Program

St. Jude Children’s Research Hospital

Jason Rosch received his Ph.D. in molecular microbiology and microbial pathogenesis from Washington University in St. Louis. He subsequently moved to St. Jude Children’s Research Hospital as a postdoctoral fellow, focusing on host-pathogen interactions in the context of the Streptococcus pneumoniae and high-risk hosts. He remained at St. Jude as faculty and has progressively risen through the ranks to his current appointment as full member in the Department of Host-Microbe Interactions.

Rosch has more than 20 years of experience working with pathogenic streptococci, as well as several other pathogens, from studying basic biology to modeling host-pathogen interactions. His current research focuses on polymicrobial respiratory infections, specifically influenza A virus and the pneumococcus, and the evolutionary constraints of antibiotic resistance development and spread using the pneumococcus as a model. The primary focus of these 2 areas of research is to understand how virulence strategies and development of antibiotic resistance differ in high-risk patient populations, through translational projects in collaboration with clinicians, as well as basic research into fundamental biological processes underlying these aspects of host-pathogen interactions. The research has encompassed lab-based experimental modeling of host-pathogen interactions and resistance development, and also detailed analysis of clinical isolates from high-risk pediatric patient populations.

Rosch’s background in bacterial genetics and pathogenesis modeling has allowed him to achieve mechanistic insights into host-pathogen interactions in such context and have revealed a number of important insights into these processes in several areas, including viral-bacterial co-infections, recalcitrant bacterial infections in immune-compromised patients and mechanistic dissection and targeted therapies to mitigate infectious complications in high-risk patient populations.

P1-001

Halophiles and Copper Resistance in the Great Salt Lake

Rylee Wamhoff, Michele Culumber

Weber State University

The Great Salt Lake is one of Utah’s most hypersaline and ecologically distinct environments, supporting communities that range from brine shrimp and birds to halophilic microbes such as Salinivibrio and Halomonas. As lake levels decline and sediments become exposed, heavy metals (including copper) are increasingly concentrated, raising questions about how native halophiles persist under metal stress. This project investigates the copper tolerance of culturable halophilic bacteria from Ladyfinger Point to better understand microbial resilience in a changing lake. Water and sediment samples were collected and plated on halophile agar (HA; 12% NaCl) to isolate distinct colonies. Fifteen pure cultures were then tested for growth on HA supplemented with 0.5, 1, and 2 mM copper sulfate to assess tolerance thresholds. Of the fifteen isolates obtained, ten demonstrated robust growth up to 2 mM, indicating unexpectedly high resistance across multiple morphotypes. Minimum inhibitory concentration testing will be performed to determine the upper limits of copper tolerance in these isolates. These findings suggest that Great Salt Lake halophiles may possess diverse and effective copper-resistance mechanisms, with potential implications for sediment stability and natural bioremediation. 16S rRNA sequencing will identify the taxa involved and clarify whether known or novel species exhibit this resilience.

P1-002

Two Novel A1 Subcluster Myobacterium Phages Whitspit and Blackflame

Abigail Pace, Jesse Henrie, Cade Brink, Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Jake W. Williams, Donald P. Breakwell, Brett E. Pickett​

Brigham Young University

With over 1031 phages thought to exist in the biosphere and fewer than 7000 sequenced genomes in GenBank, there is a paucity of annotated phage genomes. To contribute to the nationwide effort to expand the phage database, we isolated two novel mycobacteriophages, Whitspit and Blackflame, from compost samples and identified them as members of the A1 subcluster. The phages were isolated using Mycobacterium smegmatis mc² 155 and seven rounds of purification. Both phages were sequenced using an Illumina platform and assembled and annotated using Newbler, PECAAN, NCBI, HHPRED and Phamerator. We analyzed and compared Whitspit and Blackflame to other phages in this subcluster by examining the phylogeny of both phages, comparing morphology and characteristics, and generating an average nucleotide identity matrix. Our phages Whitspit and Blackflame span 48,679 base pairs with 82 predicted ORFs and  52,641 base pairs with 85 predicted ORFs, respectively. In comparison with typical A1 phages, Blackflame and Whitspit have typical lengths, total number of ORFs, GC content, and morphology. Blackflame is atypical due to a hypothetical protein located between the Lysin A/Lysin B proteins. An average nucleotide identity (ANI) matrix quantified both sequences vs others. Whitspit shows high divergence from the A1 subcluster in its large subunit terminase gene, as shown in the phylogenetic tree. Overall, the data indicate that Whitspit and Blackflame are similar in genome characteristics, yet display several novel features. The annotation and archiving of both phages will contribute to further research and understanding of A1 phages.

P1-003

Toxin/Antitoxin HicAB systems in Mycobacteriophages

Grant A. Stock, Jake W. Williams, Cade Brink, Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Brett E. Pickett, Donald P. Breakwell

Brigham Young University

Mycobacteriophages represent a large pool of genetic diversity and knowledge of their genomic data is increasing. Mycobacteriophage Qhanda was isolated from chicken compost in Hooper, Utah. Isolation was achieved using aMycobacterium smegmatis mc2 155. Following three rounds of purification, high-titer lysates (>108 pfu/ml) were prepared, and scanning electron micrographs were taken. Paired-end sequencing reads were generated using an Illumina platform, and the genomes were assembled. Annotation data were collected using PECAAN, NCBI Blast, HHpred, AlphaFold, Host-trained GeneMark and Phamerator. We observed an open reading frame in the genome of the mycobacteriophage Qhanda that is also found in other bacteriophages including Bobquesha, Malthus, MissDaisy, Patt, and Ruthiejr. This open reading frame contains a gene encoding a toxin/antitoxin HicA gene which, when found in bacteria, encodes an anti-bacteriophage mechanism. Included is a web logo of the open reading frame, indicating highly conserved segments of amino acid sequences for the gene aligned with the Qhanda amino acid sequence. Phylogenetic tree reconstruction showed relationships between closely-related K4 phages. AlphaFold comparison of a bacterial association of a Toxin/antitoxing HicAB complex with the phage protein of Toxin HicA. Qhanda and similar phages Comparison of Genomes.

P1-004

Mapping Antibiotic Resistance: A Survey of Microbial Contamination on the UVU Campus

Ben Van Noy, Alex Hayex, Kelvin Morton, Levi Brown, Lauren Brooks

Utah Valley University

Antibiotic-resistant Staphylococcus aureus poses a growing public health concern, particularly strains such as Methicillin-Resistant Staphylococcus aureus (MRSA) and Vancomycin-Resistant Staphylococcus aureus (VRSA). While MRSA infections can often be treated with alternative antibiotics, VRSA is especially concerning because vancomycin is typically used as a last-resort treatment. Our research aims to determine whether antibiotic-resistant Staphylococcus species exist on commonly touched surfaces across the UVU campus. Environmental samples will be collected from multiple campus buildings and plated onto Mannitol Salt Agar (MSA) with and without antibiotic, (methicillin or vancomycin) to select for Staphylococcus species and identify antibiotic-resistant isolates. After incubation, colony-forming units (CFUs) will be counted to evaluate bacterial growth and survival in the presence of antibiotics. Identifying the presence of antibiotic-resistant Staphylococcus in our community will increase awareness of antibiotic resistance and provide a foundation for future analysis of resistance genes.

P1-005

Isolation and characterization of Xanthomonas bacteriophages with biocontrol potential in agriculture

Natalie A. Olsen, Lydia A. Borg, Julianne H. Grose Ph.D.

Brigham Young University

Xanthomonas euvesicatoria is the causative agent of bacterial spot in tomato and pepper plants, rendering the plant and fruit unmarketable. This causes economic stress for growers, with reported losses of over 60% crop yield and over $8 million in losses for the U.S. southwestern states alone. Most control attempts for this disease are preventative, including crop rotation, antibiotic sprays, and copper sprays. However, antibiotic and copper resistance have been increasingly noted in X. euvesicatoria strains, strongly limiting the effectiveness of these pre-treatments. Bacteriophages, or phages, offer an innovative solution. Phages are highly selective viruses that infect and lyse specific strains of bacteria. Through discovery and analysis, phages have the potential to be combined as a targeted treatment, enabling a curative treatment which can rescue diseased plants. Our project aims to collect phages from sewage and test them against a collection of 37 Xanthomonas strains isolated from farms in South Carolina. Our long-term goal focuses on creating a combined cocktail therapy for infected plants. We have isolated and sequenced 11 unique phages which infect X. euvesicatoria and are in the processes of isolating more. The phages are raised to a high titer and sequenced for characterization as well as analyzed for the presence of genes involved in bacterial virulence. They will also be tested in host range assays to determine their scope against field isolates. Creating an environmentally-friendly and effective biocontrol treatment will be beneficial to growers, increasing food security and profit as well as rescuing diseased plants.

P1-006

Bioelectrochemical Nitrogen Upcycling from Bioprocessing Waste for Protein Synthesis

Lukas Keller1,2, Dr. Jia Zeng2, Dr. Luguang Wang1

1Utah State University, 2ThermoFisher Scientific

            Industrial biomanufacturing leads to the production of large quantities of bioprocessing waste, raising concerns about sustainability and nutrient loss. There are significant environmental incentives to recover these nutrients and upcycle them for other industrial uses. Bioprocessing waste often contains high concentrations of cell byproducts like ammonium, which is a toxin in high concentrations and has a known inhibitory effect on mammalian cell growth and production. One potential method of utilizing ammonia-laden waste and recycling media protein is in a bioelectrochemical system (BCS). This system utilizes the native metabolism of wastewater-derived microbes like exoelectrogens, methanogens, and acetogens to produce biogas in anaerobic conditions. BCSs can recycle culture media components by facilitating the production of single-cell protein (SCP). Bioprocessing waste, rich in ammonium, can be used as a nitrogen source for reactor growth. The SCP microbes grown from waste components can later be hydrolyzed and used for industrial processes like manufacturing feedstock for cultured meat or animal feed. This system relies on the establishment of a micro-ecosystem where the H2 and CO2 gases are produced from electrogens. Ammonium and materials from bioprocessing waste are used by homoacetogens to propagate SCP. SCP synthesis can be increased through the creation of a hypothesized ‘hydrogen production-consumption loop’ in the BCS. Homoacetogens can then become the dominant cathodic and planktonic species following methanogen inhibition and proliferate into SCP using the Wood-Ljungdahl Pathway from the produced H2 and CO2 gas.

P1-007

Calcium Recovery from CaCO3-Rich Wastes Using Fungal Produced Organic Acids

 Allondra Woods, Erika Espinosa-Ortiz

Utah State University

Achieving a circular bioeconomy requires urban mining strategies to recover resources from alkaline, calcium-rich waste streams such as concrete and industrial slag, which contain high concentrations of calcium carbonate (CaCO3). Traditional recovery relies on synthetic acids, like HCl and HNO3, to mobilize calcium for reuse in high-value applications, including fertilizers and building materials; however, these approaches are not eco-friendly. The fungus Aspergillus niger is widely used industrially due to its ability to produce large quantities of organic acids, including citric and oxalic acid. Through citric acid production A. niger offers a sustainable alternative to traditional calcium recovery, by converting insoluble calcium minerals like CaCO3 into dissolved, recoverable forms. In this study, citric acid production by A. niger was investigated as a sustainable approach for calcium solubilization from CaCO-rich waste streams. A. niger was cultivated under varying media compositions, pH, and durations at 30°C and 150-250 rpm. Organic acid production was monitored via pH and liquid chromatography. Following biomass removal, the spent acidified medium was applied to pure CaCO3 to model calcium solubilization from waste streams. The resulting leachate was analyzed for organic acids composition, pH, and dissolved calcium concentration. This study advances fungal-based leaching as a sustainable urban mining technique by establishing a strategy to produce calcium-rich leachates for use in downstream material synthesis.

P1-008

Sequencing and Analysis of the Bacteriome of Puno Bay, Lake Titicaca

Cori Bailie, Dr. Lauren Brooks

Department of Biology, Utah Valley University

Puno Bay, a distinct region of Lake Titicaca, serves as a tourist destination while providing tap and agricultural water to the surrounding regions. Contamination from mining and wastewater pollution occurs along the rivers that feed into the bay, leading to immediate concerns such as elevated levels of arsenic and pathogens, as well as secondary concerns such as algal blooms resulting from elevated nutrients. However, detailed information on how this affects the microbial community is lacking. Our study characterizes the bacteria from 7 sampling locations within Puno Bay. Triplicate samples were collected at each site and filtered through a 0.2-micron filter to capture cellular organisms. Filters were preserved in DNA Shield and brought back to Utah Valley University for DNA extraction. Samples were quantified and mailed to SeqCenter for 16S short-read Illumina sequencing. Sequence data was processed through a custom bioinformatics pipeline to evaluate common microbial ecology metrics. We found that microbial composition generally varied between collection sites, with some samples dominated by taxa associated with fecal contamination, while others contained elevated levels of cyanobacteria or chloroplasts. This suggests contamination from fecal matter is directly contributing to the microbial communities in portions of the lake. While no algal bloom was observed at the time of sampling, elevated cyanobacterial levels in other portions of the bay suggest a baseline presence of these taxa that could contribute to the frequently observed blooms.

P1-009

Investigating the impact of sequential treatment of herbicide mixes on the soil microbiome

Emily Bradley, Eric Christen, Anirban Chakraborty

Idaho State University

Land management agencies in the United States frequently employ herbicides to manage non-native invasive plants along roadways. While herbicides are highly effective in controlling weeds and invasive plants, they have been shown to adversely impact soil microbial communities and their multifaceted functions. Soil microbial communities drive nutrient cycling that impacts ecosystem health, stability, and functionality. Therefore, it is important to understand the effects herbicide applications may have on these communities. To address this knowledge gap, we investigated whether the application of an herbicide mix during a roadside revegetation experimental project impacted the soil microbial community structure using surveillance of phylogenetic marker genes. Additionally, we also monitored legacy effects of herbicide on the microbial biomass. Experimental plots situated at rest areas along the I-15 interstate highway, with each site located within a distinct level IV EPA ecoregion, were treated with varying mixes of herbicides over a three-year period. Soil was collected each summer annually at each site to perform DNA-based microbial community surveillance and microbial biomass carbon analysis. Community analysis revealed significant differences in communities across sites along with small community dissimilarity between herbicide and non-herbicide plots. Changes in biomass carbon due to legacy showed how applied herbicides affected the total biomass. These observations cumulatively suggest that herbicide applications have the potential to alter soil microbial communities.

P1-010

Nanoinjection: Bacterial Delivery into Insect Eggs using a Charged Lance

Ella Petersen, Natalie Garlock, Jonah Jex, Nathan Dauk, Amy Cherpeski, Christian McCombs , Abby White, Taylor Coffman, Abe Alton, Brian Jensen , Sandra Hope

Brigham Young University

Paratransgenesis is a strategy to control vector-borne diseases by genetically modifying a symbiotic microbe within an insect rather than the insect itself. Current approaches rely on microinjection, which uses relatively large hollow needles to deliver symbionts and often fails to penetrate the hardened eggs of many insect species. An alternative method, nanoinjection, could allow for the penetration of these hardened shells using solid metal lances and electrostatic attraction to deliver material. We tested the injection of Sodalis praecaptivus into Callosobruchus maculatus (bean beetle) eggs at three injection sites (head, side, and back) to evaluate the viability of nanoinjection in delivering bacteria into insect eggs. We hypothesized that nanoinjection can successfully deliver live bacteria into insect eggs, and that back injections offer the best conditions for beetle hatch rates as they are the predominant injection location for microinjections. Nanoinjection achieved bacterial transfer into 35.8 ± 5.3% (n=891) of injected eggs, indiscriminate of injection site. Hatch rates of injected eggs were 1.2 ± 0.8% (n=655) with bacterial integration occurring in 62.5 ± 33.6% (n=8) of hatched beetles. These beetles represent the first documented case of bacterial integration into an insect model lacking a native bacterial symbiont and underscores nanoinjection’s versatility in introducing symbionts across various insect species. Further work will focus on the physiological effects and inheritance patterns of Sodalis praecaptivus within a bean beetle model, as well as applying nanoinjection to mosquitoes, potentially establishing paratransgenic strains capable of countering vector-bone diseases. 

P1-011

Title of your poster: Quantification of Antimicrobial Reactive Oxygen and Nitrogen Species (RONS) Generated by a Non-Thermal Plasma Array

Stephanie Rood, Hassan Mohamed, Marlon Villatoro, Laney Dew1 Neil Benavente, Kahlan Carpenter, Hanna Wallin, Luke Rodriguez

Boise State University

Non-thermal plasma (NTP) has emerged as a promising non-thermal technology for microbial inactivation, largely due to its ability to generate reactive oxygen and nitrogen species (RONS). However, quantitative relationships between RONS production rates and antimicrobial efficacy remain insufficiently characterized. The reactive species hydrogen peroxide (H₂O₂), ozone (O3), and hydroxyl radicals (•OH) were measured using spectrophotometric-based assays. In this study, we developed and optimized experimental protocols to quantify the production rates of key RONS generated by a 10×10 cm electrode array operated in air under controlled conditions. Our findings correlated antimicrobial outcomes to the concentration and temporal dynamics of RONS production. Quantifying RONS concentrations based on specific device parameters allows for industrial application by understanding the amount of RONS produced at each setting. This work provides a quantitative framework for linking NTP-generated reactive species to antimicrobial effects, supporting the rational design and optimization of plasma-based sterilization technologies.

P1-012

Phylogenetic patterns of pediatric invasive pneumococcal serotype decline, persistence, and emergence in Utah during the covid-19 pandemic.

William J. Hunckler, MD, MPH1,  Nicole L. Pershing, MD, PhD1, Aurélie Kapusta, PhD1,2, Shannon Nielsen, BS1, Hillary Crandall, MD, PhD1, E. Kent Korgenski, MS1,3, Carrie L. Byington, MD4, Krow Ampofo, MD1, Anne J. Blaschke, MD, PhD1
1Department of Pediatrics, University of Utah, Salt Lake City, Utah

2IDbyDNA, Salt Lake City, UT;

3Pediatric Clinical Program, Intermountain Healthcare, Salt Lake City, UT

4University of California Health, Oakland, CA

Background: Incidence of invasive pneumococcal disease (IPD) decreased following the introduction of infection prevention efforts in the COVID 19 pandemic, followed by an increase following relaxation of precautions. As with shifts following introduction of new Pneumococcal Vaccines (PCV), uneven pressure was applied by the pandemic to different serotypes (STs). Methods: We performed Illumina sequencing, de novo genome assembly/annotation and in silico serotyping of clinical sterile site pneumococcal isolates from pediatric patients at Primary Children’s Hospital (Salt Lake City, UT) in three eras: Pre (2018-2019), Early (2020-2021), and Late Pandemic (2022-2023).  Results: 169 pneumococcal isolates from 39 STs were analyzed. From the Pre to Early to Late Pandemic, ST3 shifted from 13.9% to 20.5% to 20.7%; ST8 1.4% to 7.7% to 0%; ST11A 9.7% to 0% to 3.5%; ST15B 4.2%, to 7.7% to 1.7%;  ST19F 15.3% to 7.69% to 20.7%, ST22F; 6.9% to 12.8% to 12.0%; ST23A 4.2% to 5.1% to 0%, and ST23B1; 0% to 0% to 5.17%.  All other STs shifted less than 2.8%. Conclusions: There were significant ST shifts between the three eras, but the largest and most persistent increases were seen in STs 3 and 19F, long covered by PCVs 13 and 7. A newly covered ST paradoxically worsened, and non vaccine STs extinguished and emerged respectively. These shifts associated with non-vaccine pressures and escape from current vaccines imply the need for dynamic approaches to pneumococcal prevention.

