Microbiology Commons^™

Isolation, Enumeration And Antibiotic Profiling Of Vibrio Vulnificus And V. Parahaemolyticus From Coastal Virginia, Stanley E. Peyton Jr.

Masters Theses, 2020-current

Vibrio vulnificus and V. parahaemolyticus are gram-negative, halophilic bacteria that reside in estuarine waters and are associated with infections in humans. These bacteria can cause gastroenteritis through their presence in raw fish and shellfish consumed by humans. V. vulnificus can also produce wound infections leading to severe septicemia, and in some cases, death if not treated promptly. With increasing incidence of infections due to these two organisms, research efforts have focused on potential reservoirs and environmental conditions that can increase human exposure to, and infection, with these species of bacteria. This study was conducted in order to examine the role …

Lmea, A Conserved Cell-Envelope Protein In Mycobacteria, Is Important For Antibiotic Resistance And Cell Envelope Permeability, Sarah Hassan Osman

Masters Theses

The cell envelope of mycobacteria is critical for the survival and virulence of pathogenic species during infection, and its biosynthesis has been a proven drug target. Therefore, finding new targets in the biosynthetic pathway of cell envelope components is of great interest. Mycobacterium smegmatis is a model organism for the study of the devastating pathogen Mycobacterium tuberculosis. Previously, lipomannan elongation factor A (LmeA) has been identified as a cell envelope protein that is critical for the control of mannan chain length of lipomannan (LM) and lipoarabinomannan (LAM), lipoglycan components of the cell envelope. The deletion mutant, ∆lmeA, accumulates abnormal LM/LAM …

Contributions Of Gyra And Parc Mutations And Qnrs2 Acquisition To Ciprofloxacin Resistance In Aeromonas Veronii Hm21, Daniel J. Silverstein

Honors Scholar Theses

In recent years, ciprofloxacin resistant (CpR) Aeromonas veronii and A. hydrophila strains have been isolated from the wounds of patients receiving leech therapy. Genome comparisons of these CpR isolates revealed the presence of chromosomal mutations in gyrA and parC as well as the gain of qnrS2 on either a large, 34 kb, conjugatable, low-copy plasmid, pAv42, or on a small, 6.8 kb, high-copy plasmid, pAh1471. The minimum inhibitory concentration, MIC, for Cp of these clinical isolates ranged from 1 to ≥32 µg/mL and some harbored a qnrS2 containing plasmid. We wanted to assess the contributions of these factors in an …

Whole Genome Sequence Analysis Of A Transmissible Multidrug-Resistance Plasmid Captured Without Cultivation From Poultry Litter, Emma C. Eisemann

Senior Honors Projects, 2020-current

Use of antibiotics in the agricultural industry introduces selective pressure and, consequently, could increase the presence of antibiotic resistant organisms in surrounding environments. One such environment is litter (manure and bedding) produced during large-scale poultry production in the Shenandoah Valley. Litter, with its microorganisms, is commonly applied to fields within the Shenandoah River watershed. Antibiotic resistance (AR) and virulence genes are potentially transmissible between organisms through horizontal gene transfer of genetic mobile elements, for which poultry litter could be a reservoir. The typical, culture-based approach to detecting and analyzing AR plasmids and other mobile genetic elements is limited due to …

The Effects Of Farnesol, A Quorum Sensing Molecule From Candida Albicans, On Alcaligenes Faecalis, Savannah Hutson

Undergraduate Honors Theses

Quorum sensing molecules have become a recent focus of study to learn if and how they can be used, both on their own and in conjecture with current antimicrobial methods, as a means of bacterial control. One such quorum sensing molecule is the sesquiterpene alcohol, Farnesol, which is synthesized and released by the fungus, Candida albicans. In most in-vivo cases, our laboratory has shown that Alcaligenes faecalis overtakes C. albicans, preventing its growth. However, as a way to counteract this inhibitory effect, Farnesol may be one way that Candida has found to fight back. In this study, we …

Characterization Of The Broad-Spectrum Inhibitory Capability Of Alcaligenes Faecalis And A. Viscolactis Against Potential Pathogenic Microorganisms, Andrew Fuqua

