trees; and Northern Harriers (Circus cyaneus) were only seen in flight other than when consuming prey. Additionally, all species preferred areas with little to no presence of man-made structures other than utility poles and fencing around fields. Species’ differences in hunting methods and perch types may mitigate competition for common prey items (rodents) during migration and wintering seasons, and the presence of utility poles may increase the number of perch types available enough to encourage increased abundances of some species’ populations beyond what would otherwise be possible. These preliminary data indicate that both kestrels and Red-tailed Hawks may benefit greatly from the presence of utility poles and lines.
Mikaela Wilson, St. Edward's University
1:15-3:15 in LSR 4
Evaluating the Effects of Inducing Autophagy in unc-33 Mutants
Mikaela Wilson, Nick Ashley, Hailey Trombley, and Andrea Holgado
Autophagy, a mechanism used by cells to recycle and catabolize protein aggregates and cellular waste, is highly conserved from yeast to mammals. Recent reports indicate that UNC-33/CRMP-2/Dpysl2, a neuronal protein essential for early axonal development, is regulated during conditions that induce autophagy in the mammalian neurons. To further investigate the interplay between UNC-33 and autophagy, we constructed a C. elegans double mutant strain containing unc-33 and daf-2(e1370) mutations and characterized their phenotype. daf-2(e1370) encodes for an insulin-like IGF-1 receptor (IIR) and mutations in this gene result in induced autophagy and constitutive dauer formation. Analysis of our double mutant strain denotes that our unc-33; daf-2 double mutants grown at the restricted temperature result in a synthetic lethal phenotype. Quantification of double mutant nematodes surviving at the restrictive temperature shows that the synthetic lethality is partially penetrant, killing 78% of unc-33(e204); daf-2(e1370) and 72% of unc-33(mn407);daf-2(e1370) during embryonic stages. Observations of surviving double mutants demonstrate that these animals form abnormal dauers. The resulting abnormal dauer phenotypes are being characterized by evaluating survival in 1% SDS, pharyngeal length, life stage arrest, and proper sinusoidal movement.
Rebecca Wood, University of Central Oklahoma
1:15-3:15 in LSR 3
Serologic Evidence of Viral Reactivation and Increased Disease Activity in Patients with Systemic Lupus Erythematosus
Rebecca A. Wood, Lauren Guthridge, Emma Thurmond, Carla Guthridge, Hua Chen, Wade deJager, Susan Macwana, Neelakshi Jog, Rebecka Bourn, Stan Kamp, Rufei Lu, Cristina Arriens, Eliza Chakravarty, Katherine Thanou, Joan T. Merrill, Joel M. Guthridge, and Jud
Systemic lupus erythematosus (SLE) is a complex autoimmune disease which mainly afflicts young women. SLE can attack any organ in the body, with the disease fluctuating from active, multi-organ involvement (flares) to minimal disease. What causes flares is unknown. Our objective was to examine serologic evidence of viral exposures and re-activation with respect to SLE disease activity and cytokine levels. Our hypothesis was that reactivation of common latent viruses would increase lupus disease activity. Plasma from 184 SLE patients (n=290) and 49 controls was collected and disease activity measured. Viral responses were detected by ELISAs for EBV-Viral Capsid Antigen (VCA) (IgG and IgA), EBV-Early Antigen (EA) (IgG), Cytomegalovirus (CMV) (IgG), and Herpes Simplex Virus (HSV-1) (IgG). Plasma soluble mediators were measured by multiplex, bead-based assays and ELISAs. Serologic evidence of EBV reactivation was more common in SLE patients, per anti-EA (40% vs 13%; OR=4.57, p=0.0006) or EBV-VCA IgA (36% vs 17%; OR=2.70, p=0.019). Concentrations of EBV-VCA IgG responses were higher in SLE patients compared to controls (EBV VCA ISR = 4.44 vs 3.52; p=0.0021), as were IgA VCA and IgG EA. Cross sectional analysis of SLE patients at high (SLEDAI > 6) (n=126) vs lower (n=166) disease timepoints showed increased IgG EA responses with elevated disease activity (0.822 vs 0.540; p=0.033). SLE patients with anti-EA responses had higher IP10 (p=3.4 X 10-14), BLyS (5.5 X 10-5), and IL-10 (p=0.00013). HSV1 IgG positive SLE patients also had higher IP10 (2.2 X 10-7). This study provides serologic evidence supporting roles for viral reactivation and SLE disease flares.
1:15-3:15 in LSR 4
Pseudomonas syringae motility appendages: roles in surface colonization and survival
Jennifer Abshire and Regina McGrane
Pseudomonas syringae is a bacterial phytopathogen that causes disease in economically important crops. P. syringae is found in many different environments, each with varying levels of moisture. Therefore, the appendage it relies on for motility varies. The removal of genes encoding for these motility factors significantly impairs pathogenicity. The objective of this project was to evaluate the impact of P. syringae motility related appendages on colonization of varying environments. To characterize the role of flagella and pili, movement of deletion mutants was observed in saturated, liquid, and natural surface environments. Flagella mutants had significantly reduced motility in both saturated and liquid conditions, whereas pili mutants had significantly reduced motility in saturated conditions only. This indicates that flagella are the most relied upon appendage for motility and that pili have minor impacts. Similarly, on simulated natural surfaces flagella were the primary movement appendage. To characterize the impact of flagella and pili on colonization of plants, attachment to leaves and seeds as well as growth on leaves were evaluated. Mutants lacking flagella or both flagella and pili were impaired in leaf colonization but showed seed attachment similar to the wild type. However, mutants lacking both the flagella and pili had greater levels of attachment to leaves. To characterize the impact of flagella and pili on antibiotic sensitivity, mutants and the wild type were evaluated using a Kirby-Bauer sensitivity assay. Mutants were more sensitive to ciprofloxacin than the wild type, which is opposite to observations made in a different P. syringae strain. These results indicate that removal of the genes encoding for motility factors such as pili and flagella, directly impacts pathogenicity. Limiting the motility of P. syringae could halt or severely lessen the symptoms associated with this disease-causing pathogen and increase crop yields of host plants.
Neha Amberkar, Texas A&M University - Texarkana
1:15-3:15 in LSR 3
Microplastic Pollution in East Texas Freshwaters and Water Treatment Plants
Neha Amberkar and Vaishnavi Ramachar
Usage of plastic in modern society generates many benefits, yet there is the issue of emerging concern regarding the accumulation and pollution of plastics in aquatic environments as well as its effects in the ecosystem. Microplastics are plastic fragments smaller than 5 mm. They are especially harmful as they can easily be ingested by wildlife, affecting the food web. As this waste is discarded, the breakdown of larger plastics results in excessive accumulation in the environment as microscopic particles. This study focuses on identifying microplastic pollution in the inflow and outflow of freshwaters around water treatment plants in East Texas. To analyze the amount of microplastic fragments as well as corresponding size, water samples were collected from the surface, middle, and sediment layers. To understand microplastic pollution in raw unfiltered water as well as the effect a standard treatment plant has on eliminating these wastes before re-release into the ecosystem, samples were collected from the water that passes in and out of the sewage treatment plant. Once collected, the water samples were then divided into portions, filtered, and remaining microplastics were then observed under a microscope. Size, composition, and overall density of the microplastic fragments were analyzed. Densities were also observed corresponding to the surface, middle, or sediment layer of the waters to determine where these microplastics aggregate and have the strongest presence. Conclusions were then hypothesized for the implications of this pollution in the local ecosystem and on the larger environmental scale.
Nana Amponsah, Angelo State University
1:15-3:15 in TXH 2
Predator Avoidance Behavior in Physa acuta
Nana Amponsah and Ben Skipper
The senses of prey animals have been finely tuned by natural selection to detect potential predators before a predation attempt is begun. For aquatic invertebrates, chemosensation is the primary means by which predators are detected. However, in polluted waters, the chemosensory abilities of prey animals may be negatively impacted through interference. In such cases, prey animals may not be able to sense predators before a predation attempt. This study investigates predator avoidance behaviors of Physa acuta, a freshwater snail, exposed to two potential predators and scent of a distressed conspecific in both clean and polluted waters. Snails were observed in isolation for a period of thirty minutes. The first ten minutes allowed snails to acclimate to the new tank. In the second ten minutes we recorded the snail’s movement in the absence of predator information. Prior to the third ten minutes, we added scent of a predaceous beetle, crayfish, or water (control) and then continued recording snail behavior. We observed no significant difference in movement and behavior after the addition of beetle scent and water. However, there was significant decrease in movement after addition of crayfish scent, with some snails escaping to dry areas in the test tank. Tests in polluted water, where Physa acuta may be unable to use chemosensation (effectively), are forthcoming.
Kia Baeza, St. Edward's University
1:15-3:15 in TXH 2
Commonly Used Agricultural Pesticides and their Effect on the Immunological Function of Honeybee Populations
Kia Baeza
Colony collapse disorder (CCD) is a current phenomenon which is causing rapid declines in honeybee populations. While researchers have yet to find a direct cause of CCD, it is hypothesized that it may be the result of a combination of multiple factors such as global warming, parasitic Varroa mites, and the use of pesticides. The goal of this study was to identify the effect of four commonly used pesticides in agriculture (imidacloprid, glyphosate, thiamethoxam, and clothianidin) on the immune function of honeybees. For each pesticide, 150 honeybees were collected randomly from a set of hives in the Austin area and starved for four to six hours. A pesticide sugar water solution with biological relevant concentrations was then fed to each bee with a micropipette. Bees were then left for zero (control), two, or four hours after feeding before hemolymph was extracted. Hemolymph was added to buffer, lysed with glass beads, and placed at a -80oC to stop protein activity. Samples are currently undergoing several colorimetric assays utilizing a spectrophotometer to determine immunological protein activity including a RED 660 assay for overall protein concentrations, prophenoloxidase, catalase, and peroxidase. Anecdotally, bees subjected to the pesticide glyphosate appear to have higher mortality over time compared to the other pesticides tested. Bees that ingested glyphosate had significantly lower protein concentrations than all other pesticides tested. Prophenoloxidase activity appears highest when bees are exposed to clothianidin, while imidacloprid significantly decreases activity four hours after exposure. Results from this study will help determine what effects, if any, pesticides contribute to the onset of CCD.
Anna Bailey, Oral Roberts University
11:15-11:30 in DBU
Gene Annotation of the Hypothetical Protein-Coding Genes of Chlamydia caviae
Anna Bailey
Genetic information for a multitude of microorganisms has become readily available due to technological innovations in the field of genome sequencing. However, several of these novel genome sequences remain unexplored and require manual annotation to uncover their hypothetical protein-coding genes. Through the use of publicly available online bioinformatics programs developed by the United States government as well as researchers across the globe, primary amino acid sequences derived from hypothetical gene sequences can be used to predict their function. Examples of properties identified by these programs include protein families, conserved domains, signal peptides, transmembrane regions, open reading frames, and protein localization. The purpose of this research project is to predict the functions of five unannotated hypothetical protein-coding genes in the bacterium Chlamydia caviae, which infects the mucosal epithelium of rodents. The genes CCA_RS04585, CCA_RS04590, CCA_RS04595, CCA_RS04600, and CCA_RS04605, were analyzed and are predicted to code for the following: glycerol-3-phosphate dehydrogenase (NAD(P) (+)), a membrane protein, two type-III secretion system family proteins, and phosphoenolpyruvate carboxykinase, respectively. These predictions provide some insight into the proteome of Chlamydia caviae. Ultimately, the proposed gene annotations must be validated through molecular cloning and biochemical methods to determine whether Chlamydia caviae expresses these proteins and whether they perform their bioinformatically predicted functions.
Kaylee Ballard, The University of Tulsa
1:15-3:15 in LSR 3
Next Generation Sequencing of Staphylococcus Species Isolated from Oklahoma Retail Mushrooms
Kaylee Ballard, Madison Bickle, Leena Neyaz, Anand B. Karki, and Mohamed K. Fakhr
Staphylococcus species are important pathogens and are responsible for various infections associated with serious community and hospital acquired diseases. Most studies characterizing Staphylococcus spp. are biased toward clinical strains vs. those isolated from food. Extended study on the genomic structure and virulence potential of various species of Staphylococcus from retail mushroom is lacking in the literature. In a preliminary study conducted in our laboratory, the prevalence of Staphylococcus spp. isolated from Oklahoma retail mushrooms was significant, and several strains were phenotypically resistant to a wide range of antibiotics. Most of these strains harbor antibiotic resistance genes like mecA gene in addition to some enterotoxin genes. The objective of the current study was to determine the genomic sequence using next generation sequencing (NGS) technology of 12 various Staphylococcus species previously isolated from Oklahoma retail mushrooms. Twelve strains of Staphylococcus species (3 S. fleurettii, 2 S. sciuri, 1 S. saprophyticus, 1 S. vitulinus, 1 S. xylosus, 1 S. succinus, 1 S. pasteuri, 1 S. warneri and 1 S. haemolyticus) were streaked on TSA plates from stock cultures (-70oC) and then grown as liquid cultures in TSB media overnight. Genomic DNA was extracted from the overnight cultures using DNeasy blood and tissue kit (Qiagen) according to the manufacturer instructions. The twelve strains were also screened for the presence of plasmids using the Qiagen MediPrep plasmid isolation kit. Plasmids with various sizes appeared to be prevalent among the tested strains. Next generation sequencing library preparation are currently underway using the Illumina Nextera XT DNA library preparation kit and it will be run on the Illumina MiSeq Sequencer. Genome assembly will be performed using the CLC Genomics Workbench and the microbial genome finishing modules. Comparative genomics among the twelve sequenced Staphylococcus strains will be explored to detect genes responsible for virulence and antimicrobial resistance.