P1-013

Antibiotic Treatment and Microbial Discordance of Paired CNS-Sinus Cultures in Pediatric Sinogenic Intracranial Infections 

Taylor A. Boyd1, Isabella McNamara MD MPH2, Jonathan Race PhD1, Anne J. Blaschke MD PhD1, Jared Olson PharmD1, Nicole Pershing MD PhD1

1University of Utah, Salt Lake City, UT, USA, 2Tufts Medical Center, Boston, MA, USA

Background: Intracranial extension of infectious sinusitis is a rare but potentially life-threatening childhood disease. Streptococcus anginosus group (SAG) species are the most frequently identified pathogens, but infections are often polymicrobial. Intravenous antibiotics are the mainstay of treatment, but optimal antimicrobials remain unknown. Methods: We performed a retrospective chart review of patients admitted to Primary Children’s Hospital with ICD9 and ICD10 diagnoses of sinusitis and intracranial infection between 2004 and 2019. We compared microbiologic data, disease course, and antibiotic treatment over the study period. Results : Disease incidence significantly increased across the study period (p<0.0001). We identified 83 patients.  Surgical cultures from the sinus (n=60) and central nervous system (CNS, n=58) were reviewed.  Most patients (63%) had polymicrobial infections. SAG were the most commonly identified pathogen (57% of patients), followed by anaerobic organisms (31%). Paired surgical cultures from both the sinus and CNS were obtained for 48% of patients.  Microbiologic results were discordant between sites in 85% of these cases; Staphylococcus aureus was detected almost exclusively from sinus cultures (n=10/11).  Treatment antibiotics included ceftriaxone/metronidazole (n=35) and carbapenems (n=31), including ertapenem (n=21).  Vancomycin was frequently started empirically, with decreased duration over time (p<0.0001). All tested SAG isolates were susceptible to meropenem (n=33), ertapenem (n=3), and ceftriaxone (N=36).  Conclusion: Sinogenic intracranial infections are typically polymicrobial including SAG and anaerobes.  Sinus culture data may not accurately reflect CNS pathogens.  Ertapenem may be considered for treatment when contraindications to other antimicrobials exist.  Vancomycin is rarely required. Additional data are needed to inform generalizability of these findings.

P1-014

Chimeric autoantibody receptor (CAAR) T cells selectively target autoreactive B cells in antiphospholipid syndrome

David Carter, Brianna Davis, Abby Cheever, Kimball Demars, Scott Weber, Kim O'Neill

Department of Microbiology and Molecular Biology, Brigham Young University

Introduction: Current chimeric antigen receptor (CAR) T cell therapies effectively treat B cell malignancies but cause widespread immunosuppression through the elimination of all B cells. Antiphospholipid syndrome (APS) is a B-cell-mediated autoimmune disease that causes thrombosis and recurrent miscarriages, and which typically requires indefinite anticoagulation treatment with no cure. Chimeric autoantibody receptor (CAAR) T cells represent a more precise immunotherapy approach by redirecting T cells to target only autoreactive B cells while preserving the healthy B cell population. Methods: While CAR T cells use a single-chain variable fragment as their binding domain, CAAR T cells use the autoantigen to act as bait for autoreactive B-cell receptors. In APS, B cells produce antibodies directed against the clotting factor β2-glycoprotein I (β2GP1), which binds to phospholipids on cells. Using patient-derived epitopes, we engineered two distinct autoreactive B cell populations and two distinct β2GP1-expressing CAAR T cells that attract and eliminate target B cells. Results: One of the β2GP1 CAAR T cell populations achieved complete depletion of autoreactive B cells without exhibiting cytotoxicity against healthy B cells. A second β2GP1CAAR design killed significantly more pathogenic B cells than healthy B cells (p-value < 0.0001) but did not successfully eliminate the autoreactive population. Continuing forward, we plan to assess the cytokine secretion and activation of our β2GP1 CAAR T cells in the presence of soluble antibody and patient serum. These preliminary results demonstrate that CAAR T cell therapy can achieve selective depletion in APS, potentially offering a disease-modifying therapy that preserves protective immunity.

P1-015

Comparative Colon Transcriptomics of Crohn’s Disease and Ulcerative Colitis Reveals Shared Therapeutic Targets and Distinct Molecular Pathway Perturbations

Matt Reall, Brett Pickett

Brigham Young University

Inflammatory bowel diseases (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), involve chronic intestinal inflammation, yet the molecular pathways driving epithelial dysfunction remain incompletely understood. RNA-sequencing datasets from the Gene Expression Omnibus were analyzed on the BYU supercomputing cluster using a bioinformatics pipeline that included ARMOR-based preprocessing and differential expression analysis, SPIA pathway enrichment, and Pathway2Targets drug-target prioritization to compare colon tissue gene expression among healthy controls (n=340), CD patients (n=641), and UC patients (n=766) in R. In the CD vs control comparison, 14005 differentially expressed genes (DEGs) were identified, while 13623 DEGs were identified in the UC vs control comparison. The main transcriptional changes included strong upregulation of inflammatory and epithelial stress genes such as SERPINB3 and KLK6 in both Crohn’s disease and ulcerative colitis, with S100A12 appearing especially prominent in ulcerative colitis, alongside downregulation of genes involved in epithelial metabolism and ion transport, including ATP5MGL in both conditions and KCNQ1 in Crohn’s disease, suggesting disrupted epithelial energy metabolism and barrier integrity across both major forms of inflammatory bowel disease. These findings highlight convergent inflammatory and epithelial stress programs in CD and UC and identify potential biomarkers and therapeutic targets for understanding and treating chronic intestinal inflammation.

P1-016

Secondary Analysis of Bulk RNA-Sequencing Data from Patients Infected with Dengue Virus

Annelise K. Eldredge, Nathan D. Walker, Brett E. Pickett

Brigham Young University

Dengue virus (DENV) infects an estimated 100-400 million people annually, yet no approved therapeutic treatments exist. Characterizing the host transcriptional response in clinical samples may reveal viable drug targets and illuminate which immune cell populations drive pathology. We analyzed 65 bulk RNA-sequencing samples from two independent clinical studies (GEO/NCBI) comprising healthy controls, dengue cases, and severity-stratified dengue patients. Read preprocessing used Trim Galore and Salmon (GRCh38), with differential expression assessed via edgeR. Significantly perturbed pathways were identified using the SPIA algorithm against the KEGG and Reactome databases, and potential drug targets were prioritized using Pathway2Targets. Digital cytometry was performed with CIBERSORTx to impute immune cell-type proportions from bulk counts; differences across groups were evaluated using Kruskal-Wallis with Dunn post-hoc testing (4-group analysis) and Wilcoxon rank-sum tests (control vs. all dengue cases collapsed). We identified 5,922 differentially expressed genes between DENV-infected and healthy patients. Pathway enrichment revealed ten significantly perturbed pathways, most prominently mTOR signaling (activated) and insulin signaling (suppressed), alongside immune-related pathways. TNF and IL-6 pathway antagonists emerged as priority candidates for therapeutic repurposing. Consistent with known dengue immunopathology, CIBERSORTx deconvolution revealed robust mast cell activation specifically in severe dengue and differential M1 macrophage responses in non-severe disease, with effect sizes that may exceed previously reported magnitudes. Together, these results support TNF and IL-6 antagonism as repurposing candidates and highlight severity-stratified immune cell shifts as potential biomarkers of dengue disease progression."

P1-017

Scalable, Automated Primer Design: A Snakemake-Based Pipeline for Sensitive Bacterial Pathogen Detection       

Katelynn B. Owens, Zach Fears, Brett E. Pickett

Brigham Young University

Developing qPCR primer and probe sets to detect and distinguish between closely related bacterial pathogens can be time-consuming and costly when performed manually, often requiring extensive experimental screening to identify primers that are both highly sensitive and species-specific. This project aimed to develop an automated computational workflow to streamline the prediction of high-performing candidate PCR primer and probe sequences to differentiate and diagnose clinically relevant bacterial species. Assembled genomes from multiple strains of a target bacterial species and related members of the same genus were collected and analyzed using a Snakemake-based bioinformatics pipeline. Genomes were first annotated using Prokka, followed by pan-genome analysis with Roary to determine gene presence and absence across strains. A custom R script was then used to identify genes present in at least 99% of the target species while absent or rare in other genus members. Alleles corresponding to these candidate genes were extracted and aligned using MAFFT to generate multi-sequence alignments and identify species-conserved regions for primer binding that can sufficiently differentiate species. The existing Primer3 program was subsequently used to design candidate primer and probe sets based on predicted thermodynamic properties and sequence conservation for these target regions. This workflow generates a ranked list of candidate genes and primer configurations, providing an efficient framework for developing sensitive and species-specific molecular diagnostics using a scalable big data approach to identify target pathogens.

P1-018

Cold Atmospheric-Pressure Plasma: A Solution for Microbial Inactivation on Crop Seeds

Stephanie Rood, Hassan Mohamed, Marlon Villatoro, Briana Rojas, Hayden Cleere, Rin Komma, Teo Giannotta, Angela Komma, Catherina Htoo, Jim Browning Ph.D., Ken Cornell Ph.D.

Boise State University

Plant pathogens, including Pseudomonas syringae and Fusarium graminearum, impose an annual $21 billion burden on U.S. agriculture, while foodborne contaminants such as E. coli O157:H7 and Salmonella enterica cause millions of illnesses and thousands of deaths each year. These pathogens compromise both crop viability and the safety of raw sprouts for human consumption. This study evaluates cold atmospheric-pressure plasma (CAP) as a non-thermal intervention for decontaminating a diverse range of seeds, including sweet corn, cucumber, and mung bean. Results indicate that a 15-minute CAP exposure achieves up to a 3-log reduction (99.9%) across tested pathogens without compromising germination rates or seedling vigor. Mechanistic analysis reveals lipid peroxidation as a primary driver of microbial inactivation. These findings position CAP as a sustainable, chemical-free alternative to traditional seed treatments, offering a dual solution for crop protection and public health.

P1-019

Trypanosoma cruzi: interference of host cell lysosomal functioning via excess cholesterol to prevent parasitic invasion

Aspen E. Acuña, Dr. Daniel Clark

Weber State University

Trypanosoma cruzi (T. cruzi) is a single celled parasite found in the Americas and is the cause of the affliction known as Chagas’ disease. The life cycle of T. cruzi is indirect and relatively complex, involving three different life stages and requiring two hosts: an invertebrate (kissing bugs) and a vertebrate, the most relevant of which is humans. In humans, two distinct life stages can be found. The stage that is infective to humans and penetrates host cells is known as a trypomastigote, a motile form with an undulating membrane. Once inside a host cell, trypomastigotes differentiate into amastigotes, a nonmotile stage through which binary fission takes place. T. cruzi has a very specific and predictable way that it enters host cells. The parasite hijacks the host cells’ membrane repair process and it has been suggested that for most methods of entry, lysosomal exocytosis is required, a process that is impaired by excess intracellular cholesterol levels. The project utilizes cultured mammalian cells (Vero) to create a cell infection model that measures the effects that increased cholesterol have on the entry of T. cruzi into the cell. Infected cells will be qualitatively evaluated based on cytopathic changes and the presence of the parasite in the cultured cells. Parasite load and viability were quantified through the use of a genetically modified strain of T. cruzi which expresses β-galactosidase. This enzyme reacts with the β-Glo assay to produce measurable luminescence from which a time course of parasite life-cycle changes was established.

P1-020

150-fold enhancement in DNA assembly in an engineered E. coli strain

Misha Iqbal, Samuel Scott, Joel S. Griffitts

Brigham Young University

Construction of DNA molecules in synthetic biology typically relies on in vitro cloning methods, which can be time-consuming and labor-intensive. To overcome this challenge, we aim to develop an efficient system to assemble plasmids from linear fragments directly in the E. coli cytoplasm using short regions of sequence similarity. E. coli naturally possesses a short-homology-based repair pathway that can join DNA fragments. This pathway depends on the native exonuclease machinery, which generates single-stranded overhangs required for complementary strand annealing and subsequent ligation. To enhance in vivo DNA assembly, we aimed to (i) optimize the cytoplasmic exonucleases environment by targeted knockouts, (ii) optimize the length of DNA homology between fragments (10–24 bp), and (iii) assess the effect of fragment number (2-4 fragments) on assembly efficiency. We observed that the double knockout ΔrecD ΔxseA resulted in >150-fold increase in assembly efficiency compared to wild-type cells. In contrast, ΔxthA and Δrnt decreased efficiency, suggesting their role in generating single-stranded overhangs. Variation of overlap lengths showed that 24 bp overlap provided highest efficiency, while increasing the number of fragments reduced assembly efficiency. These findings demonstrate that simple changes to the exonuclease environment in E. coli cells can dramatically alter the efficiency of in vivo cloning, opening possibilities for efficient construction of large plasmids or complex plasmid libraries by simply transforming cells with linear DNA components.

P1-021

Evaluating Genomic Pipelines for Predicting Bacteriophage Host Range in Salmonella enterica

Noah Rogers, Bailey Jannuzzi, Morgan Johnston, Alaya Lopez, Lauren Brooks

Department of Biology, Utah Valley University

Bacteriophages infect bacteria through highly specific interactions with host receptors, making host range a key determinant of phage activity. Accurately predicting host range is crucial for applications such as phage therapy, which has been increasingly studied as an alternative to antibiotics. Current predictive tools remain limited to broad taxonomic levels, generally failing to resolve interactions below the genus or species level. This is particularly problematic for pathogens such as Salmonella enterica, which consists of 2,600 serovars with diverse surface structures and receptor profiles that influence phage susceptibility. This work aims to identify genes that are consistent with phage susceptibility through comparative genomic methods. Publicly available genomes from 13 Salmonella enterica serovars were evaluated for quality and completeness using BUSCO and QUAST, with 9 genomes passing thresholds for further analysis. Annotated assemblies were then compared using various core and pan-genome pipelines to identify differences in gene distributions between susceptible and non-susceptible strains. Results were variable across methods, reflecting differences in genome quality, content, and pipeline methodology. These results highlight the challenges of working with older or incomplete genome assemblies and emphasize the need for increasing the sequencing and availability of high-quality genomes and for experimental validation against characterized phages, such as STV18, which was isolated from wastewater and screened against the same serovar panel.

P1-022

Genomic and Comparative Analysis of EtoileNova, a Novel A3 Mycobacteriophage

 Emilie C. Barrick, Cade Brink, Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Jake W. Williams, Donald P. Breakwell, Brett E. Pickett

Brigham Young University

Mycobacteriophage EtoileNova was isolated from a soil sample containing compost using Mycobacterium smegmatis mc²155. After several rounds of purification, a high-titer lysate was prepared, and the phage was imaged using scanning electron microscopy. DNA was purified, and paired-end sequencing reads were generated on an Illumina platform. Genome annotation was performed using PECAAN, with gene prediction facilitated by GeneMark and Glimmer, while protein function was determined using HHPred and NCBI BLAST. Comparative genomic analysis was conducted using Phamerator. Electron microscopy revealed that EtoileNova is a siphovirus. The genome spans 50,081 base pairs and contains 82 predicted open reading frames (ORFs), of which 45.1% have assigned functions, including structural and DNA replication genes. Comparative analysis indicates that EtoileNova belongs to the A3 subcluster and shares the greatest similarity with phages Popcicle and Norbert, supporting close evolutionary relationships within this group.

P1-023

Effect of CD6KO on CD8 T Cell Metabolism

Jonah Christensen

Brigham Young University

Cluster of differentiation 5 (CD5) and cluster of differentiation 6 (CD6) are structurally related coreceptors that associate with the T cell receptor (TCR) and modulate T cell activation. Deletion of CD5 has previously been shown to increase basal and maximal respiration in naïve helper T cells, suggesting a role in metabolic regulation. CD6 is known to be involved in certain cancers and is also upregulated in autoimmune diseases such as psoriasis and multiple sclerosis, which indicated that it is important to the T-cell's effects in the immune system. However, its role in T cell metabolism remains unclear. To investigate this, we performed Seahorse extracellular flux analysis on CD8- T cells isolated from CD6 knockout (CD6KO) male mice. Oxygen consumption rate (OCR) was measured following sequential administration of mitochondrial inhibitors to assess basal and maximal respiratory capacity. CD8- T cells from CD6KO mice exhibited significantly increased basal and maximal respiration compared to controls. These findings suggest that CD6 may function as a negative regulator of mitochondrial respiration in CD8- T cells. Further studies are needed to determine how enhanced metabolic activity influences effector function and to evaluate the impact of CD6 deficiency on mitochondrial structure and metabolite composition.

P1-024

Analysis of F1 Mycobacteriophage Proteins and tRNA's

Lauren H. Forsberg, Sophia K. Dufour, Kyden C. Wilson, Micah D. Abrams, Joshua E. Pearce, Hunter E. Hudgins, Cade Brink, Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Jake W. Williams, Don P. Breakwell, Brett E. Pickett

Brigham Young University

Bacteriophages are abundant throughout the world, are genetically diverse, and provide insight into viral evolution. PatoPoderoso, SeaLumen, and Enzomatic are novel bacteriophages that infect Mycobacterium smegmatis mc²155 and belong to the F1 cluster within the Actinobacteria.  These phages were isolated from compost samples, collected in Provo, Utah, West Linn, Oregon, and Windsor, Vermont, respectively. Following three rounds of purification, high titer lysates (>108 pfu/ml) were prepared, and scanning electron micrographs were taken. From extracted DNA paired-end sequencing reads were generated using an Illumina platform, and the genomes were assembled, and annotated using bioinformatic tools including NCBI BLAST, HHPred, PhagesDB, Phamerator, AlphaFold, PhageScope and SEA PHAGES resources. Comparative genomic analysis was used to evaluate gene content and predict protein function. Transmission electron microscopy revealed that all three phages display Siphovirus morphology with long, noncontractile tails. Through the comparison of protein structures throughout different phage clusters, we can predict the structure of F1 major capsid proteins and see the overall conserved structure of essential proteins. A tRNA gene was found in both SeaLumen and Enzomatic, but not in PatoPoderoso. It was discovered that phages with larger genomes tend to have more tRNA genes, and phages that do not have tRNA genes may already have that gene adjusted to the host environment. tRNA is also thought to help with the ability of phages to infect a broader range of hosts. This work contributes to the growing understanding of F1 cluster mycobacteriophages and expands current bioinformatics databases.

