Environmental Microbiology and Microbial Ecology Commons^™

Gene And Protein Sequence Optimization For High-Level Production Of Fully Active And Aglycosylated Lysostaphin In Pichia Pastoris, Hongliang Zhao, Kristina Blazanovic, Yoonjoo Choi, Chris Bailey-Kellogg, Karl E. Griswold

Dartmouth Scholarship

Lysostaphin represents a promising therapeutic agent for the treatment of staphylococcal infections, in particular those of methicillin-resistant Staphylococcus aureus (MRSA). However, conventional expression systems for the enzyme suffer from various limitations, and there remains a need for an efficient and cost-effective production process to facilitate clinical translation and the development of nonmedical applications. While Pichia pastoris is widely used for high-level production of recombinant proteins, there are two major barriers to the production of lysostaphin in this industrially relevant host: lack of expression from the wild-type lysostaphin gene and aberrant glycosylation of the wild-type protein sequence. The first barrier can …

Abundance And Distribution Of Major And Understudied Archaeal Lineages At Globally Distributed Deep-Sea Hydrothermal Vents, Alexander Fenner Rutherford

Dissertations and Theses

Deep-sea hydrothermal vents are some of the most biologically productive ecosystems on Earth, yet receive little to no input of photosynthetically derived organic matter. The trophic system at hydrothermal vents is based primarily on the reduction-oxidation (redox) of inorganic chemicals by Bacteria and Archaea. However, the distributional patterns of the microorganisms that colonize deep-sea hydrothermal vent deposits and their link to the geologic setting are still not deeply understood.

The goal of the studies presented in this thesis was to quantify the abundance, and distribution of major and understudied vent colonizing archaeal groups from globally distributed and geochemically distinct hydrothermal …

Phages Of Non-Diary Lactococci: Isolation And Characterization Of Phi L47, A Phage Infecting The Grass Isolate Lactococcus Lactis Ssp Cremoris Dpc6860, Daniel Cavanagh, Caitríona M. Guinane, Horst Neve, Aidan Coffey, R. Paul Ross, Gerald F. Fitzgerald, Olivia Mcauliffe

Department of Biological Sciences Publications

Lactococci isolated from non-dairy sources have been found to possess enhanced metabolic activity when compared to dairy strains. These capabilities may be harnessed through the use of these strains as starter or adjunct cultures to produce more diverse flavor profiles in cheese and other dairy products. To understand the interactions between these organisms and the phages that infect them, a number of phages were isolated against lactococcal strains of non-dairy origin. One such phage, ΦL47, was isolated from a sewage sample using the grass isolate L. lactis ssp. cremoris DPC6860 as a host. Visualization of phage virions by transmission electron …

A Glutathione-Independent Glyoxalase Of The Dj-1 Superfamily Plays An Important Role In Managing Metabolically Generated Methylglyoxal In Candida Albicans, Sahar Hasim, Nur Ahmad Hussin, Fadhel Alomar, Keshore R. Bidasee, Kenneth W. Nickerson, Mark A. Wilson

Kenneth Nickerson Papers

Methylglyoxal is a cytotoxic reactive carbonyl compound produced by central metabolism. Dedicated glyoxalases convert methylglyoxal to D-lactate using multiple catalytic strategies. In this study, the DJ-1 superfamily member ORF 19.251/GLX3 from Candida albicans is shown to possess glyoxalase activity, making this the first demonstrated glutathione-independent glyoxalase in fungi. The crystal structure of Glx3p indicates that the protein is a monomer containing the catalytic triad Cys¹³⁶- His¹³⁷-Glu¹⁶⁸. Purified Glx3p has an in vitro methylglyoxalase activity (K_m = 5.5 mM andk_cat = 7.8 s^-1) that is significantly greater than that of …

How Clean Is The Clean Room?, Erik J. Mcfarland

STAR Program Research Presentations

In order to limit contamination and protect the integrity of search-for-life missions, spacecraft assembly clean rooms should be free of microorganisms to the extent that planetary protection guidelines require of the mission. This study monitored the bioburden of the clean room in which the Mars-destined InSight lander was being assembled. Samples of ten locations inside the clean room and one location outside were taken at monthly intervals, starting in March 2014 and scheduled to end in September 2014.