Kaylee Ballard, The University of Tulsa
9:30-9:45 in DBU
Influence of Staphylococcus aureus Co-Contamination On Survival of Campylobacter Strains At Low Temperature, Biofild Formation, and Aerotolerance
Kaylee Ballard, Anand B. Karki, Claudia Harper, and Mohamed K. Fakhr
Campylobacter, which causes the diarrheal illness Campylobacteriosis, is one of the leading bacterial foodborne pathogens in the United States. High prevalence of the bacteria has been found in retail meat and liver products, most of which are also contaminated with Staphylococcus aureus. In adverse conditions, the presence of Staphylococcus aureus has shown to have an influence on the survival and biofilm formation of Campylobacter. This study investigates the effect of Staphylococcus aureus on the survival of Campylobacter at low temperatures commonly used in retail meat storage, its biofilm formation, and aerotolerance. Six strains of Campylobacter [C. jejuni (T1-21, NCTC11168, OD2-67) and C. coli (HC2-48, WAS-33, ZV1-224)] were incubated in MHB media as well as filter sterilized S. aureus grown MHB media from two strains of S. aureus (B4-59C and B6-55A). Assays for survival with viable count, biofilm formation and aertolerance assays were performed. Results showed there was a higher survival rate for prolonged time at low temperatures in the Campylobacter strains grown in MHB media containing S. aureus metabolites versus the MHB media control. Aertolerance was also shown to be enhanced with the presence of S. aureus metabolites. This indicates that the extracellular metabolites of S. aureus influence the survival of Campylobacter. There were no consistent results on the influence of S. aureus grown media on Campylobacter biofilm formation. Similar experiments including S. aureus cells instead of the filtrate media were also conducted. Presence of S. aureus cells enhanced survival of strains of Campylobacter at low temperature as well as biofilm formation.
Emily Bedea, Southwestern Oklahoma State University
1:15-3:15 in TXH 1
In-silico analysis and homology modeling of β-carbonic anhydrase from Streptococcus sanguinis, an opportunistic pathogen involved in subacute infective endocarditis
Emily Bedea and Vijayakumar Somalinga
Streptococcus sanguinis is one of the leading causes of infective endocarditis (IE) in a susceptible population. Although several virulence factors involved in IE have been identified, the exact mechanism by which S. sanguinis colonizes endocardium remains unclear. In addition to some well-characterized virulence factors, S. sanguinis also contains virulence-associated proteins that are not well characterized. One such protein that is associated with virulence is carbonic anhydrase (CA) that catalyzes the reversible hydration of CO2 to bicarbonate. The goal of this study is to identify and characterize carbonic anhydrase from S. sanguinis. We have identified a β-CA from S. sanguinis (SsaCanB) using BLAST analysis that shows high homology to other well-studied β-carbonic anhydrases. SsCanB is an 18.2 kDa protein with no N-terminal signal peptide indicating its cytoplasmic localization. Sequence alignment of S. sanguinis with closely related homologs shows conserved domains typical of β-CA including residues important for metal coordination and protein-protein interactions. Homology modeling and subsequent structure analysis of SsCanB reveals that both proteins have α/β fold typical of β-CA’s. Analysis of the active site of SsaCanB shows conserved residues involved in zinc ion coordination similar to other well-studied β-CA’s. Furthermore, the catalytic dyad (Asp/Arg) required for the activation of water molecule coordinated with zinc ion is also conserved in SsaCanB.
Alexa Benavides, Houston Baptist University
1:15-3:15 in LSR 4
The Very Hungry Superworm: Superworms' ability to digest Polystyrene
Alexa Benavides, Julia Ta, Daisy Lopez and Agnieszka Czopik
Polystyrene (PS) is a synthetic polymer made from styrene and is commonly used in commercial packaging due to its ability to repel water and resist corrosive substances. However, the same chemical stability that makes PS so useful also poses a threat to the environment, as PS is non-biodegradable and consequently, amasses in landfills. Research published by Stanford University in 2015 provides a possible solution to this environmental crisis, since it showed that the larvae of mealworms (Tenebrio molitor) host a specific strain of gut bacteria (Exiguobacterium sp. strain YT2) that are able to break down PS into biodegradable compounds, such as water and carbon dioxide. To further expand on this idea, we fed mealworms that were maintained on only polystyrene, for seven weeks, to one group of superworms, and another group of superworms were fed mealworms that were maintained on only oatmeal for the same amount of time. We then fed PS to both groups of superworms and calculated the amount of PS eaten per worm in order to analyze if changing the worm microbiota will have an impact on their ability to digest PS. We plan to further our studies by analyzing crumbs from the PS that was fed to the superworms to determine whether there are enzymes remaining on it that will help it degrade.
Jillian Blake, Oral Roberts University
9:00-9:15 in TXH
Attraction of swarming paper wasp sexuals (Polistes spp.) to the chemical attractant N-3-methylbutyl acetamide
Jillian Blake
Before the beginning of the experiment, common passageways used by the paper wasps to enter inside the ORU dormitories were sealed in hopes of decreasing the number of wasps entering the buildings. This study investigated the numbers of paper wasps (Polistes spp.) swarming atop dormitories after the limitation of dormitory access in comparison to data from previous years. The objective was to make a weekly observation counts of swarming paper wasps on the roofs of the Francis and Michael Cardone dormitories at Oral Roberts University. With the new changes in dorm accessibility, it was predicted that the overall wasp count would be lower than that of previous years. These counts found that two distinct peaks of wasp activity occurred in early and late October due to variable weather temperature. As expected, the overall wasp count was lower than total wasp counts in years dating back to 2011. However, this reduction may be due to the second study, trapping wasps atop the dorms. Three sets of wasp traps were placed: two sets atop the Francis dormitory and one set along the fence lining the west side of Fred Creek. The traps collected a total of 680 wasps, but the effectiveness of the N-3-methylbutyl acetamide attractant compared to the control was not significant. Thus, it was concluded that further experimentation would be necessary to gather more significant data.
Hannah Butterfield, Austin College
11:30-11:45 in TXH
Investigating Synergy Between PA28γ and IKKε
Hannah Butterfield and Lance Barton
Cancer is characterized by excessive cellular proliferation and by dysregulation of signaling pathways related to growth. PA28γ is a proteasome activator frequently increased in cancer and involved in regulation of multiple cellular pathways that affect proliferation, including the Nuclear Factor κB (NFκB) pathway. Inhibitor of Nuclear Factor κB Kinase ε (IKKε) is a breast cancer oncogene that affects multiple pathways that implicate PA28γ, including the activation of NFκB. This study aims to elucidate whether PA28γ is required for normal IKKε activity. I hypothesize that PA28γ enhances the overall expression and nuclear export of IKKε, and aids IKKε-mediated cell signaling. I am assessing this through evaluating differences in IKKε nuclear, cytosolic, and overall concentration in PA28γ+/+ and PA28γ-/- murine embryonic fibroblast (MEF) cells. I am also evaluating differences in IKKε-implicated cellular processes including transcriptional activation and animal weight gain. The data does not support a role for PA28γ in regulation of IKKε.
Constance Byers, Texas A&M University - Texarkana
11:00-11:15 in DBU
Microplastic Pollution in East Texas Freshwaters and Water Treatment Plants
Constance Byers and Roya Morakabian
Usage of plastic in modern society generates many benefits, yet there is the issue of emerging concern regarding the accumulation and pollution of plastics in aquatic environments as well as its effects in the ecosystem. Microplastics are plastic fragments smaller than 5 mm. They are especially harmful as they can easily be ingested by wildlife, affecting the food web. As this waste is discarded, the breakdown of larger plastics results in excessive accumulation in the environment as microscopic particles. This study focuses on identifying microplastic pollution in the inflow and outflow of freshwaters around water treatment plants in East Texas. To analyze the amount of microplastic fragments as well as corresponding size, water samples were collected from the surface, middle, and sediment layers. To understand microplastic pollution in raw unfiltered water as well as the effect a standard treatment plant has on eliminating these wastes before re-release into the ecosystem, samples were collected from the water that passes in and out of the sewage treatment plant. Once collected, the water samples were then divided into portions, filtered, and remaining microplastics were then observed under a microscope. Size, composition, and overall density of the microplastic fragments were analyzed. Densities were also observed corresponding to the surface, middle, or sediment layer of the waters to determine where these microplastics aggregate and have the strongest presence. Conclusions were then hypothesized for the implications of this pollution in the local ecosystem and on the larger environmental scale.
Jailene Canales, University of Central Oklahoma
11:15-11:30 in TXH
Characterizing Early Developmental Defects in an Avian Model of Maternal PKU
Jailene Canales, Austin McDonough, Morgan Massey, and Nikki J Seagraves
Maternal phenylketonuria [MPKU] is a syndrome of multiple congenital anomalies including cardiovascular malformations [CVMs], brain and growth restriction when a mother with Phenylketonuria [PKU] does not control her dietary intake of Phenylalanine [Phe]. In this study, we aim to establish and characterize an avian model of MPKU. We focused on early developmental defects. We investigated the effect of 2500μM Phe exposure by in-ovo yolk injection. Following the injection, the embryos underwent further development for 48 hours until dissection was performed. At HH14-17, India ink was injected into the yolk as a contrast dye. Images were taken of embryos and they were scored based on Drake et. al (2006.). Embryos exposed to high Phe displayed gross morphological changes including developmental and growth delays, anterior and posterior abnormalities, and torsion defects. Histological analysis is underway to determine changes in heart development. Currently there is no data interrogating the mechanism by which Phe causes heart defects. We plan to utilize this model to define the mechanism of Phe cardiac teratogenicity which is critical for improving MPKU treatments and outcomes.
John Cantley, University of Texas at Tyler
1:15-3:15 in LSR 3
Pseudomonas aeruginosa Antibiotic Resistance Gene Profile in East Texas Clinical Isolates
John Cantley, Shandis Fancher, and Ali Azghani
Pseudomonas aeruginosa is an opportunistic, Gram-negative bacterium and causative agent of acute nosocomial, community-acquired pneumonia, or chronic infections. Clinical isolates of this organism harbor intrinsic and acquired antibiotic resistant genes. Improper antibiotic prescription or use may select for mutations in genes coded for ß lactamase, porins, or efflux pumps. These mutations confer resistance across multiple drugs, make the treatment of infections more challenging, and increase the rate of infection occurrence and mortality. The goal of this project is to identify antibiotic resistant genes carried by clinical isolates of P. aeruginosa in East Texas. We utilized quantitative real-time PCR on recently collected clinical isolates of P. aeruginosa in order to determine the presence of targeted antibiotic resistant genes. Bacterial subcultures were adjusted to an optical density of 0.8 (OD600), and total DNA was isolated using the boiling method for 10 min at 100 ºC. Cold ethyl alcohol was used to enhance precipitation of a salt free DNA pellet. Samples were checked for concentration and purity using a NanoDrop spectrophotometer. Quantitative real-time PCR was performed using targeted primers on the DNA extracts using 40 thermal cycles of 20 sec at 95ºC, 20 sec of 60ºC, and 30 sec of 72ºC. We are in the process of applying the primers for aforementioned genes on each of these isolates. Our far-reaching goal is to compare the rate of antibiotic resistant strains of P. aeruginosa in East Texas to global database in the context of clinicians standard care and patients compliance.
Angela Chen, Angelo State University
9:00-9:15 in LSR
Atypical Expression of Cell Cycle Genes in a Drosophila Model of Alzheimer’s Disease
Angela Chen
Understanding the genetics of the cell cycle is important for studying normal development and physiology as it guides development, function, and maintenance in our bodies. Atypical gene expression in cell cycle genes can lead to pathology such as developmental disorders and neurological diseases. Alzheimer’s disease is the most common cause of dementia and is a significant financial burden on the United States economy. Atypical expression of Cdk4 has been studied in the brain of Alzheimer’s disease patients. Asp and Mcph1 are genes that are related to the dysregulation of the cell cycle, and mutations in these genes can result in microcephaly, a human neurodevelopmental disease. However, there have been no studies so far to determine the gene expression of Mcph1, Asp, and Cdk4 in the Drosophila model of Alzheimer’s disease. The goal of this research was to develop primers to allow neurodevelopmental gene expression to be studied both qualitatively and quantitatively for neurological diseases such as microcephaly and Alzheimer’s disease. Functional primer sets were developed and expression levels of Asp, Mcph1, and Cdk4 were tested using qualitative and quantitative RT-PCR. Aberrant expression of these genes was observed in the disease models compared to the wild type strain. Characterizing the role of these genes may reveal potential use in inducing adult neural stem cells to proliferate for use in future in novel therapeutics and regenerative medicine for Alzheimer’s disease.
Clay Clarkson, Texas A&M University - Corpus Christi
1:15-3:15 in TXH 1
Quantifying the relationship between methane concentration and mcrA genes from a cultured Methanolobus spp. isolated from surfaces sediments of Tranquitas Creek, Kingsville Texas
Clay Clarkson and Brandi Kiel Reese
Methanogens are a diverse taxonomic group within the archaeal domain that are capable of methanogenesis. Methanogenesis was once thought to be unique to the phylum Euryarchaeota, however; the essential gene for methanogenesis, methyl-coenzyme M reductase (mcrA), has been discovered amongst other phyla including the renamed miscellaneous Crenarchaeota group: Bathyarchaeota. Members of Bathyarchaeota are widespread amongst environments and contain variants of mcrA, suggesting a potential ability to use other methylated alkanes other than those known today. Though, little is known about this phyla’s contribution to global methane fluxes and the exact substrates for methanogenesis. In attempt to identify and quantify methane fluxes of potential methanogenic members outside the Euryarchaeota phyla, an enrichment approach followed by subsequent transfers was employed to obtain a methanogen isolate in culture. Surface sediments of a freshwater stream system was used for the enrichment phase. Taxonomic identification of the pure cultures was done by sequencing on the 16S rRNA gene. Methane flux per individual cell was calculated by using the relationship between methane concentration and mcrA gene abundances during exponential growth phases. Quantification of methane and mcrA genes in the pure cultures was determined by gas chromatography and droplet-digital PCR. This study aims to demonstrate a potential use of mcrA gene counts as a proxy for quantifying methane fluxes.
