Theses and Dissertations at Montana State University (MSU)
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Item A mess of rotten eggs from the Two Medicine Formation, Montana(Montana State University - Bozeman, College of Letters & Science, 2024) Tucker, Katherine Lydia; Chairperson, Graduate Committee: David J. VarricchioThe Two Medicine Formation of Montana is known for producing many dinosaur eggs in varying states of preservation. An unusual clutch of eggs MOR 11881 was recently found in the formation that exhibits some distinctive preservation features. The purpose of this research is to describe this clutch using multiple techniques to identify and better understand its taphonomic history. To do this the clutch was prepared by removing sediment from around the eggs, then examined using light microscopy, scanning electron microscopy (SEM), cathodoluminescence (CL), energy dispersive x-ray spectroscopy (EDX), and electron backscatter diffraction analysis (EBSD). Preparation uncovered eleven partially compressed ellipsoidal eggs in the clutch. Microscopy of the eggshell revealed that it is made of two structurally distinct layers. The lower layer at the base of the eggshell is here referred to as the blocky layer based on the blocky texture of the crystals in SEM images. This layer is sometimes interrupted by a phosphatic dark region. The second layer is the lobed layer found along the outer surface of the eggshell. It is characterized by lobes of blocky crystals surrounded by smaller radiating crystals. In addition to the eggshell the eggs also preserve a phosphatized eggshell membrane, two distinct types of pellets, large masses of collophane crystals and hyphae like structures. Unaltered eggshell is found in the sediment surrounding the clutch. The eggshell of the MOR 11881 clutch does not match any described ootaxon but is similar to eggshell found at the Egg Mountain quarry which was never identified. The unusual structure of the eggshell could be the result of multiple stages of alteration. Several structures in the eggshell including the lobes and the dark region are most likely the result of alteration. The presence of unaltered eggshell near the eggs indicates that the source of alteration was the decay of the eggs as recorded by their contents. This decay also produced the conditions necessary for the exceptional preservation of the eggshell membrane.Item The paradoxical role of ATP in viral infections and cellular defense(Montana State University - Bozeman, College of Agriculture, 2024) Zahl, Trevor Ronald; Chairperson, Graduate Committee: Blake Wiedenheft; This is a manuscript style paper that includes co-authored chapters.Conflict between prokaryotes and viruses has driven the evolution of diverse defense systems. The study of these systems has not only expanded our knowledge of the mechanisms underlying genetic conflict, but has also revolutionized biotechnologies. I joined the Wiedenheft lab in early 2021 during the height of the SARS-CoV-2 pandemic. As such, the lab was focused on developing technologies to aid in the identification and study of SARS-CoV-2. I joined a team that was actively repurposing a type III CRISPR system as a molecular diagnostic. Type III CRISPRs are RNA targeting systems that unleash antiviral signaling cascades upon recognition of a target. Previous work in the lab had shown that this activity could be repurposed as a molecular diagnostic, but the first generation technology was not sensitive enough for clinical use. To address this need, we developed a technology termed Capture and Concentrate, which utilized 'CRISPR magnets' to concentrate target RNAs from complex mixtures. Using this technology, we increased diagnostic sensitivity by 1,000 fold. In addition to my work at the bench, I developed an in silico model of the diagnostic to inform future development efforts. While type III CRISPRs are unique due to their ability to generate antiviral signaling cascades, they also contain intrinsic endonuclease activity. In 2023 we demonstrated that this activity could be repurposed as an RNA editing technology. Using the type III system from Streptococcus thermophilus, we introduced precise genomic modifications in a recombinant sindbis virus. This technology removed the need for reverse genetics systems, which hampered the study of the rapidly evolving SARS-CoV-2 virus. In the final year of my graduate studies, I worked with a team focused on determining how, PARIS, a bacterial innate immune system, induced host death upon viral infection. PARIS is a two-component system comprised of the ATPase, AriA, and a nuclease, AriB. Previous studies had shown that PARIS induced abortive infection in response to viral proteins, but the mechanism was not known. We determined that upon antigen recognition, AriB is activated and cleaves host tRNAs to induce cell death and prevent community spread.Item Biorecovery of rare earth elements and critical minerals via Gluconobacter oxydans(Montana State University - Bozeman, College of Engineering, 2024) Kessler, Kendall Rhea; Co-chairs, Graduate Committee: Ross Carlson and Ryan AndersonThe depletion of high-grade ore deposits, accumulation of electronic waste, and the geopolitical challenges in sourcing critical materials have emphasized the need for sustainable metal recovery methods and recycling efforts in the United States. Conventional metal recovery approaches, including pyrometallurgy and hydrometallurgy, are not only environmentally unsustainable but also inadequate for the retrieval of metals from low-grade deposits. Biorecovery, defined by microorganism-mediated metal recovery, provides an advantageous alternative to traditional recovery methods due to increased sustainability, lower operational costs, and high efficiencies observed for the recovery