P1-025

Metabolism of CD6 Knockout T Cells

Spencer Hess, Miranda Sharp, Scott Weber

Brigham Young University

T cells play an important role in the adaptive immune response, producing immune regulating molecules and fighting pathogens. T cells must be activated in order to effectively respond to infection, and when activated they switch their metabolic processes from cellular respiration and glycolysis to mostly glycolysis. This metabolic change helps meet energy needs for proliferation, cytokine production, and DNA replication. A cellular coreceptor called CD5 (cluster of differentiation 5) inhibits T cell differentiation in the thymus. Previous work in our lab has shown that CD5 knockout CD4+ cells rely more on cellular respiration and glycolysis than wild type (WT) cells. CD6 (cluster of differentiation 6) has similar structure and function to CD5, and affects T cell differentiation, stabilizing the TCR and MHC, and migration of T cells from blood into tissues. The goal of our research is to determine if CD6 knockout CD4+ T cells have metabolic changes compared to WT. We harvested and isolated CD4+ T cells from CD6 knockout mouse spleens, ran Seahorse metabolic profiling assays and found that CD6 knockout mice generally have higher basal oxygen consumption rate compared to WT, with a couple of exceptions that will be elaborated upon. We also did analysis of metabolite levels in the serum from the same mice, the results of which will be available by the time of the conference. They will be ready by the time of the conference. Understanding and influencing T cell activation and metabolism could help develop treatments that modulate immune responses to treat autoimmunity.

P1-026

Secondary Analysis of Bulk RNA-Sequencing Samples Reveals Mechanistic Features and  Molecular Markers Across Multiple Phenotypic Endometriosis Comparisons

Jackson Keele, Matt Reall, Isabella Irwin, Matthew Jackson, Brett Pickett

Brigham Young University

Endometriosis is a chronic disorder characterized by abnormal (ectopic) growth of endometrial-like tissue outside of the uterus, affecting an estimated 10% of reproductive-age women worldwide. Endometriotic lesions cause chronic inflammation and scarring on intraperitoneal pelvic organs, resulting in dysmenorrhea, pelvic pain, and infertility. Individuals suffering from endometriosis lack effective, accessible diagnostic and treatment options. Ambiguous symptom presentation and a lack of reliable diagnostic tests necessitate invasive laparoscopic exploration and tissue biopsy for definitive diagnosis. The standard of care for endometriosis involves surgical excision and ablation of ectopic lesions, but the long-term effectiveness of such surgical procedures is limited given the high rate of disease recurrence. Further research is required to better understand what cellular pathways are involved in endometriosis in order to identify potential therapeutic targets and biomarkers for improved diagnostics and treatments. To better understand the molecular mechanism(s) of the disease, we collected 202 endometrial samples from 11 studies on the Gene Expression Omnibus (GEO) and preprocessed them using the ARMOR bioinformatics workflow for bulk RNA-Seq data analysis and Pathways2Targets. We quantified differentially expressed genes (DEG) across three biologically relevant tissue contrasts: ectopic vs. eutopic endometrial tissue, eutopic vs. healthy controls, and ectopic vs. healthy controls. We found widespread transcriptional remodeling both within diseased individuals and between diseased and healthy endometrial environments, consistent with previous transcriptomic characterizations of endometrial pathology. Using these DEGs and pathway enrichment analysis through SPIA and Pathways2Targets, we then identified the top existing therapeutic targets and drugs that can be repurposed for endometriosis.

P1-027

Exploring the Diversity of The E. coli Bacteriophage Collection at BYU-Idaho

Gabriel Koch, Angel Cody, Reggie Hardman, Jared Bingham, Michael Saline, Daniel Palma, Nekayla Morgan, Porter Schlerf, Joseph Layton, Andrew Field, Schuyler Allen, Weston Jones, Gabby Chavez, Dallin Nelson, Kai Mangrum, Dr. Seth Ririe PhD

Brigham Young University - Idaho

Bacteriophages, the most abundant biological entities on Earth, are being explored as alternatives to antibiotics in combating drug-resistant bacteria. In this study, twenty new bacteriophages targeting Escherichia coli were isolated from various aquatic environments such as puddles, rivers, lakes, and wastewater. The E. coli host strains used exhibited distinct antibiotic resistance profiles, representing diverse ecological sources. The researchers hypothesized that wastewater, with its high microbial diversity, would yield the most promising phages due to its concentration of bacteria and their associated viruses.

P1-028

A journey into the unknown: Discovery and Genome Annotation of Novel Bacteriophage Shroomer

Saul Labra

Idaho State University

Shroomer is a novel Microbacterium foliorum NRRL B-24224 bacteriophage isolated from a soil sample collected by a patch of unidentified mushrooms in Pocatello, ID, USA. This phage was purified then imaged via transmission electron microscopy, revealing siphoviridae morphology. Shroomer was sequenced by the University of Pittsburgh Bacteriophage Institute. Based on genetic content similarity, Shroomer was classified as an ED2 subcluster phage. Shroomer’s genome was annotated using the PECAAN (Phage Evidence Collection and Annotation Network) platform and protein coding genes were identified using Glimmer and GeneMark. Gene start sites were refined through Starterator and ribosomal binding site (RBS) analysis. Gene functions were assigned from protein-level comparisons utilizing HHPRED and NCBI BLAST. Our results show that Shroomer has unique “tool” genes that may help it successfully infect and manipulate its host. These genes include a DNA helicase, DNA polymerase, RNA ligase, Acb2, glycosidase hydrolase, and oxidoreductase. Phages utilize host DNA helicases and DNA polymerases to replicate their genomes during infection; Shroomer possesses its own DNA helicase and DNA polymerase which improves efficiency of DNA replication. Shroomer’s RNA ligase helps repair damaged tRNA. Additionally, Shroomer’s glycosidase hydrolase, allows it to break down bacterial cell walls. Of particular interest to us, is Shroomer’s oxidoreductase gene, which may help keep the host cell functioning during infection. Together, these nonessential “tool” genes improve Shroomer’s fitness.

P1-029

Two F Phages Without Subclusters

Eliza Leonard

Brigham Young University

There are 5945 mycobacteriophages in PhagesDB that have been organized into 35 different clusters, of which there are nine subclusters in cluster F (F, F1-F8). Prior to this study, there were only two known F cluster mycobacteriophages. We isolated two additional phages from local soil samples using M. smegmatis mc2155 as a host. Following three rounds of purification, high titer lysates (>108 pfu/ml) were collected. These samples were prepared and scanning electron micrographs were taken, and we named the phages Clank and Doomslug. An Illumina platform was used to generate paired-end sequencing reads. The genomes were then assembled and annotated using Newbler, PECAAN, NCBI, Phamerator, ARAGORN, and HHPRED. We hypothesized that our phages would be similar to other F cluster phages, despite not sharing a subcluster. We compared these phages to the F1 cluster, expecting to find similarity between these genes and also several key differences that may explain why our phages were not a member of a subcluster. Only 2.47% of F1 phages have a tRNA gene. Both phages lack a tRNA gene. F1 phages are temperate, containing lysin A, lysin B, and holin. Clank and Doomslug contain lysis genes. They also have forwardly-transcribed tyrosine integrases. Clank and Doomslug also lack excise genes, contrasting them with F1 phages. These differences may play a role in why Clank and Doomslug do not fall into a subcluster.

P1-030

CD6-Knockout CAR-T Cells Display Enhanced Cytotoxicity Against Lymphoma Cell Line

Collin Mickelson, Alka Gaur, Annie Larson, Nate Neyman, K. Scott Weber

 Brigham Young University

Chimeric antigen receptor (CAR)-T cells are genetically engineered T cells that target surface antigens on cancer cells, showing remarkable effectiveness in treating blood cancers. However, CAR-T cells are currently not effective in treating solid tumors, as the tumor microenvironment poses physical and metabolic barriers to effective CAR-T cell function. A notable strategy for increasing CAR-T cell cytotoxicity in a tumor microenvironment is knocking out or overexpressing some genes that regulate T cell activation. CD5 and CD6 are T cell coreceptors. Previous studies have found that CD5 is a negative regulator of TCR. CD5-deficient T cells exhibit greater proliferation and cytokine release when compared to wild type T cells, making CD5 an attractive target for cancer immunotherapy. The T cell surface protein CD6 is structurally and genetically similar to CD5, indicating that it may also function as a negative regulator. Indeed, a 2021 study found that blocking CD6 with a monoclonal antibody increased production of cytokines such as perforin and granzyme B, ultimately making these T cells better cancer killers. Work done in our lab has identified improved cytotoxicity in CD6 knockout T cells. As such, CD6 could serve as another immunotherapeutic target to enhance the action of CAR-T cells against not only leukemias and lymphomas, but potentially against solid tumors.

P1-031

Context-Dependent Effects of the HIV-1 Vpr R77Q Variant on Apoptosis

Peterson, David J.; Sithole, Sidney T.; Berges, Bradford K.

Department of Microbiology & Molecular Biology, Brigham Young University

Human Immunodeficiency Virus type 1 (HIV-1) is a well-known retrovirus that causes AIDS by depleting CD4⁺ T cells. While some infections progress rapidly, others remain slow or never advance to AIDS. Vpr (viral protein R) is an accessory protein that contributes to apoptosis. Specific genetic variants of Vpr have been correlated with slower disease progression. Of these variants, the R77Q, is one of the most studied, yet its role in apoptosis remains unclear. Some studies report that R77Q increases apoptosis, while others show little or inverse effect. Because these studies utilized different wild‑type reference sequences as their experimental baselines, we hypothesize that additional mutations in the vpr gene beyond R77Q shape how Vpr influences apoptosis. To resolve the conflicting findings, we will construct a series of HIV plasmids containing specific amino acid sequences encoding the polymorphisms, S29N, I38T, and V83I each in the background of both wild-type and R77Q. These three single point mutations were identified by comparing different baseline sequences used in studies that resulted in conflicting findings. This approach will generate a controlled panel of mutant plasmids that can be compared directly. We will then infect separate samples of HEK-293 cells to assess the apoptotic effects of the different polymorphisms by means of flow cytometry, using Annexin V and Fixable Viability Dye. This research will clarify why R77Q exhibits variable phenotypes across studies and may discover molecular features of Vpr that contribute to disease progression.

P1-032

The Effects of Quaternary Ammonium Compounds on Antibiotic Resistance in Pseudomonas aeruginosa

Xander Beagley, Dallin Pratt, Stuart Buttars, Rich Robison, Ph.D

Brigham Young University

Quaternary ammonium compounds (QACs) are disinfectants widely used in healthcare, industrial, and public settings, with usage increasing substantially during the COVID-19 pandemic due to their effectiveness against SARS-CoV-2. However, their extensive use has raised concerns that repeated or sublethal exposure may promote adaptive responses that contribute to antibiotic resistance in Pseudomonas aeruginosa, an opportunistic pathogen associated with chronic infections and multidrug resistance. Some strains of P. aeruginosa are up to 200-fold more resistant to QACs than most other pathogens. Mechanisms of QAC resistance in P. aeruginosa frequently overlap with antibiotic resistance pathways, including efflux pump overexpression and membrane modifications. Although several studies report increased antibiotic resistance following QAC exposure, findings remain inconsistent, with some describing increased class-specific resistance and others reporting broader multidrug resistance. To address these contradictions, we investigated how serial exposure to a commercial QAC  influences antibiotic resistance in laboratory strains of P. aeruginosa. Following exposure across defined concentration gradients and time intervals, minimum inhibitory concentrations (MICs) were determined for antibiotics representing multiple classes using standardized broth microdilution assays. We discovered that QAC-exposed populations had no increase in antibiotic MICs but rather showed an increase in susceptibility. This suggests that adaptation to disinfectant stress in P. aeruginosa may involve physiological trade-offs that reduce tolerance to other antimicrobial agents. Further work is being done to determine the genotypic factors behind the observed phenomena.

P1-033

Removal of CD5 on T cells alters their differentiation and cytokine production in a periodontal disease in vitro model

Alina Svitlana Rodriguez Bezruchko, Carlos Moreno, Elizabeth Boeree, Jessica Townsend, Yuko Sperry, Mahler K, Cody McStraw, Dallin Cardon, Claudia M. Tellez Freitas, K. Scott Weber

Brigham Young University

Periodontal disease is a highly prevalent oral inflammatory disease that affects nearly half of adults 30 years or older in the United States. It is characterized by excessive inflammation within the periodontal pockets typically in response to bacterial challenge and is characterized by inflamed gums, destruction of periodontal ligaments, alveolar bone loss, and tooth loss if left untreated. T cells are adaptive immune cells which play important roles in driving inflammation and alveolar bone loss during severe periodontitis. Additionally, several studies have reported associations between periodontal pathogens and chronic inflammation within the oral cavity to several systemic diseases, including inflammatory bowel disease, diabetes mellitus, cardiovascular diseases, cognitive decline and Alzheimer’s disease, chronic obstructive pulmonary disease, and certain cancers. CD5 is a glycoprotein receptor found on the surface of T cells that regulates their activation and function, and its immunoregulatory role has yet to be investigated in the context of periodontitis. Here, we characterize the functional differences between CD5 knockout T cells and wildtype T cells in an in vitro periodontal disease model, including T cell activation, differentiation, and cytokine production. Here we report that removal of CD5 increases T cell activation, effector/memory formation, and alters cytokine production. Together, these findings provide insights into the role of CD5 in modulating inflammation during periodontal disease.

P1-034

Characterization of the phage PaL41501, a potential therapeutic agent for treating multidrug-resistant Pseudomonas aeruginosa strains

Jacob Wallin, Brock Sheehan, Jeremy Wilson, William Acero Jeffries, Sicely Bybee, Kiara Whitely, Ph.D., & Ruhul Kuddus, Ph.D.

Department of Biology, Utah Valley University

Background: Pseudomonas aeruginosa is a primary pathogen that infects diverse tissues, including the ear, nose, urethra, soft tissues, lungs, and burn wounds. Some strains are multidrug-resistant (MDR). We previously isolated and partially characterized a Litunavirus, L41501, using P. aeruginosa Boston 41501 as the host. Here, we present further virological findings regarding its potential as a phage therapy agent. Methods: Bacterial strains, including MDR strains, were obtained from ATCC/BEI (Manassas, VA). The phage host range, minimum inhibitory multiplicity of infection (MOI), phage-antibiotic synergy, and inhibition of host biofilm formation were assessed using standard virological methods. Results: L41501 infected five MDR and two non-MDR strains but not eight others. It remained biologically active after 75 minutes at pH 3–11 and 37°C–67°C but was inactivated at 77°C. It causes lysogenic infections in strain Boston 41501, and lysogens are resistant to reinfection. L41501 did not completely inhibit host growth at MOI 1–1000. However, it reduced the minimum bactericidal concentration (MBC) of neomycin by 4-fold at MOI 1 and decreased biofilm formation by 6-fold at MOI 10. Conclusions: L41501 is stable across a broad pH and temperature range and inhibits biofilm formation in P. aeruginosa Boston 41501. It cannot be used alone for phage therapy due to lysogenic infection but may be combined with antibiotics. We are investigating lysogenic mechanisms to develop a mutant lacking lysogeny.

P1-035

Potential Role of ATR in Slower AIDS Progression Associated with HIV-1 Vpr mutant R77Q

Ava Wilstead, Sidney T. Sithole, Bradford K. Berges

Brigham Young University

HIV-1 typically induces necrotic T cell death, resulting in chronic inflammation that drives Acquired Immunodeficiency Syndrome (AIDS) progression. However, mutations in viral protein R (Vpr) are associated with altered disease outcomes. The HIV-1 Vpr mutant R77Q is linked to individuals that do not progress to AIDS without treatment, termed a long-term non-progressor (LTNP). The Berges lab has shown that R77Q infection results in reduced overall cell death compared to wild-type (WT) virus, while inducing increased apoptotic cell death. Apoptosis is mechanistically linked to cell cycle arrest. This project aims to define the mechanism by which R77Q induces G2 cell cycle arrest and promotes apoptosis. Ataxia telangiectasia and Rad3-related (ATR) kinase regulates G2 arrest in response to DNA damage and can promote apoptotic signaling when damage is irreversible. We hypothesize that R77Q Vpr activates ATR, driving excessive G2 arrest and apoptosis. By inhibiting ATR with caffeine, we can investigate whether ATR is a key mediator of the LTNP phenotype associated with the R77Q HIV-1 mutant.

P1-036

Expression of the Streptococcus pneumoniae small RNA sr-78 is modulated by manganese

Shannon M. Hendricks and Dr. Julia Martin 

Idaho State University

Streptococcus pneumoniae is an opportunistic bacterial pathogen that is prone to develop multidrug resistance, and remains particularly dangerous in young children and older adults. Emerging evidence suggests that small non-coding RNAs (sRNAs) may play an important role in S. pneumoniae virulence by modulating the expression of bacterial virulence factors. Over 100 sRNAs have been identified in S. pneumoniae, while their functional role is largely unexplored. In this study, we investigate the sRNA sr-78 previously identified in S. pneumoniae. This sRNA is located within the 5’ region of the mgtA gene, which encodes the Mn efflux transporter MntE. The sRNA also overlaps a cis-acting manganese (Mn)-sensing riboswitch. Based on this and published data, we hypothesized that the sRNA sr-78 is down-regulated under Mn-replete growth conditions. Using Northern blot analysis, we show that sr-78 is expressed in wild-type S. pneumoniae cells and that sr-78 expression is not impacted by treatment with metal cations, as the wild-type strain is able to regulate metal homeostasis. In contrast, sr-78 expression was significantly reduced in the mntE-null S. pneumoniae mutant impaired in Mn export when grown with excess Mn; other metal cations did not affect sr-78 expression. We also note that the expression of a larger transcript was observed during Mn-replete growth in the mntE-null mutant, which became more pronounced with increased Mn exposure. Together, these preliminary data suggest that the sRNA sr-78 may serve a role during periods of Mn limitation by modulating the expression of proteins not needed during periods of Mn intoxication.

