Life Sciences Commons^™

Natural And Lignocellulose-Enriched Microbial Communities In Great Boiling Spring, Nv, Jessica Cole

UNLV Theses, Dissertations, Professional Papers, and Capstones

The natural microbial communities present in Great Boiling Spring were investigated and contrasted against those present after in situ enrichment with lignocellulose. High-throughput cultivation-independent DNA sequencing of the V8 region of the small subunit (SSU) rRNA gene generated a total of 274,119 quality-filtered pyrosequencing fragments. Twelve natural spring samples were analyzed, including four high-temperature water samples and eight sediment samples ranging from 87 - 62 °C. Eight lignocellulosic enrichments incubated in the spring sediment and water at two high-temperature sites were analyzed. The natural water communities were found to be extremely uneven but relatively constant throughout time. The natural sediment …

Microbial Effects On The Production And Transformation Of Surfactants Within The Microlayer And Subsurface Waters In Application To Remote Sensing Techniques, Katie E. Vella

HCNSO Student Theses and Dissertations

The sea surface microlayer is a millimeter-scale interfacial layer between the atmosphere and the ocean. A number of studies have suggested that there is a unique ecosystem for marine bacteria in the sea surface microlayer, but little information exists on the microbial community composition of this ecosystem due to sampling complexities. In this work, we present an improved method to sample and compare the bacterial diversity of the sea surface microlayer with that of subsurface water at the same site. Bacterial samples were collected from the sea surface microlayer with a sampling method, which minimized sample contamination from the research …

Soil Nematodes And Their Prokaryotic Prey Along An Elevation Gradient In The Mojave Desert (Death Valley National Park, California, Usa), Amy M. Treonis, Kelsey Sutton, Brendan Kavanaugh, Archana Narla, Timothy Mcllarky, Jasmine Felder, Cecilia O'Leary, Megan Riley, Alyxandr Pikus, Sarah Thomas

Biology Faculty Publications

We characterized soil communities in the Mojave Desert across an elevation gradient. Our goal was to test the hypothesis that as soil quality improved with increasing elevation (due to increased productivity), the diversity of soil prokaryotes and nematodes would also increase. Soil organic matter and soil moisture content increased with elevation as predicted. Soil salinity did not correlate to elevation, but was highest at a mid-gradient, alluvial site. Soil nematode density, community trophic structure, and diversity did not show patterns related to elevation. Similar results were obtained for diversity of bacteria and archaea. Relationships between soil properties, nematode communities, and …

Spatial And Temporal Immune Response In House Flies In Response To Ingestion Of Bacillus Cereus And Eschericha Coli 0157-H7, Adam Fleming

Electronic Theses and Dissertations

House flies (Musca domestica L.) feed and breed on septic substrates, putting them in direct contact with a multitude of disease causing agents and can act as a bridge for those agents to humans. The house fly has previously been shown to carry many different species of bacteria that are pathogenic. Escherichia coli O157:H7 is a pathogenic enterohemorrhagic serotype of E. coli that can be vectored by the house fly. Bacillus cereus is a foodborne pathogen that has also been isolated from the house fly in previous studies. To examine vector potential for these pathogens, house flies were fed green …

Characterization Of Xylan Utilization And Discovery Of A New Endoxylanase In Thermoanaerobacterium Saccharolyticum Through Targeted Gene Deletions, Kara K. Podkaminer, Adam M. Guss, Heather L. Trajano, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

The economical production of fuels and commodity chemicals from lignocellulose requires the utilization of both the cellulose and hemicellulose fractions. Xylanase enzymes allow greater utilization of hemicellulose while also increasing cellulose hydrolysis. Recent metabolic engineering efforts have resulted in a strain of Thermoanaerobacterium saccharolyticum that can convert C5 and C6 sugars, as well as insoluble xylan, into ethanol at high yield. To better understand the process of xylan solubilization in this organism, a series of targeted deletions were constructed in the homoethanologenic T. saccharolyticum strain M0355 to characterize xylan hydrolysis and xylose utilization in this organism. While the deletion of …