of low-grade feedstocks. This study investigates the potential of bioleaching as an eco-friendly alternative in the recycling of two distinct waste feedstocks: magnetic swarf and lithium-ion batteries (LIBs). Cultivation of Gluconobacter oxydans was investigated under varying growth medium compositions, wherein increased concentrations of yeast extract were substituted for KH 2PO 4, to determine the subsequent impact on the base metal and rare earth element recovery through the application of the cell-free biolixiviant. This substitution resulted in increased growth yields and enhanced recovery with respect to magnetic swarf, whereas negligible improvement was observed for LIBs. Biorecovery has also been demonstrated for the recovery of metals from ore, where yields are a function of comminution and concentration efficiencies. Typical compressive comminution practices account for the largest proportion of energy expenditures in a mining process. Transcritical CO 2 (tCO 2) comminution, wherein ore is fractured through overcoming a rock's tensile strength, was examined to determine whether physical differences in particle generation were present as compared to traditional fracture techniques. An ore deposit in British Columbia, rich in a nickel-iron alloy mineral phase called awaruite, was examined through scanning electron microscopy, backscatter electron imaging, and energy dispersive spectroscopy to determine the impact of comminution method on awaruite recovery. Image processing was used to investigate shape factors for the individual particles. Preliminary evidence indicates despite a lack of distinct particle differences, tCO 2 comminution resulted in increased liberation and recovery of awaruite ore. Abiotic leaching studies were conducted to determine whether the method of comminution impacted leaching efficiencies. Although samples could not be quantitatively measured, initial qualitative results indicate tCO 2 comminution provides increased yields.Item Impact of kinship navigator program on rural kin caregiver wellbeing(Montana State University - Bozeman, College of Education, Health & Human Development, 2024) Foluso, Ayomide Adefunke; Chairperson, Graduate Committee: Brianna Routh; This is a manuscript style paper that includes co-authored chapters.More than 2.6 million children live in kinship families without their parents. Kinship caregivers provide care for children whose parents are unable to care for them, yet often face significant challenges that can impact their wellbeing and the caregiving situation. In line with the passing of the Family First Prevention Services Act in 2018, Kinship Navigator Programs are required to submit for evaluation in the Title IV-E Prevention Services Clearinghouse to seek federal reimbursement. This study evaluated the efficacy of the Montana Kinship Navigator Program in improving caregiver wellbeing through its intervention (Enhanced Case Management) compared to a service-as-usual control (Information and Referral only). The quasi- experimental design compared caregivers in the intervention group (n=59) to the control group (n=33) across measures of wellbeing, services accessed, and navigator assistance at case closure. Data was collected through surveys and program records. The preliminary findings support wider implementation of enhanced case management services. Analysis showed significantly higher wellbeing in the intervention group across domains of coping, enjoying life, and average wellbeing compared to controls. Navigator assistance was significantly higher for the intervention group and positively correlated with multiple wellbeing domains across all participants. Financial and health service accessed showed positive associations with improved health wellbeing across participants. Financial resources emerged as particularly important for improving wellbeing for rural kinship caregivers. White non-Hispanic caregivers were more likely to access more services and navigator support compared to Black, Indigenous, and other People of Color, who in turn were more likely report higher stress. This demonstrates the need for training and culturally informed navigator support to promote equitable access. Implementing enhanced intervention could better address the complex needs of kinship families, particularly in underserved rural communities where resources are limited. MTKNP's recognition by the Clearinghouse and state legislators can result in a more sustainably funded program and positive wellbeing outcomes for kin caregivers and their kin children.Item Structural control of the Norris Hot Springs and associated geothermal system(Montana State University - Bozeman, College of Letters & Science, 2023) Wafer, Mia Peyton; Chairperson, Graduate Committee: Andrew K. LaskowskiMany active and productive geothermal systems are structurally controlled, suggesting that certain structural configurations have higher potentials for geothermal energy. Analyzing potential productive geothermal wells is known as geothermal play fairway analysis (PFA). Identifying these high potential structural configurations is another way to locate geothermal systems without surface expressions, which represent an unquantified natural resource for geothermal energy. By studying geothermal systems with surface expressions, I move closer to being able to locate additional natural resources. The Norris Hot Springs, located in the foothills of the Tobacco Root Mountains, Southwestern Montana, presents a novel study area to test structural control on geothermal system development. I combine geologic mapping, UAV-based photogrammetry, zircon U-Pb geochronology, seismic data, and 3D geologic modeling to date and interpret the multistage development of local structures and their influence on the geothermal system