Microbiology Commons^™

The Role Of Seed Mix Characteristics And Soil Amendments During Post-Agricultural Prairie Restoration, Valerie N. Thurston, Michaela J. Woods, Madelaine Gregory, Mary C. Lloyd, Ryan W. Mcewan

Biology Faculty Publications

Introduction: Prairie restoration on degraded lands may be influenced by seed mix characteristics, including species richness and legume establishment. Soil amendments may facilitate restoration; however, the practical usefulness of techniques such as applications of whole soil and leaf mulch remains relatively unclear.

Objectives: We assessed whether increasing species richness and legume content in seed mixes enhances prairie restoration success and examined how soil amendments interact with seed mix composition to influence cover, species richness, diversity, and legume establishment.

Methods: We conducted a field experiment on a 12-ha post-agricultural site in Ohio, U.S.A. Twenty 50 × 50 m plots received one …

Hydrogels For Local Drug Delivery In Biofilm-Associated Periprosthetic Joint Infection: Current Progress And Future Directions, Karolina Kraus, Pawel Mikziński, Bindu Subhadra, Emil Paluch

Faculty Publications - Biological Sciences

Periprosthetic joint infection (PJI) remains one of the most serious complications of arthroplasty, largely due to the formation of microbial biofilms on implant surfaces. Biofilm-associated infections exhibit increased tolerance to antimicrobial therapy and host immune responses, making eradication difficult and often requiring repeated surgical interventions. Consequently, there is a growing need for effective local therapeutic strategies capable of delivering high concentrations of antimicrobial agents directly to the site of infection while minimizing systemic toxicity. Hydrogels have emerged as promising drug delivery platforms for the management of biofilm-associated PJI. Their biocompatibility, injectability, high water content, and tunable physicochemical properties enable controlled …

Tissue-Specific And Sex-Dependent Microbiome Structure In East African Cattle Ticks (Amblyomma Gemma And Rhipicephalus Pulchellus) And Their Bovine Hosts, Dennis Getange, Samson Mukaratirwa, Rua Khogali, Epaphrus Yuko, James Kabii, Joel L. Bargul, Jandouwe Villinger

All Peer-Reviewed Publications

Ticks and their vertebrate hosts share intimate ecological interactions that shape microbial community composition, with implications for tick fitness and pathogen transmission. However, the extent and specificity of tick–host microbial interactions remain poorly understood, particularly in East African tick species. Here, we characterised microbial communities across multiple tissue types from two economically important tick species (Amblyomma gemma and Rhipicephalus pulchellus) and their shared bovine host using 16S rRNA V1-V2 gene sequencing. We profiled haemolymph, midgut, salivary glands, and gonads from individual ticks (25 Am. gemma and 8 Rh. pulchellus), alongside 12 bovine blood samples. Microbiota were dominated by Proteobacteria, Firmicutes, …

Bridging The Gap In Science Research: Preparing Students For College- Level Research In The Sciences, Victoria Helwig, Yaoguang Li

UConn Library Presentations

Many first-year students enter college unprepared to fully engage with scientific literature and academic library resources. Drawing on experiences from academic librarians working with STEM students, this session explores common gaps in science research skills, including navigating databases, identifying scholarly sources, and understanding the structure of scientific literature. Participants will be introduced to practical strategies and classroom-ready activities to better prepare students for college-level research. Discussion questions: What science research skills do students struggle with most when entering college? How can high school instruction better support these skills? Where can librarians and educators collaborate to support student success?

Analysis Of The Atypical Temperature Dependence And Conformational Changes During Turnover Of A Lactobacillus Chlorogenic Acid Esterase, Nathaniel Carl, Yianni Tsigaris, Dan Ji, Nora K. Anpree, Kellie K. Omori, Cedric P. Owens

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Many bacterial chlorogenic acid esterases (ChlEs) exhibit atypical temperature behavior, featuring activities that barely change with temperature and activity maxima that fall below the thermal denaturation point. This work focuses on a ChlE from Lactobacillus helveticus (Lh–ChlE), which has a flat temperature dependence. First, it was determined that conformational changes during Lh–ChlE turnover are not rate-limiting and that the overall rate depends on the chemical step at all temperatures. Next, Lh–ChlE’s temperature dependence was investigated using a conformational equilibrium model that assumes the existence of a temperature-dependent equilibrium between an active and an inactive conformation and an activation heat capacity …

