2023 Undergraduate Research Symposium
Note: Presentations are grouped by the student’s area of research (based on the faculty mentor’s academic department), not the student’s academic major.
Poster Session A:
8:00 - 9:00 am
Animal and Food Science; Integrative Biology; Natural Resource Ecology and Management; and Veterinary Pathobiology (29 posters)
A-01 Jaxon Allen
Research Collaborators: Gabriela Lamberti
Research Presentation Title: Generating bison-cattle hybrids to evaluate parental contributions to embryonic development using in vitro procedures
Faculty Research Mentor: Joao Moraes, Animal and Food Sciences
Embryonic death is a leading factor limiting reproductive efficiency affecting humans and the livestock industry. A better understanding of embryogenesis and the contributions made by the sire and the dam can be instrumental in guiding our decisions pertaining to genomic selection. Our objectives were 1) to evaluate if hybrid embryos (Bos indicus x Bison bison) could be generated using in vitro embryo procedures and 2) compare the embryo developmental metrics between control (bovine) and hybrid embryos. In vitro embryos were produced following the media supplier protocols (IVF Bioscience) with minimal modifications.
Briefly, a total of 523 cumulus oocyte complexes (COC) were aspirated from bovine ovaries collected from an area slaughterhouse. From those 523 COC, 170 COC were fertilized using B. bison semen, and 353 COC were fertilized using semen from a proven high fertility Bos indicus sire. In vitro culture of bovine and hybrid zygotes was carried out for 8 days. Cleavage and blastocyst rates were assessed on days 3 and 8, respectively. All blastocysts produced from control and hybrid groups were cryopreserved for future assessment of embryonic viability and pregnancy rates upon transfer into recipient cows. Data was analyzed with linear mixed-effects models using SAS (version 9.4; SAS). Continuous data was assessed for normality using the UNIVARIATE procedure and log-transformed if not normally distributed. All models included the fixed effect sire and the random effect of the replicate, and data are reported as mean ± SEM. Statistical significance was defined as P ≤ 0.05, and statistical tendencies as 0.05 < P ≤ 0.10. Cleavage rates were not different (P = 0.97) between the control (84.54% ± 6.22) and hybrid (84.90% ± 8.79) groups. Likewise, blastocyst rates were similar (P = 0.77) in control (25.41% ± 3.01) and hybrid (23.85 ± 4.25) embryos. These results demonstrate our ability to generate hybrid embryos (Bos indicus x Bison bison) in vitro for the study of parental contributions to embryonic development. Future research using the generated hybrids will provide novel information that will be leveraged to reduce the impact of early embryonic mortality in the cattle industry.
A-02 Maria Julia Araujo Lopes
Research Collaborators: Dan Stein and Gabriela Lamberti
Research Presentation Title: First assessment of in vitro embryo production metrics using Angus semen after 29 and 39 years of cryopreservation
Faculty Research Mentor: Joao Moraes, Animal and Food Sciences
Cryopreservation techniques are considered the most efficient method for long-term preservation of germ cells, such as sperm. However, little is known about possible downstream consequences of storing bovine sperm cells in liquid nitrogen (LN2) over several years. The objective was to evaluate the effect of long-term semen storage in LN2 on in vitro embryo development metrics following in vitro fertilization (IVF) using semen from two Angus bulls stored for either 29 (Sire 1) or 39 (Sire 2) years. Procedures for in vitro embryo production were performed following the media supplier protocols (IVF Bioscience) with minimal modifications. Briefly, 379 cumulus oocyte complexes (COC) were aspirated from bovine ovaries supplied by a local slaughterhouse, maturated for 24h, and randomly assigned for fertilization using semen from Sire 1, Sire 2, or a Control bull (a proven high fertility Bos indicus sire). One hundred fifty-six presumptive zygotes were cultured from the Control bull, 88 from Sire 1, and 83 from Sire 2. In vitro embryo culture was carried out for 8 days. Cleavage and blastocyst rates were assessed on days 3 and 8, respectively. Data were analyzed with linear mixed-effects models using SAS (version 9.4; SAS). All models included the fixed effect of sire and the random effect of replicate. Data are reported as mean ± SEM. Statistical significance was defined as P ≤ 0.05 and statistical tendencies as 0.05 < P ≤ 0.10. Cleavage rate was reduced (P = 0.02) in embryos from the Control (58.27% ± 3.28) compared to Sire 1 (76.27% ± 4.64), and Sire 2 (77.21% ± 4.64). However, blastocyst rates from embryos that cleaved tended (P = 0.08) to be higher in Control (41.10% ± 5.80) compared to embryos from Sire 1(16.60% ± 8.20). Blastocyst rates however were not different between Control and Sire 2 (30.09% ± 8.20), or between Sires 1 and 2 (P = 0.53). These preliminary results indicate a possible negative consequence of long-term storage on embryo growth from 8 cells to the blastocyst stage, as the semen subjected to the longest storage time (39 years) had the lowest blastocyst rates from embryos that cleaved.
A-03 Kate Buckmaster
Research Collaborators: Gabriela Lamberti, Sam Melton
Research Presentation Title: The Role of Interferon-Stimulated Genes in Early Recognition of Pregnancy in Cattle
Faculty Research Mentor: Joao Moraes, Animal and Food Sciences
Nonpregnant cattle represent a financial and logistical problem for the industry; thus, early pregnancy detection is essential for the economic progression of livestock operations. The estimated cost of an open dairy cow per day is about $5. Thus, diagnosing pregnancy at day 19 (D19) vs. day 28 (D28) could result in $45 savings per cow. For a dairy farm with 5,000 cows, early pregnancy detection at D19 vs. D28 could save $225,000 every year. Interferon-tau (IFNT) is a protein secreted by the developing bovine conceptus, which is essential for maternal recognition and pregnancy establishment in cattle. IFNT secretion increases around D16 of gestation and stimulates the expression of interferon-stimulated genes (ISGs) in circulating white blood cells (WBCs). Thus, the objective of this study is to evaluate the expression of ISGs in WBCs for pregnancy detection at D19 of gestation using real-time PCR technology. Heifers (n = 55) from the North Cow Research Range were synchronized for timed artificial insemination (AI) and bled on D0 (day of insemination), D19, and D30 post-insemination. Pregnancy on D30 was assessed using a commercially available blood test (BioPRYN). Blood samples were also collected for isolating WBCs for gene expression analyses. Briefly, blood samples were centrifuged at 12,000 G for 20 min at 4OC, and the buffy coat (containing WBC) was transferred to a RNase-free microcentrifuge tube. Total RNA was isolated from the buffy coat using the Trizol reagent (Invitrogen), according to the manufacturer’s protocol. Concentrations and purity of the extracted total RNA were determined using a NanoDrop, and RNA integrity was evaluated using a 1% agarose gel. We were successful in extracting RNA from the buffy coat. Our next step will be the conversion of mRNA into cDNA, so real-time PCR can be performed for measuring the expression of a variety of ISGs, including ISG15, MX2, and OAS1. SDHA and GAPDH will be used as housekeeping genes. Results from this study will be used to evaluate the sensitivity and specificity of measuring ISG in WBC for pregnancy diagnosis at day 19 of gestation.