beneath the Norris Hot Springs. A comprehensive structural model suggests the geothermal system associated with the Norris Hot Springs is hosted by a deep network of faults and fractures in Precambrian gneiss basement. A nearby fault system primarily comprised of high angle NNW-SSE trending normal faults, which often display pervasive alteration and polymetallic vein mineralization, likely connects surficial features with deeper reactivated Laramide-era structures. A primary goal of this study was to investigate the benefits of supplementing traditional field work with novel methods. Results from this study demonstrate the utility of combining geologic mapping, 3D modeling, structural analysis, 3D modeling and UAV surveys for geothermal exploration and constrain the timing and influence of local faulting on geothermal activity at the Norris Hot Springs.Item Ewe undernutrition alters cotyledon development and function(Montana State University - Bozeman, College of Agriculture, 2023) Welter, Francesca Marie; Chairperson, Graduate Committee: Sarah R. McCoskiUndernutrition is common for animals maintained on rangelands, particularly during periods of drought. Undernutrition during gestation can be detrimental to offspring development. The placenta is a known mediator for maternal-fetal nutrient exchange, and developmental impacts to the placenta during nutritional stress contribute to poor offspring outcomes. The objective of this study was to determine the impacts of maternal nutrient restriction on the development and function of placental cotyledons. Targhee ewes were allocated into two dietary treatment groups for the entirety of gestation. The control group (CON; n = 10) was fed to meet 100% of NRC requirements and the nutrient restricted group (NR; n = 11) was fed to meet 60% of NRC requirements, to mimic the nutritional stress experienced by range ewes during drought. At the time of lambing total placental weight, total cotyledon number, and lamb birth weight was recorded. Cotyledons (n = 3 / placenta) were removed from multiple locations, weighed, and processed for RNA-sequencing. Total placental weight, total cotyledon number, and lamb birth weight were not affected by maternal diet restriction (P > 0.05). Average cotyledon weight was greater in the CON group compared to the NR group (P < 0.05). Gene ontology (GO) and Kyoto encyclopedia of genes and genomes pathways (KEGG) analyses were performed to identify biological pathways involved in placental function and nutrient transfer. Cotyledons from NR singletons vs CON singletons expressed 470 differentially expressed genes (DEGs; P < 0.05; 187 up-regulated, 283 down-regulated). Gene ontology terms represented by DEGs include hormone activity (P = 0.0003) and antigen processing and presentation (P = 0.0004 ). KEGG pathways of interest include pyruvate metabolism, estrogen signaling, extracellular matrix receptor interaction, and antigen processing and presentation. There were also 426 DEGs (P < 0.05; 195 up-regulated, 231 down-regulated) in cotyledons of NR twins vs CON twins. Represented GO categories include heme binding (P = 0.003) and oxidoreductase activity (P = 0.03). These data show that prolonged nutrient restriction alters cotyledon development and gene expression indicating impacts to placental function. These changes in placental function likely mediate poor offspring developmental outcomes observed following gestational under nutrition.Item Going beyond the voice of God: unconventional voiceover narration and the importance of point of view in documentary films(Montana State University - Bozeman, College of Arts & Architecture, 2023) Lobao, Ashley Irene; Chairperson, Graduate Committee: Hugo R. SindelarDocumentary film narration styles have evolved over the years adjusting to the changes in the medium's trends. An unconventional narration style, one that is more personal and intimate, has evolved and become the newest narration trend. This paper evaluates four films: Conservation International's 'Nature is Speaking' (2014), Raoul Peck's 'I Am Not Your Negro' (2016), Chris Marker's 'Sans Soleil' (1983), and finally, Ashley Lobao's 'Vineas Salutis: Vineyards of Salvation' (2023) to explore how an unconventional narration can establish a films point of view and deliver a deeper meaning to the viewer.Item Late gestation maternal overnutrition alters gene expression and histomorphology in neonatal foal testes(Montana State University - Bozeman, College of Agriculture, 2023) Trauner, Alexis Maria; Chairperson, Graduate Committee: Amanda N. BradberyWhile obesity has deleterious effects on metabolism, performance, lameness, and health in mature horses, little is known about the impacts to reproductive health in foals from overfed dams. The objective of this study was to determine the effects of maternal overnutrition on neonatal foal testicular development. To test this, 16 Quarter horse mares were used in a completely randomized design and stratified by expected foaling date, body weight, and body condition score into one of two dietary treatments offered twice daily beginning on gestational d 235: control (CON; n = 8) fed to meet nutrient requirements and overfed (HIGH; n = 8) fed 40% above CON. At 5 h postpartum, foals were humanely euthanized, and testes were harvested, weighed, and stored for analyses. RNA was extracted and submitted for RNA sequencing. Histomorphometric evaluations were performed using a hematoxylin & eosin stain. ImageJ software was used to evaluate individual seminiferous tubule (ST) area, ST number, and % area of STs relative to interstitial tissue, as well as Sertoli cell numbers per ST. Data were analyzed using the MIXED procedure in SAS. Testes from HIGH male foals (n = 4) were 61% larger than testes from CON male foals (n = 4; P < 0.01). RNA sequencing