P1-037

From Slippery Sequences to Tail Length: Linking Translational Regulation and Structural Conservation in Bacteriophages

Daniel A. Hemmert, Anthony L. Jeppson, Jayden S. Longhurst, Katelynn B. Owens, Zachary M. Hunsaker, Cade Brink, Payson C. Danielson, Truman D. Stohl, Jake W. Williams, Don P. Breakwell, Brett E. Pickett

Brigham Young University

Bacteriophages rely on tightly regulated gene expression and conserved structural components to successfully infect bacterial hosts. Understanding variation and conservation within these genomic and structural features provides a foundation for understanding phage evolution, assembly mechanisms, and translational regulation. This study investigated multiple characteristics across 19 bacteriophages BYU Phage Hunters students isolated using Mycobacterium smegmatis mc2155, including tail assembly chaperone slippery sequence variation, structural protein conservation, and the relationship between tail length and tape measure proteins. Comparative genomic analyses were performed to identify and align tail assembly chaperone genes, which are produced through programmed ribosomal frameshifting that generates two overlapping proteins from a single transcript. The heptanucleotide slippery sequences and surrounding regions were analyzed to assess sequence variation, frameshift efficiency, and protein stoichiometry. Synteny of phage proteins (specifically capsid and tail structural proteins, DNA packaging proteins, and head-to-tail connector proteins) was compared to assess conservation across the phages. Tape measure protein sequence lengths were also compared to observed phage tail lengths to evaluate their correlation and role in phage morphology. Results reveal strong conservation among key structural proteins, the variability in slippery sequence composition, and correlation between tape measure protein length and tail length across phages. These findings can provide insight into phage assembly, translational regulation, and evolutionary relationships.

P1-038

Scaling Antibiotic Discovery: Leveraging Undergraduate Education Across a Tiered Research Pipeline

Chelsea Bordon, Miguel Robes, Anna Perkins, Joshua Steffen, Aaron Puri, Rebeccah Kurzhals

University of Utah

Addressing the global antibiotic discovery crisis demands scalable, scientifically rigorous approaches. Using a tiered undergraduate pipeline that spans a large-enrollment CURE course and smaller cohort-based research experiences through the Science Research Initiative (SRI), we attempt to identify novel antimicrobial compounds within a single academic year while providing experiential learning opportunities for large groups of first year students. Aligned with the Tiny Earth Initiative, the first tier engages approximately 185 students in screening soil samples for novel antibiotic-producing methylotrophic bacteria. These bacteria grow on reduced one carbon compounds such as methanol that were therefore overlooked in past antimicrobial discovery efforts. This first step generates a broad pool of putative antibiotic producing bacteria. In a year, a pool of 242 antibiotic producing bacteria has been generated. A secondary liquid co-culture assay conducted by an SRI cohort provides a critical quality- control step. Over the past year, 122 strains have been confirmed as producing potential antimicrobial activity. Rigorously vetted isolates are transferred to a third group of novice undergraduates for tertiary biochemical purification and biosynthetic pathway analysis, including IC50 measurements against a panel of target strains and mass spectrometry-based molecular networking. To date, 40 crude extracts have been purified, with several indicating potentially novel antimicrobial compounds.

P1-039

Yeast Saccharomyces cerevisiae UTR1 NAD Kinase is regulated by Phosphorylation from PAS kinase

Sakhawat Ali, Julianne H Grose

Brigham Young University

NAD kinase converts NAD+ (or its reduced form NADH) to NADP+ (or NADPH) and thus has a key role in maintaining the appropriate cellular levels of NADP(H). NADPH is required in many essential biosynthetic reactions including biosynthesis of proteins, lipids and nucleic acids as well as reductive repair reactions for neutralizing reactive oxygen species (ROS). The yeast Saccharomyces cerevisiae contains three NAD kinase homologues named Utr1, Yef1 (both cytosolic) and Pos5 (mitochondrial). Our research aims to understand the regulation of yeast Utr1 NAD kinase by PAS kinase, a serine/threonine protein kinase conserved from yeast to humans, phosphorylation. Our data on yeast shows that phosphorylation of Utr1 NAD kinase by PAS kinase inhibits its activity, resulting in decreased cell growth and NADP(H) levels, and increased sensitivity to peroxide (H2O2) stress and ROS levels. In the long-term, our research will focus on understanding the regulation of human NAD kinase by human PAS kinase and its downstream effects on cell growth and proliferation and cellular levels of NADP(H) and ROS in healthy versus cancerous cells. Our long-term goal is to better understand the human NAD kinase and solidify it as a potential therapeutic target for cancer treatment.

P1-040

Detection of Free-Living and Plant-Parasitic Nematodes in Commercial Legumes

Abbie A. Butkovich-Autry¹, Trey B. Smith¹, Dylan T. Smith¹, Faye S. Calvert¹, Lanee Farr¹, Yeeun Kim¹, Teagan M. Reese¹, Elle R. Genor¹ Shayeri Noor Gani¹, Taylor C. Boone², Weston L. Hall¹, Melodie L. Weller¹,²

¹University of Utah, School of Dentistry, Salt Lake City, UT 84108; ²University of Utah, Department of Pathology, Division of Microbiology and Immunology, Salt Lake City, UT 84112

Background: This study evaluated the presence of plant-parasitic and free-living nematodes in commercially available legumes in the U.S. food supply and assessed their viability under germination conditions. Methods: Dried legumes (soybean, pinto, black, kidney, adzuki, and navy beans) were purchased, surface sterilized, and germinated for seven days. Samples were homogenized, filtered through graded sieves, and the finest fraction was collected. Nematode eggs were isolated using sucrose flotation, counted, and analyzed via microscopy and molecular methods, including DNA/RNA extraction and PCR. Viability was assessed by incubating eggs on soil and recovering hatched nematodes using a Baermann funnel. Results: Nematode eggs were consistently detected in soybean samples across multiple brands but not in other legumes. Multiple species were identified, and viable nematodes successfully developed and were recovered after incubation. Egg abundance varied among soybean samples. Conclusion: Viable nematodes are present in commercially available soybeans, but were not detected in other tested legumes. These findings suggest variability in contamination and highlight the need for further investigation into potential implications of nematode presence in food products for public health and disease transmission.

P1-041

The BRC Program: Connecting Undergraduate Students with Research Opportunities and Beyond

 Bella Edwin, Ryan Bonson, KC Serve

Idaho State University

The Bridges to Research Careers (BRC) Program is a grant-funded organization that connects undergraduate students with the foundations of research by offering two courses labeled “Bacteriophage Discovery & Characterization Lab” and “Bacteriophage Genome Annotation & Bioinformatics Lab”. These courses provide students with the opportunity to be a part of a team-based student-led research experience and to publish their findings in a scientific paper. This all began with a 5-year grant from the NIH (National Institutes of Health) given to Dr. Michael Thomas to provide materials, publication fees, conference trips, and offer scholarship money to the students. The goals of the program are to increase participation in STEM career paths by students from all backgrounds. Now, in our second year, the program has almost tripled in size. With the help of those who joined last year, they will discuss their experience in the program and provide insight into their present research opportunities.

P1-042

The novel NRAMP-type transporter in pneumococcus is repressed during iron replete growth

David W. Burnett and Julia E. Martin

Idaho State University

Manganese, iron, and other transition metals have emerged as important trace nutrients in bacterial physiology. During an infection, bacteria must acquire these metals for survival from the host environment for use as enzyme cofactors in many metabolic processes. Although metals are essential for survival, their accumulation can be toxic. This duality has forced bacteria to evolve selection strategies that maintain optimum intracellular metal levels by sensing, acquiring, storing or, when needed, exporting metals properly. The impact of metal availability on pathogenesis is particularly exemplified by the human respiratory pathogen Streptococcus pneumoniae. This bacterium typically asymptomatically colonizes the nasopharynx but can also cause life-threatening invasive infections such as pneumonia, septicemia, and meningitis. Research has shown that manganese acquisition is important for S. pneumoniae pathogenesis. Deletion of genes encoding manganese-transporters leads to decreased virulence in model infections. The role of manganese and its potential interplay with iron metabolism remains unclear in this bacterial pathogen. Using bioinformatics, gene deletion, and expression tests we identified a Streptococcus conserved bacterial manganese resistance factor MntH (spd_0161). The predicted structure suggests that MntH is an NRAMP-type transporter, although it lacks the conserved amino acid DPGN motif required for exporting manganese. Using a FLAG-tagged MntH, we show that MntH protein expression is repressed during iron replete growth; manganese and other metals do not significantly alter its expression. This work begins to establish the function of MntH in S. pneumoniae as putative transporter involved in iron homeostasis and potential interplay with manganese.

P1-043

From Sequence to Structure: AlphaFold-Assisted Annotation of Phage MiloMuff’s Minor Tail Proteins Reveals their Specific Functions

A. Granander, E. Githuku, M. Nations, M. Thomas 

Idaho State University

Bacteriophage minor tail proteins (MTPs) play critical roles in phage infection including organization of tail polymerization, facilitation of DNA injection, host recognition, and attachment. Sequence-based annotation tools frequently fail to identify functions of novel MTPs due to the scarcity of experimentally solved structures in sequence databases. We addressed this limitation using MiloMuff, a novel EB1 cluster phage with syphovirus morphology that infects Microbacterium foliorum NRRL B-24224. While standard genomic annotation of MiloMuff identified four putative MTPs, their functions remained uncharacterized. To bridge this gap, we employed AlphaFold3, ChimeraX, and Foldseek to resolve the structures and multimeric stoichiometry of MiloMuff’s MTPs, using the experimentally characterized Mycobacterium phage Bxb1 as a structural benchmark. By using ChimeraX to compare AlphaFold3-generated 3-D protein models against experimentally determined MTP structures from Bxb1, we were able to identify structural homologs for the baseplate hub and the distal tail (DIT) minor tail proteins. Furthermore, we identified a putative tail spike protein containing a peptidase domain and a putative tail wing base containing a fibronectin-binding domain, suggesting a specialized mechanism for host interaction and extracellular matrix degradation. These results demonstrate that AlphaFold-assisted structural annotation can identify MTP functions when standard sequence-based tools fail to. This approach is essential for deciphering phage-host interactions at a molecular level and directly informs the development of phage therapeutics.

P2-001

Biophysical Characterization of Lactoferrin under Environmental Stressors

Grace Clother

Boise State University

Bovine lactoferrin (LF) is an iron-binding glycoprotein with antimicrobial activity, whose function is closely linked to its physicochemical properties and structural stability. In this study, we investigated the physicochemical behavior of LF under varying environmental conditions, focusing on temperature-dependent aggregation, ionic effects, and freeze-thaw stability. Thermal transitions were characterized using calorimetry, structural changes were monitored by circular dichroism (CD) spectroscopy, and aggregation behavior was assessed by dynamic light scattering (DLS). Native LF exhibits a compact globular structure with a hydrodynamic diameter of approximately 8-10 nm. Upon heating, a transition near the denaturation temperature is accompanied by changes in secondary structure and a pronounced increase in particle size, indicating aggregation. These aggregates persist after cooling. The presence of different ionic conditions modulates aggregation behavior, affecting both size distribution and dispersity. In addition, repeated freeze-thaw cycles induce measurable changes in particle size, which may impact the functional performance of lactoferrin as an antibacterial compound.

P2-002

Identification of Plant Viruses in Utah Gardens

Laureana Lazarte, Alma Laney PhD

Biology Department, Utah Valley University

This study aims to identify the viral infections present in common garden variety plants, with a focus on those cultivated at the Utah Valley University GRIT Garden. Viral infections in garden plants are transmitted through multiple modes, including seed transmission, mechanical transmission, and vector transmission, each of which presents unique challenges for garden management and plant health. Infected plants displayed a wide range of symptoms, including chlorosis, mottling, leaf puckering, and leaf rolling, affected crop yield and growth of the plant, all of which can significantly reduce plant strength and crop yield. Understanding and identifying the diversity of viruses present in community garden settings is important for developing effective disease management strategies, such as pest management. Leaf samples were taken from symptomatic garden plants and RNA extracted using the Plant RNA kit (IBI Scientific) and subjected to high-throughput sequencing using Illumina NovaSeq. Detection of viruses was verified with RT-PCR and ELISA. Among the genera of viruses discovered in these garden varieties are Bromovirus, Potyvirus, Caulimovirus, Badnavirus, Orthotospovirus, Curtovirus, and Tobamovirus. Data analysis is ongoing, with publication to follow.

P2-003

A Comparative Analysis of Wolbachia Strains Present in Flies Around CEI Campus

Hayden Leavitt, Janelle Sagawa, Dr. Benjamin A. Burrows

College of Eastern Idaho

Wolbachia pipientis is an obligate intracellular bacterium that infects a wide range of arthropods, including many Diptera species. Because Wolbachia can influence host reproduction, fitness, and population structure, strain level variation within local insect populations can provide insight into microbial transmission patterns and potential gene flow. In this work, we investigated Wolbachia infection in the cluster fly Pollenia pediculata across the College of Eastern Idaho campus. The objective was to determine whether flies collected from different campus buildings carry genetically distinct Wolbachia strains, which could indicate limited interaction or movement between subpopulations. Cluster flies were collected from every accessible building on campus, and genomic DNA was extracted from each specimen. Initial screening for Wolbachia was performed using PCR targeting the 16S rRNA gene. The 16S rRNA sequences were found to be identical across all samples, consistent with the highly conserved nature of this gene. To achieve higher resolution, we selected the Wolbachia surface protein gene (wsp), which contains hypervariable regions. PCR amplification of wsp was performed using primers designed to target these variable regions, and positive amplicons were submitted for sequencing. Sequence data from wsp will allow for finer comparison of Wolbachia strains across campus buildings and may reveal patterns of interaction not detectable with 16S rRNA alone.

P2-004

Colorful Proteins: Engineering a Palette of Novel Chromoprotein Variants

Blake Olson, Anna Bowman, Kyler Fullmer, and Dr. Joel Griffitts. Anna Bowman and Kyler Fullmer,  Dr. Joel Griffitts

Brigham Young University

As naturally occurring proteins with nearly identical structures, chromoproteins exhibit a wide range of colors. Chromoproteins (CPs) are widely used in synthetic and molecular biology because their colors are visible without the use of specialized instruments. The expanding applications of CPs require a broader palette of variants with diverse and robust color phenotypes. To generate new chromoprotein variants, we used targeted, random mutagenesis on six different chromoproteins (amilCP, scOrange, aeBlue, amilGFP, fwYellow, and mScarlet) to cause mutations in areas in the CPs predicted to have a strong effect on the overall chromoprotein color. Screening mutagenized chromoproteins produced 145 chromoprotein variants. The 10 most novel chromoproteins were selected from the 145, and we suggest they be used for research and commercial purposes.

P2-005

Experimental Evaluation of Bioinformatics Tools for Bacteriophage Host Range Determination

Daniel Parker, Sarah Sylvester, Lauren Brooks

Utah Valley University

The accurate prediction of bacteriophage host range is essential for advancing phage therapy and microbial control strategies. Bioinformatic tools have been developed to infer host specificity from genomic data; however, their reliability across diverse phages remains uncertain. This study aims to systematically evaluate the accuracy of current host prediction algorithms through experimental validation using the previously isolated and characterized STV18 and bacteria strains available in the Microbiology Labs at UVU. Bioinformatics tools were selected for evaluation based on their upkeep by developers, with tools not being kept current for the last five years being disregarded. Additionally, some identified tools were not used due to computational difficulties. For each tool used predicted host ranges were generated and analyzed for specificity and sensitivity using the results from the laboratory. These results highlight discrepancies between computational predictions and empirical infectivity, suggesting current limitations in bioinformatic host prediction tools. Ongoing work will expand this analysis to include additional phages and additional predictive platforms to determine relative accuracy and identify patterns of misclassification. This research emphasizes the importance of integrating experimental validation with computational approaches and aims to inform the development of more reliable host prediction models for bacteriophage applications.

P2-006

From Singletons to Clusters: Synteny-Based Reassessment of Two Singleton Mycobacteriophages

Tanner L. Sessions, Kobe M. Wettstein, Cade Brink, Natalie A. Olsen Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Jake W. Williams, Don P. Breakwell, Brett E. Pickett

Brigham Young University

Out of 5,944 Actinobacteriophages, only 67 (~1%) have been characterized as singletons. Mycobacteriophages have demonstrated great genetic diversity and evolutionary patterns. Mycobacteriophages, Chubbello and Bluvara, were isolated from compost samples in Idaho and Oregon, respectively, using Mycobacterium smegmatis mc2 155 as the host. High titer bacteriophage lysates (>10⁸ PFU/mL) were generated following three consecutive rounds of purification. Samples were visualized using scanning electron microscopy, and genomic DNA was extracted for sequencing. Paired end reads were produced on an Illumina platform and assembled to obtain the complete genome sequences. Chubbello and Bluvara originally were characterized as unclustered mycobacteriophages; however, based on the evidence, we have reason to believe they belong to the P and I clusters, respectively. The genomes of Chubbello and Bluvara span 46,344 and 49,459 base pairs and contain 77 and 74 predicted open reading frames (ORFs), respectively. Of the ORFs, Chubbello has 50.0% known functional genes, and Bluvara has 48.6% known functional genes, which encode key processes that support bacteriophage structure and replication. One gene in Bluvara’s genome is an orpham, a gene with no known function or family. Our findings show that the Chubbello phage shows remarkable synteny to the P1 subcluster, whereas the Bluvara phage shows synteny towards the I cluster, but ambiguity remains about its subcluster. More research is needed to conclude with which subcluster, if any, the Bluvara phage is associated.

P2-007

Feeding activity of the brain-eating amoeba: blocking Naegleria fowleri with treatment drugs

Akanksha Singh, Anya Midavaine, Ian Ford, B. Kooper Carson, Khoi Dinh, Luis Montenegro Calla, Daniel N. Clark

Weber State University

What drugs will kill the brain-eating amoeba? Only four survivors out of 154 cases of Naegleria fowleri have occurred in the US, and those that survive are likely to suffer from minor brain damage, immune reactions, or adverse side effects from the drug treatment. Each survivor was treated with amphotericin B, but many fatal cases also attempted this drug. Infections are rare, but what makes N. fowleri dangerous is its speed (time to death is around four days post-exposure) and thus how difficult it is to diagnose and treat. We have some basic information about potential drugs: The Centers for Disease Control (CDC) list amphotericin B, fluconazole, rifampin, azithromycin, nitroxoline, and miltefosine. We investigated the effectiveness of combinations of these drugs on cultured human cells infected with N. fowleri. We hypothesized that each of the selected drugs would have varying levels of effectiveness at reducing the amoeba’s feeding levels. The investigational drugs were tested by first measuring toxicity on the amoeba to establish the appropriate dose of each drug. We then set up cultures of amoeba in 24-well plates and presented them with fluorescent plastic beads to eat, and treated them with the appropriate dose of each drug for 24-48 hours. Amoeba killed due to drug inhibition were unable to eat beads and remained negative for fluorescence when measured with a flow cytometer. We found that amphotericin B and azithromycin reduced the brain-eating amoeba’s ability to feed by 40%, while the other four drugs were ineffective or caused only a modest reduction in amoeba activity. We will next investigate combinations of drugs and run variations of our infection model. These findings will inform clinical practice as physicians attempt to save the lives of those infected with this killer parasite."