Undergraduate Honors Theses

The recent rise of multidrug resistant microorganisms has grown from an isolated concern to a massive public health crisis. It has become imperative that scientists look for new ways to combat this issue. Due to the selective pressures of competition, bacteria and other microbes possess a host of defenses and weapons designed to exploit vulnerabilities in other microorganisms. Consequently, the study of these systems and microbial interactions has much to reveal in the search for novel antimicrobial treatments. Previous research from our laboratory has discovered that both Alcaligenes faecalis and Alcaligenes viscolactis, two rarely studied and generally non-virulent bacteria, …

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 …

Identification Of Uncommon Antibiotic-Producing Illinois Soil Isolates, Lesly Muniz, Dr. Lori Scott

Identifying and Characterizing Novel Antibiotic Producing Microbes From the Soil

This project is a collaboration with the Tiny Earth Project Initiative (TEPI), which is a global network of educators and students focused on student sourcing antibiotic discovery from the soil. We researched tester strains B. subtilis and E. coli from the soil isolates obtained. We further verified if the isolates were common antibiotic bacteria. Unfortunately, this project heavily relied on biochemical tests, colony morphology, and Gram stains to reject or fail to reject our hypothesis. Our goal was to discover new antibiotic-producing bacteria that could be beneficial in combating ESKAPE strains. A proper PCR and DNA extraction would be required …

Isolated Antibiotic Producing Bacteria In Local Soil Samples Determined To Be Bacillus, Cassidy Potter, Dr. Lori Scott

Identifying and Characterizing Novel Antibiotic Producing Microbes From the Soil

Nosocomial pathogens are multi-drug resistant to antibiotics that fight bacterial infections posing danger to the public health, the most dangerous of them being the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.). This project is a collabortaion with the TIny Earth Project Initiative (TEPI), which is a global network of educators and students focused on studentsourcing antibiotic discovery from the soil. TEPI allows student-led research on local soil samples from Bettendorf, IA to discover potential novel antibiotic producing bacteria that could potentially treat ESKAPE pathogens and reduce public health risk. Two soil isolates …

Pseudomonas And Bacillus Soil Isolates Produce Antibiotics, Chelsea Brandt, Dr. Lori Scott

Identifying and Characterizing Novel Antibiotic Producing Microbes From the Soil

The recent emergence of antibiotic resistance bacterial strains presents a significant challenge and threat to human healthcare. While new methods of treatment such as bacteriophage therapy and combinations of existing antibiotics are being researched, the human population is in dire need of new antibiotics to replace those that are ineffective. This research addresses this need by identifying antibiotic producing bacteria in a soil sample from Davenport, IA. This project is a collaboration with the Tiny Earth Project Initiative (TEPI), which is a global network of educators and students focused on studentsourcing antibiotic discovery from soil. Microbiology lab techniques and 16S …

Identification Of Antibiotic Producing Soil Bacteria Against Bacillus Subtilis, Morgan Brockhouse, Dr. Lori Scott

Identifying and Characterizing Novel Antibiotic Producing Microbes From the Soil

This project is a collaboration with the Tiny Earth Project Initiative (TEPI), which is a global network of educators and students focused on student-sourcing antibiotic discovery from soil. Individual strains of soil bacteria were isolated and produced antibiotic against Bacillus subtilis. Two of these samples were sequenced using the 16S rRNA gene to reveal they are very closely related to the genus Pseudomonas.

Group B Streptococcus: Molecular Epidemiology, Pathogenic Profiling And Control Strategies, Katherine Mary Hayes

Theses

Group B Streptococcus (GBS) are frequent colonisers of the gastrointestinal tract of healthy adults with worldwide rates of 18% colonisation; however, they are also opportunistic pathogens capable of causing invasive disease, particularly in neonates and adults with underlying conditions. Indeed, GBS is the leading cause of invasive neonatal disease worldwide with a global incidence of 0.49 per 1000 live births and GBS disease in adults is increasing globally, with estimated rates of 10.9 cases per 100,000 people. While GBS remain largely susceptible to penicillin, there have been reports of reduced beta-lactam susceptibility in certain countries and resistance to other antibiotic …

Development Of Small Molecule Antibiotics Against A Conserved Rna Gene Regulatory Element In Gram-Positive Bacteria, Ville Yrjö Petteri Väre

Legacy Theses & Dissertations (2009 - 2024)

