Microbiology Commons^™

Elucidating The Role Of The Msaabcr Operon In Persister Cells Formation And Stress Response In Staphylococcus Aureus, Shanti Pandey

Dissertations

Staphylococcus aureus is a major human pathogen that causes lethal systemic conditions such as infective endocarditis, osteomyelitis, sepsis and pneumonia. Chronic in nature, these diseases are often refractory to the antibiotic treatment. Such recalcitrance is mainly caused due to the presence of persister cells, which are a bacterial subpopulation that exhibits extreme, yet transient antibiotic tolerance accompanied by a transient halt in the growth. However, upon cessation of antibiotic treatment, resumption in growth of persister cells causes recurrence of infections and treatment failure, displaying tremendous clinical significance. In this study, we show the involvement of the msaABCR operon in persister …

Elevated Incidences Of Antimicrobial Resistance And Multidrug Resistance In The Maumee River (Ohio, Usa), A Major Tributary Of Lake Erie, Maitreyee Mukherjee, Leah Marie, Cheyenne Liles, Nadia Mustafa, George Bullerjahn, Terry J. Gentry, John P. Brooks

Faculty Publications

Maumee River, the major tributary in the western basin of Lake Erie, serves as one of major sources of freshwater in the area, supplying potable, recreational, and industrial water. In this study we collected water samples from four sites in the Maumee River Bay between 2016–2017 and E. coli was isolated, enumerated, and analyzed for antimicrobial resistance (AMR) and multidrug resistance (MDR). Strikingly, 95% of the total isolates were found to be resistant to at least one antibiotic. A very high resistance to the drugs cephalothin (95.3%), ampicillin (38.3%), tetracycline (8.8%), gentamicin (8.2%), ciprofloxacin (4.2%), cefoperazone (4%), and sulfamethoxazole (1.5%) …

How Historic Shipwrecks Influence Dispersal Of Deep-Sea Microbiomes, Rachel Moseley

Master's Theses

This thesis investigates how historic shipwrecks potentially shape dispersal of deep-sea microbiomes. Processes impacting dispersal of microbiomes around artificial structures in the ocean and how they connect to other deep-sea habitats is unknown. Dispersal processes are explained for some macroorganisms by theories in ecology, specifically the Theory of Island Biogeography and the Wooden Stepping Stone hypothesis. These have not been investigated for microorganisms, and thus this work will probe if they are applicable to microbial biogeography in this habitat. Experiments were conducted to establish new “island-like” features in near proximity (25-125 m) to wooden-hulled historic shipwrecks in the northern Gulf …

The Impact Of Oxygen Availability And Oxidative Stress On Regulation Of Cyclic-Dimeric-Gmp In Listeria Monocytogenes Strain F2365, Anna G. Welch

Honors Theses

Listeria monocytogenes is a gram-positive foodborne pathogen that results in the infectious disease listeriosis. The second messenger molecule cyclic-dimeric-GMP has been found to be responsible for the regulation of expression of many of its virulence factors with diguanylate cyclases and phosphodiesterases regulating the intracellular concentration of cyclic-di-GMP through synthesis and degradation, respectively. This study investigated the possibility that the availability of oxygen is the environmental signal crucial to the regulation of these enzymes. Prior research in our laboratory has demonstrated that the intracellular concentration of cyclic-di-GMP is elevated when exposed to anaerobic conditions. Due to this, it was hypothesized that …

Study Of The Role Of The Msaabcr Operon In Tricarboxylic Acid Cycle Activity And Persister Cell Formation In Staphylococcus Aureus, Karsen Motter

Honors Theses

Staphylococcus aureus is an important human pathogen that causes wide arrays of infections ranging from minor skin infections to lethal systemic conditions such as infective endocarditis, osteomyelitis, sepsis and pneumonia. These systemic diseases are often difficult to treat due to the presence of persister cells. Persister cells are a phenotypic variant of the bacterial population that exhibit extreme and transient antibiotic tolerance accompanied by a transient halt in growth. Upon cessation of antibiotic treatment, however, persisters resume growth which results in recurrence of infections. This characteristic of persister cells therefore displays high clinical significance. In this study, we show the …