revealed 56 differentially expressed genes between HIGH and CON groups (P-adjusted < 0.05). These genes represent biochemical pathways involved in oxidative phosphorylation, fatty acid metabolism, and thermogenesis, which are integral for testicular development and function. Gene ontology analyses revealed differential gene expression was associated with biological processes relevant to reproductive function and spermatogenesis, including cellular respiration and mitochondrial function. Histological analyses revealed no difference in individual ST area (P = 0.76) or Sertoli cell number within STs (P = 0.57); however, testes from CON foals had 2.3x more STs per 10X field than HIGH foals (P = 0.01) along with STs representing a greater percentage of the image area relative to interstitial tissue (P = 0.05). These data from the neonatal foal indicate that late- gestation maternal overnutrition alters fetal testicular development with potential long-term implications.Item First digital forelimb reconstruction and range of motion assessment of the late Cretaceous dinosaur Troodon formosus(Montana State University - Bozeman, College of Letters & Science, 2023) Serio, Michael Anthony; Chairperson, Graduate Committee: David J. VarricchioTroodon formosus (Troodon) is a theropod dinosaur from the Late Cretaceous of North America primarily known from fragmentary fossil material. Hypothesized to hunt small game, Troodon, like many theropods, may have actively engaged its forelimbs to aid hunting behavior. Furthermore, the complexity of preserved nests associated with Troodon suggest it had sufficient range of motion and dexterity to use its forelimbs to manipulate its eggs. Additionally, Troodon's evolutionary position as one of the closest groups to living birds could indicate important information about the evolution of the avian wing. However, no complete forelimb material has yet been found for Troodon, and as such a complete reconstruction and range of motion (ROM) estimate of its forelimbs has yet to be attempted. This study aims to address these hypotheses and shortcomings by leveraging recent digital modelling technology to create the first forelimb reconstruction and ROM for Troodon. To overcome preservation limitations, surface scans from multiple Troodon fossils housed in the Museum of the Rockies (Bozeman, Montana) were digitally combined to reconstruct most of a forelimb. From this model, digital articulation in the software package Maya was compared with physical articulation ROM methods using 3D printed copies of the forelimb. The maximum angle of allowed motion for each forelimb joint until bone-on-bone contact was assessed. Results show increased flexion over extension in Troodon's joints, consistent with closely related species. However, Troodon also shows higher manual extension than close relatives. The humerus is also shorter and stockier, showing mechanical specialization for strength vs speed, and convergent features with primitive theropod species. These ROM and morphological differences imply a deviation in hunting strategy or forelimb utility and a deviation from avian forelimb morphology. Total manual ROM results cannot confirm whether Troodon was able to grasp its eggs and small prey single-handed, but two-handed apprehension remains feasible. This study utilized digital and physical ROM methods, analyzing for the first-time specimens that would have been previously overlooked. These results shed light on both the foraging and reproductive behavior of Troodon and have implications for understanding the evolution of modern avian reproductive behavior and the dinosaur-bird transition.Item Enabling real-time application streams in off-grid regions and mission critical applications through BRP, radios, and gateways with COTS components(Montana State University - Bozeman, College of Engineering, 2023) Mekiker, Batuhan; Chairperson, Graduate Committee: Mike WittieThe Internet of Things (IoT) applications require flexible and high-performance network solutions, but many IoT solutions can only support single-use case applications, which limits their performance and flexibility for real-time and streaming applications. LoRa offers a flexible physical layer but lacks the power needed in its link layer protocols to support real-time flows. The Beartooth Relay Protocol (BRP) expands the performance envelope of LoRa, making it suitable for a wide range of IoT applications, including those requiring real-time and streaming capabilities. However, the resource-limited nature of LoRa does not allow BRP to support self-healing mesh network capabilities or beyond two hops while maintaining real-time streams. To address the limitations of BRP in supporting mesh network capabilities and real- time streams beyond two hops, we move our focus to the development of the second- generation Beartooth Radios, MKII, and the first-generation Beartooth Gateways. We utilize Commercially-available Of Shelf Components (COTS) in the radios to provide a cost-effective, power-efficient, and compact solution for establishing real- time situational awareness. The self-healing mesh network provided with MKII and Gateways also enhances the reliability of the overall network, ensuring connectivity even in case of node failures. By incorporating military information brokers, such as the Tactical Assault Kit (TAK), the Beartooth Gateway establishes a hybrid network between Beartooth radios, gateways, and other TAK-capable devices, ensuring compatibility with existing IP networks. By combining Beartooth MKII radios, Gateways, and flexible link layer protocol elements in BRP, this research demonstrates a versatile and flexible solution that provides real-time application streams and critical situational awareness capabilities in mission-critical applications.