Macroalgae Decrease Growth And Alter Microbial Community Structure Of The Reef-Building Coral, Porites Astreoides, Rebecca Vega Thurber, Deron E. Burkepile, Adrienne M.S. Correa, Andrew R. Thurber, Andrew A. Schantz, Rory Welsh, Catharin Pritchard

Department of Biological Sciences

With the continued and unprecedented decline of coral reefs worldwide, evaluating the factors that contribute to coral demise is of critical importance. As coral cover declines, macroalgae are becoming more common on tropical reefs. Interactions between these macroalgae and corals may alter the coral microbiome, which is thought to play an important role in colony health and survival. Together, such changes in benthic macroalgae and in the coral microbiome may result in a feedback mechanism that contributes to additional coral cover loss. To determine if macroalgae alter the coral microbiome, we conducted a field-based experiment in which the coral Porites …

The Crispr/Cas Adaptive Immune System Of Pseudomonas Aeruginosa Mediates Resistance To Naturally Occurring And Engineered Phages, Kyle C. Cady, Joe Bondy-Denomy, Gary E. Heussler, Alan R. Davidson, George A. O'Toole

Dartmouth Scholarship

Here we report the isolation of 6 temperate bacteriophages (phages) that are prevented from replicating within the laboratory strain Pseudomonas aeruginosa PA14 by the endogenous CRISPR/Cas system of this microbe. These phages are only the second identified group of naturally occurring phages demonstrated to be blocked for replication by a nonengineered CRISPR/Cas system, and our results provide the first evidence that the P. aeruginosa type I-F CRISPR/Cas system can function in phage resistance. Previous studies have highlighted the importance of the protospacer adjacent motif (PAM) and a proximal 8-nucleotide seed sequence in mediating CRISPR/Cas-based immunity. Through engineering of a protospacer …

Ocean Acidification: Understanding The Coastal Carbon Pump In A High Co2 World, Rachel Cooper

Theses and Dissertations

Since the 1800s, carbon dioxide emissions due to human activities have contributed significantly to the amount of carbon in the atmosphere. Approximately a third of this carbon is absorbed by the ocean, through air-sea fluxes at the ocean surface (Sabine, 2004). Increased CO2 has changed the carbon chemistry of the ocean and hence the pH. pH is expected to drop by 0.4 by the year 2100. It is unclear how this lower pH will affect carbon cycling and sequestration with respect to the biological carbon pump. Most studies have focused on open ocean phytoplankton or bacterial communities in large, stationary …

Microbial Community Structure And Ecosystem Function In A Changing World, Melissa Ann Cregger

Doctoral Dissertations

Understanding the effects climate change will have on the structure and function of global ecosystems is a pressing ecological and social issue. Global change driven changes in atmospheric warming and precipitation régimes have begun to alter the distribution of plants and animals in, as well as the function of, ecosystems. Using two large-scale climate change manipulations, I assessed the effect of changing precipitation and temperature regimes on soil microbial community structure and function. Soil microbial communities regulate decomposition and nutrient cycling rates in ecosystems, thus understanding their response to climatic changes will enable scientists to better predict carbon feedbacks to …

Algorithms For Optimizing Cross-Overs In Dna Shuffling, Lu He, Alan M. Friedman, Chris Bailey-Kellogg

Dartmouth Scholarship

DNA shuffling generates combinatorial libraries of chimeric genes by stochastically recombining parent genes. The resulting libraries are subjected to large-scale genetic selection or screening to identify those chimeras with favorable properties (e.g., enhanced stability or enzymatic activity). While DNA shuffling has been applied quite successfully, it is limited by its homology-dependent, stochastic nature. Consequently, it is used only with parents of sufficient overall sequence identity, and provides no control over the resulting chimeric library.