P2-008 

Bacteriophages Retain Stability in Human Fluids

Riley Oesch, Sara Sadeghi, Dr. Michael Thomas

Idaho State University

The rise of multidrug-resistant (MDR) bacterial infections necessitates alternative therapeutic strategies. Bacteriophage (phage) therapy has emerged as a promising approach; however, the stability of phages in human body fluids remains insufficiently characterized. Understanding phage persistence is critical for the development of effective systemic phage therapies and optimized phage cocktails. In this study, ten bacteriophages and nineteen bacterial strains were isolated from wastewater sources, including the Pocatello Wastewater Treatment Plant, Blackfoot Wastewater Treatment Plant, and Portneuf Regional Medical Center. Phage stability was evaluated in simulated human saliva, urine, and serum over a 12-hour period. Phage lysates were incubated in each fluid, and stability was assessed by measuring plaquing efficiency using spot titration at defined time points. Our results demonstrate that phages retain significant stability across all tested fluids, though persistence varies among individual phages and fluid types. These findings highlight the importance of selecting phages with favorable stability profiles for specific physiological environments. Overall, this study supports the potential of bacteriophages as viable alternatives or adjuncts to antibiotics and provides insight into optimizing phage selection for therapeutic applications.

P2-009

Towards Identifying the Molecular Origin of Ice Nucleation Activity in Lichen

Sarah King

Boise State University

Lichens are symbiotic organisms that persist under extreme environmental conditions through effective cold-adaptation mechanisms. Recent evidence suggests that lichens produce ice-active biomolecules, including ice-nucleating proteins. In this study, we investigated Platismatia herrei, a lichen species associated with the formation of ice at high subzero temperatures. We report the isolation and characterization of the specific fungal organisms responsible for this activity. Dozens of distinct subcultures were subcultured from lichen samples and analyzed by ice nucleation assays and ice-shaping measurements. These experiments identified two independent fungal cultures with strong ice-nucleating activity. Beyond their local ecological relevance, the identification of these biological ice nucleators has broader implications for atmospheric science, as they may contribute to cloud glaciation, precipitation processes, and potential cloud-seeding applications.

P2-010

Indirect Effect of Soil Amendments Targeting Roadside Revegetation on Soil Microbial

Eric Christen, Erika Stewart, Joshua B. Grinath, Anirban Chakraborty

Idaho State University

Communities Invasive plant species are a major threat to roadside ecosystems. To manage invasive plants, land management agencies often utilize soil amendments to revegetate native plant species. Commonly used soil amendments may have indirect effects on resident soil microbial communities when applied individually and in combination with each other, yet these indirect effects remain underexplored. In this study, we investigated the indirect effects of applications of a commercial microbial inoculant with a non-synthetic micronutrient fertilizer, a wool-pellet-based fertilizer, or in combination of two fertilizers. Soil amendment applications were conducted at three experimental sites situated along the sides of a major interstate highway spanning the eastern corridor of Idaho, USA; with each site located within a distinct Level IV EPA ecoregion. Soil samples were collected annually across sites during three consecutive summers post-application for assessing soil chemical properties and to conduct DNA-based microbial community surveillance. The sites were found to host significantly different microbial communities while no noticeable community dissimilarity within samples representing individual or combinations of soil amendments were observed. However, specific genus-level clades became differentially abundant in samples treated with different soil amendments, such as Janthinobacterium, a common soil bacterium known for producing bio-active metabolites. Correlation analyses of soil chemical properties with differentially abundant taxa further revealed pH and organic matter as principal drivers of shifts in taxa abundance. Our results showed that a single application of combinations of soil amendments for land management purposes triggered taxon-level responses to treatments, even if a community-level response was not exhibited.

P2-011

Heterologous Expression of Soluble Fungal Ice-Nucleating Proteins in S. cerevisiae

Ronan Lanam, Dr. Konrad Meister

Boise State University

Heterogeneous ice nucleation at high subzero temperatures is driven by specialized ice-nucleating proteins (INPs) that catalyze the water-to-ice phase transition by lowering the free energy barrier for ice formation. While decades of work have focused on membrane-anchored bacterial proteins such as InaZ from Pseudomonas syringae, a new class of soluble, membrane-independent INPs has recently been identified in the Mortierellaceae family [1]. These fungal INPs, likely acquired through horizontal gene transfer, exhibit distinct structural adaptations that enable activity without membrane support, including a C-terminal disulfide capping motif that stabilizes the β-solenoid architecture. Here, we report on the development of a galactose-inducible expression system in Saccharomyces cerevisiae for the production of His-tagged INPs from Entomortierella parvispora. Droplet freezing assays of yeast lysates confirm gain-of-function, with freezing temperatures reaching −5 °C, comparable to native extracts (-20°C) and significantly improved over E. coli expression. These results indicate that the eukaryotic host preserves the structural integrity and assembly of highly active protein aggregates required for efficient ice nucleation. This work positions S. cerevisiae as a robust platform for producing functional, soluble INPs and provides a foundation for detailed biophysical studies and the development of next-generation technologies in cryopreservation and controlled ice formation.

[1] Eufemio et al. Science Advances 2026

P2-012

The commensal relationship between Akkermansia and Blautia species in the infant gut

Drew Allred, JB Lubin

University of Utah, Department of Pathology, Division of Microbiology and Immunology 

Akkermansia is a genus of commensal gut bacteria that has recently been found to play a significant role in the development of a healthy microbiome in infants. In silico analysis of eight previously published human datasets revealed a positive correlation between the abundance of Akkermansia and the abundance of Bluatia species, another commensal microbe. In vitro co-culture assays revealed a mutualistic relationship between these two bacteria: both grew better in co-culture with each other than in monoculture. Metabolomic analysis on growth media spent by each microbe reveals potential key mechanisms that will be tested in future in vivo studies. We expect that these studies will demonstrate the feasibility of probiotic seeding to improve the development of the infant gut microbiome.

P2-013

Characterizing Proteus mirabilis Phages for Clinical Applications

Madison E. Duffy, Dalton K. Kutzen, Julianne H. Grose

Brigham Young University

Antibiotic resistant infections are set to be the leading cause of death by the year 2050, with patients undergoing cancer treatment, the immunocompromised, and the elderly being the highest risk. Prolonged hospital stays and the use of medical devices such as catheters increase infection risk. Bacteriophages (phages), viruses that specifically infect bacteria, offer a promising alternative to traditional antibiotic treatments due to their high infection specificity and few off target effects. However, current phage therapies are developed on an individual basis, taking several months to go through the process of identification and safety testing. This project aims to enable rapid assembly of phage therapeutic cocktails, reducing the time needed for development and saving lives. My long term goal is to create phage therapies that infect a majority of the clinical antibiotic-resistant isolates. In the short term I am focusing on clinical Proteus mirabilis, a pathogenic Enterobacteriaceae. Our lab has isolated 15 novel Proteus phages. These phages will be sequenced, annotated, and genomically analyzed to confirm a lytic life cycle and the absence of any virulence factors. The phages shown to be safe via genomic analysis will then undergo host-range testing against various antibiotic-resistant isolates obtained from the CDC Antibiotic Resistance Isolate Bank. Next, they will be evaluated for safety in human therapeutic uses via toxin assays and cytokine profiling utilizing human cell lines to evaluate potential immune activation. Phages that pass through this screening will be made readily available for patient use, aiding in the fight against antibiotic resistant infections.

P2-014

Immunomodulatory Effects of Tianeptine In Vivo using Galleria mellonella Larvae Model  

Ben Laughter, Travis George, Dhruvan Gopinath, Trevor Tarter, Simon Nguyen, Victor M. Jimenez Jr., Ph.D

Noorda COM

Purpose: Tianeptine is an atypical tricyclic antidepressant with mu-opioid receptor activity, therapeutically prescribed in Europe. The US has banned it due to its high potential for abuse.  Research suggests it has anti-inflammatory effects that might explain its antidepressive properties. The focus of this study is to explore the anti-inflammatory effects of tianeptine in vivo by infecting Galleria mellonella with bacteria. G. mellonella has a similar innate immune system making it a suitable immune model. Methods: Tianeptine G. mellonella, larvae were injected with 10µL of tianeptine dissolved in PBS at five doses:16.00, 4.00, 1.00, 0.25, and 0.06 µg/mL. Non-injection and PBS controls were included. Larvae were selected based on activity, color, and weight, and observed every day for five days. Different Larve were injected with E. coli and S. epidermidis at doses of 102, 103, 104, 105, and 106 CFU to determine the LD50 prior use with tianeptine. Results: The survival of G. mellonella with tianeptine was 60% (16 µg/mL), 72% (4 µg/mL), 68% (1 µg/mL), 56% (0.25 µg/mL), and 48% (0.06 µg/mL). This demonstrates an inverse, U-shaped curve. Larvae injected with E. coli had a survival of 60% (103 CFU) and 28% (104 CFU) while S. epidermidis was 72% (105 CFU) and 28% (106 CFU). Conclusion: The inverse U-shaped survival curve of tianeptine in G. mellonella larvae is like previous results generated with macrophages in vitro. This demonstrates G. mellonella as an effective immune model. Ultimately, this work adds more context to the immunological effects of psychiatric medications. 

P2-015

What’s in the water? Proposing an Alternative Media for Standardized Fecal Contamination Testing on Peruvian Drinking Water.     

Amanda Lee1, Jacob Andrew1, Dr. Lauren Brooks2

Utah Valley University

Access to safe drinking water is an essential component of a functioning society. To ensure this need is met, we must reliably monitor for dangerous contaminants, such as fecal matter. Monitoring for fecal contamination typically uses Escherichia coli as a proxy for fecal pathogens. The United States Environmental Protection Agency (US EPA) provides standardized water quality monitoring protocols, including Method 1603, which uses Modified Membrane-thermotolerant Escherichia coli (Modified mTEC) culture media to select for and quantify Escherichia coli. However, this procedure is very costly, making it unrealistic for large-scale monitoring in certain regions, such as in low- and middle-income countries (LMIC). Aquatest, an alternative medium previously developed for monitoring E. coli in Bangalore, would reduce the cost per test from $20.32 USD to $0.26 USD. However, this methodology has not yet been standardized and implemented on a wide scale. This past December, in an ongoing collaboration with scientists and government officials across various municipalities in Peru, we examined over 100 samples in less than a week, at a cost of over $ 2,000 to process. In the long term, it is infeasible to monitor all of these samples using the EPA Standard Method. To address this concern, we conducted a pilot study comparing the results of Aquatest and Modified mTEC using samples collected from various regions in Peru. We found a strong correlation between results, suggesting Aquatest is a viable alternative for monitoring water quality.

P2-016

SELEX-Based Enrichment of ssDNA Aptamers Targeting the Ferric Citrate Receptor FecA in ExPEC  

Trevor Jones, David Erickson

Brigham Young University

The rise of antimicrobial resistance in extraintestinal pathogenic Escherichia coli (ExPEC) call for new strategies aimed towards targeted detection and intervention. The ferric citrate uptake protein FecA is an outer membrane receptor involved in iron acquisition and represents a potential molecular target. This study used Systematic Evolution of Ligands by EXponential enrichment (SELEX) to identify single-stranded DNA (ssDNA) aptamers with specificity for FecA. An ssDNA library containing a 40-nucleotide randomized region flanked by two 23-nucleotide primer sites was subjected to iterative selection against E. coli M12 strains. In each round, counter-selection was performed using ΔfecA mutant cells to remove non-specific binders, followed by positive selection with wild-type (WT) M12 to enrich for FecA-binding sequences. Bound DNA was recovered, amplified, purified, and treated with lambda exonuclease to regenerate ssDNA. This process was repeated for 12 rounds prior to sequencing. Analysis of enriched pools identified four candidate sequences, suggesting preferential binding to FecA. Ongoing work is focused on validating binding specificity and affinity using comparative assays between WT and ΔfecA strains. These results support the use of SELEX to generate ssDNA aptamers targeting outer membrane receptors such as FecA in ExPEC. Such aptamers may provide a foundation for sequence-specific detection and future targeting of bacterial iron acquisition systems.

P2-017

Cold Atmospheric-Pressure Plasma: A Solution for Microbial Inactivation on Crop Seeds

Stephanie Rood, Hassan Mohamed, Marlon Villatoro, Briana Rojas, Hayden Cleere, Rin Komma, Teo Giannotta, Angela Komma, Catherina Htoo, Jim Browning Ph.D., Ken Cornell Ph.D.

Boise State University

In the U.S., foodborne pathogens like E. coli O157:H7 and Salmonella enterica are responsible for approximately 48 million illnesses and 3,000 deaths each year. Additionally, the U.S. agricultural industry experiences annual losses estimated to reach a staggering $21 billion due to plant pathogens such as Pseudomonas syringae and Fusarium graminearum. Both foodborne illnesses and plant pathogens display a clear risk to the viability of seeds and safety of seeds sprouted for human consumption. This research focuses on the use of cold atmospheric-pressure plasma (CAP) as a novel method for pathogen inactivation on a variety of crop seeds (sweet corn, popcorn, cucumber, mung bean, and radish). These results demonstrate that CAP treatment can reduce pathogen populations by up to 99.9% and reductions of approximately 3-log CFU/mL were seen across plant and foodborne illness pathogens upon 15 minute treatments of CAP. Germination assays display that CAP treatment does not adversely affect germination rate or seedling vigor. Our findings show that CAP is a novel and environmentally friendly alternative to traditional harsh chemical treatments, allowing for a reduction in crop loss and improved food safety.

P2-018

Process Verification: Freeze Drying and Antimicrobial Treatment in Ground Beef

Samuel Price, Pragyan Bhattarai

Weber State University

Freeze drying (Cryodesiccation) is a way of preserving food by removing water via sublimation at low temperature and pressure. Because freeze-dried foods do not require refrigeration, have long shelf lives, weigh less (making them more efficient to transport) while maintaining favorable sensory properties, it is an attractive option for food manufacturers/distributors. This study investigates the reduction of microbial load in ground beef due to freeze-drying compared to freeze-drying with an additional antimicrobial ozone treatment step in ground beef. This antimicrobial treatment under development by Tech-Source Systems is awaiting patent. The microbial load in ground beef was determined via plate counting on NEOGEN Petrifilms: Total Aerobic Count, Coliform/ Escherichia coli counts, and Total Listeria counts were collected. Additionally, spiked ground beef (used as a positive control) was prepared by adding prepared bacterial cultures of E. coli ATCC 25922, Listeria monocytogenes and/or Salmonella typhimurium (grown in tryptic soy broth). After the microbial load of un-spiked and spiked ground beef was established, the beef was processed by freeze-drying or freeze-drying with additional microbial “kill step”.      Analysis of the data indicate that cryodesiccation results in an average log reduction of 3.1 log, total aerobic count, 4.2 log total coliform count, 2.3 in E. coli counts and 3.1 log reduction in L. monocytogenes counts. Samples treated with ozone showed a 4.3 log reduction in total aerobic counts, 5.8 log reduction in total coliforms, 4.5 log reduction in total E. coli counts and, a 3.9 log reduction in L. monocytogenes counts.

P2-019 - MOVED

P2-020

New Antibiotic Production Found in Fermenting Foods

Cannon Doncaster, Todd Kelson PhD

Brigham Young University - Idaho

Fermented foods, such as sourdough starter, sauerkraut, and Kimchi, are rich in bacteria that contribute to health not only through direct microbial interactions, but also through the production of bioactive compounds that affect digestion, metabolism, and immune function. Antibiotic resistance is a global threat in part due to the lack of novel compounds that enter the pipeline of research and development. The Tiny Earth is a network of students through-out the world investigating antibiotic-producing microbes in the soil environment. Our lab networks with the Tiny Earth. We isolated bacteria from sourdough starter and Kimchi. We tested these bacterial isolates against safe relatives of ESKAPE pathogens responsible for human infectious diseases. We found antibiotic production on LB and Nutrient agars against Staphylococcus Epidermidis, Escherichia Coli, Acinetobacter Baylyi, and Bacillus Subtilis. We are isolating DNA from these fermented food bacteria to identify them and investigating the antimicrobials they produce in hopes of them entering the pipeline of research to become new classes of antibiotics.

P2-021

Stress-Induced Changes in Bacterial Membrane Vesicles in the Oral Microbiome

Taylor Boone, Swetha Shankar, Melodie Weller

University of Utah

Bacterial extracellular vesicles (BEVs) play an important role in bacterial survival by facilitating processes such as immune evasion, nutrient acquisition, and waste disposal. BEV production is often triggered by environmental stress, and growing evidence suggests these vesicles contribute to host pathogen interactions and disease progression. Within the oral cavity, bacteria experience frequent environmental fluctuations, including changes in pH, that may influence BEV production and composition. This study investigated the effects of environmental stress on BEV production using both the oral pathogen Streptococcus mutans (S. mutans), and whole saliva samples collected from human participants. S. mutans, a major contributor to dental caries and other oral diseases, was grown in media across a range of pH conditions, alongside whole saliva samples exposed to similar conditions. BEVs were isolated and characterized using nanoparticle tracking analysis, and their RNA cargo was analyzed through RNA sequencing. Results revealed that environmental stress alters both BEV production and RNA cargo composition. Notably, small RNAs, including transfer RNAs (tRNAs), and ribosomal RNAs (rRNAs), were identified as predominant RNA species within vesicles. These findings suggest that stress-induced changes in the oral environment may influence BEV-mediated communication and inflammatory signaling. Overall, this work provides new insight into how environmental conditions shape BEV production and cargo in S. mutans and oral microbial communities, highlighting a potential mechanism linking bacterial stress responses to host microbiome interactions and oral health outcomes.

P2-022

Identification of Upstream Transcription Factor1 (USF1) genetic variants associated with metabolic disease from the AllofUs database

Kenneth K.K. Ewool, Kylee Bates, Mary Davis, Julianne H Grose.

Brigham Young University

Upstream stimulatory factor 1, (USF1), is global transcription factor known to regulate several genes involved in lipid and glucose metabolism. It is particularly implicated in the disease, familial combined hyperlipidemia which affects about 2% of the western population and is a major risk factor for coronary artery disease.  Through the AllofUs database, we have identified several SNPs of USF1 which are involved in a range of disease phenotypes including cancers and metabolic diseases such as type 1 and type 2 diabetes, hyperlipidemia etc., occurring in several ethnic populations. Our study showed that SNPs of USF1 occurred both in the exons and introns with the exons occurring in the second and eleventh exons. Our study focused on 4 SNPs, chr1:161043331:C:T, chr1:161039890:C:T, chr1:161039365:T:A and chr1:161039677:G:A.  SNP occurring at chr1:161043331:C:T occurred in the 5’ UTR whilst the other 3 occurred in the 3’ UTR. chr1:161043331:C:T had increased expression of USF1 both in qPCR and western blot analysis when compared to the other SNPs. This same SNP also had increased de novo lipogenesis when compared to the others. Our study shows that SNPs identified by PheWAS can display significant lipid production effects in vivo (SNP occurring at chr1:161043331:C:T), aiding in our understanding of how USF1 alleles contribute to disease.