A-04 Kendall Hays
Research Collaborators: Alayna Gerhardt, Federica La Manna, Ryan Reuter, Laura Goodman and Courtney Duchardt
Research Presentation Title: The “buzz” on virtual fencing: Livestock behavior in response to aural cues and electronic stimulation
Faculty Research Mentor: Ryan Reuter, Animal and Food Sciences
Before the introduction of widescale ranching and European settlement in the new world, bison would graze freely across the landscape without manmade barriers like fencing. Today, ranching depends on stable fencing systems to ensure effective grazing management. Fences keep cows from overgrazing in a sensitive area or keep them in an area that needs to be grazed. However, traditional fencing systems can be very expensive and require a lot of labor to maintain and move. Further, in some parts of the US, fences can impede large-scale migrations of native ungulates like pronghorn (Antilocapra americana). The introduction of virtual fencing systems would help to eliminate some of the problems that come with traditional fencing systems. However, when you lack a physical boundary, it is unclear whether cows will stay within target areas. Virtual fencing uses GPS-enabled collars on cattle to understand their movement and grazing habits, but this technology also functions as a deterrent for crossing over virtual boundaries, using aural cues followed by electric stimulation. In this way, they function much like perimeter collars for dogs. For this study, we will use GPS data from cattle collars as well as information about sound triggers and electrical stimulation to determine 1) how long it takes individual cows to respond to aural cues alone, 2) how much variation there is among individual cows in sensitivity to aural cues and 3) overall effectiveness of aural vs. electrical stimulation. Information on cattle behavior will allow ranchers to identify individuals that are less receptive to virtual fencing, removing these from herds. Further, this research will allow us to understand lags in responsiveness to aural vs. electric cues, and how wide these digital boundaries must be to be effective.
A-05 Abby Helbach
Research Collaborators: Punya Bule and Divya Jaroni
Research Presentation Title: Use of Lactic Acid Bacteria to Reduce E. coli O157:H7 and Salmonella Typhimurium on Spinach
Faculty Research Mentor: Divya Jaroni, Animal and Food Sciences
Introduction: Consumption of fresh produce, including leafy greens, has increased in the past few decades. However, these trends have also been accompanied by several foodborne illness outbreaks. Owing to their minimal processing, with no kill-step and limited use of antimicrobials, leafy greens could get contaminated with pathogens such as Escherichia coli O157:H7 and Salmonella enterica. It is therefore crucial to find effective antimicrobial treatments during flume-washing of leafy greens. Lactic acid bacteria (LAB) are known for their inhibition capabilities against foodborne pathogens. However, studies evaluating their application in wash-water, to reduce Salmonella and E. coli O157:H7 on leafy greens, are limited. Purpose: Efficacy of LAB against E. coli O157:H7 and S. Typhimurium on spinach. Materials and Methods: Two strains of LAB: Lactobacillus acidophilus C28 (LA-C28) and L. reuteri X-18 (LR-X-18) were evaluated against E. coli O157:H7 and S. Typhimurium on spinach. Spinach (~ 2.5 x 2.5 cm pieces) was prepared by washing with sterile distilled water and drying (30 min.) in a bio-safety cabinet under UV (254 nm) light. Prepared leaves were inoculated (5 log10 CFU/ml) with E. coli O157:H7 or S. Typhimurium, and pathogen attachment allowed for 1 h. Spinach was then washed (2 min) with LAB treatments (8 log10 CFU/ml) or PBS (control) and stored (4°C) for 3-days. Surviving bacterial populations were enumerated on day 0, 1, and 3 by plating on appropriate medium. Data were analyzed using one-way-ANOVA (P<0.05). Results: Immediate reductions of ~0.5 and 1.2 logs were observed in E. coli populations on spinach treated with LA-C28 and LR-X-18, respectively. This reduction was maintained over 3 days with LR-X-18 treatment, whereas further reductions were observed with LA-C28 treatment on day-3. An immediate reduction (~0.5 logs) was observed in Salmonella population when spinach was treated with LA-C28, which was further reduced by ~1.3 logs on day-3. Treatment with LR-X-18 reduced Salmonella populations by ~0.8 logs on day-0, with further reductions (0.7 logs) on day-3. The LAB strain LA-C28 performed better than the LR-X-18 as wash-water treatment. Significance: Lactic acid bacteria could be utilized in water used for washing spinach, to reduce E. coli O157:H7 and S. Typhimurium.