Katherine Collins, Austin College
9:15-9:30 in LSR
Barriers to plant establishment from seed in tallgrass prairie restoration
Katherine P. Collins
Conversion to agriculture during the late 19th and early 20th Centuries severely reduced the extent of the tallgrass prairies once prominent throughout the central United States. In an on-going tallgrass prairie restoration project at Austin College’s Sneed Prairie, students track restoration progress on nine fields managed three different ways. Success of grass establishment from planted seeds has varied dramatically among fields, and even among experimental replicates subjected to the same restoration techniques. Three fields have had dramatic success of grass establishment from planted seeds. Six fields have had less, and in some cases much less, success. The objective of this project is to test hypotheses with potential to explain the lack of widespread establishment from planted seeds in the latter six fields.
Cristin Connor, University of the Ozarks
11:45-12:00 in LSR
Temperature and the Variability of Vertebrae in Dasypus novemcinctus
Cristin Connor and Ashley Saunders
The nine-banded armadillo is a fascinating creature and exceptionally useful for scientific research. This armadillo always gives birth to four genetically identical offspring which minimizes the variables that must be accounted for in an experiment. Another interesting fact, and the focus of this study, is that the number of vertebrae in this species is variable. With only 40% of a studied population having the median number of vertebrae (Asher, et. al.) This is believed to be due to variation in the body temperature of the mother during gestation. 77% of the armadillos studied had the median number of vertebrae but three litters of offspring had abnormal counts. Two of these litters were part of the control group and all three were in a normal range of variation, only about 2-3 degrees between day and night. This brings up an issue with the temperature hypothesis because it did not appear to be the causative agent in this experiment. Perhaps a next step would be food stress, as would be experienced in the wild, which could cause more natural occurrence of vertebral variation.
Bailee Cummings, Southern Nazarene University
1:15-3:15 in LSR 3
Mosquito Sampling and Arbovirus Surveillance in Oklahoma
Bailee Cummings, Randall Welles, Helen Arango, Nicholas Back, and Caio Martinelle França
Vector-borne diseases are a major burden for public health. In the U.S., more than 640,000 cases of vector-borne diseases were reported from 2004 to 2016. In the past 13 years, mosquito-borne disease cases have tripled. Culex mosquitos are responsible for establishing endemic West Nile virus across the U.S. The current extended range of invasive Aedes mosquitos dramatically changes the landscape for arboviral risk in the U.S. because they can transmit diseases like Zika, Dengue, and Chikungunya. According to the Oklahoma State Department of Health, West Nile virus (WNV) was introduced to Oklahoma in 2002. Mosquitoes carrying WNV can infect persons of any age; however, those over the age of 50 are at greater risk of developing serious illness affecting the nervous system. From 2002-2018, the State of Oklahoma registered over 779 cases of WNV and 58 deaths. Three WNV outbreaks have occurred in Oklahoma in 2003, 2007, and 2012. This accelerating spread of arboviruses highlights the need for a proactive mosquito surveillance. Mosquitoes were collected from multiple counties in Oklahoma using CDC miniature light traps resulting in 7929 mosquitoes captured. Mosquitoes were identified to 15 different specifies from four main genera: Aedes, Culex, Anopheles, and Psorophora. Nucleic acid was extracted from field-collected mosquitos using DNA/RNA ShieldTM from 50 mosquito pools. Surveillance of viruses in mosquito pools was performed with RT-qPCR using universal primers for detection of flavivirus-specific viral nucleic acid. We identified three RT-PCR positive pools for the presence of the genus Flavivirus. We are currently working on whole viral genome sequencing for these positive samples in order to obtain phylogeographic information of the viruses.
Caitlyn Cutrer, Northwestern State University
9:45-10:00 in LSR
Identifying Leptospira interrogens in feral hog populations using the MALDI-TOF
Caitlyn Cutrer, Jordan Bringedahl, and William Simpson
Leptospira interrogens is a zoonotic pathogen that causes leptospirosis. Feral hogs (Sus scrofa) are known carriers of this bacterium and due to the hogs’ reproductive prowess their population is beginning to encroach into city water reservoirs creating a human health concern. Traditionally, identification of Leptospira sp. is done by using the microscopic agglutination test; however, this can be expensive and time consuming. Alternatively, by using MALDI-TOF we can identify multiple species of Leptospira using mass spectrum profiles. Blood was opportunistically collected from 23 feral hogs across Natchitoches Parish. The serum was separated via centrifugation. 0.5 mls of a respective serum was then pipetted into Fletcher’s medium modified with 0.05% hemin. Once growth was observed (~5-7 days), 1 ml of medium was aseptically pipetted out into a 1.5 ml tube and centrifuged, the supernatant was decanted, and the pellet spotted on a MSP 96 target polished steel plate and overlaid with 1 µl of HCCA. Samples were analyzed using the MALDI-TOF Biotyper software (v4.1.70). The results have yielded mixed results ranging from no growth, to the enrichment of Aeromonas hydrophila, Stenotrophomonas maltophilia, and multiple Pseudomonas sp.; however, some samples yielded mass spectra that could not be readily identified, due to a lack of Leptospira sp. profiles in the current version of the Biotyper database. Thus, we are working on constructing our own library of Leptospira sp. using pure cultures obtained from ATCC and will retroactively assess the previous non-identified spectra obtained from the various serum enrichments.
Erin Dempsey, East Central University
10:45-11:00 in TXH
Plant Pathogenic Fungal Species on Melons from Guatemala
Erin Dempsey, Patrick McDowell, Matt Broge, C. Biles, and A. Howard
Melons (Cantaloupe; Cucumis melo var. cantaloupensis) were purchased from local grocers in 2016 through 2019 and observed for post-harvest diseases. Of the 80 melons purchased in 2019, 75% developed fruit rot symptoms caused by fungi. The diseased tissue indicated that the majority of lesions were caused by Diaporthe spp. (syn; Phomopsis), and to a lesser extent lesions were caused by Alternaria and Fusarium spp. Plant pathogens such as Diaporthe spp. enter the surface of the melon fruit early in development and remain latent until fruit maturity. While ripe fruit is harvested and imported with no external evidence of disease, internal fruit rot becomes evident as the fruit matures. The objective of this study was to characterize Diaporthe spp. imported in Guatemalan melons. Fungal isolates were characterized based on culture growth characteristics, spore morphology, and DNA analysis. Guatemalan isolates were morphologically similar to D. sojae and D. curcurbitae. Deoxyribonucleic acid (DNA) was extracted from fungal hyphae and purified polymerase chain reactions (PCR) products were Eurofin, Inc. for sequencing. Sequencing analysis demonstrated that some of the isolates were a match for D. pterocarpi and species within the D. arecae complex Our finding of pathogenic Diaporthe spp. suggest that plant pathogens are carried across international borders and imported into the United States. Further analysis is being conducted on the melons collected in 2018 and 2019.
Cuishan Deng, East Central University
1:15-3:15 in LSR 4
The difference in leaf-tying caterpillar communities between forest edge and interior habitats
Cuishan Deng, Norbu Gurung, and H. George Wang
Edge effect is an important factor in shaping community patterns, particularly in forest habitats. We conducted an experiment to investigate the potential edge effect on forest arthropod communities at the Nature Conservancy’s Pontotoc Ridge Preserve. We set up artificial leaf ties on red oak trees in both edge and interior habitats, and conducted two censuses in summer 2018. We recorded the morphospecies and the number of all arthropods in the leaf ties during each census. We calculated the Simpson Diversity Index for all arthropods on each tree and compared them between the two habitats using mixed-effects modeling. We also compared the number of individuals between the two habitats using Poisson regression. There was no significant difference between the two habitats for either metrics. We then compared the densities of caterpillars (Lepidoptera) between the two habitats, and found them to occur more on trees in the interior habitat. Similarly, the number of aphids (Aphidae) was significantly higher on trees in the interior habitat.
Kade Ezell, Southwestern Oklahoma State University
1:15-3:15 in TXH 2
Antimicrobial activity of phytopathogen biosurfactants against human pathogens
Kade Ezell and Regina McGrane
Antimicrobials play a major role in the fight against bacterial growth in many areas of the modern world. Without the benefits of antimicrobials, the medical field, food industry, and the overall well-being of all life would be significantly impacted. The gram-negative, bacterial, phytopathogen Pseudomonas syringae produces the biosurfactants syringafactin and rhamnolipid while colonizing plant tissue. These biosurfactants lower surface tension, act as lubricants for bacterial swarming motility, and increase diffusion of plant nutrients. Our laboratory has observed repulsion of P. syringae deletion mutants lacking biosurfactant production and leaf colonizing bacteria when inoculated on swarming agar media in close proximity to wild type P. syringae. We hypothesize that biosurfactants may be important for P. syringae competition in the environment through repulsion of nearby bacteria. The effects of syringafactin and rhamnolipid on bacterial human pathogens have not been studied. The objective of this work is to evaluate the motility of human pathogens in the presence of P. syringae and to determine if biosurfactants are responsible for repulsion or growth inhibition. To evaluate the impact of P. syringae on the behavior of human pathogens, strains were inoculated in close proximity to P. syringae and swarming motility was observed. Results show that P. syringae repelled all but two gram-negative human pathogens tested but did not impact gram-positive human pathogens. Repelled gram-negative strains were also challenged with purified syringafactin and rhamnolipid. Results demonstrate that syringafactin plays a role in observed repulsion. Preliminary studies suggest syringafactin may also inhibit the survival of gram-negative human pathogens. Collectively, this work supports the proposal that P. syringae utilizes biosurfactants to outcompete closely related bacteria. Due to the potential antimicrobial properties possessed by syringafactin, P. syringae could be used to make antibiotics that attack gram-negative bacteria that live in competition with P. syringae.
Emily Falcon, University of Central Oklahoma
1:15-3:15 in LSR 4
Documenting the Expansion of an Invasion of Mediterranean Geckos (Hemidactylus turcicus) at the University of Central Oklahoma and its Surrounding Area
Emily C. Falcon, Samah Houmam, Gary D. Thomas, Laura B. Kimmel, and Allyson M. Fenwick
The Mediterranean gecko (Hemidactylus turcicus) is an exotic, nocturnal species characterized by slow dispersal which makes it a good model for studying invasions. These geckos were intentionally, repeatedly introduced to the University of Central Oklahoma (UCO) during 1963-1965 and 1985-1997. Surveys 2005–2010 and 2014–2018 documented the spread of geckos from seven to 30 buildings on campus, and six buildings off-campus in the surrounding community. Seven additional buildings on campus and one building off-campus were inspected but were uninhabited. We collected 213 tail tissue samples with a goal of having 20 samples from each building. We will go back to surveyed buildings where geckos were not observed during the fall. We will also survey new buildings. Based on genotyping of 16 previously published microsatellite loci, we found two subpopulations on and off campus. Using analyses with STRUCTURE and ARLEQUIN, we expect the buildings farther from the introduction site to cluster together, and to have more genetic differentiation compared to the source population. This project continues to monitor and document the geographic and genetic progress of a population of exotic species as it slowly expands. Data generated will help answer questions about other exotic and possibly harmful species and their adaptations to urban areas.
Shandis Fancher, University of Texas at Tyler
1:15-3:15 in LSR 4
Determining the Mechanism of Pseudomonas Elastase Induced Activation of EGFR
Shandis Fancher, Dustin DeLong, Dustin Patterson, and Ali Azghani
Pseudomonas aeruginosa is a Gram-negative bacterium which functions in an opportunistic manner in immunocompromised patients. Several toxins and proteases play a significant role in the pathogenesis of P. aeruginosa acute infections as well as in Cystic Fibrosis associated chronic lung inflammation. We have reported that elastase isolated from P. aeruginosa (PE) is an activator of inflammatory pathways in vitro. This study examines the mechanisms of PE-induced inflammatory responses of lung epithelial cells in an effort to determine a more effective treatment modality. Our working hypothesis for this project is that PE activates epithelial growth factor receptors (EGFR) indirectly through activation of a secondary soluble ligand. Indirect interactions between PE and EGFR expressed on human pulmonary epithelial cell line A549 were analyzed using Virus Like Particle (VLP) encapsulated elastase. Cells were maintained in DMEM/F-12 with 10% FBS and 1% antibiotic-antimycotic solution. For experiments, cells were grown to confluence in 6-well plates, semi-starved in 2% FBS overnight, and starved in HBSS for 2 hours before treatment. Target treatments included commercial PE, recombinant PE (rPE), and rPE encapsulated in Virus Like Particles (VLP/rPE) constructed in the lab. Cells were treated with HBSS and empty VLP as negative controls and EGF as a positive control. Cells were lysed post-treatment for protein extraction and concentration determination, and SDS-PAGE analysis. Western blots were performed using primary antibodies targeting phosphorylated EGFR. We found that EGF, free PE, and rPE activate EGFR while encapsulated PE failed to do so. Our preliminary data indicates that PE activates EGFR by direct ligand/receptor interactions, the nature of which will be explored in successive studies.