De Novo Pyrimidine Synthesis Controls Germinal Center B Cell And Plasma Cell Fates And Systemic Autoimmunity, Julia L. Weber, Tien Bui, Keomonyroth Nuon, Adam J. Fike, Sophia M. Crocker, Kristen N. Bricker, Anju Maharjan, Wujuan Zhang, Aaron R. Goldman, Sathi Babu Chodisetti, Ziaur S. M. Rahman

Department of Microbiology and Immunology Faculty Papers

Whether and how pyrimidine metabolites promote systemic autoimmunity is unknown. Here, metabolomics and 15N-amide glutamine tracing show enhanced flux through de novo pyrimidine synthesis in systemic lupus erythematosus (SLE)-prone B cells. Temporal inhibition of pyrimidine synthesis dampens SLE-prone but not foreign antigen-specific germinal center (GC), plasma cell (PC), and antibody responses. Uridine monophosphate synthase (UMPS) conditional deletion, however, reveals a B cell-intrinsic requirement of de novo pyrimidine synthesis in foreign antigen-driven and SLE-prone GC, PC, and antibody responses and kidney immune complex deposition. Metabolomics, mitochondrial stress test, metabolic flow cytometry, glycolytic rate assay, and RNA sequencing highlight the importance of …

Pharky, Phedro, Phractured, And Stagephright: Complete Genome Sequences Of Four Ek2 Cluster Microbacterium Phages, Susan M. R. Gurney, Medha Gupta, Deeksha Reddy, Gloria Zacharias, Aleda S. Jomy, Carly J. Smith, Matthew T. Mcdonald, Brett M. Condon

Student Papers, Posters & Projects

We report the complete genome sequence of four Microbacterium phages, Pharky, Phedro, Phractured, and StagePhright, isolated using the bacterial host Microbacterium foliorum from soil samples collected in Philadelphia. All four phages are genetically very similar and contain a gene of unknown function, which is uncommon in the EK2 cluster.

Homocysteine Thiolactone From Translation Proofreading Is An Endogenous Ligand For Cell-Cell Signaling By The Receptor Sdia, Lorenzo Eugenio Leiva, Roberto A. Zúñiga, Gregory J. Phillips, Michael Ibba

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Several widespread mechanisms enable cellular communication and coordinate the behavior of bacterial populations. The modules responsible for cellular communication are often found in matching pairs: an enzyme that synthesizes a signal molecule and a specific receptor that decodes it. A notable exception is Escherichia coli SdiA, a homolog of known N-acyl-homoserine lactone (AHL) receptors, for which no corresponding AHL synthase exists. SdiA is an orphan receptor that enables cross-talk by sensing AHLs from other bacterial species. In this work, we investigated whether homocysteine thiolactone (HTL), an AHL-like molecule arising from a proofreading reaction of methionyl tRNA synthetase (MetRS) to correct …

Birds Of A Tether Flock Together: Examining Centromere Tethering And Chromosome Segregation In Caulobacter Crescentus, Jake Dwyer, Simon Kruczak, Rodrigo Arana-Garrido, Paola Mera

MICRO-CCS student projects

Introduction:

Chromosome segregation is essential for all domains of life. Caulobacter crescentus is a key model for studying this process. It divides asymmetrically into two morphologically distinct cells. Proteins can be seen directly with fluorescent tags.

Research done on Caulobacter can apply to many different bacteria. The ParABS system is integral to chromosome segregation in Caulobacter.

ParS – DNA sequence on the chromosome

ParB – CTP-binding protein that specifically recognizes parS

ParA – ATP-binding protein that non-specifically binds DNA and provides the force for segregation.

PopZ – Polar tethering protein via interactions with ParB.

TipN - Polar protein that …

Has Plant Domestication Compromised Plant Defensive Chemistry? Aphid Induced Stress In Tomatoes, Nhi Do, Satinderpal Kaur, Minxing Zhu, Esther N. Ngumbi

MICRO-CCS student projects

Tomato plants have natural defensive mechanisms.

Direct: Phenolics are secondary metabolites that can protect the plant by intoxicating the herbivores and/or producing strong repulsive odors.

Indirect: VOCs, Volatile Organic Chemicals, are gas phase signals that attract the predators of the attacking herbivore to the plant.