Bacterial infections and the rise of antibiotic resistance, especially multidrug resistant strains, have generated a clear need for discovery of novel therapeutics. Most antibiotics in use today are derivatives of previous antibiotics to which resistance mechanisms already exist, and traditionally they have a single target: either a protein or rRNA. Gram-positive bacteria regulate the expression of several essential genes or operons using a mechanism called the T-box. The T-box is a structurally conserved riboswitch-like gene regulator in the 5’-untranslated region (UTR) of numerous essential genes of Gram-positive bacteria. T-boxes are stabilized by cognate, unacylated tRNA ligands, allowing the formation of …

Screening The Cultivable Cave Microbial Mats For The Production Of Antimicrobial Compounds And Antibiotic Resistance, Jerneja Ambrožič Avguštin, Patricia Petrič, Lejla Pašić

International Journal of Speleology

The current work extends the phenotypic characterization of a bacterial culture collection obtained from white, yellow, grey or pink microbial cave wall colonies that are common in the caves of Slovenian Karst. We have determined antibiotic resistance to 22 natural and synthetic antibiotics in 69 isolates from the microbial mats. Thirty-eight isolates (52%) were resistant to 1-5 antibiotics; another 27 isolates (37%) were resistant to 6-10 antibiotics; and 7 isolates (0.1%) were resistant to 11-17 antibiotics. We screened for production of antimicrobial compounds by growing cave isolates on five different media and overlaying individual cultures with ten Gram-positive and Gram …

Multi-Omic Understanding Of The Evolution Of Xenobiotic Tolerance In Bacterial Isolates And Communities, Tayte Paul Campbell

Arts & Sciences Electronic Theses and Dissertations

Xenobiotic compounds are any chemicals that are released into an environment by human action and that occur at concentrations higher than found naturally. Xenobiotics, including aromatic compounds and antibiotics, are recalcitrant to degradation because they are often toxic or mutagenic. Despite this toxicity, bacteria account for a large portion of xenobiotic degradation in the environment. Bacteria are able to adapt to these foreign chemicals, gaining increased levels of tolerance and increased rates of xenobiotic degradation. On the strain level, increased tolerance can be caused by mutations in individual cells or through the acquisition of genes from other cells. At the …

Assessing The Prevalence Of Multidrug Resistant Salmonella Enterica From Stream Sediment And Poultry Litter In The Shenandoah Valley Of Virginia, Noah Greenman

Masters Theses, 2010-2019

Multidrug resistant Salmonella enterica present in stream sediment and poultry litter represent a critical health concern. A small number of S. enterica serotypes are responsible for most lab-confirmed infections in the US each year. To assess the prevalence of these significant strains, we isolated 88 S. enterica from stream sediment and poultry litter. Sequence data for all isolates were generated using an Illumina® sequencing platform, with long-reads for some isolates from the Oxford Nanopore MinION™ used in a hybrid genome assembly approach. Isolates were typed according to their serotype and multi-locus sequence type using SeqSero/SISTR and Enterobase respectively. Antibiotic resistance …

Biofilm Producing Clinical Staphylococcus Aureus Isolates Augmented Prevalence Of Antibiotic Resistant Cases In Tertiary Care Hospitals Of Nepal, Sarita Manandhar, Anjana Singh, Ajit Varma, Shanti Pandey, Neeraj Shrivastava

Student Publications

Staphylococcus aureus, a notorious human pathogen, is a major cause of the community as well as healthcare associated infections. It can cause a diversity of recalcitrant infections mainly due to the acquisition of resistance to multiple drugs, its diverse range of virulence factors, and the ability to produce biofilm in indwelling medical devices. Such biofilm associated chronic infections often lead to increase in morbidity and mortality posing a high socio-economic burden, especially in developing countries. Since biofilm formation and antibiotic resistance function dependent on each other, detection of biofilm expression in clinical isolates would be advantageous in treatment decision. …

The Ability Of Novel Phage To Infect Virulent Bacillus Anthracis Isolates, Hyrum Smith Shumway

Theses and Dissertations

Bacillus anthracis is a soil dwelling microbe with pronounced pathogenic potential. Historically, anthrax has infected livestock and man. In the modern-age, anthrax is a bioterrorism concern with major incidents every decade. While the threat of large scale attacks is currently viewed as unlikely, the threat is consistent and constant. Current methods to defend against such an attack focus on antibiotics and containment of public panic. Antibiotic resistance, while not currently an issue for anthrax, could easily become so with genetically engineered weaponized strains created by rogue states or independent actors. This project evolved from collaborations between the Grose lab and …

Antibacterial Properties Of Novel Amphiphiles: Exploring Structure-Activity Relationships, Reafa Hossain