A-06 Caitlyn Quinn
Research Collaborators: Kaylee Rumbaugh
Research Presentation Title: Evaluation of Probiotic Lactic Acid Bacteria Resistance to Acid, Bile, and Antibiotics
Faculty Research Mentor: Divya Jaroni, Animal and Food Sciences
Introduction: With the emergence of antibiotic resistance (AR) as a global threat, alternatives to antibiotics are being sought in the food-animal industry. Lactic acid bacteria (LAB) are widely used as probiotics, with early reports showing alleviation and prevention of intestinal disorders, improved growth-performance, increased nutrient utilization, and decreased use of antibiotics. Although LAB are generally recognized as safe, some of them have shown intrinsic or acquired AR, which could be transferred to pathogenic microorganisms. Therefore, it is crucial to evaluate the AR of LAB that hold potential for use as probiotics. Additionally, to be effective in the GI-tract, they need to be able to survive the unfavorable GI-tract environment, such as acid and bile conditions. Purpose: Evaluation of antibiotic susceptibility and acid/bile tolerance of probiotic LAB. Methods: Strains of LAB (Lactobacillus, Lactococcus, Leuconostoc, or Streptococcus) showing excellent or very-good viability after extended frozen-storage (-80℃) were tested for acid and bile tolerance. Each strain (1x108 CFU/ml) was grown for 0, 1, 3, and 6 h (37/42℃; anaerobic incubation) in MRS broth adjusted to pH (2, 4, 5, 7) and bile (0, 0.1, 0.3, 0.5%), and absorbance measured at 620 nm (acid) and 660 nm (bile). Acid/bile tolerance was categorized as, excellent: increased growth at all pH/bile levels over 6 h; or very-good: increased growth at at-least two pH/bile levels over 6 h. These strains were then evaluated for resistance to common antibiotics (ampicillin, clindamycin, erythromycin, gentamycin, streptomycin, tetracycline, and vancomycin), using disk-diffusion assay and reading the diameter of inhibition zones. Strains showing zones >20 mm were classified as sensitive (S), between 15-19 mm as intermediately sensitive (I), and zones ≤14 mm as resistant (R).
Results: Of the 65 strains tested, 13% showed excellent, 35% very-good, and 39% good tolerance to acidic conditions, while 29% exhibited excellent, 16% very-good, and 52% good tolerance to bile. All tested LAB were sensitive (S or I) to erythromycin and clindamycin, followed by ampicillin (92%), tetracycline (67%), vancomycin (42%), gentamycin (17%), and streptomycin (8%). Significance: Selected LAB, showing excellent to very-good acid/bile tolerance and sensitivity to antibiotics could be potential candidates for a probiotic.
A-07 Natalee Richardson
Research Collaborators: Anna Goldkamp
Research Presentation Title: In vivo Assay: Bacterial Small Non-coding RNAs May Regulate Eukaryotic Gene Expression
Faculty Research Mentor: Darren Hagen, Animal and Food Sciences
Wolbachia is present in 70% of insects because of this it is widely studied. Research suggests that one or more small RNAs, made by the Wolbachia bacteria is responsible for initiating and maintaining the ability to manipulate its host’s reproduction which is a phenomenon called cytoplasmic incompatibility (CI). While if small RNAs derived from bacteria are utilized by host machinery to alter expression of host genes remains unknown. Early evidence suggests that some Wolbachia- derived small RNAs hybridize to Drosophila protein coding genes, which regulates protein expression. If the small RNAs are produced in enough volume to bind to Drosophila targets with enough strength they could regulate protein expression. The bacterial small RNA sequences have been derived from RNA library pools that we sequenced using RNA-Seq. A plasmid was designed that would essentially be built from two existing plasmids. The pHd-DsRed plasmid and the act5c plasmid each contain specific pieces that were needed to complete the designed plasmid. Each plasmid was grown in cells in order for the necessary pieces to be isolated. For the pHd-DsRed plasmid, restriction enzyme digestion was used to cut the section of the plasmid we needed and a gel electrophoresis was used to separate the piece from the rest of the plasmid. For the act5c plasmid, PCR was used to amplify and isolate a section of the plasmid.The pac5-DsRed plasmid, containing the ACT5C promoter, a fluorescence gene, and an ampicillin resistance gene, has been completed and transfected into cells. We have plated the cells on ampicillin coated plates to isolate the cells containing plasmids. The next step is to add a target sequence to the plasmid for the Wolbachia small RNAs to attach to. For the future of this project we plan to culture cells containing the pac5-DsRed plasmid with the target sequence added. There will be two separate sets of cells being grown, one infected with Wolbachia and the other uninfected.
A-08 Hannah Silensky
Research Collaborators: Ravi Jadeja and Xin Mei Tang
Research Presentation Title: Low-Temperature Antimicrobial Intervention for In-Shell Pecans
Faculty Research Mentor: Ravi Jadeja, Animal and Food Sciences
1 in 6 Americans contract a foodborne illness each year. There are over 128,000 cases resulting in hospitalization and approximately 3,000 leading to death. Over the past decade there has been a wide variety of different pecan products that have been recalled due to Salmonella contamination. Salmonella is a gram-negative bacterium that is most thought of being related to animals and animal products and is currently the #1 reason for bacterial illnesses. Symptoms typically set in 12-72 hours after being exposed presenting as diarrhea, fever, headache, and abdominal cramps. The goal of this study is to find what temperature pairs best with the best sanitization treatment to rid the contaminated pecan of Salmonella. Current interventions set in place by the USDA include thermal and nonthermal treatment. Although these interventions are successful, they have downsides that pose significant risks to the workers, machinery, and the quality of the pecans. For this experiment, a pathogenic cocktail of approximately 9 logs CFU/ml was prepared. 25 ml of the cocktail was added to 400 g of in-shell nuts into a whirpak bag and agitated for 2 minutes strait. Following that, the pecans dried in the laminar flow hood for two hours and then were left to refrigerate overnight. 85ppm peracetic acid (PAA) was heated to different temperatures ranging from 90*F to 150*F. The inoculated pecans were chosen 3 at a time to go into a treatment solution in a 600 ml beaker. A 400 ml beaker was placed on top to help maintain complete submersion for the full two minutes of agitation in the treatment solution. The nuts were removed using forceps and placed in 30ml of 10% sodium metabisulfite neutralizing buffer. In the bag with the buffer, the pecans were shaken continuously for another 2 minutes. A sample dilution was taken and plated. The plates were then counted after 24- and 48-hour incubations. Given the high growing population in Oklahoma, the results of this experiment have found that PAA treatment at 110*F is a suitable alternative to the current interventions.