Domestication of plants may chemically alter these defenses by prioritizing fruit production over defensive traits.

We hypothesize that wild/ancient tomato plants would have a greater quantity of volatiles and increased concentration of leaf total phenolics compared to cultivated tomatoes during aphid herbivory.

How Does High-Oil Sorghum Change Potential Extracellular Enzyme Activity Across Soil Depth Profiles Under Drought Conditions?, Krystal Monique Role, Rounak Patra, Wendy Yang

MICRO-CCS student projects

Transgenic oil sorghum reallocates photosynthate-carbon (C) into vegetative lipids, accumulating nonstructural lipids in both shoot and root tissues. Altered plant C allocation and root lipid accumulation may shift root exudate chemistry, belowground C inputs, and microbial nutrient demand throughout the soil profile. While aboveground productivity in high-oil sorghum is well-characterized, its belowground impacts on soil microbial enzyme activity and exchangeable cation dynamics, especially across soil horizons under drought stress remain unknown.

Project objective was to find out how transgenic high oil sorghum (Tx-HO1) influences CNP acquiring extracellular enzyme activities and soil exchangeable cation dynamics across soil horizons under drought and …

Investigation Of Biological Denitrification Inhibition By Specific Anthocyanins, Kristin Palazzo, Holly Anderson, Aislinn Geedey, Angela Kent

MICRO-CCS student projects

Nitrogen fertilizers are applied to corn fields to support high crop yields, but some nitrogen is lost through microbial denitrification, producing nitrous oxide (N2O), a potent greenhouse gas. Some plants produce certain flavonoids that can suppress microbial denitrification, a trait known as biological denitrification inhibition (BDI). Maize produces flavonoids in the form of anthocyanins, including cyanidin-3-glucoside (C3G), the major anthocyanin produced by maize.

The goals of this project were to determine whether C3G:

is produced in maize roots

inhibits microbial denitrification

Recovering Plasmid-Id Barcoded Strains Of Sinorhizobium Meliloti From A Native Soil Microbiome, Kenny Sacks, Hannah Murray, Katy Heath

MICRO-CCS student projects

Introduction

Legume-Rhizobia Mutualism: Nitrogen is an essential nutrient for plant growth. Plants can’t produce nitrogenous compounds. Rhizobia fix atmospheric nitrogen gas into bioavailable forms. Rhizobia form a nutrient exchange mutualism with leguminous plants.

Project description

Can our bacteria persist within the rhizosphere and into the nodule when faced with native microbial competitors? The Medicago-Sinorhizobium mutualism is used as a model system. Survival of strains post competition is difficult to measure due to identical 16S rRNA sequences and similar marker genes. Plasmid-ID barcoding sequences may provide a way of testing strains against a community.

Soil Microbiome Effect On Seedling Diseases And High Throughput Isolate Identification, Annalaysia Smith, Mara Krone, Santiago Mideros

MICRO-CCS student projects

The soil microbiome has a substantial effect on disease suppression. Organic farming is a practice used to create a more diverse microbiome which can create disease suppression. Different agricultural practices such as fertilization and rotation system are known to affect the microbiome (Schlatter et al 2017). Therefore, it is important to study the effects of different microbiomes on disease suppression. Furthermore, methods are necessary for quantifying the effect of disease suppression on specific pathogen populations. To do this, it is important to properly identify the pathogens.

Objectives:

1. Test if different nitrogen treatments cause variations in organic soils disease suppression. …

Effects Of Varying Ph Levels On Prophenoloxidase Activity And Hemocyte Recruitment In Daphnia Dentifera, Yusra Siddiqi, Carla Caceres

MICRO-CCS student projects

Anthropogenic activities and natural processes are responsible for changing the pH of freshwaters which can impact aquatic life. D. dentifera is a keystone species found in midwestern lakes that is susceptible to a parasitic yeast called Australozyma monospora. A. monospora spores puncture the Daphnia gut into the hemolymph and begin using the host’s resources. Invertebrate immunity is under researched.

Research objective: How will different pH impact prophenoloxidase (proPO) activity and hemocyte recruitment in Daphnia dentifera?

Screening A Random Hile Mutant Library To Characterize Hild Regulation In Salmonella, Danista Kannah, Gracious Donkor, James M. Slauch

MICRO-CCS student projects

Salmonella is a leading cause of gastrointestinal diseases globally. It utilizes a type three secretion system (T3SS) encoded on Salmonella Pathogenicity Island 1 (SPI1) to invade host intestinal epithelial cells during infection.