Results: This paper presents efficient methods to extend the scope of DNA shuffling to handle significantly more diverse parents and to generate more predictable, optimized libraries. …

Genome Sequence Of The Mesophilic Thermotogales Bacterium Mesotoga Prima Mesg1.Ag.4.2 Reveals The Largest Thermotogales Genome To Date, Olga Zhaxybayeva, Kristen S. Swithers, Julia Foght, Anna G. Green, David Bruce, Chris Detter, Shunsheng Han, Hazuki Teshima, James Han, Tanja Woyke, Sam Pitluck, Matt Nolan, Natalia Ivanova, Amrita Pati, Miriam L. Land, Marlena Dlutek, W Ford Doolittle, Kenneth M. Noll, Camilla L. Nesbo

Dartmouth Scholarship

Here we describe the genome of Mesotoga prima MesG1.Ag4.2, the first genome of a mesophilic Thermotogales bacterium. Mesotoga prima was isolated from a polychlorinated biphenyl (PCB)-dechlorinating enrichment culture from Baltimore Harbor sediments. Its 2.97 Mb genome is considerably larger than any previously sequenced Thermotogales genomes, which range between 1.86 and 2.30 Mb. This larger size is due to both higher numbers of protein-coding genes and larger intergenic regions. In particular, the M. prima genome contains more genes for proteins involved in regulatory functions, for instance those involved in regulation of transcription. Together with its closest relative, Kosmotoga olearia, it …

The Proteomics Of Lipid Droplets: Structure, Dynamics, And Functions Of The Organelle Conserved From Bacteria To Humans., Li Yang, Yunfeng Ding, Yong Chen, Shuyan Zhang, Chaoxing Huo, Yang Wang, Jinhai Yu, Peng Zhang, Huimin Na, Huina Zhang, Yanbin Ma, Pingsheng Liu

Faculty Scholarship for the College of Science & Mathematics

Lipid droplets are cellular organelles that consists of a neutral lipid core covered by a monolayer of phospholipids and many proteins. They are thought to function in the storage, transport, and metabolism of lipids, in signaling, and as a specialized microenvironment for metabolism in most types of cells from prokaryotic to eukaryotic organisms. Lipid droplets have received a lot of attention in the last 10 years as they are linked to the progression of many metabolic diseases and hold great potential for the development of neutral lipid-derived products, such as biofuels, food supplements, hormones, and medicines. Proteomic analysis of lipid …

Lapg, Required For Modulating Biofilm Formation By Pseudomonas Fluorescens Pf0-1, Is A Calcium-Dependent Protease, Chelsea D. Boyd, Debashree Chatterjee, Holger Sondermann, George A. O'Toole

Dartmouth Scholarship

Biofilm formation by Pseudomonas fluorescens Pf0-1 requires the cell surface adhesin LapA. We previously reported that LapG, a periplasmic cysteine protease of P. fluorescens, cleaves the N terminus of LapA, thus releasing this adhesin from the cell surface and resulting in loss of the ability to make a biofilm. The activity of LapG is regulated by the inner membrane-localized cyclic-di-GMP receptor LapD via direct protein-protein interactions. Here we present chelation and metal add-back studies demonstrating that calcium availability regulates biofilm formation by P. fluorescens Pf0-1. The determination that LapG is a calcium-dependent protease, based on in vivo and in vitro …

Analyzing Cellular Internalization Of Nanoparticles And Bacteria By Multi-Spectral Imaging Flow Cytometry, Yashdeep Phanse, Amanda E. Ramer-Tait, Sherree L. Friend, Brenda Carrillo-Conde, Paul Lueth, Carrie J. Oster, Gregory J. Phillips, Balaji Narasimhan, Michael J. Wannemuehler, Bryan H. Bellaire

Department of Food Science and Technology: Faculty Publications

Nanoparticulate systems have emerged as valuable tools in vaccine delivery through their ability to efficiently deliver cargo, including proteins, to antigen presenting cells^1-5. Internalization of nanoparticles (NP) by antigen presenting cells is a critical step in generating an effective immune response to the encapsulated antigen. To determine how changes in nanoparticle formulation impact function, we sought to develop a high throughput, quantitative experimental protocol that was compatible with detecting internalized nanoparticles as well as bacteria. To date, two independent techniques, microscopy and flow cytometry, have been the methods used to study the phagocytosis of nanoparticles. The high throughput …

Pyrosequencing Of Bacterial Symbionts Within Axinella Corrugata Sponges: Diversity And Seasonal Variability, James R. White, Jignasa Patel, Andrea Ottesen, Gabriela Arce, Patricia Blackwelder, Jose V. Lopez

Marine & Environmental Sciences Faculty Articles

Background: Marine sponge species are of significant interest to many scientific fields including marine ecology, conservation biology, genetics, host-microbe symbiosis and pharmacology. One of the most intriguing aspects of the sponge ‘‘holobiont’’ system is the unique physiology, interaction with microbes from the marine environment and the development of a complex commensal microbial community. However, intraspecific variability and temporal stability of sponge-associated bacterial symbionts remain relatively unknown.