A-09 Samuel van Rhijn
Research Collaborators: Gabriela Lamberti and Caleb Melton
Research Presentation Title: Unraveling the Biology of Conceptus Elongation
Faculty Research Mentor: Joao Moraes, Animal and Food Sciences
Infertility and subfertility represent pervasive problems in domestic animals and humans, and embryonic mortality is a major factor limiting reproductive efficiency. In cattle, most embryonic losses occur during the first month of gestation that involves the period of blastocyst formation, conceptus elongation, maternal recognition of pregnancy, implantation, and beginning of placentation. The genetic and epigenetic regulation of embryonic and placental development in ruminants is still poorly understood. The goal of this project is to use innovative system biology approaches to investigate conceptus elongation in cattle. Our overarching hypothesis is that the transcriptional regulation of conceptus elongation involves complex regulatory networks controlled by transcription factors and microRNAs. Specific objectives of the present experiment include: 1) generate transcriptome, 2) microRNA, and 3) epigenomic data from bovine conceptuses during elongation. For that, Angus heifers (n = 9) were superovulated and bred using semen from a proven high-fertility Bos indicus sire. Conceptuses were recovered from donor heifers on day 13 of gestation by flushing the uterus, using a standard nonsurgical flushing technique. Recovered conceptuses were individually photographed and snap frozen for further analyses. A total of 63 conceptuses were recovered from 9 donor heifers. From the 63 conceptuses recovered, 3.17% were classified as spherical (0.05 - 0.2 mm), 44.44% as ovoid (0.2 - 5mm), 44.44% as tubular (5 - 15mm), and 7.94% as filamentous (16 - 35 mm). Recovered elongating conceptuses from the four distinct developmental stages (e.g. spherical, ovoid, tubular, and filamentous) will be subjected to bulk RNA-seq, microRNA-seq, and single cell ATAC-seq analyses. The goal of this ongoing research is to generate novel data of the genetic drivers regulating conceptus elongation, a critical period for maternal recognition of pregnancy in cattle. Understanding the normal mechanisms regulating embryonic and placental development is necessary to acquire basic knowledge that can serve as a foundation to diagnose abnormal embryogenesis. The long-term goal of this research is to improve cattle production systems and reduce the impact of reproductive problems in cattle operations.
A-10 Jade Wilkinson
Research Collaborators: Kavya Gavai
Research Presentation Title: The Effect of Acid Adaptation on Pathogenic Bacteria Used as Challenge Organisms for Microbial Validation of the Biltong Process
Faculty Research Mentor: Peter Muriana, Animal and Food Sciences
Biltong is a South African dried beef product that has grown in popularity in U.S. markets. Unlike traditional beef jerky, biltong is dried at ambient temperature and humidity following marination. However, biltong processing fails to meet the requirements of the USDA-FSIS regarding meat and poultry products, therefore a microbial validation study must be provided demonstrating sufficient reductions of a ‘pathogen of concern’ can be achieved. The use of acid-adapted cultures during validation studies for acidic foods are ‘highly recommended’ by USDA-FSIS. If challenge cultures are not acid-adapted prior to acid treatment or marination, they may be overly sensitive, resulting in falsely high microbial reductions. It is believed that acid-adapting cultures for product inoculation and testing would ensure that the process is sufficiently robust to accomplish a 5-log reduction of the pathogenic organisms, however some publications have suggested otherwise. The objective is to perform and compare process validation using acid-adapted and non-adapted cultures for biltong processing to determine whether acid-adaptation is a prerequisite culture treatment for validation studies. Mixed strain “cocktails” of Salmonella spp., L. monocytogenes, and E. coli are individual prepared in TSB containing 1% glucose (acid adapted) and TSB without glucose (non-adapted). Beef cuts were inoculated with culture treatments and refrigerated to promote bacterial attachment. Inoculated samples were dipped in water or in 5% lactic acid and marinaded for 30 min with a mixture of spices, salt, and vinegar using a vacuum tumbler. The marinaded beef pieces were hung in a temperature-controlled humidity oven to dry (75oF, 55% RH). Beef was sampled for microbial enumeration after inoculation, post-dip, post-marinade, and after drying for 4-, 8-, and 10-days. Trials were performed in duplicate replication with triplicate samples tested per sampling time. Our current results obtained with Salmonella demonstrates greater reduction with acid-adapted cultures than with non-adapted cultures (the opposite of what USDA-FSIS suggests). Acid-adapting cultures before inoculating beef samples ensures that only a robust commercial process would achieve a 5-log reduction to produce safe foods. This validation study carries the potential to influence future USDA-FSIS policy and determine whether or not adaptation should be required for acidic food validation studies.
A-11 Emma Ainsworth
Research Collaborators: Allyson Degrassi
Research Presentation Title: Burned soil does not impact detection or consumption of seed by granivorous rodents
Faculty Research Mentor: Allyson Degrassi, Integrative Biology
The occurrence of intense wildfires has increased in recent years, particularly in areas plagued by drought. This increase in the frequency of wildfires has provided ecologists with the unique opportunity to investigate species survivorship and direct mortality among various taxa, but less is known about how wildfires affect organismal behavior. Particularly, the ability to locate resources required for survival, growth, and reproduction, such as food, may be diminished by wildlife. Many rodents and other granivorous animals are seed dispersers which are important for forest health and range. They rely on their ability to detect seeds in various soil types for consumption. By changing the soil composition, wildfires have the potential to impact the ability for rodents and other granivores to detect and consume seeds, which may indirectly and negatively impact forest regeneration. This experiment tested the effects of burned soil on the ability for rodents to detect seed. In hardwood dominated forest at Blandy Experimental Farm in Boyce, VA, we randomly set foraging trays with 3g of sunflower seed in 1L of soil (i.e., top-soil or sand) from four different conditions: 1) unburned topsoil, 2) unburned sand, 3) burned topsoil, and 4) burned sand. Animals were allowed to forage ad libitum for approximately 12 hours overnight before foraging trays were collected and seed was re-weighed to calculate the GUD from each treatment and control. There was no significant difference between burn state, and no significant difference between soil type. This data can be applied to fire ecology management techniques to improve the techniques used to manage native land.