Interregulation Between Msaabcr Operon And Ccpe To Determine Staphylococcal Metabolism And Virulence, Erin R. Cox

Honors Theses

Staphylococcus aureus is a complex human pathogen that causes problems in both healthcare and community settings. Staphylococcus aureus is a gram-positive, sphere shaped bacterium that usually colonizes in the nasal cavity of healthy individuals. Staphylococcus aureus infections are a growing health concern due to its ability to produce virulence factors, such as pigmentation, protease production, and capsule formation. Two regulators in S. aureus virulence factors are msaABCR, which is a newly characterized operon, and ccpE. In order to determine the interaction between these two regulators in regulating virulence and metabolism in S. aureus, ccpE and ccpE/ …

Phosphodiesterase D Is Involved In Bile Resistance In Listeria Monocytogenes, Sophia A. Ali

Honors Theses

Listeria monocytogenes is a deadly foodborne bacterium that is responsible for almost 20% of food-related deaths in the United States. Listeria monocytogenes contaminates ready-to-eat products such as cheese, deli meat, and ice cream. Once ingested, it invades the intestinal lining and can enter the bloodstream, causing listeriosis. There is a gap in the knowledge of the pathogenesis of L. monocytogenes in how it is able to survive in the gastrointestinal tract in the presence of bile, which has bactericidal properties. Previous studies have suggested that the second messenger cyclic-dimeric-GMP may be involved in the regulation of virulence factors of Listeria. …

Phenotypic Characterizations Of Msaabcr Operon Deletion In Staphylococcus Epidermidis Rp62a, Raelyn Williams

Honors Theses

Staphylococcus epidermidis is a human pathogen that is increasingly known for its role in hospital infections associated with implantable medical devices. Antibiotic resistance has become a concerning issue for these infections as this bacteria have various virulence traits that help to evade immune response and antibiotic treatment. Currently, the most effective way to treat S. epidermidis infection is removal of the implant and long-term antibiotic treatment. S. epidermidis causes infection by expressing several protein factors that induce biofilm formation, the bacteria’s primary virulence mechanism. The purpose of this study was to perform the phenotypic characterizations of the msaABCR operon in …

The Role Of Gac/Rsm Signal Transduction Pathway In The Biology Of Antagonistic Marine Bacterium Gynuella Sunshinyii 449, Anhthu Truong

Honors Theses

Gynuella sunshinyii is a newly characterized bacterium with a remarkable ability to produce multiple secondary metabolites that act as potential antibiotics for medical applications. Currently, little is known about gene regulation in this unusual microorganism. In 2018, studies conducted of rhizosphere microbiomes of tidal marsh grasses on Deer Island, MS, resulted in multiple isolates of halophilic, aerobic, Gram-negative bacteria with antagonistic properties. One of the most active antagonists was identified as Gynuella sunshinyii 449. The genome of strain 449 was sequenced with Illumina MiSeq, assembled with Unicycler and SPAdes, and annotated with RASTtk. The annotation revealed a 6.09 Mb genome …

Root Exudates Alter The Expression Of Diverse Metabolic, Transport, Regulatory, And Stress Response Genes In Rhizosphere Pseudomonas, Olga V. Mavrodi, Janiece R. Mcwilliams, Jacob O. Peter, Anna Berim, Karl A. Hassan, Liam D.H. Elbourne, Melissa K. Letourneau, David R. Gang, Ian T. Paulsen, David M. Weller, Linda S. Thomashow, Alex S. Flynt, Dmitri V. Mavrodi

Faculty Publications

Plants live in association with microorganisms that positively influence plant development, vigor, and fitness in response to pathogens and abiotic stressors. The bulk of the plant microbiome is concentrated belowground at the plant root-soil interface. Plant roots secrete carbon-rich rhizodeposits containing primary and secondary low molecular weight metabolites, lysates, and mucilages. These exudates provide nutrients for soil microorganisms and modulate their affinity to host plants, but molecular details of this process are largely unresolved. We addressed this gap by focusing on the molecular dialog between eight well-characterized beneficial strains of the Pseudomonas fluorescens group and Brachypodium distachyon, a model …