Bioburden was determined using qPCR and ATP analyses with qPCR targating 16S rRNA gene copy number and ATP targating metabolic activity of …

Nitrogen Fertilization Has A Stronger Effect On Soil Nitrogen-Fixing Bacterial Communities Than Elevated Atmospheric Co2, Sean T. Berthrong, Chris M. Yeager, Laverne Gallegos-Graves, Blaire Steven, Stephanie A. Eichorst, Robert B. Jackson, Cheryl R. Kuske

Scholarship and Professional Work - LAS

Biological nitrogen fixation is the primary supply of N to most ecosystems, yet there is considerable uncertainty about how N-fixing bacteria will respond to global change factors such as increasing atmospheric CO₂ and N deposition. Using the nifH gene as a molecular marker, we studied how the community structure of N-fixing soil bacteria from temperate pine, aspen, and sweet gum stands and a brackish tidal marsh responded to multiyear elevated CO₂ conditions. We also examined how N availability, specifically, N fertilization, interacted with elevated CO₂ to affect these communities in the temperate pine forest. Based on data …

Investigating The Interactions Between Cyanobacteria And Vibrio Parahaemolyticus, Caroline E. Ward

Honors Theses and Capstones

One well-known pathogen that has been the topic of many recent studies is Vibrio parahaemolyticus, which causes thousands of foodborne illnesses a year, mostly from the ingestion of raw or undercooked oysters. It has been shown cyanobacteria can act as a long-term reservoir of Vibrio cholerae, another pathogenic Vibrio, by encasing the cells within mucilaginous sheaths during which Vibrios enter a viable but non-culturable state. In this study we investigated the interaction of V. parahaemolyticus with cyanobacteria to determine whether cyanobacteria aid in the longevity and survival of V. parahaemolyticus. We found that non-pathogenic V. parahaemolyticus …

The Distribution Of Fitness Effects Of Spontaneous Mutations In Vibrio Fischeri, Chelsea K. Jones

Honors Theses and Capstones

Mutations are the ultimate source of the biological diversity on which natural selection acts, but the vast majority of these mutations are harmful. As such, mutations lead to disease states like cancer, extinction of small populations, and can drive pathogen evolution. Unfortunately, because mutations are rare and past studies have been subject to detection biases, very little is known about the distribution of fitness effects from naturally occurring mutations. In this study, we used mutation accumulation and full genome sequencing to capture naturally occurring mutations before they were exposed to the sieve of natural selection in Vibrio fischeri. We …

Dielectric Barrier Discharge Atmospheric Cold Plasma For Inactivation Of Pseudomonas Aeruginosa Biofilms, Dana Ziuzina, Sonal Patil, Patrick Cullen, Daniela Boehm, Paula Bourke

Articles

In recent years, atmospheric cold plasma (ACP) has been widely investigated for potential application as an alternative decontamination technology in biomedical and healthcare sectors. In this study, the antimicrobial efficacy of ACP against Pseudomonas aeruginosa biofilms was investigated. The 48-h biofilms were treated inside sealed polypropylene containers with a high-voltage dielectric barrier discharge (DBD) ACP (80 kVRMS) and subsequently stored for 24 h at room temperature. Treatment for 60 s by either the direct or indirect mode of ACP exposure (inside or outside plasma discharge, respectively) reduced bacterial populations by an average of 5.4 log cycles from an initial 6.6 …

Identification And Ss-Lactam Resistance In Aquatic Isolates Of Enterobacter Cloacae And Their Status In Microbiota Of Domica Cave In Slovak Karst (Slovakia), Barbora Gaalova, Alena Donauerova, Milan Seman, Helena Bujdakova