A-12 Maya Albert
Research Presentation Title: Neither weapon size nor body size can explain the mating patterns observed in a wild population of leaf-footed cactus bugs, Narnia femorata
Faculty Research Mentor: Zachary Emberts, Integrative Biology
Neither weapon size nor body size can explain the mating patterns observed in a wild population of leaf-footed cactus bugs, Narnia femorata. Competition is a common occurrence in the animal kingdom. Individuals may fight over food or shelter, but most often individuals fight over access to mates. For example, male bighorn sheep clash horns to gain mating opportunities with females. Several lab studies have shown that large individuals with big weapons are likely to win a given fight and ultimately gain mating opportunities. However, relatively little is known about mating behaviors outside of lab settings. To determine if the results are similar in the lab and the field, we conducted several selection analyses to assess the degree to which body and weapon size determine a male’s access to mates. We specifically conducted these analyses on the leaf-footed cactus bug, Narnia femorata, which is a species that has had its fighting and mating behavior well documented in the lab. Specifically, previous studies have shown males with larger bodies and weapon sizes outcompete smaller males and gain more mating opportunities. However, contrary to the lab studies, we found no evidence to suggest that body size nor weapon size influences a male’s mating opportunities in wild populations. Our results highlight the importance of studying animals in their natural environment.
A-13 Steven Downs
Research Collaborators: Calvin Schaefer
Research Presentation Title: Ecological and Evolutionary Correlates of Skull Morphology in Venomous Snakes
Faculty Research Mentor: Dr Guin Wogan, Integrative Biology
Venom has evolved repeatedly across the Tree of Life and is an ecologically relevant phenotypic trait that is tied directly to the fundamental need for animals to acquire sustenance. Many aspects of the morphology of the venom delivery system have been shown to vary according to the particular ecology of a species, but a broad scale study on how these factors may influence other aspects of snake morphology such as skull shape has not been attempted. The goal of this research is to examine how ecology, venom composition, and dentition can relate to variation in skull morphology in venomous snakes. To examine this question, I have sampled a subset of venomous snakes (n=93) across multiple families (n=5) and have generated high-resolution morphological data using microCT scanning, which I analyzed using Slicermorph using landmarks and 3D morhpometrics to create a general shape model of the skull for each species. To examine ecological factors, I have evaluated dietary niche breadth and foraging modes for these same species. I conducted diet analyses and calculated dietary niche breadth using Squamatabase, and I have compiled data for foraging mode. To investigate venom composition, I used the ToxCodAn bioinformatic pipeline to generate a list of venom components for each species. All of this work uses phylogenetic comparative approach in R to examine the relationship between skull shape and ecological factors, venom composition, and dentition. I found that foraging mode is highly conserved in Viperids, Elapids, and Lamprophiids, but diverse in Colubrids and Homolopsids. Therefore, I expect the skull morphology to follow the same pattern.
A-14 Vanessa Franco
Research Collaborators: Elizabeth McCullagh, Andrea Gentry, Madison Merideth
Research Presentation Title: The Proliferation of Oligodendrocytes in Fragile X Mice
Faculty Research Mentor: Elizabeth McCullagh, Integrative Biology
Autism spectrum disorder is a condition that is associated with many sensory issues, one of which is auditory hypersensitivity. Fragile X syndrome (FXS) is a common monogenic form of autism. The direct cause for auditory hypersensitivity is still relatively unknown, however it can be hypothesized that deficits in myelination and impaired oligodendrocytes (OL) may play a role. Oligodendrocytes are glial cells that are responsible for producing myelin sheath that insulates neurons in the central nervous system, resulting in fast transmission of electrical impulses. Transmission of electrical impulses is crucial for accurately processing sounds, making the auditory brainstem one of the most heavily myelinated areas of the brain. This project aims to evaluate whether there is a higher prevalence of oligodendrocyte precursor cells (OPC) to mature OLs. To answer this research question, we examined the auditory brainstem of FXS and then used immunohistochemistry to differentiate OLs & OPCs. We will compare knockout mice (Fmr1 genotype) to wild-type mice (control) oligodendrocytes in the auditory brainstem. Using a confocal microscope, we quantified the number of OPCs and OLs. This project, along with previous work done in the lab, we can see a possible correlation between auditory hypersensitivity in FXS and an increase in OPCs. This connection provides one part of the overall cause of auditory hypersensitivity in FXS. Future research of auditory processing in FXS and autism includes establishing other phenotypes as biomarkers.
A-15 Kennedy Funa
Research Collaborators: Michael Reichert
Research Presentation Title: A Comparative Study on Temporal Integration Strategies of Acoustic Signals in Cope's Gray Tree Frogs
Faculty Research Mentor: Michael Reichert, Integrative Biology
Cope’s Gray Treefrog (Hyla chrysoscelis) has a “sister species”, Hyla versicolor, that is visually
identical to itself but has a different call and is genetically incompatible when mating. Our
research question is whether H. chrysoscelis calling varies when responding to a synthetic call
with segments of heterospecific (H. versicolor) and conspecific (H. chrysoscelis) signaling with
differing temporal order and proportions. Because of the varying temporal order of H. versicolor
and H. chrysoscelis elements in the synthetic calls, we can discover the cognitive processes
behind H. chrysoscelis responses by determining if they listen to an entire call or just the
beginning to decide their response. Furthermore, due to the data from the differing proportions
of heterospecific and conspecific segments in the synthetic calls we can determine if there is a
ratio between the segments that decides how H. chrysoscelis will respond. By presenting them
with signals with conflicting information, we can test how they are putting that information
together to determine their response.To test this, we found Hyla chrysoscelis specimens near
Lake Mcpherson in Stillwater, Oklahoma and recorded their calling before they heard the
synthetic call (baseline) and after they heard the synthetic call (playback). A baseline call is the
frog’s natural calling behavior. We will compare the number of pulses in each frog call, type of
call (either advertisement/mating or aggressive), and call duration in both the baseline and the
playback call recordings. From these data, we can gain insight into how Cope’s Gray Tree Frogs process auditory signals. We are currently analyzing calls from this experiment and we predict that H. chrysoscelis should have a stronger response to calls (longer duration advertisement calls and/or more aggressive calls) when the conspecific pulses are at the beginning of the call, and when the call contains a higher proportion of conspecific pulses
A-16 Anthony Mabrey III
Research Presentation Title: Giant mesquite bugs with large weapons have more mating opportunities
Faculty Research Mentor: Zachary Emberts, Integrative Biology
Many animals compete with other members of the species over access to resources and females. Several lab studies have shown that males with larger weapons are more likely to outcompete other males and gaining access to mating opportunities. However, relatively less is known about whether this pattern that is frequently observed in the lab is also observed in the wild. To better understand mating patterns in the wild, we conducted a selection analysis on weapon size and body size to find out whether there was a correlation between these traits and the number of mates a male has using the giant mesquite bug, Thasus neocalifornicus. We found that both weapon size and body size could explain the number of mates found with any given male. These results largely corroborate previous lab studies that have shown that weapon size is an important trait for accessing mating opportunities. However, our finding that body size is important conflicts with previous lab studies conducted on this species. We hypothesize that the discrepancy between the lab and field studies could be due to a lack of predation in a lab setting, where having a larger body could dissuade some predators of T. neocalifornicus.