P2-023

Enhancing the Efficacy of CAR T cells Against Tumors Through Genetic and Metabolic Reprogramming

Alka Gaur, Annie Larson, Nate Neyman, Collin Mickelson, K. Scott Weber

Brigham Young University

CAR-T cell therapy is a powerful cancer treatment that has demonstrated significant success, particularly for hematological cancers. However, when it comes to solid tumors, CAR-T therapy still faces significant challenges. These solid tumors are surrounded by tough physical barriers like dense extracellular matrix and cancer-associated fibroblasts that make it hard for the CAR T cells to get in. Solid tumors are highly heterogeneous and composed of diverse cell populations expressing different antigens, which increases the risk of on-target off-tumor effects. To improve CAR-T therapy against these hurdles, scientists are exploring innovative approaches like knocking out or overexpressing specific genes in T cells to improve CAR-T cell signaling, activation, and cytokine production. Our approach involves knocking out the CD6 gene to boost CAR T cell performance. Cluster of differentiation 6 (CD6) is a close structural and genetic relative of CD5, and both have been shown to have a negative regulatory effect on T cell activation. They both have been generated from the process of gene duplication. Since the removal of CD5 enhances the anti-tumor activity of CAR T cells, we hypothesized that removing CD6 could similarly boost T cell activation and improve anti-tumor function. We are comparing the cytolytic function as well as metabolic differences of wild-type CAR T cells to CD6 knockout CAR T cells to check if there are significant differences in their function. Future characterization of the role of CD6 on CAR T cell function could lead to improved therapeutic strategies for solid tumors.

P2-024

Understanding the genetic basis of salt tolerance in nitrogen-fixing symbiotic bacteria

Iqra Farooq, Joel Griffitts

Brigham Young University

Soil salinity is becoming a serious global problem, as it reduces plant growth and interferes with important plant-microbe interactions. In legume crops like alfalfa, high salt levels can disrupt the relationship with Sinorhizobium species, which play a key role in nitrogen fixation. Because of this, improving the salt tolerance of these bacteria is important for maintaining crop productivity in saline environments. I am carrying out laboratory evolution of Sinorhizobium meliloti bacteria under salt stress, selecting populations with progressively higher ability to grow in medium supplemented with NaCl. I am cryopreserving samples of S. meliloti that can grow in up to 650 mM NaCl. Genomic DNA from both the original (ancestral) strains and the evolved strains will be sequenced using high-accuracy Illumina sequencing and compared to a reference genome. By looking for mutations that appear across multiple independent lineages, I will be able to identify genes most crucial for controlling salt tolerance. The evolved strains will be further tested to examine their ability to form effective symbiotic relationships with alfalfa under salt stress. Overall, this work will help improve our understanding of how microbes adapt to stressful environments and support efforts to enhance crop productivity in saline soils.

P2-025

Bovine Colostrum Supplementation Modulates Intestinal Tract Functionality in a Murine Multiple Sclerosis Model

Natalie Sofaly,  Megan Gates, Sean M. Schumacher, William J. Doyle, Javier Ochoa-Repáraz

Boise State University

Multiple sclerosis (MS) is a chronic autoimmune disease characterized by demyelination of the central nervous system (CNS). The community of microbes residing in the intestinal tract, termed the gut microbiome, has growing evidence supporting its role as an immunomodulator in MS; this bidirectional relationship is referred to as the gut-brain axis. Additionally, the intestinal barrier is essential for bidirectional homeostasis, serving as a physical barrier between the immune system and intestinal microbes, with disruption of the intestinal epithelium and detrimental changes in the gut microbiome (dysbiosis) observed in MS patients and animal models. Compared to mature milk, bovine colostrum (BC) has a higher concentration of bioactive proteins, fats, vitamins, minerals, hormones, and growth factors. Additionally, BC supplementation supports epithelial barrier integrity, inflammatory regulation, and modulation of the gastrointestinal microbiome. Despite recent interest in BC as a modulator of intestinal homeostasis, it has not been investigated in the intestinal microenvironment of MS or comparable animal models. In this study, we aim to evaluate clinical outcomes in the experimental autoimmune encephalomyelitis (EAE) C57BL/6 mouse model treated daily with oral BC or saline (as a sham control). Secreted fecal samples on day 0, 10, and 21 were analyzed using 16s rRNA sequencing, and intestinal barrier permeability was measured using a fluorescein isothiocyanate-conjugated (FIT-C) dextran permeability assay. Preliminary results indicate that BC-treatment improved intestinal barrier integrity and altered intestinal microbiome composition in EAE-induced mice.

P2-026

Deletion of TLR5 Enhances Host Survival in a Murine Model of E. coli Sepsis

O’Connor J Matthews, Alex Tran, Amanda C. Richards, Zoe Masson, William J. Brazelton, Matthew A. Mulvey

University of Utah

Sepsis is a life-threatening condition sparked by a dysregulated immune response to a perceived threat, often stemming from but not limited to bacterial infection. Worldwide, it is estimated that sepsis accounts for 10 million deaths per year, with a disproportionate impact on neonatal and elderly populations. One leading cause of sepsis is infection by Escherichia coli, a flagellated gram-negative rod commonly associated with urinary tract infection, diarrheal disease, and bacteremia. Early detection of E. coli infection is dictated by the innate immune system and includes interactions between toll-like receptor 5 (TLR5) and bacterial flagellin. We explored the contribution of TLR5 on inflammation and mortality during E. coli sepsis in a murine model of sepsis. We found that Tlr5+/+ mice faced significantly higher levels of mortality than Tlr5-/- mice during sepsis, despite harboring similar levels of bacteria in major organ systems. Transcriptomic analysis of whole blood from septic mice revealed substantial differential gene expression between Tlr5 wild-type and knockout mice, particularly in pathways related to cytokine stimulus, JAK-STAT signaling, and response to bacterial infection.

P2-027

Investigating the Role of bim and HIV-cDNA Nuclear Import in R77Q-induced apoptosis

Macey C. Call, Sidney T. Sithole, Bradford K. Berges

Brigham Young University

Human immunodeficiency virus type 1 (HIV-1) Viral protein R (Vpr) contributes to viral pathogenicity, and Vpr polymorphisms correlate with varying rates of disease progression. The R77Q mutant correlates with delayed progression to AIDS. Our lab previously demonstrated that R77Q induces apoptosis in HUT78 cells, potentially contributing to this phenotype, but the underlying mechanism of apoptotic induction remains unclear. RNA-Sequencing of HUT78 cells revealed enrichment of gene expression changes in apoptotic pathways and upregulation of the pro-apoptotic gene bim in R77Q-infected cells compared to wild-type (WT). Because cytoplasmic HIV-1 cDNA can activate DNA damage responses, we hypothesize that impaired nuclear import of viral cDNA in R77Q-infected cells triggers DNA damage signaling and bim upregulation, leading to apoptosis.

P2-028

Discovery of Short Antimicrobial Peptides in a Periplasmic Expression System

Adam Dunn, Steven Medina, Caleb Parker, Matthew Steed, Joel Griffitts

Brigham Young University

As bacteria become more resistant to the available antibiotics there is an increased demand to better understand the function and mechanism of antimicrobial peptides. However, there has not been significant research focused on the characteristics of short sequence antimicrobial peptides. These molecules are more easily synthesized and have the potential to be more stable than their larger antibiotic counterparts. In this project, we aim to discover new Antimicrobial Peptides (AMPs) using a novel peptide expression system in E. coli. We used a plasmid inducible expression system to deliver a randomized library of peptides into the periplasm. We tested a randomized library of various length containing either Isoleucine or lysine. Toxicity was then determined based on the ability of a colony to grow in the presence of inducer. The colonies that we identified as being toxic were then sequenced and further analyzed. After analysis we determined several patterns that pertain to both position and spacing within the peptide. Using this system, we have successfully identified various AMPs by testing both rationally designed peptides and randomized peptide libraries. The patterns we have found as well as the expression system we have developed have the potential to help in the prediction and discovery of new AMPs.

P2-029

BronnyJames and Nibley - Cluster J Phages

Matthew R Hatch, Cameron E Bringhurst, Porter T Danielson, Cade Brink, Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Jake W. Williams, Donald P. Breakwell, Brett E. Pickett

Brigham Young University

Bacteriophages that infect Mycobacterium smegmatis mc²155 display highly mosaic genomes, giving insight into viral diversity, genome structure, and gene function. This poster compares the genomic diversity of BronnyJames and Nibley with other cluster J phages. These phages were isolated from soil samples taken in Utah County, and following three rounds of purification, high-titer lysates (>108 pfu/ml) were prepared, and scanning electron micrographs were taken. Paired-end sequencing reads were generated using an Illumina platform, and the genomes were assembled. We analyzed tail shape and size, average nucleotide identity, HNH-associated encoding genes, and protein-based phylogenetic trees to test our hypothesis. The J-phage terminase amino acid sequences were aligned using MAFFT, and a phylogeny of the alignment was generated using IQTREE. Evolutionary relationships between Nibley, BronnyJames, and other J-phages were demonstrated. Using FASTA files from PECAAN and Phamerator, Pham maps showed notable differences between the genomic sequences of Nibley, BronnyJames, and two other typical J-phages, NihilNomen and Optimus. Using BronnyJames and Nibley genomes, along with other fully annotated phages from the J cluster, we calculated the average nucleotide identity (ANI). We analyzed the relationship of size and protein interactions between the tape measure protein and its associated tail tube, while also visualizing how the tail itself is constructed as a functional part of the phage.

P2-030

Host Range Analysis of Microbacterium Bacteriophages

Zachary Martinez, Alex Boyd, Ashton Mathews, Dr. Jack Shurley

Idaho State University

Bacteriophages are viruses that infect bacteria and play an important role in advancing our understanding of phage therapy. However, phage-host interactions are often highly specific and can be difficult to predict. In this study, 13 previously isolated bacteriophages were tested against 13 Microbacterium species to evaluate host range. Results showed that the bacteriophages Bengal, Hollow Purple, Mentos, Milo Muff, Rowlf, TicTac, Two Bits, and Steakfry exhibited relatively broad host ranges compared to the remaining isolates, with Two Bits demonstrating the broadest host range overall. Additionally, one bacterial isolate (M. liquefaciens) showed susceptibility to all 13 phages, similar to M. foliorum. These findings contribute to a better understanding of phage-host specificity and may have implications for the development of phage-based applications.

P2-031

Genome annotation & host range analysis of bacteriophage Statler. 

Daniel Puentes Navarro, Sara Sadeghi, Dr. Michael A. Thomas

Idaho State University

In fall 2023, ISU BIOL 1111 Phage Discovery Lab (part of HHMI / SEA-PHAGES program) isolated phage Statler from a thatch & soil sample. Statler has siphovirus morphology and belongs to Actinobacteriophage cluster EG. Following DNA sequencing by SeqCenter (Pittsburgh) and genome assembly by the Thomas lab, I identified 40 putative proteins out of 108 genes using bioinformatics tools. Notably, Statler contains a new function found in Microbacterium phages called cyclic oligonucleotide sequestration protein; this protein (gp57) is involved in inhibiting bacterial antiphage CBASS gene; several other EG phages encode it. A host range analysis was performed, with phage Statler, Rowlf (also cluster EG), SallyK (cluster EG), and KillerTomato (cluster EE), all found on the ISU campus by previous BIOL 1111 students. These were tested for their ability to infect hosts other than M. foliorum, using 12 Microbacterium host species and 4 other bacterial species (Staphylococcus aureus, E. coli, Bacillus subtilis & B.cereus) in a series of spot tests and plaque assays. All M. foliorum phages tested have a broad host range, infecting 9 or 10 additional Microbacterium species and none of the other genera, with little overlap among preferred hosts.

P2-032

Investigating the Role of TNF in HIV-1 Infected Cell Death      

Brennan Wells, Sidney Sithole, Bradford K. Berges

Brigham Young University

HIV-1 is the causative agent of the terminal disease AIDS. Clinical studies have determined that patients infected with a mutant of HIV-1, Vpr-R77Q, do not progress towards AIDS. Subsequently, our lab has discovered two notable differences between the Vpr-R77Q variant and WT HIV-1: increased levels of apoptosis in infected cells (which is thought to contribute to the lack of AIDS progression), and decreased levels of inflammatory cytokines, namely Tumor Necrosis Factor (TNF). The purpose of this ongoing experiment is to investigate the potential inverse correlation between TNF levels and apoptosis levels through the introduction of additional TNF cytokines to an infected system and flow cytometry analysis.

P2-033

Analysis of Genomic Conservation and Evolution in E Cluster Bacteriophages

Hunter E. Hudgins, Emily R. Hoopes, Leah P. Sim, Hunter J. Fuja, Cade Brink, Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Jake W. Williams, Donald P. Breakwell, & Brett E. Pickett

Brigham Young University

Bacteriophages that infect Mycobacterium smegmatis mc2155 provide key insights into viral evolution due to high mosaicism and gene content variability. Bacteriophages were isolated from soil samples in Oregon and Utah, using at least three rounds of purification, and amplified to high-titer lysates (>10^8 pfu/ml). Scanning electron micrographs were taken, paired-end sequencing reads were generated using an Illumina platform, and the genomes were analyzed using multiple bioinformatic tools such as NCBI, Phamerator, Starterator, HHPred, PDB, AlphaFold, SCOPE, and PECAAN. Here, we document the isolation and comparative genomic analysis of the E cluster phages Greep, Ptowny, and PaddyBaddy. In this study, we specifically analyzed the relationship between helix-turn-helix DNA binding domains with genome length, the presence and order of the lysin A and lysin B genes, the structure and anticodons of tRNA, and the presence and structure of orphams. We demonstrate here, using comparative genomic and structural analyses, variability in helix-turn-helix DNA-binding domains, the presence and functional significance of orphams, and the organization of lysis-associated genes. Comparison of the lysin A, holin, hypothetical protein, and lysin B complex may provide insight into conserved and divergent mechanisms of host cell lysis in these phages, and how the number of helix-turn-helix DNA-binding domains relates to genome length across other phage clusters.  Thus, we can conclude that there are specific regions of genetic diversity and conservation across the E cluster.

P2-034

Students vs. Viruses: 73 Novel Bacteriophages Discovered Over Three Years

Katelynn B. Owens, Zachary M. Hunsaker, Daniel A. Hemmert, Anthony L. Jeppson, Jayden S. Longhurst, Cade Brink, Payson C. Danielson, Truman D. Stohl, Jake W. Williams, Natalie A. Olsen, Don P. Breakwell, Brett E. Pickett

Brigham Young University

Over three consecutive (2024-2026), at the BYU Phage Hunters program achieved its core goal of having each student researcher discover a novel bacteriophage infecting Mycobacterium smegmatis mc^2 155. Across three cohorts, a combined total of 73 bacteriophages were isolated, purified to high-titer lysates (>10^8 pfu/mL), imaged by scanning electron microscopy, and sequenced on an Illumina paired-end platform. Genome sizes ranged from 14,822 to 156,103 bp, with average lengths increasing year-over-year reflecting growing subcluster diversity. Cluster representation expanded from 7 subclusters in 2024 to 9 in 2026, with novel additions including clusters A3, F, and the program’s first unclustered phage. Morphological analysis revealed exclusively siphoviridae morphology in the two most recent cohorts. All genomes have been computationally annotated, with the majority also manually annotated. This multi-year effort represents the largest phage sequencing initiative in BYU Phage Hunters history and demonstrates the program’s sustained ability to generate student-drive, publication-quality genomic data.

P2-035

Undergraduate-Driven Investigation of Antibiotic Persistence in Clinical ExPEC Isolates: Phenotypic Characterization and Gene Identification

Dr. Alexis Rousek 

University of Utah

Urinary tract infections (UTIs) are among the most common bacterial infections worldwide, with Extraintestinal Pathogenic Escherichia coli (ExPEC) responsible for the majority of cases. Recurrent UTIs represent a significant clinical burden, yet the mechanisms driving recurrence remain incompletely understood. Antibiotic-tolerant persister cells, a phenotypically distinct subpopulation capable of surviving lethal antibiotic concentrations, are hypothesized to contribute to treatment failure and infection recurrence, making their characterization across diverse clinical isolates a key research priority. The Science Research Initiative (SRI) here at the University of Utah places freshman undergraduates into active research labs, providing early research experience and publication opportunities to far more undergraduate students than the traditional academic research structure can accommodate. Within this framework, my lab of 17 undergraduate researchers is systematically characterizing 266 clinical ExPEC isolates. Students are currently performing swim motility, biofilm formation, and minimum inhibitory concentration (MIC) assays for seven clinically relevant antibiotics across the isolate collection. Next, strain-specific MIC data will inform persister cell kill assays to quantify antibiotic-tolerant survivors and identify high-persistence strains, with the goal of identifying genes influencing the persistence phenotypes present in our sequenced isolate library. This work aims to demonstrate that rigorous undergraduate research can generate clinically meaningful insight into ExPEC pathogenesis and antibiotic tolerance, while simultaneously training the next generation of microbiologists.

P2-036

Toxin/antitoxin HicAB Systems in Mycobacteriophages

Grant A Stock, Jake W Williams, Cade Brink, Payson C Danielson, Daniel A Hemmert, Zachary M Hunsaker, Jayden S Longhurst, Katelynn B Owens, Truman D Stohl, Brett E Pickett, Donald P Breakwell

Brigham Young University

Mycobacteriophages represent a large pool of genetic diversity and knowledge of their genomic data is increasing. Mycobacteriophage Qhanda was isolated from chicken compost in Hooper, Utah. Isolation was achieved using a Mycobacterium smegmatis mc2 155. Following three rounds of purification, high-titer lysates (>108 pfu/ml) were prepared, and scanning electron micrographs were taken. Paired-end sequencing reads were generated using an Illumina platform, and the genomes were assembled. Annotation data were collected

using PECAAN, NCBI Blast, HHpred, AlphaFold, Host-trained GeneMark and Phamerator. We observed an open reading frame in the genome of the mycobacteriophage Qhanda that is also found in other bacteriophages including Bobquesha, Juliette, Malthus, MissDaisy, Patt, and Ruthiejr. This open reading frame contains a gene encoding a toxin/antitoxin system which, when found in bacteria, encodes an anti-bacteriophage mechanism. Included is a web logo of the open reading frame, indicating highly conserved segments of amino acid sequences for the gene aligned with the Qhanda amino acid sequence. Phylogenetic tree reconstruction showed relationships between closely-related K4 phages. AlphaFold comparison of a bacterial association of a Toxin/antitoxing HicAB complex with the phage protein of Toxin HicA. Qhanda and similar phages Comparison of Genomes.

P2-037

From Singletons to Clusters: Synteny-Based Reassessment of Two Singleton Mycobacteriophages.