The Msaabcr Operon Regulates Persister Formation By Modulating Energy Metabolism In Staphylococcus Aureus, Shanti Pandey, Gyan S. Sahukhal, Mohamed O. Elasri

Faculty Publications

Staphylococcus aureus is a major human pathogen that causes chronic, systemic infections, and the recalcitrance of these infections is mainly due to the presence of persister cells, which are a bacterial subpopulation that exhibits extreme, yet transient, antibiotic tolerance accompanied by a transient halt in growth. However, upon cessation of antibiotic treatment, a resumption in growth of persister cells causes recurrence of infections and treatment failure. Previously, we reported the involvement of msaABCR in several important staphylococcal phenotypes, including the formation of persister cells. Additionally, observations of the regulation of several metabolic genes by the msaABCR operon in transcriptomics and …

Microbial Functional Responses In Marine Biofilms Exposed To Deepwater Horizon Spill Contaminants, Rachel L. Mugge, Jennifer L. Salerno, Leila J. Hamdan

Faculty Publications

Marine biofilms are essential biological components that transform built structures into artificial reefs. Anthropogenic contaminants released into the marine environment, such as crude oil and chemical dispersant from an oil spill, may disrupt the diversity and function of these foundational biofilms. To investigate the response of marine biofilm microbiomes from distinct environments to contaminants and to address microbial functional response, biofilm metagenomes were analyzed from two short-term microcosms, one using surface seawater (SSW) and the other using deep seawater (DSW). Following exposure to crude oil, chemical dispersant, and dispersed oil, taxonomically distinct communities were observed between microcosms from different source …

Review: Formation And Metabolic Function Of Coral Rubble Biofilms In The Reef Ecosystem, Andres Sanchez-Quinto, Luisa I. Falcon

Gulf and Caribbean Research

When coral dies, their calcareous skeletons constitute coral rubble in conjunction with the cementing activity of coralline algae and bacteria, creating a secondary reef structure which takes from years to decades to form. Healthy coral reefs differ from coral—rubble dominated reefs in microbial taxonomic composition and metabolic functional roles. The metabolisms of healthy reefs are dominated by autotrophic pathways, where carbon and nitrogen fixation dominate, while the metabolism of rubble—dominated reefs predominate in degradation of organic matter. Nitrogen fixation is 3 orders of magnitude lower in rubble—dominated reefs than in healthy reefs. Coral—rubble harbors a vast diversity of microbes that …

The Role Of The Msaabcr Operon In Implant-Associated Chronic Osteomyelitis In Staphylococcus Aureus Usa300 Lac, Gyan S. Sahukhal, Michelle Tucci, Hamed Benghuzzi, Gerri Wilson, Mohamed O. Elasri

Faculty Publications

The msaABCR operon regulates several staphylococcal phenotypes such as biofilm formation, capsule production, protease production, pigmentation, antibiotic resistance, and persister cells formation. The msaABCR operon is required for maintaining the cell wall integrity via affecting peptidoglycan cross-linking. The msaABCR operon also plays a role in oxidative stress defense mechanism, which is required to facilitate persistent and recurrent staphylococcal infections. Staphylococcus aureus is the most frequent cause of chronic implant-associated osteomyelitis (OM). The CA-MRSA USA300 strains are predominant in the United States and cause severe infections, including bone and joint infections.

Two Hundred And Fifty-Four Metagenome-Assembled Bacterial Genomes From The Bank Vole Gut Microbiota, Anton Lavrinienko, Eugene Tukalenko, Timothy A. Mousseau, Luke R. Thompson, Rob Knight, Tapio Mappes, Phillip C. Watts

Faculty Publications

Vertebrate gut microbiota provide many essential services to their host. To better understand the diversity of such services provided by gut microbiota in wild rodents, we assembled metagenome shotgun sequence data from a small mammal, the bank vole Myodes glareolus (Rodentia, Cricetidae). We were able to identify 254 metagenome assembled genomes (MAGs) that were at least 50% (n = 133 MAGs), 80% (n = 77 MAGs) or 95% (n = 44 MAGs) complete. As typical for a rodent gut microbiota, these MAGs are dominated by taxa assigned to the phyla Bacteroidetes (n = 132 MAGs) and …