A-17 Chace McCoy
Research Collaborators: Samantha Garza
Research Presentation Title: Does Infection by a Fungal Pathogen Affect Mate Choice in Toads? An Immunogenetic Analysis
Faculty Research Mentor: Bruce Waldman, Integrative biology
By studying the immunogenetics of threatened or declining species, we can implement conservation efforts to maximize disease resistance for susceptible groups.
It has been proven that genes within the major histocompatibility complex (MHC) can affect mate choice in many vertebrate classes of animals, from fishes to humans. However, the mechanism by which MHC variation affects mate choice is not well understood. I will focus my efforts on exploring MHC effects on mate choice in American toads, a species in which females have been shown to recognize and avoid mating with related males by properties of their advertisement calls. The extent to which mating behavior is influenced by individuals’ MHC genotype will be explored, as well as any influence caused by infection load of the the fungal pathogen Batrachochytrium dendrobatidis. The study will involve isolating DNA from toe clippings collected from calling males and toads after they have mated naturally in the field. Using MHC-specific gene polymerase chain reaction (PCR) amplification and high-resolution melting (HRM) genotyping, I will study toad mating patterns as they compare to infection load and MHC genotype. Acoustic properties of males’ advertisement calls will be analyzed for their statistical relation with MHC genotype to determine whether females may detect a males’ genotype by sound. Since the MHC molecule is present in all jawed vertebrates, research regarding MHC function is applicable in many areas of immunogenetics.
A-18 Ishani Ray
Research Collaborators: Amita Chawla
Research Presentation Title: Characterization of auditory physiology in FXS in critical developmental timepoints
Faculty Research Mentor: Elizabeth McCullagh, Integrative Biology
Autism spectrum disorders are strongly associated with auditory hypersensitivity. Fragile X syndrome (FXS), a common monogenic cause of ASD, results from transcriptional silencing of the Fmr1 gene and reduced expression of fragile X messenger ribonucleoprotein (FMRP). FMRP directly impacts myelin proteins and various brain regions show reduced/delayed myelination in FXS, suggesting deficits seen in FXS may be caused by alterations to myelination. FXS is a neurodevelopmental disorder, therefore characterizing when during development auditory dysfunction arises in addition to understanding if these changes are myelin dependent is critical to elucidating the full etiology of FXS. Auditory brainstem response (ABR) measurements record 1-4 waves, each corresponding to part of the ascending auditory pathway; the latency of which could be directly related to myelination of auditory areas. To characterize the physiology of myelination deficits in FXS at developmental time points, ABR measurements were taken for transgenic Fmr1 mice and controls before (P8-10), during (P12-14) or after (P21-23 and adult) hearing onset in mice. This allowed us to study the developmental emergence of auditory disruptions in Fmr1 transgenic mice and identify critical windows where underlying auditory pathways are established. We hypothesize that transgenic Fmr1 mice will have increased latencies in their binaural ABR waves (3, 4 & BIC) compared to the wildtype at different developmental time points. These data will aid in identifying the critical developmental windows of neural circuitry establishment in auditory sensory systems and potential myelination impairments that underly auditory dysfunction observed in patients and mice with FXS.
A-19 Rabeca Richardson
Research Collaborators: A. Valdez, L. Zhai, Bo Zhang
Research Presentation Title: Projecting Mechanical Treatment Outcomes on Invasive Grass to Improve Control Efficiency
Faculty Research Mentor: Bo Zhang, Integrative Biology
Background. With the increasing impacts of climate change, species invasions are expected to worsen, especially by drought. Meanwhile, drought could decrease the efficiency of invasion control. Given the worsened invasion and decreased control efficiency under drought, there is an urgent need to improve control methods, e.g., changing the treatment timing, to sustain the control efficiency. However, the variation in efficiency of mechanical control (e.g., clipping, mowing) by treatment timing is largely unknown. Methods. To address the knowledge gap, we conducted modeling analyses and greenhouse experiments to compare efficiency of mechanical control early-applied and evenly-applied during a growing season under two drought levels. We hypothesized that the efficiency of early applied mechanical treatments will decrease under drought conditions, making even-applied treatments more efficient in the presence of drought. The modeling analyses are based on a spatial-specific logistic equation model to capture the dynamics of an invasive species under the different treatment timings and drought levels. To validate the model, we conducted a greenhouse experiment focusing on the interactions of drought and treatment timing on the growth and functionality of Sorghum halpense. Results. Our model results indicated that applying early-season (short) interval treatments is a more efficient approach without the presence of drought, while late-season (long) interval treatments have a higher efficiency under drought. Our modeling results were further supported by preliminary observations of the ongoing greenhouse study. Under drought absence, the early and late season interval clippings showed a large difference in biomass, but under drought, this difference was much less, indicating that the efficiency of early season interval clippings decreased under drought. Conclusion. Thus, our study suggests that widely proposed early treatment could be modified to sustain the control efficiency under increasing drought. The model we developed here can potentially be extended and inform field managers in their efforts to control invasions more efficiently.