Tanner L. Sessions, Kobe M. Wettstein, Cade Brink, Natalie A. Olsen, Payson C. Danielson, Daniel A. Hemmert, Zachary M. Hunsaker, Jayden S. Longhurst, Katelynn B. Owens, Truman D. Stohl, Jake W. Williams, Donald P. Breakwell, Brett E. Pickett

Brigham Young University

Out of 5,944 Actinobacteriophages, only 67 (~1%) have been characterized as singletons. Mycobacteriophages have demonstrated great genetic diversity and evolutionary patterns. Mycobacteriophages, Chubbello and Bluvara, were isolated from compost samples in Idaho and Oregon, respectively, using Mycobacterium smegmatis mc2 155 as the host. High‑titer bacteriophage lysates (>10⁸ PFU/mL) were generated following three consecutive rounds of purification. Samples were visualized using scanning electron microscopy, and genomic DNA was extracted for sequencing. Paired‑end reads were produced on an Illumina platform and assembled to obtain the complete genome sequences. Chubbello and Bluvara originally were characterized as unclustered mycobacteriophages; however, based on the evidence, we have reason to believe they belong to the P and I clusters, respectively. The genomes of Chubbello and Bluvara span 46,344 and 49,459 base pairs and contain 77 and 74 predicted open reading frames (ORFs), respectively. Of the ORFs, Chubbello has 50.0% known functional genes, and Bluvara has 48.6% known functional genes, which encode key processes that support bacteriophage structure and replication. One gene in Bluvara’s genome is an orpham, a gene with no known function or family. Our findings show that the Chubbello phage shows remarkable synteny to the P1 subcluster, whereas the Bluvara phage shows synteny towards the I cluster, but ambiguity remains about its subcluster. More research is needed to conclude with which subcluster, if any, the Bluvara phage is associated.

P2-038

 Characterization of Metabolism in CD6 Knockout T cells

Miranda Sharp, Spencer Hess, Jonah Christensen, Scott Weber

Brigham Young University

T cells play an important role in the adaptive immune system as they help orchestrate immune responses to fight off infection. For T cells to fight infection, they need to switch their metabolic pathways from glycolysis and mitochondrial respiration to solely glycolysis. This change is important because the glycolysis process produces molecules for proliferation, cytokine production, and DNA replication. In our lab, we have been interested in looking at CD5 (cluster of differentiation 5), a surface receptor that inhibits T cell differentiation in the thymus. In recent data from the Weber lab, we have found that CD5 knockout CD4+ T cells rely more heavily on mitochondrial respiration and glycolysis when compared to the wild type control [2]. We are also interested in CD6 (cluster of differentiation 6), a protein in the same protein family as CD5. This receptor found on the membrane of T cells is involved with determining T cell lineage differentiation, stabilization of the TCR and MHC complex, and helps T cells migrate from the bloodstream into tissues. Recently, we have been looking at CD6 knockout mice and have seen that the basal oxygen consumption rate is statistically higher compared to the control. Further research is needed into how this change effects the mitochondria structure and other downstream targets in the cell.

P2-039

Bridging Annotation Methods: A Case Study of Manual and Computational Gene Calling

Olivia B. Tateoka, Natalie A. Olsen, Brett E. Pickett, PhD, Richard A. Robison PhD.

Brigham Young University

Bacteriophage genome annotation is complicated by overlapping genes, alternative start sites, and translational events such as ribosomal frameshifting. We compared the original SEA-PHAGES annotation of several mycobacteriophages with a reannotation produced using Pharokka to better understand how manual and automated approaches differ. In the comparison, it was apparent that the overall gene content was similar, but the two annotations differed in gene boundaries, start site selection and the treatment of hypothetical proteins. In direct comparison of translated hypothetical proteins, only a subset matched exactly, suggesting that differences in start codon choice and ORF prediction account for much of the disagreement. We also identified a ribosomal slippage region in certain phages that is recognized during manual curation but is not explicitly modeled by standard automated annotation. These results show that this comparison is a useful case study for understanding where manual and computational phage annotation agree and where they diverge. Ongoing work will extend this comparison to additional mycobacteriophages to identify patterns that are consistent across genomes and improve annotation consistency.

P2-040

Gut Microbial Composition and its Association with Physical Activity

Niki Diepeveen, Lauren Brooks Ph.D., Zoe Thompson Ph.D.

Utah Valley University

The microbiome of the gut is an increasingly important area of research as it plays a key role in human health, influencing not only digestion but also mood, mental health, and behavior Often, this link is referred to as the gut-brain axis. The bacteria in the gut play an important role in this connection. These organisms are responsible for production of many neurotransmitters including serotonin and dopamine. They even affect levels of endocannabinoids, which help regulate voluntary exercise and motivation (Dohnalova et al., 2022), and produce short-chain fatty acids, which stimulate sensory neurons in the gut that run to the brain's reward center. We study a mouse model of voluntary exercise where mice have been selected for high levels of wheel running over many generations of breeding. These high-runner mice voluntarily run on wheels about three times more than their non-selected control counterparts. The differences in the microbiomes of these groups of mice could help to uncover which bacteria are correlated with voluntary exercise behavior. While previous studies have examined the microbiome in weanling mice (McNamara et al., 2023), we plan to identify bacteria in the gut microbiome of adult mice. We will perform 16S ribosomal RNA sequencing on fecal samples collected from twelve high-running and twelve control female mice, followed by BLAST analysis to identify bacterial taxa. This data will be compared to known functions of bacteria, especially those involved in neurotransmitter and endocannabinoid production. We hypothesize that high-running mice have distinct gut microbiomes with a higher number of bacteria associated with neurotransmitter and endocannabinoid production, which may support their enhanced motivation for physical activity. Our research will help us understand the correlation between gut microbiome composition, voluntary exercise, and potential motivation for exercise behavior.

Dohnalova, L., Lundgren, P., Carty, J. R. E., Goldstein, N., Wenski, S. L., Nanudorn, P., Thiengmag, S., Huang, K.-P., Litichevskiy, L., Descamps, H. C., Chellappa, K., Glassman, A., Kessler, S., Kim, J., Cox, T. O., Dmitrieva-Posocco, O., Wong, A. C., Allman, E. L., Ghosh, S., Thaiss, C. A. (2022). A microbiome-dependent gut brain pathway regulates motivation for exercise. Nature, 612(7941), 739-747. https://doi.org/10.1038/s41586-022-05525-z

McNamara, M. P., Venable, E. M., Cadney, M. D., Castro, A. A., Schmill, M. P., Kazzazi, L., Carmody, R. N., & Garland, Jr, T. (2023). Weanling gut microbiota composition of a mouse model selectively bred for high voluntary wheel-running behavior. Journal of Experimental Biology, 226(4), jeb245081. https://doi.org/10.1242/jeb.245081"

P2-041

Disinfecting Baby Formula: Bacteriophage to fight Cronobactor sakazakii

Boyce Fraidenburg, Heather Hughes, Martha Moffat, J. Tanner Moeller and Julianne H Grose 

With antimicrobial resistance on the rise, the World Health Organization estimates that antimicrobial resistance in bacteria was responsible for 4.95 million deaths in 2019, and antimicrobial resistant bacteria is predicted to be responsible for over 169 million deaths by 2050. There is a rising need for alternative strategies to combat bacterial infections. Cronobacter sakazakii is a pathogenic bacterium that contaminates powdered infant formula and other low-moisture powdered products, causing severe illnesses such as sepsis, meningitis, and necrotizing enterocolitis in infants and immunocompromised individuals. Although antibiotics are commonly used to treat C. sakazakii infections, antibiotic-resistant strains are becoming more prevalent. To address this growing concern, we aim to identify bacteriophages that serve as an alternative antimicrobial strategy. Our current study investigates 23 bacteriophages (phages) –viruses that selectively infect bacteria– for their use in phage therapy. First, these phages will be fully sequenced and analyzed for safety. Then, they will then be analyzed for their ability to infect different C. sakazakii strains that are commonly found, including infection efficiency, how well a phage infects and lyses the target C. sakazakii bacteria. Preliminary host range trials indicate variability in each phage's ability to infect different C. sakazakii strains, with some phages displaying a broader host range. Completion of our study will identify which cocktail of phages will provide the safest and most effective phage therapy to fight C. sakazakii. 

Featured Presentation:

Uncovering hidden genomic variation in Mycobacterium tuberculosis    

Gabriella Veytsel1, Marco Marchetti2, Derek Warner3, Liliana Salvador4, Katharine Walter1

1University of Utah, Department of Internal Medicine, Division of Epidemiology 

2Utah Center for Genetic Discovery

3University of Utah, Health Sciences Center DNA Sequencing Core

4University of Arizona, Animal & Comparative Biomedical Sciences

Accurate recovery of bacterial genomic variation underlies phylogenetic and transmission inferences but remains difficult with widely used short-read sequencing data. In particular, studies of clonal bacteria, like M. tuberculosis, suffer sequencing biases from tandem repeats, high GC content, and homopolymers under current technologies. Novel sequencing technologies like AVITI have demonstrated superior performance in these genomic contexts but have not been validated for an application in bacterial whole genome sequencing. Using M. tuberculosis as a case study, we benchmark Element’s novel Avidite Base Chemistry sequencing in recovering variants following challenging genomic features and difficult-to-map regions. In this study, we sequenced two M. tuberculosis lineage 4 ATCC DNA standards (H37Ra and X003899) using both Illumina NovaSeq and Element’s AVITI platform to evaluate sequencing, mapping, and variant calling. We performed variant calling in the mtb-call2 pipeline and evaluated accuracy using vcfdist against the H37Ra and X003899 “truth” sets, annotating variant calls as true positives, false positives, and false negatives. In addition, we sequenced seven Mtb diagnostic cultures submitted to a national laboratory to evaluate application to real-world data. We assessed performance metrics following homopolymers and in repetitive and difficult-to-map contexts. Even at a lower sequencing depth, AVITI attained similar or higher performance metrics compared to Illumina. AVITI achieved higher quality reads and mapping rates, with lower duplication rates. Upon benchmarking on a “truth” set, AVITI variant calling precision was comparable. Together, our results indicate that new sequencing approaches can address current challenges with short read sequencing. The recovery of additional variation could be used to improve high-resolution bacterial phylogenetic inference and inform public health during an outbreak.

Standard Presentations:

Investigating Signaling Pathways of LINK CAARs in a Grave’s Disease Model

Brianna Davis1, Abigail Cheever1, Chloe Kang1, Morgan Payne2, Spencer Jefferies2, Kimball Demars1, Brandon Gassaway2, Kim O’Neill1, Scott Weber1

1 Brigham Young University, Department of Microbiology and Molecular Biology

2 Brigham Young University, Department of Biochemistry

            Chimeric autoantibody receptor (CAAR) T cells are a novel approach for treating B-cell mediated autoimmune diseases as they can specifically eliminate the autoantibody-producing B cells. To enhance the safety of this design, potential off-target effects of CAAR T cell treatments could be mitigated by using AND-gated logic CAAR T cells. LINK CAR T cells express two receptors, each specific to distinct targets and terminating in intracellular domains made of distinct proximal T cell signaling proteins (LAT or SLP76), requiring binding of both receptors to kill target cells. We designed and evaluated LINK CAAR T cells specific for Grave’s Disease B cells, using the autoantigen thyroid stimulating hormone receptor (TSHR) as one receptor (specific for anti-TSHR BCRs) and an anti-CD19 scFv (specific to B cells) as the other. In vitro assays demonstrated that our LINK CAAR T cells exhibited a more ‘OR’-gated phenotype, as binding of a single receptor type was sufficient for killing. To further investigate the underlying signaling pathway responses, stimulated LINK CAAR T cells were analyzed with phosphoproteomics. We found that in SLP76-containing LINK CAARs, SLP76 is significantly phosphorylated even in unstimulated conditions. Tonic signaling and overexpression of actin and cytoskeleton remodeling proteins were also observed in SLP76-containing LINK CAAR T cells. Together, these suggest that LINK CAAR T cells have a lowered activation threshold and are primed to kill target cells even with only one signal. Future work is needed to modify the LINK CAAR T cells for Grave’s Disease to fully establish ‘AND’-gated activation and cytotoxicity.

Cold Atmospheric Pressure Plasma for Bacterial Inactivation in a Porcine Ex Vivo Chronic Wound Model 

Travis Hudok

Department of Chemistry and Biochemistry, Boise State University

Chronic wound infections are difficult to treat due to persistent bacterial colonization and antibiotic resistance. Cold atmospheric pressure plasma (CAP) offers a potential alternative antimicrobial strategy therapy through the generation of reactive oxygen and nitrogen species (RONS). In this study, a planar dielectric barrier discharge CAP device was evaluated using an ex vivo porcine chronic wound model infected with Staphylococcus aureus and Pseudomonas aeruginosa. A single 10 min plasma treatment reduced bacterial viability by approximately 92.5% and 97%, respectively, with argon and air plasmas producing the greatest inactivation. Repeated treatments maintained low bacterial levels over 48 h while preserving host tissue metabolic activity. Plasma exposure induced marked bacterial membrane disruption and increased lipid peroxidation in infected wounds, with scavenger studies suggesting a preferential role for hydroxyl radicals in bacterial oxidation. These findings demonstrate that CAP effectively reduces bacterial burden in a physiologically relevant chronic wound model and supports its further development for chronic wound infection management.

Beyond Pathway Enrichment: Uncovering Disease-Relevant Regulatory Modules in Bulk RNA-Seq via GRN Clustering, Text Mining, and Enrichment: A Case Study in SHS-Induced Preeclampsia 

Archarlie Chou1, Juan Arroyo2, Brett Pickett1

1Brigham Young University, Department of Molecular and Microbiology

2Brigham Young University, Department of Cell Biology and Physiology

Bulk RNA-seq data pose a major analytical challenge due to the large amount of generated information, making it difficult to identify biologically relevant and meaningful patterns. Here, we report a new pipeline that enables improved and integrated analysis for a disease of interest, after which the algorithm returns the most relevant clustered gene regulatory networks (GRNs), along with advanced visualizations to help infer signaling pathways from transcription factors (TFs) and to significantly differentially expressed genes (DEGs). Briefly, we first combined a GRN (derived from KBoost) with a protein–protein interaction (PPI) network generated from DEGs, then applied greedy modularity optimization to identify subgraphs (communities). Within each community, we performed gene ontology (GO) enrichment and pathway enrichment via SPIA (performed at the GO/pathway level). Gene-disease associations were retrieved from DisGeNET, JensenLab, PubTator-based, Perplexity queries text mining. Communities were then re-evaluated using hypergeometric statistics based on the number of disease-relevant genes and the on the number of high-ranking centrality nodes they contained (performed at the communities’ level). Network visualization is powered by Force-Graph (3D). This analytical workflow generates clusters of genes based on GRNs that likely reflect true regulatory influence and can automatically prioritize biologically meaningful modules for further inspection. In a case study of a secondhand smoke–induced preeclampsia mouse model, we identified several potentially activated TFs and estimated the presence of their downstream targets using DoRothEA.

Plasma vs. Pathogens: Molecular Mechanisms of RONS-Mediated Viral Inactivation

Krystal Sosa, Razmira Hamidovic, Tessa Sprague, Dr. Jim Browning, and Dr. Ken Cornell

Boise State University

The control of viral infections is currently hindered by a limited antiviral pharmacopeia and the tissue toxicity of chemical disinfectants. Cold Atmospheric Pressure Plasma (CAP), a partially ionized gas generating Reactive Oxygen and Nitrogen Species (RONS), presents a promising non-corrosive alternative.Using a novel eight-channel linear discharge array, we demonstrated broad-spectrum biocidal activity, inactivating >50% of viral particles with brief exposure. Mechanistic investigations into enveloped (HSV-1) and non-enveloped (FCV) viruses revealed that RONS drive viral degradation through lipid peroxidation, tyrosine nitrosylation, and nucleic acid alteration. To evaluate clinical utility, we utilized an ex vivo porcine eye model, where CAP treatment significantly reduced HSV-1 infection levels and progeny virus production within corneal lesions.These findings highlight CAP’s potential to combat persistent infections like HSV-1, which causes 40,000 annual cases of blindness. By defining the biochemical interactions between plasma and viral molecules, this work establishes a framework for integrating physical-based therapies into public health infrastructure where conventional pharmacology meets its limits.

Antimicrobial peptide discovery: forging a path to clinical relevance  

Caleb Parker, Matthew Steed, Adam Dunn, Steven Medina, Joel Griffitts

Brigham Young University

Rising rates of antibiotic-resistant infections have sparked a deeper interest in alternatives to traditional small-molecule antibiotics. One promising alternative is the class of molecules known as antimicrobial peptides (AMPs), which rarely induce bacterial resistance. A limitation of AMPs is that they are often quickly degraded by proteases, preventing them from reaching and killing microbes. We are developing a new platform to discover thousands of new cyclic AMPs. Cyclization is a common strategy to increase resistance to proteolysis, but it is usually only done to one peptide at a time. Our platform will improve upon this by screening hundreds of thousands of cyclic peptides in a short amount of time. We are using an in vivo self-screening system, where E. coli expresses and responds to randomized peptides. If a particular peptide is antimicrobial, it will inhibit the growth of its producing cell. To cyclize peptides, we fused a periplasmic signal sequence to eight randomized amino acids flanked by two cysteine residues. The signal sequence exports it into the periplasm, where the cysteine residues are oxidized to form a disulfide bond. Preliminary results indicate that our system is effective at finding new AMPs. Continuing development and scaling of our platform will lead to high-throughput sequencing and testing multiple cyclization schemes in E. coli. Using our cyclization screening system, we can identify thousands of new AMPs, yielding insights into patterns of antimicrobial activity, clues regarding novel mechanisms of action, and candidates with desirable drug-like qualities.