Microplastic Abundance, Distribution And Impacts On Sargassum-Associated Juvenile Fishes In The Gulf Of Mexico, Olivia Lestrade

Master's Theses

Microplastics are a concern in marine environments because they are highly durable, ubiquitous, and can be mistaken for food and ingested by small organisms. Pelagic Sargassum, an important habitat for larval and juvenile stages of many fish species, is found in large surface aggregations, and may provide complex structure in which microplastics become trapped. This could lead to greater risk of microplastic ingestion by fish early life stages associated with Sargassum habitats. To better understand the impacts of microplastics within Sargassum communities, this study examined 1) microplastic concentrations and ingestion by juvenile fishes associated with Sargassum; 2) the microbial …

Significance Of Viral Activity For Regulating Heterotrophic Prokaryote Community Dynamics Along A Meridional Gradient Of Stratification In The Northeast Atlantic Ocean, Kristina D.A. Mojica, Corina P.D. Brussaard

Faculty Publications

How microbial populations interact influences the availability and flux of organic carbon in the ocean. Understanding how these interactions vary over broad spatial scales is therefore a fundamental aim of microbial oceanography. In this study, we assessed variations in the abundances, production, virus and grazing induced mortality of heterotrophic prokaryotes during summer along a meridional gradient in stratification in the North Atlantic Ocean. Heterotrophic prokaryote abundance and activity varied with phytoplankton biomass, while the relative distribution of prokaryotic subpopulations (ratio of high nucleic acid fluorescent (HNA) and low nucleic acid fluorescent (LNA) cells) was significantly correlated to phytoplankton mortality mode …

Tick-Borne Pathogens Shape The Native Microbiome Within Tick Vectors, Abdulsalam Adegoke, Deepak Kumar, Cailyn Bobo, Muhammad Imran Rashid, Aneela Zameer Durrani, Muhammad Sohail Sajid, Shahid Karim

Faculty Publications

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Ticks are blood-feeding arthropods and transmit a variety of medically important viral, bacterial, protozoan pathogens to animals and humans. Ticks also harbor a diverse community of microbes linked to their biological processes, such as hematophagy, and hence affect vector competence. The interactions between bacterial and/or protozoan pathogens and the tick microbiome is a black-box, and therefore we tested the hypothesis that the presence of a protozoan or bacterial pathogen will alter the microbial composition within a tick. Hence, this study was designed to define the microbial composition of two tick species, …

Draft Genome Sequences Of Six Strains Isolated From The Rhizosphere Of Wheat Grown In Cadmium-Contaminated Soil, Vira Hovorukha, Ankita Bhattacharyya, Olga Iungin, Hanna Tashyreva, Victoria Romanovska, Olesia Havyrliuk, Olena Bielikova, Claire Blackwell, Brian Burks, Cara Cothern, Jakia Elliott, Jonathan Hoover, Alexis Jones, Christian Leise, Linda Lowmiller, Ahmed Mohamed, Tiffany Mullen, Ethan Nettleton, Karshanda Polk, Benny Tran, Teresa Tran, Manuel Vega, Landon Ware, Emily Welch, Leandra Williams, Madison Woodward, Kaylin Young, Olga Mavrodi, Oleksandr Tashyrev, Dmitri Mavrodi

Faculty Publications

This study presents high-quality draft genome assemblies of six bacterial strains isolated from the roots of wheat grown in soil contaminated with cadmium. The results of this study will help to elucidate at the molecular level how heavy metals affect interactions between beneficial rhizobacteria and crop plants.