Yesica Garcia, Southwestern Oklahoma State University
1:15-3:15 in LSR 3
Isolating Rhizobacteria for Promoting Growth of Winter Wheat
Yesica Garcia and Regina McGrane
Increasing crop yields is of critical importance for feeding an ever-growing population. Plant growth promoting rhizobacteria (PGPR) have been found to increase crop yield in various ways including inducing systemic resistance, regulating plant-growth promoting hormones, and increasing nutrient intake by assimilating nutrients into useable forms. PGPR are also appealing because of the smaller amounts needed to induce benefits compared to synthetic fertilizers. Winter wheat is a major crop in Oklahoma; more than five million acres were planted across the state in 2011. It also serves as pasture for the production of millions of pounds of beef. Currently, approximately two pounds of nitrogen are used to produce one bushel of profitable winter wheat yields. In an attempt to positively impact wheat yields, we sought to isolate PGPR from soil in the Weatherford, Oklahoma area. The objective of the experiment was to identify PGPR that most evidently increased winter wheat yield. Soil samples were collected, and bacteria were isolated via serial dilution plating. The isolated strains were then introduced onto the winter wheat seeds, plants were grown for two weeks, and yield was quantified by measuring total fresh weight, stem and shoot length, and stem and shoot weight. Plants with potential PGPR were compared to control plants that were not inoculated. In this work, eighteen strains were tested; none significantly increased the measured indicators of yield. However, we observed that some plants exhibited greener pigmentation and greater leaf number, suggesting the rhizobacteria may induce alternative health benefits. In future experiments, these phenotypes will also be quantified, and rhizobacteria will be isolated from a broader variety of soil types. Collectively, this work aims to identify rhizobacteria for use in agriculture with the purpose of benefiting crop yields.
Selena Gomez, Oral Roberts University
10:00-10:15 in LSR
Canine diets in the digital age: an examination of dog owner’s feeding practices and external signs of health in dogs
Selena Gomez
Dogs have been known as man’s best friend for years, they serve practical roles in society such as police dogs and service animals but more commonly serve as family members. As dogs transitioned from pets to family members, the feeding practices of owners changed from table scraps to commercial dog food. The popularity of commercial dog food increased because dog nutrition was considered a science best left for the experts, marketing strategies used by dog food companies, and convenient packaging and storage of dog food. Today most dogs eat commercial dog foods which contain by-products and have been recalled due to chemical contamination and toxic nutrient amounts which have led to debate about their nutritional quality and increased the prevalence of alternative diets. A survey was administered to dog owners in a convenience sample to explore the dog’s role in the household, owners’ feeding practices, factors influencing owners’ choice in dog food, and external signs of health (coat, skin, teeth, energy, stool, and body condition) in dogs. This study found that the majority of owners considered their dog family. Most owners fed commercial dog food and many fed diets had aspects of alternative dog foods including real meat as the first ingredient, no by-products, no grains, and all natural ingredients. There was no significant difference (P > 0.05) between dog food manufacturers and their effects on the external signs of health. Also, there was no significant difference (P > 0.05) between the dog owners’ blind estimation of body condition score and open body condition score estimation.
Constance Green, East Central University
1:15-3:15 in TXH 1
Finding Novel Antibiotic Producing Bacteria
Constance Green and April Nesbit
The demand for new antibiotics is extremely high due to an increase in antibiotic-resistant bacteria. This is causing common infections to become untreatable. Antibiotics are naturally occurring chemicals secreted from bacteria or fungi that kill other microbes. Thus, antibiotic producing bacteria are typically found in locations with high levels of competition with other microbes, such as the soil. Our goal is to use soil samples to find bacteria that produce previously unknown antibiotics in the hopes of postponing a post-antibiotic era. After testing sixteen sites for potential antibiotic-producing bacteria, we have found four isolates, which show inhibition of Enterococcus faecium with some also inhibiting Escherichia coli. Identification of these isolates using staining, biochemical testing, and 16S sequencing indicates that they are all Gram positive, endospore-forming bacilli in the Genus Bacillus. Future studies are to determine the species and isolate the secreted chemical causing inhibition of E. faecium and E. coli.
Virginia Green, University of Dallas
1:15-3:15 in LSR 4
Mortality during Pseudomonas aeruginosa chronic infection of Drosophila melanogaster dependent on mating experience of host
Virginia Green, Eli Wallace, Deanna M. Soper, and William L. Cody
Evolutionary trade-offs are extremely significant in understanding why certain phenotypes evolve and persist in different species. Such a trade-off may exist between reproduction and bacterial resistance in Drosophila melanogaster. Recent studies suggest that reproduction may negatively affect immunity, which in turn influences the coevolution of virulence in pathogens such as Pseudomonas aeruginosa. Drosophila melanogaster males are known to produce seminal proteins that reduce female life span, and studies suggest that these proteins may reduce females’ ability to resist acute infection. Characterization of this trade- off will further illuminate the immunological cost of reproduction in fruit flies, leading to a more complete understanding of the coevolutionary dynamics of bacterial virulence and host resistance. While decreased immune function following mating has been noted previously, acute models of infection using thoracic puncture have traditionally been used rather than the chronic oral route. Given the influence of route of infection on host immune response, a thorough analysis of the effects of sexual experience must examine whether induced immunodeficiency is also observed against chronic infection. In this study, we orally infected groups of virgin and experienced flies with P. aeruginosa and compared mortality rates to determine whether either group displayed increased resistance to infection. Experienced female flies showed increased mortality after chronic infection compared to virgin females. These observations suggest that the act of mating reduces female flies’ ability to resist oral bacterial infection, possibly as a result of exposure to seminal proteins. These findings may shed light on the coevolutionary past of host-pathogen pairs and suggest that coevolution infection models may need to be expanded to account for such factors as host experience.
Joselyn Gutierrez, West Texas A&M University
10:00-10:15 in TXH
The Effects of Summer and Winter Burning on Herpetofauna in a Sand Sage Prairie Ecosystem
Joselyn M. Gutierrez, Richard T. Kazmaier, and Matthew W. Poole
Fire is a natural component in prairie ecosystems, and controlled burning is one of the most common tools used to rehabilitate prairie habitat. However, most research addressing impacts of prescribed fire have focused on short-term responses, and most studies that have addressed the impact of fire on herpetofauna have focused on burning that is conducted during the dormant season. From 2004– 2009, we explored short-term responses to seasonality of fire on small vertebrates in a sandsage prairie ecosystem. In 2018, we reinitiated research in the same study plots to begin evaluating longer-term effects in the system. Drift fence arrays with pitfall traps were used to sample herpetofauna in a randomized complete block design with 5 blocks and 3 treatments (summer-burned, winter-burned, and unburned) during early and late season sampling periods. We regressed year of sampling against abundances and community metrics to evaluate linear responses to the different burning regimes over this time period. In the early season, green toads tended to increase over time with winter-burning (P = 0.095), but six-lined racerunners, plains narrowmouths, and most community metrics tended to increase over time with summer-burning (P < 0.092). Both six-lined racerunners and plains blind snakes increased over time in both burning treatments (P < 0.004), while both species diversity and species richness trended to slightly increase over time with winter-burning (P < 0.101) in the late season. Trends detected were generally weak, perhaps because of a long-term severe drought that has persisted in the region and could be masking treatment effects. Continued monitoring of these populations should elucidate treatment effects. However, our results suggest that summer-burning may be more beneficial to herpetofaunal communities than other treatments.
Brendan Harrison, Southwestern Oklahoma State University
1:15-3:15 in LSR 4
How sex ratios & population density affect sexual conflict over pairing duration in Hyalella amphipods
Brendan Harrison and Rickey Cothran
Research on sexual conflict, i.e. different evolutionary interests between the sexes, has exploded over the past few decades, but we know relatively little about how demographics affect sexual conflict in nature. Sex ratio is a demographic parameter that can affect sexual conflict through male-female encounter rates, which in-turn affects harassment experienced by females. Population density can have similar effects on female-male encounter rates. Under such scenarios the energy necessary to resist male harassment may alter the outcome of sexual conflict over mating. We tested the effects of sex ratio and density on pairing duration in a freshwater amphipod in the genus Hyalella. Males prefer longer pairing durations than females because pairing allows males to monopolize access to receptive females. While pairing for long periods is costly for both sexes (e.g., decreased foraging efficiency and increased predation risk), the net cost is higher for females because they do not reap the benefits of increased mating success. We hypothesized that females would experience longer pairing durations in male-biased and denser populations. To test these hypotheses, we set up populations that varied in sex ratio (female-biased, male-biased, or equal sex ratios) and population density (high or low). We discovered that guarding duration increased significantly from female-biased treatments to equal sex ratio treatments and again from equal sex ratio treatments to male-biased treatments, while no significant difference was found between low and high density treatments. These results indicate that density has no effect on guarding duration whereas sex ratio does. In nature Hyalella populations are usually female-biased because males are more likely to be eaten by predators. Our results suggest that changes in predator populations could upset the natural female-bias of these populations and increase the intensity of conflict, which would have cascading effects on population health by lowering female fitness.
Emily Hernandez and Elizabeth Gwartney, Oklahoma City University
1:15-3:15 in TXH 1
The Enemy of my Enemy is my Friend: Mycobacteriophage and the Treatment of Drug-Resistant Tuberculosis
Emily Hernandez, Loren Matrone, Elizabeth Gwartney, and Greg Mullen
Drug Resistant Tuberculosis (TB) is a worldwide health crisis being faced today. According to the World Health Organization, roughly 240,000 people died from drug resistant TB in 2016 alone. In response to this global emergency, new methods of treatment are being developed and tested in the lab setting, one of which being by Mycobacteriophage. Mycobacteriophage are viruses that infect mycobacteria and spread through internal reproduction. This form of therapy will include upwards of four or five different phages to avoid issues with resistance, which calls for identification of unique types of phage that enter the bacteria through different pathways. The goal of our study is to characterize OCU1 and OCU2 phage by cloning portions of their genomes for sequencing and by also identifying possible mechanisms of resistance by isolating phage-resistant mycobacteria. We have high hopes that the research being done within the OCU Biology department will leave a mark on the journey to identifying a solution to drug resistant TB.
Fiorella Hernandez, St. Edward's University
9:15-9:30 in DBU
Examining unc-33 expression using GFP transcriptional reporter
Fiorella Hernandez, Hailey Trombley, and Andrea Holgado
An emerging avenue of research aimed towards the development of therapies for Alzheimer’s Disease (AD) includes the investigation of a Microtubule Associated Protein (MAP) called CRMP2. Recent studies demonstrate that animal models for Alzheimer’s disease have increased levels of inactivated CRMP2. Besides its role in Alzheimer’s Disease, CRMP2 and its C. elegans orthologue UNC-33 were shown to control axonal elongation and microtubule assembly. Additionally, current research suggests that CRMP2/UNC-33 functionality can be regulated by autophagy, the engulfing of cytoplasmic elements into the autophagolysosome for degradation. To study the latter, we began the characterization of a transgenic C. elegans strain that expresses GFP under the regulation of an unc-33 promoter. The first steps of this characterization consisted in the sequencing of the transgene integrated in these nematodes. Secondly, we determined the expression pattern of GFP in intact nematodes throughout their life cycle. Lastly, we investigated colocalization of the GFP reporter with known cellular markers. Findings from these studies include: the sequencing of a 2,700 base pair promoter that maps to the 5’ UTR of the unc-33 gene; a neuronal localization of the GFP reporter at all stages of the C. elegans life cycle; and a distinctive cellular expression pattern in nematodes with enhanced autophagy. These findings corroborate previous research and provide further evidence for the autophagy-based regulation of the expression profile of CRMP2/unc-33.
Fernanda Hernandez Sanchez, University of the Ozarks
10:45-11:00 in DBU
Novel Approaches for Investigating Cancer Growth Inhibition and Male Contraception by DEC-TEC and PROTAC
Fernanda Hernández-Sánchez, Zhuang Jin, and Martin M. Matzuk
The bromodomain protein 4 (BRD4) and bromodomain testis-specific protein (BRDT) are attractive therapeutic targets for cancer and male contraception, respectively. In this work, a series of dual inhibitors for BRD4 and BRDT were designed and synthesized based on DNA-Encoded Chemistry Technology (DEC-TEC) and proteolysis targeting chimera (PROTAC). In vitro activities of the inhibitors on the bromodomain 1 (BD1) and the bromodomain 2 (BD2) of the two protein targets (BRD4 and BRDT) were determined by the alphascreen assay. Cell growth inhibitory activities of the inhibitors on cell MV4;11 were also evaluated.
Minh-Nguyet Hoang, University of Dallas
10:30-10:45 in DBU
Novel Glial Cells Affect Sensitivity to Nociception in Third Instar Larvae of Drosophila melanogaster
Minh-Nguyet Hoang, Justin Patawaran, Julia Krause, Catherine Hand, Deanna M. Soper, and Drew Stenesen
Pain is a sensory stimulus with a variety of therapeutic applications. Several genes involved in nociception within Drosophila melanogaster have been identified. Beta-alanine transporter (BalaT) is known to function in glial cells supporting the adult fly visual system; however, its role in larvae has remains unexplored. Here we identify novel BalaT expressing cells at tracheal branches points of wandering third instar larvae. Sensory neurons extend processes along tracheal branches and affect the organism’s response to pain. To examine the role of these new bilateral tracheal (BLT) cells in nociception, we performed temperature-response assays with BalaT loss-of-function mutants. Our data demonstrates an increased sensitivity to pain in BalaT mutants relative to wild type controls. From our current findings, we propose that BLT cells are glial and function in support of sensory neurons transmitting information related to painful stimuli.