HilE directly regulates HilD via a direct protein-protein interaction. The Slauch lab identified two HilD variants, A141V and A141T, which are blind to HilE. These variants activate hilA expression even in the presence of HilE, indicating that the binding of HilE to these mutants is compromised.

Hypothesis: Performing random mutagenesis of hilE could lead to changes at the protein level which could compensate for the loss of interaction …

Fungal Diversity In Lespedeza Cuneata Roots, Gwendolyn Happ, Sophia Mcdade, Anthony Yannarell

MICRO-CCS student projects

Background

• Plants form a relationship with fungi that can be beneficial or harmful
• Fungi may grow on the plant surface, within plant tissues, or around roots
• Plants release compounds that invite fungi to their roots and chemicals  that can diminish unwanted fungi

Why Lespedeza cuneata? 

• Invasive that competes with native plants and damages biodiversity within plant and animal communities
• Alters soil microbiome
• Expansive seed bank and deep root system
• Understanding its microbiome can help in understanding invasive species success and ecology

Efficacy And Evolutionary Dynamics Of 222 Nm Far-Uvc Against Vibrio Cholerae, Kadicha Madakimova, Angus Wu, Nkrumah Grant

MICRO-CCS student projects

The Far-UVC light at 222 nm wavelength (UV222) is one of the emerging methods of disinfection of bacteria in public spaces such as hospitals and labs.

• Does not penetrate human skin and does not cause harm to eyes making it a safer alternative to conventional UV systems.

• Primarily targets proteins preventing regrowth and repair pathways.

However, the potential for bacterial adaptation following sublethal UV222 exposure remains largely unknown.

Central question: How does QS transcriptional state influence the efficacy and resistance profile of V. cholerae to UV222 nm irradiation?

Hypothesis: Qurom-sensing transcriptional state determines V. cholerae susceptibility to UV222 nm UV …

A Tale Of Two Symbionts: Comparative Cellular Localization Of Two Manipulative Rickettsiella Strains In A Spider, Kaitlyn Costanza, Thea Bliss, Matthew Doremus

MICRO-CCS student projects

Many arthropods harbor maternally transmitted endosymbionts that manipulate host reproduction to increase their transmission. One such manipulation is cytoplasmic incompatibility (CI), in which embryos produced by infected males and uninfected females fail to develop.

Rickettsiella has recently been discovered to cause CI in Mermessus fradeorum. Two geographically distinct Rickettsiella strains infect this spider: R1 causes strong CI, whereas R2 causes weaker CI.

Hypothesis: H1: Both Rickettsiella strains will localize within or near developing sperm cells in the testes. H2: The stronger R1 strain will exhibit more consistent localization within infected adult reproductive tissues than the weaker R2 strain.

The Deep Darkness Microbiome: Functional And Taxonomic Diversity In An Oligotrophic Temperate Cave, Philips O. Akinwole, Nina G. Shaffer '25, Emma E.C. Jacobs '25, Zoe Green '27, Thi Doan '27, Crimson Hickman '28, Kaija M. Carr '25, Kenneth L. Brown

Biology Faculty publications

Many subterranean ecosystems are chronically energy limited, yet the mechanisms governing microbial community assembly and metabolic function under extreme carbon scarcity remain poorly resolved. We combined 16S rRNA gene sequencing with community-level physiological profiling (CLPP) to examine how physicochemical gradients regulate microbial diversity and carbon-use strategies across a surface-subsurface transition into the Deep Darkness zone of Indiana Caverns. Progressive isolation from surface inputs resulted in sharp declines in microbial biomass, total organic carbon, and nitrogen, accompanied by increased water content and C:N ratios, indicating strong attenuation of allochthonous organic matter and intensified resource filtering. Microbial communities spanned 42 bacterial phyla, …

Mobile Genetic Elements Shape Microbial Diversity And Functions In Thawing Permafrost Soils, Jiarong Guo, Samuel T. N. Aroney, Guillermo Domínguez-Huerta, Derek Smith, Dean Vik, Carlos Owusu-Ansah, Akbar Adjie Pratama, Sergei Solonenko, Funing Tian, Cristina Howard-Varona, Zhi-Ping Zhong, Aminata Fofana, Garrett J. Smith, Suzanne B. Hodgkins, Dylan Cronin, Isogenie Field Teams 2010–2017, 2019, Emerge Coordinators, Ben J. Woodcroft, Gene W. Tyson, Virginia I. Rich, Matthew B. Sullivan, Simon Roux, Sarah C. Bagby, Michael Ibba