Methodology/Principal Findings: We have characterized the bacterial symbiont community biodiversity of seven different individuals of the Caribbean reef sponge Axinella corrugata, from two different Florida reef locations during variable seasons using …

Host Pathogen Interactions: Is Arabidopsis Thaliana Remembered By Its Nemesis Pseudomonas Syringae?, Daniel Z. Kreiser

Lawrence University Honors Projects

Plants contain innate immune systems that deter pathogen infection. Pattern recognition receptors bind microbe-associated molecular patterns (MAMPs), triggering immunity. MAMPs are proteins exclusive to pathogens that are typically indispensable for their survival. For this reason, MAMPs cannot be mutated or removed without causing pathogen death. However, this does not necessitate constitutive expression of MAMPs. In this study, the MAMP response of Arabidopsis thaliana was utilized to determine differential detection of MAMPs expressed by Pseudomonas syringe pv. tomato DC3000 when pretreated with A. thaliana. Results demonstrated that more MAMPs are detected when P. syringae had previously encountered A. thaliana, …

Enumeration Of Sandy Sediment Bacteria: Search For Optimal Protocol, Slava Epstein, Jeffrey Rossel

Slava Epstein

We examined and compared a variety of existing protocols for enumeration of bacteria from marine sandy sediments. The focus was on how to dislodge bacteria from sediment particles; a commercial blender, an ultrasonic cleaner, and an ultrasonic cell disrupter were tested. The ultrasonic cell disrupter was found to be the most efficient device for bacterial dislodgment. With a 5 mm microtip vibrating at 109 μm amplitude and 20 kHz, the optimal sonication time of small (≤ 0.5 cm³) samples was 180 s. Having identified the optimal dislodgment treatment, we went through other steps of sediment bacteria enumeration (use of surfactants, …

Enumeration Of Sandy Sediment Bacteria: Are The Counts Quantitative Or Relative?, Slava S. Epstein, D. Alexander, K. Cosman, A. Dompé, S. Gallagher, J. Jarsobski, E. Laning, R. Martinez, G. Panasik, C. Peluso, R. Runde, E. Timmer

Slava Epstein

Several tests were carried out to enable evaluation of the precision with which sandy sediment bacteria could be enumerated. This represented the first attempt to place direct epifluorescence counts of benthic bacteria on a quantitative, rather than relative, ground. The tests combined in situ radioisotope ([³H], [¹⁴C]) labeling of sediment bacteria, bacterial dislodgment by ultrasonic treatment, and bacterial enumeration via fluorescent staining. The results provided direct and indirect evidence that the employed protocol for bacterial enumeration accounted for 88 to 98% of all bacteria present in sediments. The identified approach thus allowed for a rather complete quantification of sediment bacteria. …

Prevalence Of Copper Resistance Among Foliar Bacterial Pathogens Of Tomato In Tennessee, Jonathon Thomas Mixon

Masters Theses

Foliar bacterial diseases are among the most important diseases of tomato. Bacterial spot is caused by four species: Xanthomonas euvesicatoria, X. vesicatoria, X. perforans, and X. gardneri, hereafter referred to as Xanthomonas spp. Bacterial speck is caused by Pseudomonas syringae pv. tomato and bacterial canker is caused by Clavibacter michiganensis subsp. michiganensis. Fixed copper products are relied upon extensively for control, due to a lack of effective and economical alternatives. Copper resistance in bacterial spot and speck pathogens has been reported worldwide. Copper resistance quite likely exists in Tennessee, but the extent has never been …