A-20 Conner Risley
Research Collaborators: Z T Emberts
Research Presentation Title: Mozena arizonensis Males with Larger Weapons have Access to More Mates
Faculty Research Mentor: Zachary Thomas Emberts, Integrative Biology
In some animals, individuals fight with other members of their species for mating opportunities. An example of this can be seen in the wild when deer clash their antlers together. Researchers often use insects to study this behavior in the lab. One well studied insect group recently receiving a lot of attention are leaf-footed bugs. However, only a fraction of the >2,000 species have been studied. Here, we conducted selection analysis in the wild on the understudied leaf-footed bug Mozena arizonensis. We found that femur width, a component of their weapon, could explain mating opportunities in this species. This result, coupled with our personal observations, suggests that males use their enlarged and spine-ridden hind legs to compete with other males over access to females. This finding is consistent with lab studies conducted on other closely related species in this group.
A-21 David Schrader
Research Collaborators: Sierra Williams, Jennifer Grindstaff
Research Presentation Title: The effect of polyinosinic-polycytidylic acid (poly I:C) on the social behavior of zebra finches (Taeniopygia guttata)
Faculty Research Mentor: Sierra Williams, Integrative Biology
Early viral immune challenge (EVIC) may alter behavioral phenotype. Inflammation caused by EVIC can lead to synaptic disorganization, as the development of neural circuitry is sensitive to adversity during this period. Subsequently, individuals exposed to EVIC exhibit altered learning, memory, activity, and sociality behaviors. Our objective was to observe the effect of EVIC on the social behavior of adult zebra finches (Taeniopygia guttata). We challenged nestling zebra finches with Polyinosinic-polycytidylic acid (Poly I:C), a synthetic dsRNA virus. For comparison, we created two control groups: nestlings injected with an equal volume of saline and non-injected nestlings that were only handled. Adult zebra finches were assessed for sociality by exposing them to an unfamiliar cage mate. We quantified behavior using an ethogram. Birds injected with Poly I:C had more close contacts than uninjected birds but less than saline injected birds. Saline injected birds had more beak wipes than the non-injected birds. This analysis demonstrates further research is needed to elucidate the interaction between EVIC and social behavior. This research contributes to our understanding of the mechanisms that cause behavior and disease exposure to covary.
A-22 Mariela Kundak and Kaitlyn Tucker
Research Collaborators: Lindsey Buehler, Courtney Duchardt
Research Presentation Title: Prairie Dogs in the Wichitas: Mysterious Decline in Colonies
Faculty Research Mentor: Courtney Duchardt, Natural Resource Ecology and Management
Wildlife declines are becoming more prevalent in the United States due to habitat loss, disease, and species competition. Black-tailed prairie dogs (Cynomys ludovicianus) are declining across the Great Plains due to both habitat loss and persecution because of perceived and real conflict with agriculture. Especially in smaller prairie dog colonies in the central US, this depletion has been recognized, as have cascading effects on associated wildlife species. The Wichita Mountains National Wildlife Refuge in Oklahoma is currently facing a similar decline; one colony at the refuge recently declined, and no prairie dogs have been observed since the summer of 2022. One factor that could cause this decline could be vegetation height, because prairie dogs prefer short vegetation. We are especially interested in analyzing if the vegetation is too tall for the prairie dogs to manage themselves in the eastern part of their range, where it is wetter, and grass grows taller. Using spatial boundary data and species counts over the years, we will attempt to determine if prairie dogs left this colony to merge with another colony, or if other factors drove these declines. We hope to mark animals and possibly use genetic testing to evaluate these patterns in the future.
A-23 Zoe Protzman
Research Collaborators: Lindsey Buehler
Research Presentation Title: Shrike a Pose! Potential Use of Trail Cameras for Demographic Sampling of the Loggerhead Shrike
Faculty Research Mentor: Courtney Duchardt, Natural Resource Ecology and Management
The Loggerhead Shrike is a small and unique songbird, as it hunts its prey like a raptor and stores the food on a thorn or barbed wire to eat later. The shrike also prefers to have a larger territory than most songbirds which can make assessing local populations difficult. Right now, according to the IUCN Red List, shrikes are seen as near threatened globally, with decreasing populations. Enough that eastern Canada has listed them as endangered and threatened in western Canada. In a study conducted by Dr. Duchardt and Lindsey Buehler in the OSU Rangeland Biodiversity Lab, they found when a trail camera was knocked by elk or cattle, Loggerhead Shrikes were sometimes observed in camera images perching on camera enclosures. This observation initiated a project to determine if trail cameras can be used to monitor shrike populations. To do this, we will look through previous camera trap data and mark every photo that has a Loggerhead Shrike. Next, we will model shrike occupancy. We believe that if shrikes are using cameras within their territory to hunt or perch, this could be an effective way to collect population data. If this is the case, it would make collecting that data easier, as field data can be difficult to conduct with their large territories.
A-24 Vanessa Rendon
Research Collaborators: Ben Kelly
Research Presentation Title: The Impacts of Intermittency on Surface Water Crayfish Communities
Faculty Research Mentor: Lindsey Bruckerhoff, Natural Resource Ecology and Management
Flow regime is a major driver of aquatic ecology. Due to increasing anthropogenic influences and altered precipitation patterns, flow regimes are changing at a rapid rate with devastating effects on ecosystems. Despite their importance to both food webs and stream structure, very little is known about crayfish communities. This is especially true in intermittent streams where collecting data is difficult due to the dramatic fluctuations in flow patterns and a lack of gauge data. We aim to assess how surface water crayfish communities shift across a gradient of intermittency to better understand how disturbances shape community structure. The study will take place in the Glover River basin of the Ouachita Mountains where the unique topography creates variations in hydrology and patterns of stream drying. Ten study sites, categorized as intermittent flashy or intermittent runoff, were chosen equidistant from the main Glover River and sampled every three months to capture changes in community structure as seasons shift. The results of this study will assist in analyzing how crayfish respond to disturbance and help inform decisions involving their management and preservation.