Development Of An Attenuated Zika Virus By Editing The 5’ Untranslated Region, E. Ashley Thompson

Master's Theses

Zika Virus (ZIKV) is a mosquito-transmitted flavivirus that usually causes no symptoms to mild febrile in humans, and it has been regarded as an insignificant pathogen to public health. However, recent outbreaks of ZIKV infection have revealed that ZIKV can cause severe neurological effects in adults, such as Guillain-Barre Syndrome (GBS), and in infants whose mothers acquired the virus during pregnancy, causing Congenital Zika Syndrome (CZS). Currently, no approved vaccine is available and there is a critical need to develop an effective and safe vaccine. While most vaccine developmental strategies target the viral prM-E protein of ZIKV, we aimed to …

Elucidating The Molecular Pathways Involved In The Fitness And Water Stress Response Of Pseudomonas Synxantha 2-79 In The Plant Rhizosphere, Clint Henry D. Pablo

Master's Theses

Pseudomonas synxantha 2-79 is a biocontrol agent that represents beneficial indigenous rhizobacteria that are broadly distributed in the Pacific Northwest, USA and flourish in the rhizosphere of commercially grown wheat under surprisingly arid conditions. The molecular adaptation of 2-79-like bacteria to plants growing in dry soils is poorly understood. We hypothesized that the ability of 2-79 to colonize and persist in the rhizosphere of water-stressed plants is underpinned by the formation of hydrating biofilms and the utilization of root exudates that contain plant-derived osmoprotectants called quaternary ammonium compounds (QACs). We tested this hypothesis by identifying waters stress response pathways in …

Regulation Of Weak Acid-Dependent Cell Death By The Msaabcr Operon During Biofilm Formation Of Staphylococcus Aureus, Sarah-Elizabeth M. Polley

Honors Theses

Staphylococcus aureus is a dangerous human pathogen and a leading cause of both nosocomial and community-associated infections. Infection by S. aureus is a growing health concern due to the emergence of antibiotic resistance and the capability to form robust biofilms. Major constituents of the biofilm extracellular matrix are demonstrated to be released via controlled autolysis of a subpopulation of the biofilm. As indicated in past studies, the programmed cell death (PCD) of S. aureus grown in a biofilm microenvironment is potentiated by the buildup of acetate, a byproduct of glucose metabolism, which causes cytoplasmic acidification. Furthermore, it has been shown …

The Effects Of Light On The Bacterial Biomass Of Decomposing Leaf Litter In Freshwater, Savannah L. Underwood

Honors Theses

Recent evidence has suggested that the presence of light (and in conjuction, periphytic algae) stimulates the growth and production of bacteria on decaying plant litter. However, the current method of measuring bacterial biomass, flow cytometry, drastically underestimates the bacterial biomass levels associated with litter samples. To determine and correct for this underestimation, the present study determined conversion factors using direct counting methods via epifluorescence microscopy. These conversion factors were then applied to flow cytometry counts for seven different studies. The studies analyzed the effects of light and other covarying factors on the bacterial biomass associated with various samples of leaf …

Antibiotic Resistant Bacteria, Antibiotic Resistance Genes And Potential Drivers In The Aquatic Environments, Shuo Shen

Dissertations

As antibiotic resistance genes in aquatic environment have been increasing across the world, affecting water quality and public health, many studies documented concentrations of antibiotic resistance genes and some studies discussed their potential drivers. However, systematic and quantitative reviews that link antibiotic resistance genes (ARGs) to anthropogenic and environmental factors are limited. Nevertheless, this information will be important for developing regulation policy on controlling antibiotic use and therefore reducing potential risks to antibiotic resistance. I conducted meta-analysis of ARGs concentration at a global scale using Bayesian inference to explore climatic and socio-economic factors as drivers. I found local-scale climatic variables …

Listeria Monocytogenes Response To Anaerobic Environments, Brandy N. Roberts, Damayanti Chakravarty, J.C. Gardner Iii, Steven C. Ricke, Janet R. Donaldson

Faculty Publications

Listeria monocytogenes is a Gram-positive facultative anaerobic bacterium that is responsible for the disease, listeriosis. It is particularly lethal in pregnant women, the fetus, the elderly and the immunocompromised. The pathogen survives and replicates over a wide range of temperatures (4 to 42 °C), pH, salt and oxygen concentrations. Because it can withstand various environments, L. monocytogenes is a major concern in food processing industries, especially in dairy products and ready-to-eat fruits, vegetables and deli meats. The environment in which the pathogen is exposed can influence the expression of virulence genes. For instance, studies have shown that variations in oxygen …

