Environmental Microbiology and Microbial Ecology Commons^™

Enhanced Microbial Utilization Of Recalcitrant Cellulose By An Ex Vivo Cellulosome-Microbe Complex, Chun You, Xiao-Zhou Zhang, Noppadon Sathitsuksanoh, Lee R. Lynd

Dartmouth Scholarship

A cellulosome-microbe complex was assembled ex vivo on the surface of Bacillus subtilis displaying a miniscaffoldin that can bind with three dockerin-containing cellulase components: the endoglucanase Cel5, the processive endoglucanase Cel9, and the cellobiohydrolase Cel48. The hydrolysis performances of the synthetic cellulosome bound to living cells, the synthetic cellulosome, a noncomplexed cellulase mixture with the same catalytic components, and a commercial fungal enzyme mixture were investigated on low-accessibility recalcitrant Avicel and high accessibility regenerated amorphous cellulose (RAC). The cellbound cellulosome exhibited 4.5- and 2.3-fold-higher hydrolysis ability than cell-free cellulosome on Avicel and RAC, respectively. The cellulosome-microbe synergy was not completely …

Exploring The Effects Of Hypoxia On Sulfate Reducing Anaerobes, Annaliese K. Jones

Senior Honors Projects

Exploring the Effects of Hypoxia on Sulfate Reducing Anaerobes

Annaliese K. Jones

Sponsor: Bethany Jenkins, Cell and Molecular Biology

As a student about to graduate with a degree in Biological Sciences, I find myself faced with the need to find my own independent research interests and scientific voice. As a result of my interests in the fields of both microbiology and ecology, I am drawn to questions surrounding the role and behavior of microorganisms in the environment. With climate change being an issue capturing the attention of a large portion of the scientific community, I have chosen to focus my …

Gravesoil Microbial Community Structure During Carcass Decomposition, Amy E. Maile

Department of Entomology: Dissertations, Theses, and Student Research

Dead bodies placed on soil represent unique challenges for investigators. Although processes in soils can be used to estimate postmortem interval, we know very little about how carcasses and insects affect gravesoil microbial communities.

To address this, the current project was composed of two experiments. Experiment one was conducted to investigate the effect of surface type on carcass decomposition and evaluate soil ecology methods. Experiment two was conducted to investigate the presence of an insect population (Lucilia sericata Meigen) on gravesoil microbial communities. Both experiments were conducted in a laboratory setting using freshly killed mouse carcasses. Mouse carcasses were …

Zap1 Control Of Cell-Cell Signaling In Candida Albicans Biofilms, Shantanu Ganguly, Andrew C. Bishop, Wenjie Xu, Suman Ghosh, Kenneth W. Nickerson, Frederick Lanni, Jana Patton-Vogt, Aaron P. Mitchell

Kenneth Nickerson Papers

Biofilms of Candida albicans include both yeast cells and hyphae. Prior studies indicated that a zap1/ mutant, defective in zinc regulator Zap1, has increased accumulation of yeast cells in biofilms. This altered yeast-hypha balance may arise from internal regulatory alterations or from an effect on the production of diffusible quorum-sensing (QS) molecules. Here, we develop biosensor reporter strains that express yeastspecific YWP1-RFP or hypha-specific HWP1-RFP, along with a constitutive TDH3-GFP normalization standard. Seeding these biosensor strains into biofilms allows a biological activity assay of the surrounding biofilm milieu. A zap1/ biofilm induces the yeast-specific YWP1-RFP reporter in a wild-type …

High Ethanol Titers From Cellulose By Using Metabolically Engineered Thermophilic, Anaerobic Microbes, D. Aaron Argyros, Shital A. Tripathi, Trisha F. Barrett, Stephen R. Rogers, Lawrence F. Feinberg, Daniel G. Olson, Justin M. Foden, Bethany B. Miller, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza

Dartmouth Scholarship

This work describes novel genetic tools for use in Clostridium thermocellum that allow creation of unmarked mutations while using a replicating plasmid. The strategy employed counter-selections developed from the native C. thermocellum hpt gene and the Thermoanaerobacterium saccharolyticum tdk gene and was used to delete the genes for both lactate dehydrogenase (Ldh) and phosphotransacetylase (Pta). The Δldh Δpta mutant was evolved for 2,000 h, resulting in a stable strain with 40:1 ethanol selectivity and a 4.2-fold increase in ethanol yield over the wild-type strain. Ethanol production from cellulose was investigated with an engineered coculture of organic acid-deficient engineered strains of …

Ecology And Genetic Structure Of A Northern Temperate Vibrio Cholerae Population Related To Toxigenic Isolates, Brian M. Schuster, Anna L. Tyzik, Rachel A. Donner, Megan J. Striplin, Salvador Almagro-Moreno

Dartmouth Scholarship

Although Vibrio cholerae is an important human pathogen, little is known about its populations in regions where the organism is endemic but where cholera disease is rare. A total of 31 independent isolates confirmed as V. cholerae were collected from water, sediment, and oysters in 2008 and 2009 from the Great Bay Estuary (GBE) in New Hampshire, a location where the organism has never been detected. Environmental analyses suggested that abundance correlates most strongly with rainfall events, as determined from data averaged over several days prior to collection. Phenotyping, genotyping, and multilocus sequence analysis (MLSA) revealed a highly diverse endemic …