Senior Honors Projects, 2010-2019

The increased cases of antibiotic resistance have large implication in hospital settings where infections by antibiotic resistant bacteria are harder to treat resulting in longer stays at the hospital, which drastically increases the costs to patients and hospitals. To address this matter, many research groups are searching for an alternative to antibiotics. One option is the development of amphiphiles, some of which have antibacterial properties. Amphiphiles contain a hydrophilic, polar head group, and a hydrophobic, nonpolar tail, which may intercalate into the cell membrane, resulting in cell lysis. Understanding the impact of amphiphile geometry on antibacterial activity allows for the …

Comparison Of Beta-Lactamase Genes In Environmental Rahnella Isolates, Katherine D'Angelo, Ray A. Larsen

Honors Projects

Antibiotic resistance is a growing problem in the field of healthcare. Antibiotics are becoming less effective as species of bacteria adapt and share resistance mechanisms. If transmission of mechanisms can be better understood at the molecular level, inhibitors could be developed to lessen the likelihood of antibiotic resistance. In this study, Rahnella spp were isolated from environmental sources on MacConkey plates containing 100 µg ml^-1 ampicillin, and confirmed by 16S rRNA gene sequencing. Whole genomic DNA was extracted from isolates and initial amplifications were performed by polymerase chain reaction (PCR) using primers specific for 16S amplification. New primers were …

Emergence Of The L Phenotype In Group B Streptococci In The South Of Ireland, Katherine Hayes, Lesley Cotter, L. Barry, Fiona O'Halloran

Department of Biological Sciences Publications

Group B Streptococcal isolates (n = 235) from the South of Ireland were characterised by serotyping, antimicrobial susceptibility and determination of the phenotypic and genotypic mechanisms of resistance. Resistance to erythromycin and clindamycin was observed in 21·3% and 20·4% of the total population, respectively. The c-MLSB phenotype was the most common phenotype detected (62%), with ermB being the predominant genetic determinant, present in 84% of resistant isolates. The rare L phenotype was observed in 2·9% (n = 7) of isolates, four of which harboured the lsaC gene responsible for clindamycin resistance. Serotypes Ia, III and II were the most common …

The Making Of A Pathogen: Implications Of Phage Domestication In Acinetobacter Baumannii, Allison Welp

Undergraduate Theses

Mutations and horizontal gene transfer have allowed for rapid evolution of many species of bacteria, allowing them to become more virulent and resistant to antibiotics. As a result of these changes, Acinetobacter baumannii has become one of the most prominent drug-resistant bacteria in hospitals. This nosocomial pathogen is capable of causing a range of infections—from pneumonia to sepsis—and is extremely difficult to eradicate from hospital settings. Despite its current status, this species has not always been apparent in healthcare. The emergence of this organism has been extremely rapid; once an innocuous environmental organism, A. baumannii is now resistant to all …

Genomic Analysis Of Factors Associated With Low Prevalence Of Antibiotic Resistance In Extraintestinal Pathogenic Escherichia Coli Sequence Type 95 Strains, Craig M. Stephens, Sheila Adams-Sapper, Manraj Sekhon, James R. Johnson, Lee W. Riley

Biology

Extraintestinal pathogenic Escherichia coli (ExPEC) strains belonging to multilocus sequence type 95 (ST95) are globally distributed and a common cause of infections in humans and domestic fowl. ST95 isolates generally show a lower prevalence of acquired antimicrobial resistance than other pandemic ExPEC lineages. We took a genomic approach to identify factors that may underlie reduced resistance. We fully assembled genomes for four ST95 isolates representing the four major fimH-based lineages within ST95 and also analyzed draft-level genomes from another 82 ST95 isolates, largely from the western United States. The fully assembled genomes of antibiotic-resistant isolates carried resistance genes exclusively on …

A Survey Of Β-Lactam Antibiotic Resistance Genes And Culturable Ampicillin Resistant Bacteria In Minnesota Soils, Emily M. Wollmuth

Departmental Honors Projects

Since the discovery of penicillin, antibiotics have been an essential tool in the treatment of bacterial infections and diseases. It is estimated that antibiotics and antibiotic resistance genes have existed for billions of years. With the increasing appearance of resistant pathogenic bacteria, there has been growing concern. β-lactam antibiotics make up the largest portion of the global market, so resistance to these antibiotics is especially alarming. It has been theorized that frequency and type of antibiotic resistance genes vary by area. Previous studies suggest that these differences may be related to antibiotic use in agricultural and urban areas. To survey …