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Ecosystems are shaped by communities of microorganisms whose niches and impacts depend on functional profiles influenced by gene gains and losses. Culture-based experiments demonstrate that mobile genetic elements (MGEs) can mediate gene flux, but quantitative understanding of these dynamics in natural systems remains limited. Here we develop and apply a systematic, meta-omic framework to investigate MGEs in a complex natural system using an 8-year soil time series collected at Stordalen Mire, in Sweden’s thawing permafrost margin. In this climate-critical peatland, we identify ~2.1 million MGE recombinases across 89 microbial phyla and assess ecological distributions, affected functions, past mobility and current …

Biofilm Formation By Histophilus Somni On 3d Bovine Respiratory Tissue Cultures, Bindu Subhadra, Neeti Gandhi, Dianjun Cao, Yue-Jia Lee, Padmavathy Rajagopalan, Thomas J. Inzana

Faculty Publications - Biological Sciences

The development of bacterial biofilms following infection involves bacterial attachment to tissues and maturation of the biofilm matrix (BM). However, studying the development of bacterial biofilms in vivo is difficult to follow. An established example of biofilm formation during host infection is bovine respiratory disease (BRD) due to Histophilus somni. To more accurately investigate in vivo biofilm development, three-dimensional bovine airway organoids comprised of bovine turbinate epithelial (BE) and pulmonary artery endothelial (BAE) cells embedded in type I collagen were developed. Cell-free concentrated culture supernatant (CCS) from biofilm-grown H. somni suppressed proliferation of epithelial cells more than endothelial cells, affected …

Promiscuous Mutualisms: John Dupré And The Centrality Of Symbiosis In The Living World, Thomas Pradeu

Presidential Fellows Articles and Research

My goal in this chapter is to show that symbiosis, especially symbiotic interactions between ‘macrobes’ (non-microbial organisms) and microbes, is a central topic in John Dupré’s work. This topic is undeniably one of the topics by which Dupré has gained wide recognition among practicing scientists, in addition to his recognition within the philosophical community. I spell out why symbiosis has turned out to be a sort of ideal trope for Dupré, as it encapsulates many of his main personal themes—from promiscuous realism to process ontology. I then show that immunology offers one interesting and complementary entry point into microbial symbiosis …

Hiv-1 Tat-Driven Glutamate Dysregulation: Implications For Cognitive Impairment In Hand, Brenna Duffy, Michael Nonnemacher, Sandhya Kortagere

Kimmel Cancer Center Faculty Papers

PURPOSE OF REVIEW: HIV-1-associated neurocognitive disorders (HAND) manifest in 15% to 50% of people with HIV, impairing learning and memory and executive function. The chronic generation of HIV-1 Transactivator of transcription (Tat) likely contributes to HAND via direct neuronal toxicity and glial-mediated toxicity. This review summarizes our current understanding of how chronic Tat generation from microglia and astrocytes promote glutamate excitotoxicity.

RECENT FINDINGS: In recent years, the indirect effects of Tat through the activation of glial cells have gained significant interest. This review highlights microglia and astrocytes as HIV-1 reservoirs that release Tat protein in the central nervous system. Specific …

Mapping The Tar Vrna Interaction With Hiv-1 Integrase, Jian Sun, Rahul Yadav, Tolga Catmakas, Luke Fisher, Nicholas C. Fitzkee, Jacques J. Kessl

Faculty Publications

A series of critical interactions within the viral core between viral RNA (vRNA) and HIV-1 Integrase (IN) has previously been reported. In these studies, contact points between vRNA and IN were identified using RNA-seq and MS-based protein foot-printing approaches. Several IN amino acids located in its C-terminal domain (CTD) were found to be essential for vRNA binding, and their alanine substitution severely impacted the correct morphogenesis of the mature viral core. Here, we have used the TAR element to extend these studies by performing a comprehensive mapping of the interaction by deploying RNA crosslinking and NMR methodologies. Together, these approaches …

Synergies Of Microecology And Macroecology Elicit Insights For Creation Care, David Warners, John T. Wertz