International Journal of Speleology

Domica Cave is located in the Slovak Karst National Park in southern Slovakia. Heterotrophic cultivable psychrophilic and mesophilic microbiota were confirmed to be prevalent in this cave. Escherichia coli was the most abundant bacterium, followed by Enterobacter species and Serrratia species. Enterobacter cloacae isolates belong to the group of faecal contaminants (coliforms) of concern in water. Their status in cave ecosystem is questionable, but we observed E. cloacae in water samples all years round suggesting an autochthonous origin. We were concerned with possible contamination of the cave water with resistant E. cloacae from animal farms located 2-3 km from the …

The Unseen World: Environmental Microbial Sequencing And Identification Methods For Ecologists, Naupaka B. Zimmerman, J. Izard, C. Klatt, J. Zhou, E. Aronson

Biology Faculty Publications

Microorganisms inhabit almost every environment, comprise the majority of diversity on Earth, are important in biogeochemical cycling, and may be vital to ecosystem responses to large-scale climatic change. In recent years, ecologists have begun to use rapidly advancing molecular techniques to address questions about microbial diversity, biogeography, and responses to environmental change. Studies of microbes in the environment generally focus on three broad objectives: determining which organisms are present, what their functional capabilities are, and which are active at any given time. However, comprehending the range of methodologies currently in use can be daunting. To provide an overview of environmental …

Investigating The Persistence And "Naturalization" Potential Of Salmonella In Non-Host Environments Using Culture-Based And Molecular-Based Fingerprinting Techniques, Robyn L. Morrison

Theses and Dissertations (Comprehensive)

Recent studies suggest the potential exists for enteric microorganisms to become “naturalized” to the complex natural environment through maintenance of their populations via replication and adaptation to the stress imposed by their external surroundings. The aim of this research was to better understand the potential for antimicrobial resistant (AMR) Salmonella to become “naturalized” and the physiological adaptations that allow for non-host survivability as reflected by AMR and community profiling. To better elucidate the prominent factors leading to a “naturalized” condition, two approaches were taken: 1) survey-based field research to gain perspective on the prevalence of AMR Salmonella isolated from variously …

Impacts Of Freshwater Flushing On Anammox Community Structure And Activities In The New River Estuary, Usa, Ja Lisa, B Song, Cr Tobias, Ka Duernberger

VIMS Articles

Anaerobic ammonium oxidation (anammox) and denitrification are 2 microbial nitrogen removal processes that may play an important role in controlling the intensity and duration of estuarine and coastal eutrophication. Sediment communities in the New River Estuary, North Carolina were investigated to determine the dynamics of anammox activity and community structure in conjunction with environmental conditions. N-15 tracer incubation experiments with sediment slurries were used to measure anammox and denitrification rates and estimate anammox contribution to total N-2 production. Molecular analyses targeting the hydrazine oxidoreductase (hzo) gene were conducted to examine the structure of anammox communities and quantify the abundance of …

Effects Of Elevated Salinity And Oxidative Stress On The Physiology Of The Toxigenic Cyanobacterium Microcystis Aeruginosa, Billy Christopher Warhurst

UNF Graduate Theses and Dissertations

Harmful algal blooms (HABs) are found worldwide, particularly in places where warm, well-lit, and stagnant waters are common. HABs can have negative effects on aquatic plants and wildlife due to the reduction in light availability associated with turbidity, decrease in O₂ availability, and the production of secondary metabolites that can harm or even prove lethal. Aquatic ecosystems are regularly being affected by elevated salinity because of recent water management strategies, episodes of drought, and salt water intrusion. This research focused on how salinity levels ranging from 0-10ppt affected physiological attributes such as cellular growth and abundance, cell mortality, toxin …

The Pleiotropic Effects Of Beneficial Mutations Of Adapted Escherichia Coli Populations, Brian Scott Van Dam