The Antimicrobial Activity And Cellular Pathways Targeted By P-Anisaldehyde And Epigallocatechin Gallate In The Opportunistic Human Pathogen Pseudomonas Aeruginosa, Yetunde Adewumni, Sanchirmaa Namjilsuren, William D. Walker, Dahlia N. Amato, Douglas V. Amato, Olga V. Mavrodi, Derek L. Patton, Dmitri V. Mavrodi

Faculty Publications

Plant-derived aldehydes are constituents of essential oils that possess broad-spectrum antimicrobial activity and kill microorganisms without promoting resistance. In our previous study, we incorporated p-anisaldehyde from star anise into a polymer network called PANDAs (Pro-Antimicrobial Networks via Degradable Acetals) and used it as a novel drug delivery platform. PANDAs released p-anisaldehyde upon a change in pH and humidity, and controlled growth of the multi-drug resistant pathogen Pseudomonas aeruginosa PAO1. In this study, we identified cellular pathways targeted by p-anisaldehyde, by generating 10,000 transposon mutants of PAO1 and screened them for hypersensitivity to p-anisaldehyde. To improve the …

The Histoplasma Capsulatum Ddr48 Gene Is Required For Survival Within Macrophages And Resistance To Oxidative Stress And Antifungal Drugs, Logan Blancett

Dissertations

Histoplasma capsulatum(Hc)is a systemic, dimorphic fungal pathogen that affects upwards of 500,000 individuals in the United States annually. Hc grows as a multicellular mold at environmental temperatures; whereas, upon inhalation into a human or other mammalian host, it transforms into a unicellular, pathogenic yeast. The research presented in this dissertation is focused on characterizing the DNA damage-responsive gene HcDDR48. HcDDR48was originally isolated via a subtractive DNA library enriched for transcripts enriched in the mold-phase of Hcgrowth. Upon further analysis we found that HcDDR48is not just expressed in the mold morphotype, but both growth programs …

The Antimicrobial Activity And Cellular Targets Of Plant Derived Aldehydes And Degradable Pro-Antimicrobial Networks In Pseudomonas Aeruginosa, Yetunde Adewunmi

Dissertations

Essential oils (EOs) are plant-derived products that have been long exploited for their antimicrobial activities in medicine, agriculture, and food preservation. EOs represent a promising alternative to conventional antibiotics due to the broad-range antimicrobial activity, low toxicity to human commensal bacteria, and the capacity to kill microorganisms without promoting resistance. Despite the progress in the understanding of the biological activity of EOs, many aspects of their mode of action remain inconclusive. The overarching aim of this work was to address these gaps by studying molecular interactions between antimicrobial plant aldehydes and the opportunistic human pathogen Pseudomonas aeruginosa. We initiated …

The Effect Of Historic Shipwrecks On Sediment Microbiomes In The Northern Gulf Of Mexico, Melissa Brock

Master's Theses

Microorganisms are ubiquitously distributed across all aquatic habitats, but it is the environmental conditions of a habitat that determine which microorganisms can thrive in terms of abundance or metabolic activity. Habitats that experience consistent physical and chemical environmental conditions repeatedly favor specific taxonomic groups which may result in a microbial assemblage that is commonly associated with that habitat (i.e., a core microbiome). Core microbiomes have been identified for a variety of natural marine habitats including methane seeps, wood falls, octocoral, and deep-sea sediments. However, it was unknown if the presence of man-made structures on the seafloor, including historic shipwrecks, also …

The Thiol Specific Antioxidant (Tsa1) Gene Is Required For Survival In Macrophages And Oxidative Stress Resistance In Histoplasma Capsulatum, Lauren Kennedy

Master's Theses

Histoplasma capsulatum(Hc) is a pathogenic fungus that causes one of the most common invasive fungal respiratory diseases, Histoplasmosis. Histoplasmaundergoes a dimorphic shift from mold to yeast which is crucial to pathogenesis of the organism.

The thiol specific antioxidant gene, Tsa1,is strongly upregulated in the yeast (pathogenic) morphotype. This data led to the hypothesis that this gene plays a role in protecting Hcfrom host mediated oxidative attack. To characterize Tsa1 function, a knockdown strain (tsa1-RNAi) was created by RNAi gene silencing. Expression of Tsa1in the tsa1-RNAistrain was reduced to 10% that …