A-25 Paola Salazar Salazar
Research Collaborators: Courtney Duchardt
Research Presentation Title: The complex predator-prey relationships between the American Badger (Taxidea Taxus) and Black-Tailed Prairie Dog (Cynomys Ludovicianus) population dynamics
Faculty Research Mentor: Courtney Duchardt, Natural Resource Ecology and Management
To better manage and protect species and their habitats, it’s important to understand predator- prey relationships, regulating the populations of prey species to maintain ecological balance, managing agricultural practices to control the populations of prairie dogs, and provide advance species ecology scientific knowledge to inform the future research and conservationists about mesocarnivores and the prey species. Prairie dogs are a keystone species in their native grasslands. In these same grasslands there are also predators like mesocarnivores who play an important role shaping these ecosystems such as badgers, swift foxes, and black-footed ferrets, and these species can be impacted by fluctuations in prairie dog numbers. Conversely, predators may negatively impact prairie dog numbers in certain situations. We are using data from long-term camera trapping monitoring program to investigate the relationship between American badger and prairie dogs in their native habitats to investigate the relationship between these two species. The results will help guide conservation efforts for the black-tailed prairie dogs and managing the grasslands they thrive in. In the future, studies should continue to explore and understand the complex interactions the American badger and their prey have to gain knowledge of the ecological dynamic of these ecosystems.
A-26 Maddie Watts
Research Collaborators: Aaron Russell, Saroj Adhikari
Research Presentation Title: Factors affecting prescribed burn cost in the Great Plains
Faculty Research Mentor: Omkar Joshi, Natural Resource Ecology and Management
In the Great Plains, fire is an important land management tool essential to maintaining and preserving the region’s ecosystems. Prescribed fire is critical in perpetuating regular fire return intervals and enhancing land characteristics. Since private landowners are the primary stewards of the forest and rangeland in this region, their decision to conduct burning is important for the sustainable management of natural resources. Subsequently, the cost is one of the major obstacles in the implementation of prescribed fire. Therefore, our study objective is to understand the factors that play a role in determining the operational expenses while conducting a burn. In this project, we performed a multivariate regression analysis of cost factors using data from a survey of prescribed burn professionals in the Great Plains region. The average cost of prescribed fire in the study region was found to be $11.23. The results suggest that firebreak types, number of burns, and acres burned may play a role in cost. Identification of the cost of prescribed fire and the factors influencing it allows landowners, prescribed burn professionals, and government agencies in the Great Plains to better understand, implement, and facilitate prescribed burns as part of land management plans.
A-27 Cody Whitley
Research Collaborators: Rakshya Shrestha, Roshan Ghimire, Debarati Chanda, Chakravarthi Rajamohan
Research Presentation Title: Utilizing exogenous IL-10 as a possible therapeutic for inflammation caused by SARS-CoV-2
Faculty Research Mentor: Rudra Channappanavar, Veterinary Pathobiology
Inflammation is one of the leading causes of morbidity for COVID-19 caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Previous literature has attempted to utilize Interleukin-10 (IL-10) in treating inflammation. Although it has not shown effectiveness in treating systemic inflammation, other studies have shown that IL-10 is an effective regulator for pulmonary inflammation. Here, we seek to determine the viability of using IL-10 as an inflammatory regulator, to effectively clear the SARS-CoV-2 infection without inducing more cellular damage. In our studies we treated bone marrow derived macrophages with recombinant murine IL-10. We saw a significant decrease in levels of pro-inflammatory cytokines such as Tumor Necrosis Factor-α (TNF-α) and Interleukin-6 (IL-6) with stimulation of TLR 3 and 7 agonists. With IL-10 treatment, Monocyte Chemotactic Protein 1(MCP-1) has shown slight decreases when stimulated with a TLR 3 agonist. Thus far, initial experiments have shown implications for inflammatory regulation and potential therapeutic applications in response to SARS-CoV-2 envelope protein as well as TLR agonists. Future directions will be to determine whether IL-10 will produce comparable results with in-vivo experiments.
A-28 Raygan Kyeremateng
Research Collaborators: Molly Koeck and Colter Chitwood
Research Presentation Title: Feral Pig Population in Southeast Oklahoma
Faculty Research Mentor: Colter Chitwood, Natural Resource Ecology and Management
The Oklahoma Department of Wildlife Conservation wants to understand the biodiversity and wildlife in multiple Wildlife Management Areas (WMA) to maintain and manage the wildlife in Oklahoma. This research is part of a larger project that surveys the biodiversity of James Collins and the San Bois Wildlife Management Area. This project is about finding the frequency of feral pigs individuals and groups of feral pigs (sounders) in the WMAs. Knowing the number and distribution of feral pigs is important because feral pigs can reproduce rapidly, reduce plant diversity and replacement, and have other negative effects on the natural and unnatural world, if unmanaged. The project is currently still processing the data stage, which is preventing me from making a final conclusion on the research.
A-29 Ethan Hood
Research Collaborators: Dan Shoup, Andy Dzialowski, Jason Schooley, Casey Pennock
Research Presentation Title: Evaluating the effect of turbidity on foraging by post-larval Paddlefish
Faculty Research Mentor: Jim Long, Natural Resource Ecology and Management
Previous studies evaluating restoration success of Paddlefish (Polyodon spathula) suggested excessive turbidity in rivers may inhibit foraging by juveniles prior to the development of the rostrum, which contains electroreceptors to help find zooplankton prey. To evaluate this hypothesis, we conducted a laboratory experiment in 10-gallon aquaria to test the effect of turbidity on foraging by post-larval Paddlefish. Aquaria were each assigned a water clarity treatment level of clear (tapwater clarity), 40, 20, 10, or 5cm of Secchi depth (clearer water has a greater Secchi depth than turbid water). Water clarity levels were created by suspending bentonite clay in water until the desired Secchi depth was created. For each trial, approximately 300 zooplankton were added to the aquaria followed by three post-larval Paddlefish, which were allowed to feed on zooplankton for approximately 24 hours. After the 24-hr period, Paddlefish were removed from the aquaria, euthanized with MS-222, and preserved in 70% ethanol. In the lab, Paddlefish were dissected and zooplankton in the gut were counted. Three trials of each turbidity treatment were conducted on each of three dates (total of nine replicates/treatment). Enumeration of consumed zooplankton is still ongoing, but gut contents of two-thirds of experimental fish have been examined to date. Preliminary results indicate a weak, but positive relationship between number of zooplankton consumed and Secchi depth transparency, suggesting turbidity may inhibit foraging by juvenile Paddlefish before the rostrum is fully developed.