Honors Theses and Capstones

Mutations that improve fitness in one environment can often be beneficial, deleterious, or neutral in alternative environments. When a single mutation effects fitness in multiple environments, it is said to be a pleiotropic, which can have important consequences for niche specialization, niche expansion, speciation, and even extinction in the face of environmental change. While previous studies have revealed that pleiotropy is nearly universal, the role of adaptive history in the spectrum of pleiotropic effects has yet to undergo detailed experimental observation. Using experimental evolution we gathered beneficial mutations in a previously adapted strain of Escherichia coli growing in the same …

Ether Bridge Formation And Chemical Diversification In Loline Alkaloid Biosynthesis, Juan Pan

Theses and Dissertations--Plant Pathology

Loline alkaloids, found in many grass-Epichloë symbiota, are toxic or feeding deterrent to invertebrates. The loline alkaloids all share a saturated pyrrolizidine ring with a 1-amine group and an ether bridge linking C2 and C7. The steps in biosynthesis of loline alkaloids are catalyzed by enzymes encoded by a gene cluster, designated LOL, in the Epichloë genome. This dissertation addresses the enzymatic, genetic and evolutionary basis for diversification of these alkaloids, focusing on ether bridge formation and the subsequent modifications of the 1-amine to form different loline alkaloids.

Through gene complementation of a natural lolO mutant and comparison …

The Role Of Surface Chemistry In The Toxicity Of Manufactured Cerium Dioxide Nanomaterials To Caenorhabditis Elegans, Emily Kay Oostveen

Theses and Dissertations--Plant and Soil Sciences

Manufactured CeO₂ nanomaterials (CeO₂-MNMs) are used for a wide variety of applications including diesel fuel additives and chemical/mechanical planarization media. To test the effects of CeO₂-MNM surface coating charge on to model organism Caenorhabditis elegans, we synthesized 4 nm CeO₂ with cationic (DEAE-), anionic (CM-), and neutral (DEX) coatings. In L3 nematodes exposed for 24 hours, DEAE-CeO₂ induced lethality at lower concentrations than CM- or DEX-CeO₂. Feeding slightly decreased CeO₂ toxicity, regardless of coating. In L2 nematodes exposed for 48 hours with feeding, DEAE-CeO₂ caused lethality at the …

Soil Microbial Community Distributions And Disease Suppressiveness In The Coastal Plain Of Georgia, Michael J. Sabula

Electronic Theses and Dissertations

This study compared the soil microbial communities of three vegetation types in the coastal plain of Georgia: 1. crop land actively in use for agricultural production, 2. transitional grassland in early stages of secondary succession, and 3. pristine unmanaged forest land. Microbial species diversity and quantities of microbial DNA were determined from each of these vegetation types at three separate locations near Statesboro, Georgia. Length heterogeneity PCR(LH-PCR) methods and subsequent analysis of fungal, bacterial, and metazoan communities by analysis of similarity (ANOSIM) revealed high within-group similarity by vegetation type, indicating land management intensity and vegetation cover is a strong determining …

Bacteria In Bioethanol Fermentations, Qing Li

Theses and Dissertations--Plant and Soil Sciences

To gain a better understanding of contaminating bacteria in bioethanol industry, we profiled the bacterial community structure in corn-based bioethanol fermentations and evaluated its correlation to environmental variables. Twenty-three batches of corn-mash sample were collected from six bioethanol facilities. The V4 region of the collective bacterial 16S rRNA genes was analyzed by Illumina Miseq sequencing to investigate the bacterial community structure. Non-metric multidimensional scaling (NMDS) ordination plots were constructed to visualize bacterial community structure groupings among different samples, as well as the effects of multiple environmental variables on community structure variation. Our results suggest that bacterial community structure is facility-specific, …

Purification Of An Inducible Dnase From A Thermophilic Fungus, Kyle Landry, Andrea Vu, Robert Levin

Kyle S Landry