Pyrosequencing The Bemisia Tabaci Transcriptome Reveals A Highly Diverse Bacterial Community And A Robust System For Insecticide Resistance, Wen Xie, Qing-Shu Meng, Qing-Jun Wu, Shao-Li Wang, Xin Yang, Ni-Na Yang, Ru-Mei Li, Xiao-Guo Jiao, Hui-Peng Pan, Bai-Ming Liu, Qi Su, Bao-Yun Xu, Song-Nian Hu, Xuguo Zhou, You-Jun Zhang

Entomology Faculty Publications

BACKGROUND: Bemisia tabaci (Gennadius) is a phloem-feeding insect poised to become one of the major insect pests in open field and greenhouse production systems throughout the world. The high level of resistance to insecticides is a main factor that hinders continued use of insecticides for suppression of B. tabaci. Despite its prevalence, little is known about B. tabaci at the genome level. To fill this gap, an invasive B. tabaci B biotype was subjected to pyrosequencing-based transcriptome analysis to identify genes and gene networks putatively involved in various physiological and toxicological processes.

METHODOLOGY AND PRINCIPAL FINDINGS: Using Roche 454 pyrosequencing, …

Toxin-Antitoxic Loci Vapbc-1 And Vapxd Contribute To Survival And Virulence In Nontypeable Haemophilus Influenzae, Dabin Ren, Anna N. Walker, Dayle A. Daines

Biological Sciences Faculty Publications

Background: Nontypeable Haemophilus influenzae (NTHi) is a significant human pathogen responsible for respiratory tract infections and the most common cause of recurrent otitis media. Type II toxin-antitoxin (TA) systems are genetic elements that code for a stable protein toxin and a labile antitoxin that are thought to be involved in metabolic regulation of bacteria by enabling a switch to a dormant state under stress conditions. The contribution to infection persistence of the NTHi TA loci vapBC-1 and vapXD was examined in this study.

Results: Deletions in vapBC-1, vapXD and vapBC-1 vapXD significantly decreased the survival of NTHi co-cultured with …

Bacterial Community Structure Of Contrasting Soils Underlying Bornean Rain Forests: Inferences From Microarray And Next-Generation Sequencing Methods, Sabrina E. Russo, Ryan Legge, Karrie A. Weber, Eoin L. Brodie, Katherine C. Goldfarb, Andrew K. Benson, Sylvester Tan

School of Biological Sciences: Faculty Publications

Soil microbial diversity is vast, and we lack even basic understanding of how this diversity is distributed ecologically. Using pyrosequencing and microarray methods, we quantified the structure of bacterial communities in two contrasting soils underlying Bornean rain forest (clay and sandy loam) that differ markedly in soil properties, aboveground tree flora, and leaf litter decomposition rates. We found significant soil-related taxonomic and phylogenetic differences between communities that, due to their proximity, are independent of climate. Bacterial communities showed distinct compositional and taxon-abundance distributions that were significantly correlated with the structure of the overlying tree community. Richness of bacteria was greater …

Diversity Of The Ohio River Bacterial Communities Using Next-Generation Sequencing Techniques, Emily Michele Anneken

Theses, Dissertations and Capstones

The ability to sequence bacterial genetic material directly from environmental samples has unlocked the field of metagenomics. Next-generation sequencing techniques, such as pyrosequencing, have been instrumental in providing knowledge about bacterial communities. This study focused on identifying the bacterial diversity of the Ohio River via pyrosequencing and compared the diversity of cultivable bacteria versus the entire river community. To ensure the maximum number of cultivable bacteria was obtained, cultures were grown on nonselective media and incubated at river temperature. Bacterial DNA was extracted, sequenced, and classified. The dominant phyla for the Ohio River included Cyanobacteria (38-66% of the total), Actinobacteria …

Structural Characterization Of A Conserved, Calcium-Dependent Periplasmic Protease From Legionella Pneumophila, Debashree Chatterjee, Chelsea D. Boyd, George A. O'Toole, Holger Sondermann