Group Size And Nest Spacing Affect Buggy Creek Virus (Togaviridae: Alphavirus) Infection In Nestling House Sparrows, Valerie A. Brown, Charles R. Brown

Papers in Ornithology

The transmission of parasites and pathogens among vertebrates often depends on host population size, host species diversity, and the extent of crowding among potential hosts, but little is known about how these variables apply to most vector-borne pathogens such as the arboviruses (arthropod-borne viruses). Buggy Creek virus (BCRV; Togaviridae: Alphavirus) is an RNA arbovirus transmitted by the swallow bug (Oeciacus vicarius) to the cliff swallow (Petrochelidon pyrrhonota) and the introduced house sparrow (Passer domesticus) that has recently invaded swallow nesting colonies. The virus has little impact on cliff swallows, but house sparrows are seriously …

Human Pathogen Shown To Cause Disease In The Threatened Elkhorn Coral Acropora Palmata, Kathryn Patterson Sutherland, Sameera Shaban, Jessica L. Joyner, James W. Porter, Erin K. Lipp

Faculty Publications

Coral reefs are in severe decline. Infections by the human pathogen Serratia marcescens have contributed to precipitous losses in the common Caribbean elkhorn coral, Acropora palmata, culminating in its listing under the United States Endangered Species Act. During a 2003 outbreak of this coral disease, called acroporid serratiosis (APS), a unique strain of the pathogen, Serratia marcescens strain PDR60, was identified from diseased A. palmata, human wastewater, the non-host coral Siderastrea siderea and the corallivorous snail Coralliophila abbreviata. In order to examine humans as a source and other marine invertebrates as vectors and/or reservoirs of the APS pathogen, challenge experiments …

Identification Of Nitrifying Bacteria Contained In A Commercial Inoculant Using Molecular Biology Techniques, Anthony Harrington, John Perry, Penny S. Amy

Undergraduate Research Opportunities Program (UROP)

Nitrifying bacteria play an important role in aquatic and terrestrial environments through the nitrogen cycle. Nitrification, one of the processes of the nitrogen cycle, refers to the oxidation of ammonia to nitrate. This process requires two types of chemoautotrophic bacteria, ammonia-oxidizing bacteria (AOB), and nitrite-oxidizing bacteria (NOB). These bacteria are essential in maintaining an optimal environment for plants and aquatic organisms, such as fish. Current applications of nitrifiers include: inoculants for aquariums, biofertilizers, and nitrogen removal in wastewater treatment plants. This study wants to identify a consortium of nitrifers that can be used to produce sufficient nitrate for plants in …

Biological Effects On Serpentinite Weathering, Mary H. Evert, Julie Baumeister, Elisabeth Hausrath

Undergraduate Research Opportunities Program (UROP)

Serpentinites, perhaps more than any other rock type, control the composition and evolution of the development of the surrounding ecosystems. The bulk chemistry of serpentinite rocks, high in Mg and trace elements, and low in nutrients such as Ca, K, P, and N, causes an extreme and stressful environment for ecosystems. However, the role that those serpentine ecosystems play in development of serpentine soils has not been examined.

Due to the unusual chemistry of serpentine soils, serpentine ecosystems have deeper and better-developed root systems than other ecosystems. The rhizosphere of serpentine systems, documented to produce abundant organic acids and siderophores, …

A Spatial And Temporal Analysis Of Microbial Communities In Great Boiling Spring, Nevada, U.S.A., Jessica K. Guy, Joseph P. Peacock, Jeremy A. Dodsworth, Tanja Woyke, Tijana G. Del Rio, Brian P. Hedlund

Undergraduate Research Opportunities Program (UROP)

Great Boiling Spring (GBS) is a large, circumneutral, long residence time geothermal spring in the US Great Basin. Twelve samples were taken from four different sediment sites and the planktonic community in the bulk water of GBS on up to four different dates. Microbial community composition and diversity was assessed by using a barcoded, improved universal primer set targeting the V8 portion of the 16S rRNA gene and PCR. Over 200,000 products were sequenced using the Roche 454 GS FLX Titanium System. Sediment and planktonic microbial communities were distinct with very little overlap, regardless of the sampling location or temperature. …

Growth And Hydrogen Production Of Rubrivivax Gelatinosus Cbs, Shannon A. Swanson, Karen Wawrousek

STAR Program Research Presentations

Rubrivivax gelatinosus CBS is a purple non-sulfur photosynthetic bacterium that has been found to produce substantial amounts of Hydrogen (H2) which could be used as a clean burning biofuel. CBS has been known to grow on a variety of carbon sources, but it is unknown as to which carbon sources it has the greatest potential to use, and this understanding could be helpful in engineering future strains of the bacteria. Hydrogen production in CBS occurs when carbon monoxide (CO) is present and is presumed to be under the control of the RcoM transcription factor. When CBS wild-type strains were grown …