University Faculty Publications and Creative Works

This article reports on a collection of microecological and macroecological insights that emerged from extended conversations between Calvin University professors David Warners, a botanist concentrating in restoration ecology, and John Wertz, a micro-biologist specializing in microbial ecology. Classic descriptions of ecology identify biological interactions as happening at the ecosystem, community, or population level. And yet, ecological interactions are very much occurring at the microbial level as well. Furthermore, microbial-level interactions frequently influence the quality and character of macroecological relationships and vice versa. Allowing ecological principles to be applied across all life forms broadens our comprehension of interactions that are happening …

Urobiome Composition Varies With Vesicoureteral Reflux Grade And History Of Febrile Urinary Tract Infection, Theresa C. Barrett, Kelvin Li, Adam Fitch, Asha Patel, Bhanu Sharma, Jacqueline Holden, Kermina Abdelsayed, Nina N. Kowalewski, Sridhar Narla, Daniel Bushnell, Barbara A. Methé, Catherine S. Forster

Department of Medicine Faculty Papers

Vesicoureteral reflux (VUR) is a risk factor for urinary tract infection (UTI). However, data describing the urobiome in children with VUR are limited. Here, we characterized the urobiome in children with and without VUR to better understand its potential role in VUR pathogenesis. Catheterized urine samples were obtained from children who presented for voiding cystourethrogram (VCUG) between 12/1/2020 and 10/3/2023. Children were excluded if they had any surgical procedures of the genitourinary tract in the 30 days prior to VCUG or were on treatment antibiotics at the time of VCUG. 16S rRNA gene sequencing and taxonomical classification identified the urobiome. …

Acquisition Behavior Response: Investigating The Ability Of Leptoglossus Phyllopus To Choose Between A “Good” Versus “Sub-Optimal” Strain Of Caballeronia, Irene L. Pham

2026 Spring Honors Capstones Projects

Leptoglossus phyllopus is a polyphagous pest that primarily feasts on citrus, tomatoes, and legumes. They must form a mutualistic symbiosis with Caballeronia every generation by ingesting the bacterium from their environment. However, not all species of Caballeronia confer the same fitness on the L. phyllopus host, raising the question of whether L. phyllopus possesses a partner choice mechanism that would allow it to distinguish between a good (GAOX1) and a suboptimal (Lep1P3) symbiosis partner. This was examined by conducting a choice experiment in which the two symbionts (GAOX1 vs. Lep1P3) were placed in a plastic box, and the bug’s roaming …

Understanding Tlr3 Immune Response Activation In Cryptosporidium Parvum And Cryspovirus Pathogenesis, Surayya Hudayberganova

Biochemistry and Molecular Biology

Cryptosporidiosis is a serious gastrointestinal illness caused by a protozoan parasite known as Cryptosporidium. It is particularly dangerous for malnourished children and those with compromised immune systems. My research focuses on the Toll-like receptor 3 (TLR3) response to the Cryptosporidium parvum infection, specifically in the presence of the parasite’s viral mutualist named cryspovirus (CSpV1). Results using research methodologies like ELISA and RT-qPCR have indicated that TLR3 expression increases during the infection process of Cryptosporidium parvum (C. parvum). Furthermore, one intriguing observation concluded during this study suggests that administering the TLR3-specific inhibitor CU-CPT4a to infected cells drastically decreased this activation. This …

Validation Of A Real-Time Pcr Laboratory Developed Test For The Detection Of Pneumocystis Jirovecii In Bronchoalveolar Lavage Samples, Caitlyn Moon

Honors Scholar Theses

Pneumocystis jirovecii is an opportunistic, host-obligate fungal pathogen responsible for causing P. jirovecii pneumonia (PJP) in humans. PJP is treated with trimethoprim-sulfamethoxazole (TMP-SMX), a strong broad-spectrum antibiotic that carries risk of renal and hepatotoxicity. Therefore, rapid diagnosis or exclusion of PJP is crucial to ensure PJP-positive patients receive effective treatment and PJP-negative patients avoid unnecessary antibiotics. Unlike most microorganisms, P. jirovecii cannot be cultured, presenting complications for the typical methods of diagnosing fungal pathogens. Historically, the gold standard for diagnosis was direct microscopic examination with stains such as methenamine silver and calcofluor white, but this diagnostic technique requires highly skilled …