Dartmouth Scholarship

The bacterial dinucleotide second messenger c-di-GMP has emerged as a central molecule in regulating bacterial behavior, including motility and biofilm formation. Proteins for the synthesis and degradation of c-di-GMP and effectors for its signal transmission are widely used in the bacterial domain. Previous work established the GGDEF-EAL domain-containing receptor LapD as a central switch in Pseudomonas fluorescens cell adhesion. LapD senses c-di-GMP inside the cytosol and relays this signal to the outside by the differential recruitment of the periplasmic protease LapG. Here we identify the core components of an orthologous system in Legionella pneumophila. Despite only moderate sequence conservation at …

Larval Parasitoids And Larval Diseases Of Malacosoma Neustria L. (Lepidoptera: Lasiocampidae) Detected In Erzurum Province, Turkey, Hi̇kmet Özbek, Sali̇ha Çoruh

Turkish Journal of Zoology

The larval parasitoids and larval diseases of Malacosoma neustria (L.) (Lepidoptera: Lasiocampidae) in various host plants were investigated in Erzurum Province, Turkey, during 2004 and 2005. The larval parasitoids were reared in a laboratory, and a total of 29 parasitoid species were established in association with the larvae of M. neustria in the families Tachinidae (Diptera), Braconidae, Ichneumonidae, and Chalcidoidea (Hymenoptera). Of the 6 tachinid parasitoids, Pales pavida (Meigen) was the most prevalent species and accounted for 25% of the tachinid records. Among the tachinids, there were 2 new records for the Turkish fauna: Exorista amoena Mesnil and Palesisa nudioculata …

Emerging Microbial And Viral Infections Of The Central Nervous System, Brian J. Balin, Christine J. Hammond

PCOM Scholarly Papers

This review focuses on emerging viral and bacterial infections in the human central nervous system (CNS) that are responsible for significant global morbidity and mortality. These infections include those responsible for acute neurological disease such as meningitis and encephalitis as well those associated with chronic neurodegenerative conditions. Recent changes in climate conditions and pollution have been precipitating factors leading to the emergence of many of these pathogenic organisms. In addition, increased urbanization, global travel, life span, and exposure to new vectors have promoted the organisms’ spread across the globe. Categorization of many of these organisms includes identification of new species, …

Effects Of Sugar-Amendment On Bacterial & Fungal Abundance In Native Vs. Nonnative-Dominated Soils Of A Puget Lowland Prairie, Jessica Wong

Summer Research

Scotch broom is a nonnative plant that has invaded the plant communities of the Pacific Northwest in the United States. It associates with bacteria in the soil to fix nitrogen from the atmosphere, thereby elevating soil nitrogen levels and encouraging the invasion of the community by other nonnative plants like itself. Researchers have used sugar-amendment to decrease soil nitrogen and restore native plant growth. Our study took place in Glacial Heritage Preserve, a Puget lowland prairie that has been invaded by broom in several areas. We aimed to investigate whether sugar-amendment increased or decreased bacterial and fungal abundance in native …

The Role Of Clay Minerals In The Transformation Of E. Coli, Ekaterini Porttellou

Dissertations and Theses

No abstract provided.

Novel Phenol Soluble Modulin Derivatives In Community-Associated Methicillin- Resistant Staphylococcus Aureus Identified Through Imaging Mass Spectrometry., D. Gonzalez, Cheryl Okumura, A. Hollands, R. Kersten, K. Akong-Moore, M. Pence, C. Malone, J. Derieux, B. Moore, A. Horswill, J. Dixon, P. Dorrestein, V. Nizet

Cheryl Okumura

Staphylococcus aureus causes a wide range of human disease ranging from localized skin and soft tissue infections to potentially lethal systemic infections. S. aureus has the biosynthetic ability to generate numerous virulence factors that assist in circumventing the innate immune system during disease pathogenesis. Recent studies have uncovered a set of extracellular peptides produced by community-associated methicillin-resistant S. aureus (CA-MRSA) with homology to the phenol-soluble modulins (PSMs) from Staphylococcus epidermidis. CA-MRSA PSMs contribute to skin infection and recruit and lyse neutrophils, and truncated versions of these peptides possess antimicrobial activity. In this study, novel CA-MRSA PSM derivatives were discovered by …