Joshua Holsing, Oral Roberts University
11:30-11:45 in DBU
Antimicrobial Effects of Electrically Modified Lyophilic Silver Hydrosol in vitro and in vivo
Joshua Holsing
Silver has been known for many medicinal uses in the forms of colloidal silver and silver nanoparticles, both of which pose possible health risks when improperly applied. Silver in the form of an electrically modified hydrosol bypasses the adverse effects of prior forms, yet still maintains potent antimicrobial properties. This study focuses on the efficacy of this electrically modified silver hydrosol against A. faecalis, E. coli, MRSA, P. aeruginosa and S. epidermidis by utilizing a modified Kirby-Bauer disk diffusion method to measure the zone of inhibition (ZOI) against the concentration of the hydrosol; and against S. aureus, S. epidermidis, and P. aeruginosa by measuring dosage contingent percent decrease on live epithelial tissue to determine if efficacy is the result of concentration or dosage based treatment. The concentration dependent test results concluded statistically significant ZOI (all p < 0.01), yet all but S. epidermidis treatments failed post hoc test significance. The dosage dependent test concluded a near total eradication of cultured bacteria up to a 100% decrease in bacterial growth (p < 0.0463) with no change between treated and untreated epithelial cells. These results indicate safe usage of the silver hydrosol in vivo and support the hypothesis of high efficacy dose dependent treatment on multiple nosocomial gram negative and gram positive bacterial strains. Possible future research could fixate on the silver hydrosol’s interaction within a living system with exhaustive testing on the hepatocytes for presence of silver.
Aubrey Howard, West Texas A&M University
1:15-3:15 in TXH 1
Microscopic Characterization of the Aeroallergen and Reduction in Reduction Index on Using Air Purifiers With AHPCO and Plasma Nanotechnology
Aubrey Howard, Nabarun Ghosh, Jon Bennert, and Jeff Bennert
Allergies are a hypersensitivity of the immune system to normally benign substances. The Panhandle of Texas sees diagnosis rates typically double that of the state average. Seasonal allergies can have a strong impact on the quality of everyday life due to the presence of aeroallergens in the air such as pollen. In order to aid diagnosis of seasonal allergies and characterize the aeroallergens in the Texas Panhandle, research must be conducted to establish a foundation for determining the trends of data and provide a reference for identification of allergenic weed species. Similar studies have been conducted, however they have not included native angiosperms specific to the Texas Panhandle, making it more difficult for individuals to identify plant origins of pollen. In order to address this concern, a set of observations must be made analyzing the morphological characterization of pollen structure to aid in diagnosis and establish a reference for researchers for identification of pollen grains to the plant they have originated. We have recorded the GPS location, collected the whole plant where possible for making herbaria and the flowers for the pollen analysis. Flowering plants have been collected and their pollen analyzed and characterized. Micrographs were taken using an Olympus DP74 digital camera attached to a Olympus BX40 microscope and analyzed using cellSense digital software. The most significant aeroallergens recorded were the pollens like grass pollen (Poaceae), Short Ragweed (Ambrosia artemisiifolia), Common Sunflower (Helianthus annuus), Hairy Sunflower (Helianthus hirsutus), Silverleaf Nightshade (Solanum elaeagnifolium) and Lamb’s Quarters (Chenopodium album) as well as fungal spores like Alternaria, ascospores from Pezizales, Drechslera, Stachybotrys, Cladosporium and Curvularia. Further analyses of the gathered data are needed for prediction on the distribution of pollen and pollen seasons. We aim to incorporate the AHPCO and Plasma Nanotechnology for developing an advanced air purification system to reduce the indoor particulate matters including all forms of aeroallergen. This data collecting stage of research will help establish a foundation for future research and is the first crucial step for the hypothetico-deductive approach.
Nancy Huynh, Texas Wesleyan University
1:15-3:15 in LSR 4
The effect of active ingredients in mouthwash on Streptococcus sobrinus biofilm formation
Nancy Huynh and Christopher Parker
A collection of microorganisms in the human body can either be healthy or cause diseases depending on the bacterial organism. Microbes that adhere to surfaces in the mouth that form biofilms can negatively affect overall health. The microbes, Streptococcus mutans and Streptococcus sobrinus, may be associated with cavities and lead to “rampant” tooth decay. We tested the effect of ethanol in mouthwash on the formation of S. sobrinus biofilm. This research can provide insight into whether alcohol in mouthwash is effective on inhibiting S. sobrinus biofilm formation. In this study, S. sobrinus biofilm formation capabilities were investigated under varying levels of ethanol concentration in Listerine® Zero mouthwash. The results of the One-Way Analysis of Variance (ANOVA) statistical test showed that there was no difference between the varying levels of ethanol concentrations. Thus, ethanol levels in mouthwash may not inhibit S. sobrinus biofilm formation, and additional research is required to test the active ingredients from essential oils and antibacterial activities of mouthwashes.
Mary Kadatska, Oral Roberts University
9:15-9:30 in TXH
Effects of trinitrobenzene induced colitis on vasoactive intestinal peptide, calcitonin gene-related peptide and morphology of colon in rats
Mary Kadatska
The primary pain medications used today are opioid based. However, they have caused a decrease in life expectancy in years as recent as 2015-2017 leading to the opioid crisis because they are widely known to cause development of tolerance and to promote substance abuse disorders with opioids as well as other drugs, (Koenig 2018, Dai et al. 2018). These circumstances call for a search for a replacement for opioid medications. In this study, we looked primarily at vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP), and at basic morphology of the epithelial lining of the intestinal mucosa. VIP is known to have certain motile, secretory, and anti-inflammatory functions in the colon (Del Valle-Pinero et al. 2015, Abad et al. 2005). CGRP is known to have vasodilatory, nociceptive, secretory, and motor functions which can be linked to pain and inflammation (Cottrell et al. 2012). The experimental rat was treated with 2,4,6-trinitrobenzene sulfonic acid (TNBS), and its CGRP/VIP colon levels were measured by measuring mean grey intensity (MGI) of antibody stains particular to CGRP and VIP and compared with naïve and control (ethanol treated) colons. Our results showed a significant increase in VIP MGI per nerve fiber (p0.05).
Jack Koszykowski, Oral Roberts University
10:30-10:45 in LSR
The Effects of Azadirachta indica (neem) Extract on RP62A Staphylococcus epidermidis Biofilm Formation
Jack Koszykowski
Biofilms are dense microbial communities of bacteria. In biofilms, bacteria possess a higher defense than their planktonic forms. Within these close-knit communities, bacteria transformation is abundant due to a communication process referred to as quorum sensing. Because of this, bacteria within biofilms often possess antibiotic resistance traits. Due to the increased pathogenicity of the bacteria, biofilms form a severe threat to humans with indwelling medical devices on which biofilms can form. Thus, the search for novel compounds that can inhibit or deter biofilm growth has been a frequented area of study. In this study, varying concentrations of leaf extracts from Azadirachta indica, or neem, were tested on RP62A, a biofilm forming strain of Staphylococcus epidermidis. Neem was chosen due to its wide variety of medicinal properties. The goal was to inhibit biofilm formation. The Crystal Violet Assay used to determine biofilm levels showed that as the concentrations of neem increased the absorption reading increased significantly. The dead/alive assay used to determine bacterial viability showed that as the concentration of neem increased, bacteria viability decreased, with the exception of neem that wasn’t dissolved in DMSO. The results were largely inconclusive. While the neem showed to possess antibacterial activity, the assay used to determine biofilm levels did not yield any definitive data due to the characters of the neem within the wells of the 96-well microtiter plate. The assay to determine bacteria viability showed that 1/2 concentration of neem had the highest effectiveness in preventing both biofilm and planktonic growth.
BreAnna Le, Southwestern Oklahoma State University
1:15-3:15 in TXH 1
Investigating effects of innate immune agonists on co-receptor B7-H4 in B lymphocytes
BreAnna Le and Christopher G. Horton
The immune system depends on communication from innate immune cells, which act quickly upon pathogenic microbes through stimulation of their pattern recognition receptors, to adaptive immune cells, which respond accordingly to signals such as cytokines and cell surface proteins (co-receptors). Co-receptors can help activate or inhibit the functions of the adaptive immune response and can also lead to specific adaptive immune cell differentiation. B7-H4 is predominantly viewed as an inhibitory co-receptor expressed in non-lymphoid tissues, especially among tumors where it functions to impair anti-tumor immunity. We have observed a mild, agonist-specific increase in B7-H4 expression in mouse primary B cells in previous studies. From these results, we hypothesized that exposing B lymphocytes to different degrees of pathogen-like stimuli will alter the expression of B7-H4. In our experiments, while maintaining actively growing cultures of an A20 cell line, we stimulated B lymphocytes with varying concentrations of imiquimod, LPS, DMXAA, and polyIC for 6-72 hours, and observed their expression of B7-H4 by flow cytometry. Our results demonstrated that stimulation with these agonists, particularly imiquimod, LPS, and DMXAA after a 72 hour incubation, lead to significant alterations in B7-H4’s expression, while cells stimulated with polyIC showed no significant expression when compared to unstimulated cells. Although all three concentrations used for imiquimod, LPS, and DMXAA on average showed significance, the most dramatic effects were observed using imiquimod. Future studies such as using qRT-PCR to insure the presence of the gene will be needed for more in-depth characterization, but our results thus far begin to shed light on the possible mechanism of how B7-H4 expression is initiated. Most importantly, B7-H4 studies such as these can lead to possible cancer/tumor treatments and vaccinations, in which inhibiting B7-H4’s expression will hinder its interference with anti-tumor immunity.
Sidney Lyons, East Central University
1:15-3:15 in TXH 2
The effect of Iron Oxide Nano particles on development, and fluctuating asymmetry of Zophobas morio larvae
Sidney Lyons, Randall Maples, and George Wang
Nanoparticles are used in a variety of ways to introduce large materials on the molecular level. We examined the potential effect of nano particles on darkling beetle (Zophobas morio) larvae development. We hypothesized that the iron oxide nano particles would stunt the growth of the beetle larvae, and cause fluctuating asymmetry in the antenna and limbs of adults. We believe this because fluctuating asymmetry is an indicator of stress on an organism. We reared a total of 40 Z. morio larvae divided into two groups. We fed the larvae in the treatment group carrots coated with a small amount of iron oxide nano particles, while larvae in the control group were fed organic carrots. The larvae were allowed to pupate and enclose in the lab. We recorded the mass and length of the pupae, and measured the antenna length, and leg length of adults.
Monica A. Martinez, Austin College
10:00-10:15 in DBU
Estradiol Effects on Hibernating Physiology of Ground Squirrels
Monica A. Vania Martinez and Jessica Healy
Estradiol is a sex hormone with anorexigenic effects. Previous literature reports that exogenous estradiol reduces food intake and body fat in non-hibernating rodents, yet few studies confirm whether this pattern is applicable to the Thirteen-Lined Ground Squirrel (Ictidomys tridecemlineatus). In 2018, 5 individuals were implanted with capsules that contained either vehicle (control) or exogenous estradiol (Est+) and food intake, body composition, and metabolic rate were measured during the summer active period. Preliminary analysis suggests that Est+ animals may have increased metabolic rate compared with controls during the implant period, but it is unknown if estradiol implantation has long term effects on torpid metabolic rate. Individuals were aroused from an initial torpor at 18, 15, and 5°C and their oxygen consumption was recorded as a proxy for metabolic rate. Observing the effects of exogenous estradiol on torpid metabolic rate will give more insight into the physiological mechanisms underlying energy balance and allocation within Ictidomys tridecemlineatus.
Susan McGuire, Texas A&M University - Corpus Christi
1:15-3:15 in TXH 2
Investigating Nutrient and Organic Carbon Distribution in Wetland Ecosystems
Susan McGuire, Rachel Weisend, and Brandi Kiel Reese
Wetlands provide many ecosystem services for both animals and humans. Wetland communities in the Gulf Coast are changing as mangroves are expanding throughout the region and into previous northern limits. This project will investigate the carbon and nutrient distribution within the sediment of the three vegetation types (mangrove-dominated, Spartina-dominated, and seagrass-dominated) that can be found within coastal wetlands. We measured nitrate, nitrite, ammonium and carbon in order to understand the changes in geochemical signals and sediment properties with respect to depth, seasonality, and vegetation in a wetland system. This will help understand ecosystem shifts across a spatial and temporal scale. We found that there were no significant differences between different seasons within the same vegetation type. However, mangrove-dominated sediment was significantly different from Spartina-dominated sediment (p value < 0.05) and seagrass-dominated sediment (p value < 0.05), but when comparing Spartina-dominated sediment to seagrass-dominated sediment there was no significant difference. Also, all November samples were significantly different than all August samples (p value < 0.05), but there was no significant difference when comparing November samples to June sample, or June to August. This information will help us asses changes in macrofauna, meiofauna, and the microbial community within a wetland ecosystem.
Shayla Miller, Southwestern Oklahoma State University
1:15-3:15 in TXH 2
The Battle of the Sexes: How female condition affects conflict over mate guarding duration in Hyalella amphipods
Shayla Miller and Rickey Cothran
Sexual conflict, different evolutionary interests between females and males, is common in nature and is expected to shape the evolution of mating traits and affect the productivity of populations. Most research has focused on male “offense” traits with relatively less research on female “defense” traits. We explored how female condition affects conflict over mate guarding duration in a freshwater amphipod species in the genus Hyalella. We hypothesized that females in poor condition would experience longer mate guarding periods because they are unable to resist male pairing attempts. We tested this hypothesis by varying the amount of spirulina, a high nutrient protein powder, in female’s diets. Females received either a high-quality diet or a low quality-diet. Females from each diet treatment were then exposed to either one or three males representing a low and high harassment treatment, respectively. We found a significant effect of harassment treatment but no significant effect of female diet on pairing duration. It is possible that our diet treatment did not significantly affect female condition or other female traits, e.g., morphological traits are important in resolving conflict over pairing duration. Our results show that harassment levels do have an effect on pairing duration. Longer pairing durations could affect female amphipod fecundity by reducing foraging efficiency, which could have a negative impact on the overall health of an amphipod population.
