Life Sciences Commons^™

The Response Of Microcystis And The Microbiome To Exogenous Hydrogen Peroxide, Bryan A. Puma

Masters Theses, 2020-current

Microcystis spp. produce prolific annual blooms in freshwater systems worldwide. The success of these blooms depends heavily on the Microcystis spp. overcoming environmental factors such as oxidative stress caused by reactive oxygen species (ROS) like hydrogen peroxide. Most Microcystis genomes do not contain many genes for responding to oxidative stress, including those that encode the enzyme catalase. Other cyanobacteria such as Prochlorococcus that also lack hydrogen peroxide degrading enzymes have been shown to benefit from partnerships with heterotrophic bacteria. Microcystis spp. may also receive similar benefits from heterotrophic bacteria in the phycosphere. To test this hypothesis, we examined the …

The Impact Of Iaa And Tryptophan On Microcystis Bloom Community Dynamics, Hunter R. Baylous

Masters Theses, 2020-current

Microcystis aeruginosa is a freshwater cyanobacterial species that degrades freshwater and brackish ecosystems due to its capacity to form toxic cyanobacterial harmful algal blooms (cyanoHABs). There both abiotic and the biotic factors that exacerbate the presence of cyanoHABs. Current research is looking at the microbial interactions that occur between microbes and phytoplankton. Microbes interact with cyanobacteria in the phycosphere, where nutrients are exchanged between the two. Understanding the chemical currencies exchanged can help to show interactions that are beneficial for cyanoHAB formation A key player in the growth promotion of cyanoHABs is hypothesized to be auxins, which are synthesized by …

A Stable-Isotope Probing Approach To Modeling Bidirectional Nutrient Exchange In The Microcystis Phycosphere, Malia Gardner

Masters Theses, 2020-current

Cyanobacterial harmful algal blooms (cHABs) plague freshwater systems worldwide and are projected to increase in intensity in the coming decades. cHABs damage aquatic ecosystems by blocking light penetration into the water column, creating hypoxic conditions, and releasing toxins. One of the most prolific cHAB formers is the cosmopolitan genus of cyanobacteria Microcystis. Global climate change and anthropogenic loading of nutrients such as nitrogen (N) and phosphorus (P) fuel Microcystis bloom formation. Increasing global temperatures favor Microcystis because of its high optimal growth temperature. N input is of particular importance for Microcystis because it is unable to fix atmospheric N, …

Ecophysiology Of Toxic Bloom-Forming Cyanobacteria And Their Symbionts, Dominique J. Lockwood

Masters Theses, 2020-current

Currently, there is a pernicious microbe in the genus Microcystis that is putting the world’s freshwater resources at risk. A member of the phylum Cyanobacteria, otherwise referred to as blue-green algae, Microcystis forms massive algal blooms in lakes and has the potential to produce a toxin, which does not only harm humans, but also pets, livestock, and aquatic life. As recently as 2016, in the city of Toledo, Ohio, nearly 1 million residents went without clean drinking water for an entire weekend due to one of these Microcystis blooms. However, Microcystis does not only bloom in Ohio. In fact, …

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 …

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 …

Exploring The Microbiome: Diversity Of The Microbial Community Of Three Foam Nesting Frogs, Genus: Polypedates, Across A Developmental Gradient, Sarah Mcgrath

Masters Theses, 2010-2019

Characterization of microbial biodiversity, including that of the amphibian skin-associated microbiome, is a frontier of research recently made accessible through advances in sequencing technology. Microbial interaction with a host has been determined to have profound influences on host health across a wide range of macroscopic organisms. For amphibians, the influence of the skin-associated microbiome has been found to have particular importance, as amphibians are currently one of the fastest disappearing vertebrate groups on the planet, largely in part to skin-associated diseases caused by pathogenic microbes. Therefore, it is important to characterize the amphibian skin-associated microbiome, particularly for species with no …

Isolation, Sequencing, And Characterization Of Four Transmissible Antibiotic Resistance Plasmids Captured From Bacteria In Stream Sediments, Curtis J. Kapsak

Masters Theses, 2010-2019

Self-transmissible plasmids are key vectors in the transfer of resistance, catabolic, and other genes among bacteria native to environments such as streams and wetlands. The evolution of antibiotic resistance in particular is known to be powerfully affected by conjugative plasmid transfer due to the ease in which some plasmids can be horizontally transferred into a broad range of host bacteria and their ability to exchange mobile genetic elements that often contain antibiotic resistance genes.