Featured Presentation

Mechanisms of Mycobacterium tuberculosis survival under vaccine-induced immunity

Kimra S. James1, Neharika Jain2, Kelly Witzl1, Nico Cicchetti1, Sarah M. Fortune3, Thomas R. Ioerger4, *Amanda J. Martinot2, *Allison F. Carey1

*Corresponding authors

1Division of Microbiology & Immunology, Department of Pathology, University of Utah

2Department of Infectious Disease and Global Health, Tufts University Cummings School of Veterinary Medicine

3Department of Immunology & Infectious Diseases, Harvard T.H. Chan School of Public Health

4Department of Computer Science and Engineering, Texas A&M University

            Tuberculosis (TB), remains a persistent global health challenge due to the lack of an effective vaccine. The only licensed TB vaccine, Bacille Calmette-Guerin (BCG), is a live attenuated strain of Mycobacterium bovis that protects young children from severe disease but fails to provide protection through adulthood. It is unclear why BCG provides incomplete protection despite inducing a robust Th1 immune response. We set out to interrogate mycobacterial determinants of vaccine escape using a functional genomics approach, TnSeq, to define bacterial genes required for survival in mice vaccinated with BCG, the live attenuated Mtb vaccine strain, LprG, and in mice with Mtb immunity conferred by prior infection. We find that critical virulence genes associated with acute infection and exponential growth are less essential in hosts with adaptive immunity, including genes encoding the Esx-1 and Mce1 systems. Genetic requirements for Mtb growth in vaccinated and previously Mtb-infected hosts mirror the genetic requirements reported for bacteria under in vitro conditions that reflect aspects of the adaptive immune response. Across distinct immunization conditions, differences in genetic requirements between live attenuated vaccines and vaccination routes are observed, suggesting that different immunization strategies impose distinct bacterial stressors. Collectively, these data support the idea that Mtb requires genes that enable stress adaptation and growth arrest upon encountering the restrictive host environment induced by the adaptive immune response. We demonstrate that TnSeq can be used to understand the bacterial genetic requirements for survival in vaccinated hosts across pre-clinical live attenuated vaccines and therefore may be applied to other vaccine modalities. Understanding how Mtb survives vaccine-induced immunity has the potential to inform the development of new vaccines or adjuvant therapies.

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Silent Killer: how the innate immune receptor Clec12a defends against potentially harmful microbes

Morgan Greenewood

University of Utah, Pathology, June Round Lab

We are constantly exposed to a vast range of microbes starting at birth, some beneficial, others benign, and still others pathogenic. Our immune system must not only maintain tolerance to this complex community of microorganisms that becomes our microbiota but also defend against any pathogens. How our immune system does this without excess inflammation remains vastly unknown. We have determined that the innate immune receptor Clec12a, functions at the cusp of these two functions within the intestine. This pattern recognition receptor (PRR) is unique in that unlike other PRRs that mount pro-inflammatory responses, Clec12a does the opposite. It inhibits pro-inflammatory signaling downstream of other PRRs. Mice lacking Clec12a get worsened colitis, and upon analysis of the Clec12a-/- intestinal microbiota composition, we see an outgrowth of the bacteria Faecalibaculum rodentium, which, if administered to WT mice, can drive worsened colitis, suggesting that it has pathogenic qualities. We find that Clec12a can bind specific intestinal bacteria and that, upon binding is involved in phagocytosis, resulting in microbiota compositional changes. Further, when Clec12a-/- macrophages are stimulated with F. rodentium, these macrophages exhibit significantly more pro-inflammatory cytokine production. Together, these data suggest that Clec12a acts as a silent killer within the intestine, binding potentially harmful microbes and removing them without eliciting excessive inflammation.

Oxygen-dependent metabolism affects virulence traits in Pectobacterium carotovorum

Carlton Collins, Ethan Shreeve, Foxx Naylor, Mailon Odell, Jeffrey Schachterle

Brigham Young University, Department of Microbiology and Molecular Biology

Pectobacterium carotovorum (Pcc) is a phytopathogen that causes soft rot disease by releasing plant cell wall-degrading enzymes (PCWDEs). Soft rot affects potatoes and other vegetables, resulting in annual losses exceeding $400 million. Pcc, a facultative anaerobe able to switch between aerobic and anaerobic metabolism depending on oxygen availability, is more virulent in low-oxygen environments. Recent studies in V. cholerae, another facultative anaerobe, suggest that aerobic metabolism machinery is essential for proliferation, even in low-oxygen environments. Understanding the impact of aerobic and anaerobic metabolism on virulence-related traits in Pcc provides insights into storage and handling, and in the identification of novel targets for chemical and biological control. In many enterobacteria, the Pyruvate Dehydrogenase Complex (PDH) converts pyruvate into acetyl-CoA in aerobic environments, and Pyruvate Formate Lyase converts pyruvate to acetyl-CoA in anaerobic environments. To investigate the role of metabolism in virulence-related traits, we removed genes essential to PDH (aceE) and PFL (pflB) by creating knockout mutants using lambda red recombination. Genes were cloned into pBBR1 for complementation, and phenotypes associated with virulence were measured on colonies grown aerobically and in an anaerobic chamber. Preliminary results indicate that disrupting PDH and PFL decreases swimming motility, and that disrupting PDH increases biofilm formation when compared to wild type. This provides a direct link between the oxygen-dependent modulation of metabolism and virulence in Pectobacterium carotovorum. Further investigations are underway to assess additional virulence-related traits and genes.

Genetic control of Group 3 (K96) capsule synthesis and complement resistance in extraintestinal pathogenic Escherichia coli

Rachael David Prince, Rajesh Bogati, Titan Lytle, Andres Collado Silva, Rachael LeBaron, Tiana Sorge, Michael Olson, Eric Wilson, and David Erickson

Department of Microbiology and Molecular Biology, Brigham Young University

Extraintestinal pathogenic Escherichia coli (ExPEC) frequently produce Group 2 or 3 capsules that enhance virulence, yet genetic regulation and specific roles of Group 3 capsules remain poorly understood. Here we demonstrate that the K96 Group 3 capsule in ExPEC strain M12 confers resistance to human serum complement. Using genetic screens, we identified phase-variable mutations in kpsC and essential roles for transcriptional regulators OxyR and RfaH in activating capsule synthesis via a distant promoter. Mutants lacking OxyR or RfaH failed to produce capsule, exhibited complement sensitivity, and showed attenuated virulence in Galleria mellonella. These findings reveal novel mechanisms controlling Group 3 capsule expression and underscore the capsule’s contribution to immune evasion.

Characterization of CD5/CD6 Double Knockout Mice Reveals Effects on Immune Cell Differentiation and T Cell Activation

Alina Rodriguez

Brigham Young University

CD5 and CD6 are closely related T cell receptors that regulate TCR signaling and influence T cell activation, differentiation, and function. While individual deletion of CD5 or CD6 has been shown to enhance T cell responses, the combined impact of their simultaneous deletion remains unknown. Here, we characterize CD5/CD6 double knockout (DKO) mice, alongside CD5 knockout, CD6 knockout, and wild-type controls, including both male and female mice. Immunophenotyping of peripheral white blood cells, including T cells, B cells, monocytes, macrophages, and granulocytes, along with T cell subsets, has shown how single and double deletions affect immune cell composition. Additionally, we measured calcium influx with flow cytometry to evaluate T cell activation as a functional readout of TCR signaling. Comparisons across genotypes will clarify whether CD5 and CD6 act redundantly, compensatorily, or independently in regulating T cell activation and immune cell differentiation. This study provides a detailed characterization of CD5/CD6 double knockout mice and establishes a foundation for understanding how these homologous co-receptors shape immune differentiation and T cell function. These insights may inform strategies to enhance T cell activation, persistence, and efficacy for future immunotherapy applications.

Mechanistic Insights into Slower AIDS Progression Associated with the HIV-1 Vpr R77Q Variant

Sidney T. Sithole, Joshua Ramsey, Macey Call, Brian D. Poole, Brett E. Pickett, Bradford K. Berges

Department of Microbiology and Molecular Biology, Brigham Young University, Provo, Utah, USA

HIV-1 viral protein R (Vpr) is a multifunctional regulator of viral pathogenesis, and natural polymorphisms in vpr have been linked to differences in disease progression. The HIV-1 vpr R77Q mutation is associated with delayed progression to AIDS. We previously showed that R77Q-infected CD4+ T cells not only undergo less cell death overall, but also die via a non-inflammatory apoptotic pathway. This qualitative shift toward non-inflammatory apoptosis may help explain the slower clinical progression seen in individuals harboring this mutant. To investigate the mechanism driving this phenotype, we performed RNA-seq on HUT78 cells infected with either wild-type NL4-3 or the R77Q mutant. At 72 hours post-infection, transcriptomes were heterogeneous, and differential expression analysis identified 289 significantly altered genes in the R77Q vs. WT comparison. Enrichment analysis and Gene Ontology terms highlighted apoptosis-related pathways. While both strains induced pro-apoptotic genes, the R77Q mutant showed a striking failure to upregulate key anti-apoptotic regulators, most notably bcl-2, whereas WT infection resulted in strong bcl-2 induction. Protein–protein interaction analysis of the Bcl-2 family network suggested disrupted or weakened interactions consistent with a loss-of-function phenotype. Follow-up assays confirmed reduced Bcl-2 expression at the protein level in R77Q-infected cells, supporting the transcriptional findings and the observed increase in non-inflammatory apoptosis. Together, these results provide a mechanistic explanation for how the R77Q mutant may contribute to delayed AIDS progression. By promoting a controlled, apoptotic cell-death program through impaired upregulation of anti-apoptotic pathways, the R77Q mutant elicits a host response that may reduce inflammation and slow disease progression.

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Rhizosphere bacteria promote seedling growth and suggest non-canonical nitrogen fixation strategies

Johnathon Brim, Sierra Grandy, Joseph Richardson, Jeffrey Schachterle

Brigham Young University Microbiology and Molecular Biology Department

Synthetic nitrogen fertilizers have revolutionized agriculture, but their overuse has led to severe environmental consequences, including waterway eutrophication, greenhouse gas emissions, and soil degradation. Biological nitrogen fixation (BNF) by rhizosphere bacteria offers a sustainable alternative, yet options for BNF systems remain limited by environmental constraints and plant host range. Here, we isolated and characterized rhizosphere bacteria capable of growth on nitrogen-free media and promoting seedling growth.  The majority of isolates belong to the genera Siccibacter and Stenotrophomonas.  Genome sequencing revealed that a Siccibacter turicensis isolate exhibited robust growth in nitrogen-free medium, consistent with nitrogen fixation activity, despite lacking canonical nitrogenase genes, suggesting a potentially novel nitrogen fixation mechanism.  The S. turicensis genome also contains a type III secretion system which could facilitate interactions with host plants.  Importantly, hypersensitive response assays using Nicotiana benthamiana produced negative results, indicating no pathogenic potential.  Several isolates displayed levansucrase activity, producing the exopolysaccharide levan, a common trait of plant-associated bacteria. In seedling inoculation assays, plants treated with these bacteria had significantly better germination and growth compared to uninoculated controls.  Together, these findings identify novel rhizosphere bacteria with beneficial effects on early plant development and provide genomic resources for future work into alternative nitrogen fixation strategies and plant-microbe interactions.

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From Structure to Activity: A Preliminary Look at Fungal Networks Under Stress

Erika Espinosa-Ortiz1, Bradley Lawson1, Venkatraman Siddharthan2

1Utah State University - Biological Engineering Department

2Utah State University - Animal, Dairy, & Veterinary Sciences Department

Fungi play critical roles in applied and environmental microbiology, contributing to nutrient cycling, biodegradation, soil structure, and interactions with plants and other microorganisms. Many of these functions are mediated by filamentous fungal networks whose morphology and physiological state dynamically respond to environmental stress. However, linking changes in network structure to functional activity remains challenging. Fluorescent reporters such as green fluorescent protein (GFP) offer a promising approach to bridge this gap by enabling non-invasive visualization of fungal activity within living networks. In this preliminary study, we explore the combined use of GFP fluorescence and morphological observations to examine how fungal networks respond to different stress conditions commonly encountered in environmental and applied settings. Fluorescence imaging provides a spatially resolved view of activity within the mycelium, while concurrent assessment of network morphology captures broader structural responses at the network level. Rather than focusing on specific pathways or mechanisms, this work aims to evaluate the feasibility and potential insight gained from integrating these complementary perspectives. Initial observations suggest that stress can induce heterogeneous responses across fungal networks, with variations in fluorescence and structure emerging in different regions of the mycelium. These patterns indicate that functional responses relevant to environmental performance - such as stress tolerance, persistence, or resource allocation - may not be uniformly distributed and may therefore be overlooked by bulk or morphology-only measurements. Overall, this exploratory study establishes a foundation for future work linking fungal network structure to function under environmentally relevant conditions. Such an integrated approach has potential applications in understanding fungal behavior in soils, engineered systems, and biotechnological processes, where stress responses strongly influence performance and ecological impact.

The Plant Virome of Grand Gulch Utah

Alma G. Laney1,Megan Frisby1, Elise Bennett1, Rob Hess1, Jaylee Montague1, Caleb Sawyer1, Max Taylor2, Geoff Zahn1,3, and Erin Riggs1,4

1Utah Valley University, 2Hopi Nation, 3William and Mary Computational & Applied Mathematics & Statistics program, 4Oregon Coast Community College

Grand Gulch is located in the Southeastern corner of Utah on the San Juan Rivier within the Bears Ears National Monument. Historically, this was a site of ancient agriculture and human settlement by several groups of people including the Basket Makers (ca, 200 AD). Repeated droughts over the last 1000 years led to people dispersing from the area. Due to the remoteness of the location, little is known about plant pathogens in the location. Samples were collected various locations including the San Juan River-Grand Gulch convergence and head waters (2022), Water Canyon (2023), and Deer Canyon and Three Kiva Pueblo just outside of the canyon system (2024). Preference was given to collecting plants with typical plant viral symptoms, though other samples were also collected. Samples were returned to the lab and immediately processed for storage at -80C. RNA was extracted using the Plant Total RNA extraction kit (IBI Scientific) and subjected to paired end reads on the NovaSeq to a 3 GB coverage for 14 samples. Once the adaptors were trimmed and the paired end files concatenated, the samples were analyzed using VirFind. Several new plant viruses were detected including a novel potyvirus and allexivirus that were found in two divergent plant hosts, Brassica oleracea and Datura wrightii, from different portions of the canyon suggesting transmission of these viruses within the canyon. This was further supported by the presence of unidentified aphids on the B. oleracea samples collected. Beet curly top virus was also detected within several hosts within the canyon system. Given that these viruses are agriculturally important, these could pose a risk to agriculture in the region as well as to native plants in the region.

Microplastics as Physical Hubs Amplifying Antibiotic Resistance Under Mixed-Contaminant Selection in Wastewater Systems 

Tahira Rahman1, Akib Al Mahir1, Dr. Liyuan (Joanna) Hou2

1Department of Civil and Environmental Engineering, Utah State University; Utah Water Research Laboratory (UWRL)

2Department of Biological Engineering, Utah State University, Utah Water Research Laboratory (UWRL)

Microplastics (MPs) are increasingly recognized as potential hotspots for antibiotic resistance in wastewater systems; however, their interactions with co-occurring contaminants and treatment processes remain insufficiently understood. This study combined laboratory-scale experiments and full-scale wastewater treatment plant (WWTP) investigations to elucidate the roles of MPs, pharmaceuticals and personal care products (PPCPs), and heavy metals in shaping the antibiotic resistome. Drip flow reactor experiments demonstrated that MP-associated biofilms accumulated significantly higher abundances of ARGs and MGEs than non-plastic controls, despite comparable biofilm biomass. Polymer type structured distinct plastisphere communities, indicating selective microbial assembly on different MP surfaces. Copper (Cu) and sulfamethoxazole (SFX) imposed gene-specific selective pressures on ARG profiles, whereas amitriptyline (AMP) exhibited minimal effects at environmentally relevant concentrations. No significant synergistic interactions among MPs and chemical contaminants were observed. Complementary analyses across treatment stages in two WWTPs revealed that biological treatment reorganized microbial interaction networks, concentrating ARGs and MGEs into highly connected network hubs and increasing the potential for horizontal gene transfer. Statistical modeling identified dominant MP polymers as central physical drivers of ARG enrichment, while PPCPs and heavy metals primarily acted as chemical selective factors, and microbial community composition defined the available host reservoir. Together, these findings demonstrate that microplastics function as physical hubs that facilitate ARG accumulation by supporting dense biofilms, whereas co-occurring contaminants impose gene-specific selection. These results underscore the need to integrate physical substrates, chemical exposures, and microbial ecology when assessing antibiotic resistance risks in wastewater systems.

Fusarium venenatum as a biosorbent for cadmium removal 

Bradley Lawson1, Allondra Woods1, Joan Mclean2, Erika Espinosa-Ortiz1

1Utah State University - Biological Engineering Department

2Utah State University - Utah Water Research Laboratory

Heavy metals are environmental pollutants of increasing concern that can be introduced into aquatic ecosystems predominately through industrial waste, mining and smelting activities, and the burning of fossil fuels. Cadmium is a known cancer-causing agent in humans and poses a significant risk to aquatic health as it can cause acute mortality and long-term complications in aquatic life. As the rise of cadmium pollution continues it is imperative to develop accessible and sustainable methods for remediation to ensure clean water across the globe. Fungi offer a potential solution for the removal of cadmium from contaminated aquatic environments. Fungi are diverse in their ability to remediate heavy metal contamination and contain multiple pathways for cadmium removal, such as adsorption, absorption, and transformation. This study proposes the use of the fungus Fusarium venenatum for the biosorption of cadmium from contaminated solutions. F. venenatum predominantly removes cadmium through adsorption to functional groups (carboxylic acid and amides). This study tested the effects of pH (4-8) and initial biomass dosage (0.25-2.5 g/L) on the effects of cadmium removal and found that the optimal pH range was from 5-8 and the optimal biomass dosage was 2 g/L. Under these conditions, the fungus could remove 92% of cadmium from a 10 mg/L solution in under 4 hours highlighting its capability of being used as a biosorbent for cadmium in contaminated aquatic environments. Future work will look at the desorption of Cd from the fungal biomass to assess the mechanisms of Cd removal.

Ecophysiology of Dormant Bacterial Endospores in Seafloor Sediments Impacted by Continuous Emission of Warm Fluids from Deep Oceanic Crusts  

Rhys Ellis, Anirban Chakraborty

Idaho State University Department of Biological Sciences

Dormant endospores of thermophilic bacteria are abundant within the deep subsurface and have also been detected globally throughout the cold seabed. Distribution patterns of these subsurface bacteria may be explained by a dispersal history originating in warm subsurface habitats, e.g., the deep crustal biosphere. The Juan de Fuca ridge, located off the coast of the Pacific Northwest, presents an accessible crustal system with multiple sampling observatories permanently drilled into underlying crusts. To assess the potential of endospore dispersal originating from warm ocean crusts, we examined endospore distribution in seafloor sediments near a non-operational sampling platform (U1301A), located on the eastern flank of the Juan de Fuca ridge, where a consistent discharge of warm crustal fluids into the surrounding water column has been observed for almost two decades. 25-cm-deep sediment cores were collected by ROV Jason from three sites near U1301A along a transect moving away from the borehole. Cores were also collected from a nearby operational, non-leaking borehole (U1362B). Endospore germination in pasteurized surface sediments was stimulated by combining sediments with artificial seawater medium amended with sulfate and a mix of volatile fatty acids (VFAs), followed by anoxic incubations at 50 and 65F for six weeks. Sulfate and VFA depletion patterns in subsamples collected at different time points indicated activity of germinated endospores, while community analyses using 16S rRNA gene amplicon sequencing showed noticeable variation in endospore communities at different coring sites. Additionally, we observed distinct endospore lineages associated with specific incubation temperatures within sediments from the same sediment core.