Taylor Miller, Oral Roberts University
11:45-12:00 in DBU
Anti-cancer effects of compounds secreted by Flammulina velutipes
Taylor Miller
Compounds collected from various mushroom species, including Flammulina velutipes, have been shown to have immunomodulatory and antitumor effects (Seklic et al. 2016). This experiment was performed to investigate the cytotoxic activity of F. velutipes secretions on MCF10A breast epithelial cells and MDA MB 468 breast adenocarcinoma cells. It was hypothesized that there would be a significant difference in cell viability between the two cell lines when treated with the mushroom secretions. The first preliminary cytotoxicity assay revealed a decrease in cell viability in both the MCF10A cells and in the MDA MB 468 cells. A student’s t-test also revealed a significant difference in cell viability between the two populations, the tumorigenic cells having the higher average percent viability. The next cytotoxicity assay measured the average percent viability of the cell lines at varying concentrations. A significant difference was found at 1X and 0.5X concentrations, the highest difference being at the 0.5X concentration. The third and final cytotoxicity assay tested only the 0.5X concentration of compound on the cell lines. This showed that the non-tumorigenic MCF10A cells had a significantly higher average cell viability percentage than the tumorigenic MDA MB 468 cells. The results support the hypothesis that F. velutipes secretions exhibit cytotoxic activity in cancer cells. There is evidence to support the idea of selective cytotoxicity between normal epithelial tissue and cancerous tissue.
Julio Molina-Pineda, University of the Ozarks
1:15-3:15 in TXH 1
Antioxidant Carbon Nanoparticle Results in Novel Auditory Response in Mice
Julio A. Molina-Pineda and Fred A. Pereira
Deafness is a common side effect in people undergoing chemotherapeutic treatment with Cisplatin, a chemical proven to be ototoxic due to its oxidative damage that results in disturbance of the endocochlear potential. An antioxidant carbon nanoparticle has been found to protect the ear from such damage and to enhance auditory response as well. Two experiments were performed: one in which the nanoparticle dosage and its effects were investigated by studying the effect of three continuous doses. And a second experiment in which the inner ear and brain’s potassium channels mRNA expression was compared between nanoparticle treated and untreated mice. Preliminary findings suggest that the nanoparticle increases and delays auditory response, and that higher dosage helps in maintaining the enhancement rather than increasing it. Furthermore, the mRNA analysis suggests a possible mechanism for the enhancement caused by the nanoparticle.
Julio Molina-Pineda, University of the Ozarks
11:15-11:30 in LSR
Maternal Commensal Bacteria Inhibits Intrauterine ZIKV Infection and Fetal Growth Restriction in a Mouse Model
Julio Molina-Pineda, Maxim Seferovic, Gregory Valentine, and Kjersti Aagaard
ZIKV has been associated with congenital infection, microcephaly, and pregnancy loss due to viral mutations that facilitate maternal to fetal viral spread. Commensal bacterial microbes have been demonstrated to modulate host immunology and viral infection. We therefore hypothesized that commensal bacteria would influence maternal to fetal transmission of ZIKA virus. Employing a mouse model of Zika infection, we compared ZIKV transmission rate and pup infectivity between normal conventional (C) and germ free (GF) gnotobiotic mice. Pregnant Swiss-Webster mice were given four doses of 1x104 PFU of a contemporaneous ZIKA strain (ZIKV) or a mock injection (M) on embryonic days 4 through 7. Conventionalized (GF exposed to bacteria at weaning) or immune inhibiting anti-IFNI1 antibody were negative and positive controls, respectively. Tissues were recovered at e18.5 just prior to delivery and TRIzol extraction performed for genomic viral RNA. TaqMan based qPCR was performed from cDNA and quantified against ZIKV standard curves. ZIKA infected placentas were positive in 82% of demised fetus compared to 52% of live pups (p=0.04). No virus was found in either maternal or fetal brains, however, virus was recovered from maternal spleen and uterus, and was highest by an order of magnitude in fetal placental tissue overall.
Nicole Mountz, Oral Roberts University
9:30-9:45 in TXH
Cytotoxic Effects of Inonotus obliquus Secretions on Non-Tumor and Tumor Cells
Nicole Mountz
The recent development of targeted cancer therapies allows for treatment that is selective of molecular expressions unique to cancer cells. Extractions of Inonotus obliquus have shown antineoplastic properties that could provide naturally-derived targeted therapies with reduced harm to normal cells. This study aimed to investigate the cytotoxic activity of I. obliquus secretions against the proliferation of human breast carcinoma cells and compared to normal human mammary epithelial cells. I. obliquus mycelia were grown and passaged on a potato/dextrose/yeast agar supplemented with woodchips (20 ºC). Secretions were harvested from mycelia with Opti-MEM cell medium (48 hours; 37 ºC; 0% CO2), purified and stored at 4 °C. Purified secretions were tested on MDA-MB-468 cells and MCF 10A cells with an MTT cytotoxicity assay (24 hours; 5% CO2; 37 ºC). Absorbance values were measured at a ratio of OD570 to OD605 and used to calculate cell viability percentage. MTT cytotoxicity assay results showed a significantly greater cell viability of normal mammary epithelial cells than of breast carcinoma cells after 24 hours of exposure (22.1% vs. 2.26% respectively; P < 0.01). Results indicate the presence of cancer-specific cytotoxic activity within I. obliquus secretions. Furthermore, these results suggest the potential use of I. obliquus secretions as a cancer-targeted treatment.
McKayla Muse, University of Central Oklahoma
11:30-11:45 in LSR
Analysis of Proliferation and Migration in Phenylalanine, Retinoic Acid, and 4-diethylaminobenzaldehyde Treated Cells
McKayla Muse, Kayley Pate, Gabriel Rucci, and Nikki J Seagraves
Maternal phenylketonuria [MPKU] is a syndrome of multiple congenital anomalies including cardiovascular malformations [CVMs], and brain and growth restriction when a mother with Phenylketonuria [PKU] does not control her dietary intake of Phenylalanine [Phe]. However, the mechanisms responsible for Phe-induced CVMs are poorly understood. Our lab has preliminary evidence that high levels of Phe could inhibit Retinoic Acid [RA] signaling, which typically promotes the expression of genes such as proliferation, migration, and differentiation. Proliferation and migration of the neural crest cells are important in formation of the outflow tract (OFT) and aortic arch arteries (AAA). We hypothesize that Phe inhibits migration and proliferation, which may contribute to the defects seen in MPKU. We also looked at the effects of exposure to RA and 4-diethylaminobenzaldehyde [DEAB], a known RA inhibitor. We conducted in-vitro proliferation and migration assays on several cell types to determine if proliferation and migration was affected by Phe, RA, and DEAB exposure. Images were analyzed with ImageJ and GraphPad Prism. Present research suggests that Phe exposure causes a significant decrease in proliferation of cells. It is also shown that RA increases or does not affect proliferation, and that DEAB decreases cell proliferation. In this way, Phe is similar to DEAB, which suggests that it also acts as an RA inhibitor. This could contribute to the CVMs observed in MPKU. The results of migration studies showed that Phe acted similar to DEAB, where the rate of migration was slower than that of RA, which is known to increase migration. This suggests that Phe may act as an inhibitor of migration. This is significant because it eludes to a potential mechanism that Phe could affect RA signaling, thus resulting in the types of defects observed in human MPKU.
Lois Nguapa, Oral Roberts University
10:45-11:00 in LSR
Bioinformatic Prediction of Gene Function in Streptobacillus moniliformis
Lois Nguapa
Since the completion of the Human Genome Project, DNA sequencing technologies have made vast advancements in the sequencing of organismal genomes. Determining function, on the other hand, still proves challenging, as this requires the time-consuming investigation of a single gene and its protein product in vitro and in vivo. In the last couple of decades, the amount of genomic and proteomic data, filled with thousands of hypothetical genes, has increased to a large degree. This information, deposited in publicly available databases, can be used to predict gene function from a DNA sequence alone using online bioinformatics programs developed by the United States government and researchers around the globe. The purpose of this research is to predict the functions of hypothetical bacterial protein-coding genes that have been identified in the organism Streptobacillus moniliformis. Five hypothetical genes were analyzed from the Streptobacillus moniliformis DSM12112 genome: SMON_RS07405, SMON_RS07640, SMON_RS07645, SMON_RS07655, and SMON_RS07660. Various bioinformatic methods were applied to both the raw nucleotide sequence and amino acid sequences of these genes and compared against those of other well-characterized genes from other organisms. SMON_RS07405 was predicted to a methyltransferase, SMON_RS07640 and SMON_RS07645 were predicted to be ABC permeases, SMON_RS07655 was predicted to be an alpha/beta hydrolase, and SMON_RS07660 was predicted to be a tetratricopeptide (TPR) repeat. These findings cannot be confirmed until the proteins have been expressed in model organisms and tested in the laboratory biochemically.
Toi Nguyen, Texas Wesleyan University
1:15-3:15 in TXH 1
Identification and Distribution of Wolbachia in Natural Culex quinquefasciatus Populations in Tarrant County, TX.
Toi Nguyen
Wolbachia, the most prevalent genus of endosymbiont in invertebrates, has been utilized successfully as a biological control agent against mosquito-borne viruses. Therefore, knowing the distribution of Wolbachia in the natural mosquito populations is essential to implement the control programs. Culex quinquefasciatus is the primary vector transmitting West Nile Virus in Tarrant County, TX. In this project, polymerase chain reactions (PCRs) using general Wolbachia surface protein (wsp) primers were performed to detect Wolbachia in one hundred mosquito samples from the natural Cx. quinquefasciatus populations in six zip code areas of Tarrant County, TX. Overall, sixty-four mosquito samples were found infected with Wolbachia. Further contingency analysis using the estimated odds ratios and their 95% CI’s revealed that the natural Cx. quinquefasciatus population at one zip code (76051) was more likely to be detected with Wolbachia than at the other five zip codes. In addition, we will present the results from PCRs using group- and strain-specific wsp primers as well as phylogenetic analysis.
Sophia Ostertag, University of Dallas
1:15-3:15 in TXH 1
Coevolution of host-pathogen interactions in a Pseudomonas aeruginosa chronic infection model of Drosophila melanogaster
Sophia Ostertag, Sarah Tripplehorn, Natalie Sandlin, Ana Henriquez, Deanna Soper, and William Cody
Pseudomonas aeruginosa is an antibiotic resistant, opportunistic pathogen that causes both acute and chronic infections in a variety of hosts. P. aeruginosa is a major cause of acute human infections like UTIs, ventilator-associated pneumonia infections, and surgical site infections. Notably, chronic P. aeruginosa infections are the leading cause of morbidity and mortality in cystic fibrosis patients. The innate immune system in Drosophila melanogaster makes it a suitable host for examining these acute and chronic infections. This study will provide insight into the evolutionary response of antibiotic resistant bacterial strains, with the intent of understanding the implications of host-pathogen interactions. With repeated coexposure of these species, virulence, immunity, and reproductive traits alter in a dynamic coevolutionary relationship. The Red Queen hypothesis postulates that pathogenic species serve to increase the level of genetic diversity within a population. With pathogenic selection pressures, the host population may develop random mutations that allow for its survival under that environmental circumstance. The pathogenic population may similarly develop adaptive mutations. Three experimentally enforced evolutionary treatment groups will be used: host evolution, pathogen evolution, and coevolution. In the host evolution treatment group, successive generations of D. melanogaster are exposed to stock samples of PAO1. In the pathogen evolution treatment group, successive generations of lethal PAO1 are extracted from deceased flies and used to infect a stock population of D. melanogaster. In the coevolution treatment group, successive generations of offspring are infected with the lethal strain of PAO1, which is extracted from deceased flies of the previous generation. The variation in mortality among the host species will be characterized, with the goal to provide insight on the coevolutionary host-pathogen interaction, virulence factors, and the survivability of the host.
Amanda Pang, East Central University
1:15-3:15 in LSR 3
Growth Inhibition of Chaetomium globosum After Exposure to NSAIDs
Amanda Min Xing Pang, Leah Dudley, Hannah Taff, Tyler Rowsey, and Charles Biles.
Molds (fungi) are primary allergens in homes and businesses that cause a range of respiratory problems. Chaetomium globosum is one of the most common fungal species that causes water-damage in buildings. This research was conducted to determine the inhibitory activity of NSAIDs on C. globosum. Previous experiments indicated that low levels of NSAIDs inhibited growth of C. globosum when applied to sheetrock. Recent experiments have tested the hypothesis that NSAIDs enter the fungal cells and shut down respiratory channels of the mitochondria. This research was conducted in order to understand whether NSAID solutions must be present on the substrate before inoculation, or are able to penetrate the cell wall of existing spores and prevent germination. Subcultures of C. globosum ascospores were quantified using a hematocytometer with a light microscope, then placed in different concentrations of various NSAID solutions for 24 hours. The ascospores were washed twice by centrifugation at 15,000 rpm in sterile water. After washing, the ascospores were pipetted onto 9cm Petri media. In the initial experiment after the first centrifugation, 900 μL of the supernatant was removed and also placed on PDA media in 9cm Petri dishes. This was to determine if the ascospores were centrifuged to the bottom 100 μL of the solution. Data from the supernatant experiment showed only 1-3 ascospores were present in the extracted supernatant. Fungi undergo vacuolation under stress, which results in enlarged vacuoles in the hyphal cells. Fungi on control and treatment plates were observed under light microscopy. Acetylsalicylic acid (Aspirin) completely inhibit growth at 10 mM concentration after washing the ascospores. In contrast, Naproxen (Aleve™) displayed no inhibitory effects on C. globosum growth.