Occurrence, Antibiotic Resistance, And Survival Of Fecal Enterococci In Turkey Litter, Steven Glynn Mcbride Ii

Masters Theses, 2010-2019

The United States Environmental Protection Agency’s National Water Quality Inventory and the Commonwealth of Virginia’s 305(b)/303(d) Water Quality Assessment Integrated Report show fecal bacteria to be the most common cause of impairment for both streams and estuaries. Human and animal sources have both been identified as significant contributors of pathogenic bacteria to surface waters. In this study, turkey litter from a farm in Shenandoah County, VA was surveyed for total culturable bacteria and total culturable enterococci before and after a transition to organic rearing practices. The enterococci were identified to species phenotypically using the Biolog Microbial Identification System and resistance …

Whole Genome Sequencing As A Tool For Identifying Phenotypic Properties And Underlying Genetic Mechanisms In Staphylococcus Pseudintermedius, Matthew C. Riley

Doctoral Dissertations

Staphylococcus pseudintermedius is a Gram-positive bacterial opportunistic pathogen commonly associated with dermal infections in canines, but capable of causing serious disease in other species. Reports of human infections caused by S. pseudintermedius along with an increase in resistance to multiple antibiotics highlights the importance of this organism. Whole genome sequencing can allow large scale investigation of genetic mechanisms underlying phenotypic properties that contribute to the expansion of successful S. pseudintermedius clonal lineages.

The increase in multidrug and methicillin-resistant S. pseudintermedius (MRSP) may result from horizontal transfer of genetic material between bacterial isolates, yet is thought to be rare in Staphylococci …

Next-Generation Sequencing Of A Multi-Drug Resistance Plasmid Captured From Stream Sediment, Kevin G. Libuit

Masters Theses, 2010-2019

Plasmids in agriculturally-impacted bodies of water may play a significant role in the dissemination of antibiotic resistance. Previously, Erika Gehr, as part of her M.S. thesis work in our laboratory, captured environmental plasmids without cultivation of host bacteria from stream sediment into Escherichia coli. Individual plasmids were capable of conferring resistance to a surprising array of antibiotics including aminoglycosides and extended-spectrum β-lactams. In this study, we developed a method to sequence multi-drug resistance plasmids using both Oxford Nanopore MinION and Ion Torrent Personal Genome Machine sequencers. Plasmid pEG1-1 was sequenced on both platforms and a hybrid assembly utilizing data …

The Efficacy Of Zinc And Manganese In Controlling Methicillin-Resistant Staphylococcus Aureus Wound Infections In Vitro, Patrick B. Lau

Honors Scholar Theses

The emergence of Methicillin-Resistant Stapylococcus aureus has triggered an increased interest in finding alternative natural antimicrobials to control the pathogen and combat growing antibiotic resistance. This study investigated the antimicrobial effect of two naturally occurring essential minerals, zinc (Zn) and manganese (Mn) on MRSA for potential application in wound infections. The sub inhibitory concentration (SIC) and minimum inhibitory concentration (MIC) of Zn and Mn against MRSA were determined. The effect of MIC and 2x MIC of Zn and Mn in increasing MRSA susceptibility to oxacillin, and the effect of SIC and MIC of these minerals on MRSA cell adherence and …

Isolation And Characterization Of Microbial Communities From Hydraulic Fracturing Fluids, Sheridan S. Brewer 6284984

Chancellor’s Honors Program Projects

No abstract provided.

Isolation And Epidemiological Analysis Of Verotoxigenic E. Coli From Patients With Acute Gastroenteritis: A Prospective Study Aiming To Reduce The Burden Of Infection In The Cork Region., Shirley Ross

Masters

Verotoxigenic Escherichia coli (VTEC) is a serious gastrointestinal pathogen associated with morbidity and mortality in the first world, as accentuated by the O104 VTEC outbreak in Germany in 2011. Molecular detection of Verotoxin (VT) has increased the detection of VTEC across Europe however, Ireland still has the highest incidence and isolation is still a key step in the confirmation of infection and the control and prevention of outbreaks. Currently no standard protocol exists for the detection, isolation and characterisation of non-O157 VTEC and it is feared that due to this that the prevalence of VTEC is under-reported. This body of …