In this study, we captured tetracycline resistance plasmids from stream sediments impacted by agricultural runoff. We selected for resistance plasmids using tetracycline, an antibiotic commonly used …

A Cure For Salmonella: Engaging Students In Pathogen Microbiology And Bioinformatics, Sophie Jurgensen

Senior Honors Projects, 2010-2019

Advances in Next Generation Sequencing (NGS) technology have generated a vast amount of publicly available genomic data, creating a need for students with training in computational analysis. This laboratory lesson is a course-based undergraduate research experience (CURE) focusing on environmental Salmonella, a common foodborne pathogen that is of great interest to public health laboratories but is relatively less virulent than most other such pathogens. As discovery is a central tenet of CUREs, students isolate novel Salmonella enterica and related strains from stream sediment, poultry litter, or other sources in the first half of the lesson (Module 1). They also …

Survey Of Microbial Urea Degrader Diversity In Two Freshwater Ecosystems: Lake Shenandoah And The Shenandoah River, Naomi E. Gilbert

Senior Honors Projects, 2010-2019

One of the primary drivers of cyanobacterial harmful algal blooms (cHABs) in freshwater systems is nutrient loading, particularly of nitrogen and phosphorus. There has been an increased focus on assessing the role of nitrogen (N) in freshwater lakes and rivers that suffer cHABs. Urea, a widely-used, N-rich fertilizer, is a source of interest due to its abundance in freshwater ecosystems, primarily caused by anthropogenic nutrient loading. While recent work has shown that cHAB population succession may favor the toxic cyanobacterium Microcystis in urea-rich waters, the diversity of the associated bacterial community capable of degrading urea has yet to be determined. …

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 …

Isolation And Enumeration Of Vibrio Vulnificus And Vibrio Parahaemolyticus From Coastal Virginia, Zackary A. Zayakosky

Senior Honors Projects, 2010-2019

Vibrio vulnificus and V. parahaemolyticus are gram-negative, halophilic bacteria that are found throughout estuarial waters during the summer months, and are commonly associated with human infection. Gastroenteritis and other related symptoms can occur following infection from either organism, which most often occurs as a result of consumption of raw oysters or other seafood. V. vulnificus is particularly virulent, and can also produce wound infections that lead to severe septicemia and death. Due to the increasing rates of infection for these two organisms, recent research efforts have focused on potential environmental conditions and reservoirs that would be indicative of increased Vibrio …

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 …

Comparative Analysis Of Anti-Bd Bacteria From Six Malagasy Frog Species Of Ranomafana National Park, Kelsey Savage

Senior Honors Projects, 2010-2019

As Malagasy amphibians are facing an impending extinction crisis from the lethal skin fungus Batrachochytrium dendrobatidis (Bd), it has become imperative to proactively mitigate the threat. Bd sporangia develop in the skin of infected amphibians and cause the skin to thicken, leading to ionic imbalance and eventual heart failure. It has been shown that certain bacterial species are able to inhibit Bd growth on amphibians by producing antifungal metabolites. Community-based probiotics are one approach used to combat chytridomycosis by inoculating an environment with Bd-inhibitory bacteria so that many amphibian species are treated at once. With this method, it is important …

Transduction As The Method Of Horizontal Gene Transfer Of The Staphylococcal Chromosomal Cassette Mec (Sccmec), Amber B. Sauder

Senior Honors Projects, 2010-2019

Methicillin-resistant Staphylococcus aureus (MRSA) gains resistance to β-lactam antibiotics through a mutated penicillin binding protein (PBP2a) encoded on the SCCmec element. In combination with the recombinase encoded by ccr, these two genes are used as markers of the mobile genetic element (SCCmec). Due to recent increases in community acquired MRSA infections, the mechanisms of antibiotic resistance gene transfer have gained attention. Transduction, a method of horizontal gene transfer mediated by bacteriophage, is believed to be responsible for the movement of the SCCmec element. Recent studies have shown the transduction of the SCCmec element in clinical isolates; however, this study is …