Kayle Patatanian, University of Central Oklahoma
1:15-3:15 in TXH 1
Analysis of cardiac teratogenicity of phenylalanine using the avian model: the role of the focal adhesion pathway Analysis of cardiac teratogenicity of phenylalanine using the avian model: the role of the focal adhesion pathway
Kayle Patatanian, Jamie Watson, and Nikki Seagraves
Maternal phenylketonuria (MPKU) is characterized by the teratogenic effects of phenylalanine (Phe). High levels of intrauterine Phe cause cardiovascular malformations, intellectual impairment, microcephaly, and low gestation mass. Mothers with PKU must maintain a strict diet to insure serum Phe concentrations remain below 360 µM to avoid these congenital defects. Previous functional genomic studies done by our lab have determined that the focal adhesion pathway (FAP) is significantly enriched under Phe treatment. Six genes associated with this pathway are variably expressed. Our project aims to visualize their expression during stages HH10, 12, and14 in chicken embryo heart and out flow tract tissue using in-situ hybridization. Hepatocyte growth factor is one of the six genes under investigation and has been shown to promote angiogenesis through its effects on cell survival, differentiation, and motility.
Blake Pate, Dallas Baptist University
9:30-9:45 in LSR
Concordance of HPV genotyping between histopathology and trigger cytology
C. Blake Pate
The human papillomavirus is the most sexually transmitted virus in the United States with over 14 million individuals being infected by the virus each year. The most oncogenic HPV genotypes, 16 and 18, give rise to over 70% of all cervical cancers, while other genotypes can cause genital warts and other problematic conditions. It is for this reason that research be conducted to improve the effectiveness the HPV vaccine and the current methods of genotyping in order to aid the public in the prevention of such a debilitating virus. In this study, HPV genotyping results acquired from clinical routine pap smear (cytological) results from patients enrolled in the HPV Vaccine Effectiveness Study at Yale University were compared to the biopsy (histopathological) results in order to determine the sensitivity, specificity, and accuracy of the cytological reports. The concordant analysis confirmed our hypothesis that the HPV genotyping from the trigger cytology report would be greater than 90% concordant with that obtained from the biopsy when collected within 12 months apart. These findings open the door for increasing interest in developing more accurate and dependable HPV genotyping techniques in cytological specimens to allow for greater ease in acquiring HPV test results. In addition, these results are currently allowing researchers in the HPV Vaccine Effectiveness Study to enroll patients into the study more rapidly using cytological genotyping results rather than the more time-intensive histopathological specimens. A prospective four year study that aims to confirm these findings with a larger number of specimens is now underway to establish HPV surveillance tools that are most cost and resource efficient.
Kayley Pate, University of Central Oklahoma
1:15-3:15 in TXH 2
Analysis of Proliferation and Migration in Phenylalanine, Retinoic Acid, and 4-diethylaminobenzaldehyde Treated Cells
Kayley Pate and Mckayla Muse
Maternal phenylketonuria [MPKU] is a syndrome of multiple congenital anomalies including cardiovascular malformations [CVMs], and brain and growth restriction when a mother with Phenylketonuria [PKU] does not control her dietary intake of Phenylalanine [Phe]. However, the mechanisms responsible for Phe-induced CVMs are poorly understood. Our lab has preliminary evidence that high levels of Phe could inhibit Retinoic Acid [RA] signaling, which typically promotes the expression of genes such as proliferation, migration, and differentiation. Proliferation and migration of the neural crest cells are important in formation of the outflow tract (OFT) and aortic arch arteries (AAA). We hypothesize that Phe inhibits migration and proliferation, which may contribute to the defects seen in MPKU. We also looked at the effects of exposure to RA and 4-diethylaminobenzaldehyde [DEAB], a known RA inhibitor. We conducted in-vitro proliferation and migration assays on several cell types to determine if proliferation and migration was affected by Phe, RA, and DEAB exposure. Images were analyzed with ImageJ and GraphPad Prism. Present research suggests that Phe exposure causes a significant decrease in proliferation of cells. It is also shown that RA increases or does not affect proliferation, and that DEAB decreases cell proliferation. In this way, Phe is similar to DEAB, which suggests that it also acts as an RA inhibitor. This could contribute to the CVMs observed in MPKU. The results of migration studies showed that Phe acted similar to DEAB, where the rate of migration was slower than that of RA, which is known to increase migration. This suggests that Phe may act as an inhibitor of migration. This is significant because it eludes to a potential mechanism that Phe could affect RA signaling, thus resulting in the types of defects observed in human MPKU.
Vincent Phan, University of Dallas
1:15-3:15 in TXH 2
Novel Therapeutics for Head and Neck Cancer
Vincent Q.D. Phan, Caroline R. Delma, and Mohan Natarajan
Head and Neck Cancer is one of the most potent cancers today, and treatment for it hasn’t improved over the last 2 decades. Head and Neck Squamous Cell Carcinoma (HNSCC) has a 5-year survival rate of 50%. This means that people may not even make it past 5 years after they put their cancer into remission. This is motivation for us to try to find better alternatives that can preserve life. We experimented with EC359, which is a pipeline drug that was made to treat mainly pancreatic cancer but can inhibit the group of Head and Neck Cancer. We also used the cancer cell lines HSC-3 (tongue cancer) and CAL-27 (salivary gland cancer) because they had the most metastatic potential against the body. We did several different in-vitro tests with EC359 against the cancer cells. We would grow cancer cells in T-flasks until we had enough cells to start experiments with. We did Fold Growth and MTT assays to see the cytotoxicity of EC359 against the cancer cell lines. We followed that with Fluorescence Microscopy to see whether apoptosis or necrosis was occurring within the cells. We followed this with western blotting to try to understand which apoptotic pathway EC359 took. Then we finished with a Scratch Assay to see if EC359 had any effect on the metastatic capabilities of CAL-27 and HSC-3. EC359 showed cytotoxicity in CAL-27 and HSC-3 at low Nano-molar range. EC359 showed the ability to suppress the viability of CAL-27 and HSC-3 cells and inhibited their proliferation. EC359 induced apoptosis in CAL-27 and HSC-3 cells by initiating the intrinsic apoptotic pathway. EC359 exhibited slight inhibition on the migration of CAL-27 but seemed ineffective against HSC-3 cells.
Jennifer Prophet, Southwestern Oklahoma State University
1:15-3:15 in LSR 3
Timing of Avian Migration Onset Through the Oklahoma City Area 1995-2017 Using NOAA Weather Data
Jennifer Prophet, Amy Hofeld, and Zach Jones
Avian migration timing and duration may be linked to large-scale climate patterns and reflect shifts in average global temperatures. The National Centers for Environmental Information (NCEI) provides an archive of NOAA weather radar data scans occurring every ten minutes dating back to 1995. Our primary data of interest are 1) reflectivity and 2) radial velocity. Aerial density and flight direction were collected for available years and compared to global temperatures to examine seasonal migration effects. We used KTLX (OKC) weather radar data to determine time, duration, intensity and direction of seasonal avian migrations. Fall migrations began in September and spring migrations in March between 1995-2017, while the global average temperature steadily increased by 2.358 ̊C. Fall migration trended toward later onset dates, while Spring migration trended toward earlier. Fall migration onset exhibits a stronger positive relationship with temperature than spring, potentially because inter-year March temperature patterns vary more than those of September. Ground truthing of radar data using eBird sightings revealed large-scale migratory events in Oklahoma during the second week of the fall migratory period. Comparatively, in spring migration there was a continuous migratory event occurring throughout the first 3 weeks of migration. This continuous rate over time may also contribute to the weaker relationship between migratory onset and temperature in the spring.
Rebecca Romero, West Texas A&M University
1:15-3:15 in TXH 2
Current aspects of PM 2.5 and analyzing the aeroallergen of the Texas panhandle
Rebecca Romero and Nabarun Ghosh
PM 2.5 stands for particulate matter 2.5, also called particulate pollution and is air pollutant particles not visible with the naked eye due to their microscopic size of 2.5 microns. Hazardous Aeroallergen particles of this size are dangerous because they pass the body’s defenses into the lungs and bloodstream. Hazardous PM 2.5 have the potential to lead to death. Data shows out of 6 people who die from air pollution, one person dies from PM 2.5. In this study, I’ll analyze PM 2.5 and Aeroallergens in the Texas panhandle and conduct studies on how these factors affect this area. One concern the Texas panhandle encounters is the air pollution coming from the feedlots here. Also, humans spend most of their time indoors; and using air purifiers inside can help maintain cleaner air indoors. I will use the BX Olympus Digital microscope to analyze the PM 2.5 and the aeroallergen that are prevalent in the Texas panhandle. I will follow the standard procedure to collect dust particles floating in the air as designed by AAAAI (American Academy of Allergy, Asthma, and Immunology). I will use clean glass slides on which I shall place double sticky tape and save them in a clean slide box. I will put double sided sticky tape on a clean glass slide and place the slide on the roof. I will allow the slides to be exposed to the air for 24 hours, 48 hours, and 72 hours. The prepared slides will be viewed using a BX 40 Digital Olympus Microscope. Being able to go cleaner indoors using the Advanced Hydrated Photo Catalytic Oxidation (AHPCO) Nanotechnology to purify the air and going greener outdoors, will assist the world in decreasing the steady incline of harmful air pollution and create a better environment for our health.
Ruqea Saheb, Texas Wesleyan University
1:15-3:15 in TXH 2
The Change in Oral Microflora in Response to Coffee Intake
Ruqea M. Saheb
The human microbiome presents a diverse array of microorganisms. Specifically, the human oral microflora is composed of 600 taxa at the species level. The distinct habitats of mouth influence the colonization of microorganisms due to varying environmental conditions such as pH levels, oxygen exposure, temperature, and nutrients. Therefore, the disruption of a specific colonized habitat may lead to disruption in the oral microflora. Coffee exerts antibacterial activity due to its rich composition of phenolic compounds, which has been studied to inhibit the growth of the common decay-causing bacteria, Streptococcus mutans. In this study, genomic DNA was collected from students at Texas Wesleyan University and represented coffee consumers and non-coffee consumers. The genomic DNA was sequenced in order to characterize the oral microflora based on the 16s rRNA sequence. We hypothesized that the oral microflora decreases in species richness and relative abundance in response to coffee intake. In addition, we hypothesized that there will be significant differences in metabolic pathways present in these two groups. Our preliminary findings support these hypotheses in that the two groups differ in species richness, species abundance, and metabolic pathways.
Anthony Sanchez, St. Edward's University
11:00-11:15 in TXH
UNC-33, a microtubule associated protein, plays a key role in neuronal autophagy
Anthony Sanchez and Andrea Holgado
Autophagy is a self-degradative mechanism used within cells to maintain homeostasis in recycling cellular waste and protein aggregates through an autophagosomal-lysosomal system. The regulation of autophagy has been implicated in axonal development via CRMP-2/UNC-33, a microtubule associated protein (MAP) that serves a key role in neuronal development. To better evaluate the relationship between CRMP-2/UNC-33 and autophagy, we studied autophagy in unc-33 mutants by monitoring autophagosome maturation and pathway flux. Autophagy was induced via starvation and monitored using double fluorescently tagged autophagosomal proteins (dFP). The cleavage of the dFP into monomeric fluorescent protein (mFP) upon arrival at the lysosome was used as a representation of flux through the pathway. Our findings show that unc-33 mutants have defects in autophagic flux under starved conditions. Further analysis of autophagosome trafficking is currently underway to elucidate a potential mechanism for the link between MAPs, axonal development and homeostatic mechanisms such as autophagy.
Faith Sawyer, Dallas Baptist University
9:00-9:15 in DBU
MRSA: The Missing Link
Faith Sawyer
The emergence of antibiotic-resistant bacteria has multiplied the danger of Staphylococcus aureus infections, which are the most common cause of skin and soft tissue infections (Alam, et al., 2015). When the infecting S. aureus bacteria acquires antibiotic resistance factors, complications increase and difficulty in patient treatment rises. In an attempt to understand the bacteria’s process of evolving from non-antibiotic resistant staphylococci bacteria, scientists have examined the DNA of various Methicillin-resistant Staphylococcus aureus strains, focusing mainly on the mobile genetic elements (MGEs) contained within the genome. While MRSA’s evolution from non-resistant strains through MGEs may reveal answers, examination of horizontally transferred genetic material through the lens of absence—an area that has not been subjected to much in-depth research—may provide substantial clues. When compared to antibiotic-susceptible strains, are there genes missing from the MRSA genome? To investigate this possibility, the NCBI database was used to obtain the complete genomes of an antibiotic-susceptible S. aureus strain (MSSA), and two different isolates of MRSA from the USA300 strain. Using the bioinformatics software UGENE, the MRSA isolates were individually compared to the MSSA strain by creating dot plots of their genomes. Any notable interruptions in nucleotide base pair matches on the dot plots—indicating a genetic sequence that was found in the MSSA strain but not found in the MRSA isolate—was analyzed with NCBI nucleotide BLAST to predict its function. Findings showed that specific enzymes and bacteriophage proteins were absent from a prophage region in the MRSA isolates.
Abigail Sequeira, University of Dallas
9:45-10:00 in TXH
Parasitic infection and exposure alters mate choice in Potamopyrgus antipodarum
Abigail Sequeira and Deanna M. Soper
Parasites can manipulate host traits in a variety of ways, such as alterations to host behavior, morphology, and physiology. Most research in this field examines parasitic alterations on post-infected hosts, not under parasite exposure. Potamopyrgus antipodarum, a freshwater snail from New Zealand, is known to coevolve with a sterilizing trematode parasite in glacial lakes. In one previous study, Potamopyrgus antipodarum increased mating rates and number of partners upon parasitic exposure alone. In this study, we exposed two different field-collected populations of P. antipodarum to the parasite Microphallus and observed the number of mating and rejecting events when placed with males from sympatric and allopatric populations. Use of field-collected snails meant that a proportion of female snails were already infected, and post-test dissections revealed this to be the case. We found that females altered mating behavior, but only when exposed and infected. This data indicates that exposure to parasites alone can alter female behavior, but only when females are infected. These results highlight the importance of examining host behavioral alterations upon exposure to parasites, but that future tests should include both infected and uninfected hosts.
Max Shaw, West Texas A&M University
1:15-3:15 in LSR 3
Analysis of the amino acid content for beef, chicken and turkey bone broths
Max Shaw and Nick Flynn
Amino acids provide essential building blocks for the proteins that provide structural support and help control signals within our bodies thus allowing us to live and function normally on a daily basis. Quantifying key amino acids in various health products may help us better understand their contribution to general health and support or repudiate claims made by supplement companies. For this project we quantified 19 individual amino acids in bone broth from beef, chicken and turkey. Amino acids from commercial samples of each broth were analyzed using an HPLC method with ninhydrin post column derivatization for quantification. Norleucine was used as the internal standard for analysis. The comparison of the broth concentrations to meat and PAP concentrations allowed us to see that broths may have an ability to contribute vital amino acids to one’s diet, but do not appear to be a better source.
Theresa Stein, Southwestern Oklahoma State University
1:15-3:15 in LSR 4
The effects of stream order, season, and drainage on the abundance of two caddisfly families
Theresa Stein, Amy Hofeld, and Skyler Mills
Rivers are constantly flowing and vary in different locations. Therefore, one area of a river or stream may differ from another in abundance of organisms. However, streams remain connected, as changes upstream may affect areas downstream. We hypothesized that stream order, season, and drainage would affect the abundance of two caddisfly families (Leptoceridae and Hydroptilidae). Species within each of these families have been identified as species of conservation concern by the Oklahoma Department of Wildlife and Conservation. We collected adult caddisflies in the Kiamichi River, Little River, and Spring Creek drainages using manned and unmanned light traps. Samples were collected during both spring and summer months. For both families, we found no difference in abundance between seasons or among drainages. We found that the abundance of leptocerids was significantly higher in third order streams than first or second order streams. However, stream order did not have a significant effect on hydroptilid abundance. We conclude that stream order affects the abundance of Leptoceridae in eastern Oklahoma streams. This information can be used to optimize sampling effort to detect species of conservation concern in this family of caddisflies.
Zoey Stormes, Angelo State University
1:15-3:15 in LSR 3
The Effects of HEPA Air Filters on Airborne Mold Spores in College Dormitories
Bethany Chapman, Lily Ellzey, Zoey Stormes, and Nicholas Negovetich
Every year thousands of students across the nation are required to live in university campus dorms, residence buildings maintained by the university that may have been in operation for years. After an article discussing the death of a University of Maryland immunocompromised student had mold implicated in her cause of death and fellow students at our university who are mold sensitive complained of their symptoms bothering them since arriving on campus, we became intrigued on methods universities or students could employ in individual dorm rooms to decrease overall mold in their dorm. One easy fix is to replace the standard air filter with a better one. This study investigates the effects of changing air filters to a higher MERV quality in dorm rooms on airborne mold spores. Through exposing SDA agar plates to the ambient air in dorm rooms, the mold in two different buildings of the same residence hall was assessed. A second set of agar plates was exposed after changing the air filters in each room, allowing for measurement of the effect new filters have on mold spore counts. Assessment of the before (x=2.05 spores/plate) and after plates (x=1.78 spores/plate) revealed no significant difference between the two, indicating that changing air filters does not reduce the number of mold spores in the tested dorm. However, there appears to be significant differences in the quantity of mold spores between the individual dorms.
Emma Thornburg, Austin College
11:45-12:00 in TXH
Investigating the Positive and Negative Regulators of PA28γ in Cancer
Emma Thornburg and Lance Barton
PA28γ is an ATP and ubiquitin-independent proteasome activator implicated in the development of cancer. Many of the downstream targets of PA28γ can benefit cancer development, and multiple cancers demonstrate increased expression of PA28γ. While the benefits of elevated PA28γ activity have been studied, the mechanisms by which PA28γ expression is regulated have not been fully elucidated. Two key regulators of PA28γ expression, miRNA-7 and p38, may be involved in the increased expression of PA28γ in cancer cells. I hypothesize that in cell lines with increasing amounts of PA28γ, p38 activity will be increased concomitantly, while miRNA-7 expression will be decreased. I also hypothesize that the decrease in miRNA-7 could have a synergistic effect on p38 signaling, leading to an overall larger increase in PA28γ in cancer cell lines. Results confirm a role for both p38 and miRNA-7 signaling in the increased expression of PA28γ in a tumorigenic cell line and a metastasized cancer cell line. Interestingly, p38 appears to also play a role in the regulation of miRNA-7 in PA28γ +/+ cells. Finally, results show that p38 inhibition decreases cellular proliferation while miRNA-7 inhibition increases cellular proliferation. Overall, these results demonstrate that the regulation of PA28γ by p38 and miRNA-7 may play a role in cancer development through increased proliferation of cells.
Keith Tittle, Texas Wesleyan University
11:00-11:15 in LSR
Species-area Relationships across Conserved lands in Texas: Does iNaturalist Work?
Keith Allen Tittle
Species-area relationships provide insights into ecological communities that in turn can be used to determine where conservation efforts are needed. Observations on iNaturalist provide a basis for estimating species richness using citizen science. iNaturalist has been used to collect species richness data from more than 60 parks and conserved areas near 3 major urban areas of North and Central Texas. According to the theory of island biogeography, area size predicts species richness. Species-area relationships are being assessed using linear regression following log transformation of area size and species number. The models for plant species reveal that richness increases as area size increases. However, much of species richness remains unexplained by area size alone. Human disturbance can accelerate extinction rates in conserved lands across Texas. iNaturalist can lessen the impact of this disturbance.
Rachel Vargas, St. Edward's University
1:15-3:15 in LSR 3
Hemocyte Protein Analysis as an Indicator of Immunological Response in Tigrosa helluo Following Leg Autotomy
Rachel Vargas
Autotomy is a survival mechanism utilized by various organism, in which an individual voluntarily loses a body part in response to an external stressor, often in the form of a predator attack. While this capability is a benefit towards immediate survival, there are notable impediments towards the overall survival of an autotomized individual. Behavioral and physiological alterations are commonly present following the autotomy event. The molecular components aiding in the recuperation of an organism after the loss of a leg have been of particular interest as they are indicative of the organism’s overall immunological activity. In invertebrates, the hemocytes incorporated within hemolymph are important factors in cellular immunity. Within reponses to leg autotomy, hemocyte interactions with foreign matter entering through an open wound can be assessed through the various proteins involved with the hemocyte activity. By analyzing the proteins present in the hemolymph of the wolf spider Tigrosa helluo after autotomy, we were able to identify the overall presence and function of the proteins associated with this stress response. The cost of this survival technique was indirectly inferred through the quantification of different proteins released during autotomy as a function of the immunological response. Protein concentration and prophenoloxidase activity, an precursor for the organisms encapsulation response, was measured in both the controlled and autotomized female Tigrosa helluo and the control sample of the males. While there was no significant differences between either of the parameters across any of the sample groups, the observed patterns of this preliminary data suggests a possible difference between protein activity in females and males, and a difference in the prophenoloxidase activity in autotomize individuals compared to non-autotomized individuals.
Brent Wesley, Angelo State University
9:45-10:00 in DBU
Genetic Barcoding Analysis of Agave
Brent Wesley, Loren Ammerman, and Bonnie Amos
Century plants, or Agave, grow in the southwestern United States and three species are known from the Big Bend National Park region. At least one of these species, Agave havardiana, provides a food source for and is pollinated by Mexican long-nosed bats (Leptonycteris nivalis). It can be difficult to determine which species of plant a bat has visited from morphological analysis of the pollen that clings to the fur. Our goal was to take a molecular approach and identify a barcoding gene that would be useful to be able to distinguish species of Agave plants from their pollen. Another goal was to determine how genetically different two species that grow in Big Bend National Park are from each other (A. havardiana and A. lechuguilla). We collected leaves from 36 plants (16 A. havardiana and 13 A. lechuguilla) and extracted DNA. PCR was performed with forward and reverse primers for 5 genes (4 chloroplast genes and 1 nuclear gene region) that have been useful for distinguishing other species of plants. PCR products were purified and sequenced. We analyzed the similarities and differences between the genes from each species, calculated divergent values, and determined the best primer set for pollen barcoding viability. The best primer set, or gene, will be the one with consistent, species-specific DNA differences to clearly distinguish between species of Agave in the region. We found that 3 of the genes were identical among the samples of the two species. One nuclear gene, ITS2, showed a low level of sequence divergence with a pairwise divergence value of 1.38% using a Tamura 3 parameter with gamma distribution. We conclude that the best gene for species level barcoding of agave is ITS2 because of the evidence of sequence divergence.
Dani Whiting, Southwestern Oklahoma State University
1:15-3:15 in TXH 1
Raptor Perch Locations and Types in Southwestern Oklahoma
Dani Whiting and Zach Jones
Migrating and overwintering raptor species use a variety of hunting methods and perching types to search for and initiate prey captures from rural and agricultural landscapes surrounding Weatherford, OK. The primary objective of this study was to identify species preferences for perch types (primarily a range of power transmission line pole sizes and materials, as well as pole density) and landscape context (amount of tilled, non-tilled pasture, woodland, man-made structures other than power poles, etc.) near chosen hunting locations. Two 48-km routes were sampled beginning in fall 2018, one north and one south of Weatherford, OK. Preliminary data show the following trends: American Kestrels (Falco sparverius) primarily used wires between poles; Red-Tailed Hawks (Buteo jamaicensis) indicated a preference for trees; and Northern Harriers (Circus cyaneus) were only seen in flight other than when consuming prey. Additionally, all species preferred areas with little to no presence of man-made structures other than utility poles and fencing around fields. Species’ differences in hunting methods and perch types may mitigate competition for common prey items (rodents) during migration and wintering seasons, and the presence of utility poles may increase the number of perch types available enough to encourage increased abundances of some species’ populations beyond what would otherwise be possible. These preliminary data indicate that both kestrels and Red-tailed Hawks may benefit greatly from the presence of utility poles and lines.
Mikaela Wilson, St. Edward's University
1:15-3:15 in LSR 4
Evaluating the Effects of Inducing Autophagy in unc-33 Mutants
Mikaela Wilson, Nick Ashley, Hailey Trombley, and Andrea Holgado
Autophagy, a mechanism used by cells to recycle and catabolize protein aggregates and cellular waste, is highly conserved from yeast to mammals. Recent reports indicate that UNC-33/CRMP-2/Dpysl2, a neuronal protein essential for early axonal development, is regulated during conditions that induce autophagy in the mammalian neurons. To further investigate the interplay between UNC-33 and autophagy, we constructed a C. elegans double mutant strain containing unc-33 and daf-2(e1370) mutations and characterized their phenotype. daf-2(e1370) encodes for an insulin-like IGF-1 receptor (IIR) and mutations in this gene result in induced autophagy and constitutive dauer formation. Analysis of our double mutant strain denotes that our unc-33; daf-2 double mutants grown at the restricted temperature result in a synthetic lethal phenotype. Quantification of double mutant nematodes surviving at the restrictive temperature shows that the synthetic lethality is partially penetrant, killing 78% of unc-33(e204); daf-2(e1370) and 72% of unc-33(mn407);daf-2(e1370) during embryonic stages. Observations of surviving double mutants demonstrate that these animals form abnormal dauers. The resulting abnormal dauer phenotypes are being characterized by evaluating survival in 1% SDS, pharyngeal length, life stage arrest, and proper sinusoidal movement.
Rebecca Wood, University of Central Oklahoma
1:15-3:15 in LSR 3
Serologic Evidence of Viral Reactivation and Increased Disease Activity in Patients with Systemic Lupus Erythematosus
Rebecca A. Wood, Lauren Guthridge, Emma Thurmond, Carla Guthridge, Hua Chen, Wade deJager, Susan Macwana, Neelakshi Jog, Rebecka Bourn, Stan Kamp, Rufei Lu, Cristina Arriens, Eliza Chakravarty, Katherine Thanou, Joan T. Merrill, Joel M. Guthridge, and Jud
Systemic lupus erythematosus (SLE) is a complex autoimmune disease which mainly afflicts young women. SLE can attack any organ in the body, with the disease fluctuating from active, multi-organ involvement (flares) to minimal disease. What causes flares is unknown. Our objective was to examine serologic evidence of viral exposures and re-activation with respect to SLE disease activity and cytokine levels. Our hypothesis was that reactivation of common latent viruses would increase lupus disease activity. Plasma from 184 SLE patients (n=290) and 49 controls was collected and disease activity measured. Viral responses were detected by ELISAs for EBV-Viral Capsid Antigen (VCA) (IgG and IgA), EBV-Early Antigen (EA) (IgG), Cytomegalovirus (CMV) (IgG), and Herpes Simplex Virus (HSV-1) (IgG). Plasma soluble mediators were measured by multiplex, bead-based assays and ELISAs. Serologic evidence of EBV reactivation was more common in SLE patients, per anti-EA (40% vs 13%; OR=4.57, p=0.0006) or EBV-VCA IgA (36% vs 17%; OR=2.70, p=0.019). Concentrations of EBV-VCA IgG responses were higher in SLE patients compared to controls (EBV VCA ISR = 4.44 vs 3.52; p=0.0021), as were IgA VCA and IgG EA. Cross sectional analysis of SLE patients at high (SLEDAI > 6) (n=126) vs lower (n=166) disease timepoints showed increased IgG EA responses with elevated disease activity (0.822 vs 0.540; p=0.033). SLE patients with anti-EA responses had higher IP10 (p=3.4 X 10-14), BLyS (5.5 X 10-5), and IL-10 (p=0.00013). HSV1 IgG positive SLE patients also had higher IP10 (2.2 X 10-7). This study provides serologic evidence supporting roles for viral reactivation and SLE disease flares.