Applied And Mechanistic Studies Of Microbial 17beta-Estradiol Degradation, Zhongtian Li

Department of Civil and Environmental Engineering: Dissertations, Theses, and Student Research

The presence of natural estrogens, a class of endocrine disrupting compounds, in water has caused increasing concerns over their adverse impacts on the health of aquatic eco-systems and human beings. In this study, adsorption characteristics of two natural estrogens, 17β-estradiol (E2) and estrone (E1), on granular activated carbon (GAC) were investigated in isotherm tests and in a GAC column. The GAC column was then converted to a biologically active carbon (BAC) column and the removal efficiency of E2 and its primary biodegradation intermediate E1 were monitored. During BAC operation, the impacts of various reactor operation parameters, such as the carbon …

In Situ Enrichment Of Ocean Crust Microbes On Igneous Minerals And Glasses Using An Osmotic Flow‐Through Device, Amy Renee Smith, Martin Fisk, Radu Popa, Mark Nielsen, C. Geoffrey Wheat, Hans W. Jannasch, Andrew T. Fisher, Keir Becker, Stefan M. Sievert, Gilberto Flores

Biology Faculty Publications and Presentations

The Integrated Ocean Drilling Program (IODP) Hole 1301A on the eastern flank of Juan de Fuca Ridge was used in the first long-term deployment of microbial enrichment flow cells using osmotically driven pumps in a subseafloor borehole. Three novel osmotically driven colonization systems with unidirectional flow were deployed in the borehole and incubated for 4 years to determine the microbial colonization preferences for 12 minerals and glasses present in igneous rocks. Following recovery of the colonization systems, we measured cell density on the minerals and glasses by fluorescent staining and direct counting and found some significant differences between mineral samples. …

Novel Model For Multispecies Biofilms That Uses Rigid Gas-Permeable Lenses, Rebecca Peyyala, Sreenatha S. Kirakodu, Jeffrey L. Ebersole, Karen F. Novak

Center for Oral Health Research Faculty Publications

Oral biofilms comprise complex multispecies consortia aided by specific inter- and intraspecies interactions occurring among commensals and pathogenic bacterial species. Oral biofilms are primary initiating factors of periodontal disease, although complex multifactorial biological influences, including host cell responses, contribute to the individual outcome of the disease. To provide a system to study initial stages of interaction between oral biofilms and the host cells that contribute to the disease process, we developed a novel in vitro model system to grow biofilms on rigid gas-permeable contact lenses (RGPLs), which enable oxygen to permeate through the lens material. Bacterial species belonging to early- …

An Investigation Of The Composition, Microbial Digestion And Potential Applications Of An Industrial By-Product Of Seaweed Extraction, Juncal Nogales Esteban

Theses

Commercial preparation of seaweed extract from Ascophyllum nodosum, for use as fertilizer and soil improver, produces a residue which requires remediation. This residue is rich in nutrients and offers potential for other added-value products. The residue’s composition and microbial flora were studied, a microbial digestion system was developed, and extracts were screened for anti-hyaluronidase and anti-elastase activities.

The residue had a pH of 8.61±0.39, 16% (w/w) TS which comprised 40.6% minerals, 29.5% fibre, 20.9% lipid, 4.9% protein and 0.48% polyphenols. The microbial digestion included an initial 3-day anaerobic phase during which pH decreased from 9.12 to 7.89. At day 3, …

Pcr Primers For The Detection Of Propane And Butane-Oxidizing Microorganisms, Brian Jeremy Chan

Master's Theses

In an increasingly energy-hungry world, our capacity to meet the heightened energy demands of the future has become a pressing matter. The most urgent of these concerns are tied to the accessibility of petroleum. Various experts have proselytized both the imminent arrival of peak oil production rates and the ensuing decline of those rates thereafter. And to that end, the development of novel and advanced oil exploration methodologies has become almost as important as finding the sources of oil themselves.

The soils above petroleum reservoirs play host to various communities of alkane- oxidizing bacteria that can utilize the natural gas …

Cellulose- And Xylan-Degrading Thermophilic Anaerobic Bacteria From Biocompost, M. V. Sizova, J. A. Izquierdo, N. S. Panikov, L. R. Lynd

Dartmouth Scholarship

Nine thermophilic cellulolytic clostridial isolates and four other noncellulolytic bacterial isolates were isolated from self-heated biocompost via preliminary enrichment culture on microcrystalline cellulose. All cellulolytic isolates grew vigorously on cellulose, with the formation of either ethanol and acetate or acetate and formate as principal fermentation products as well as lactate and glycerol as minor products. In addition, two out of nine cellulolytic strains were able to utilize xylan and pretreated wood with roughly the same efficiency as for cellulose. The major products of xylan fermentation were acetate and formate, with minor contributions of lactate and ethanol. Phylogenetic analyses of 16S …

Potential Fossil Endoliths In Vesicular Pillow Basalt, Coral Patch Seamount, Eastern North Atlantic Ocean, Barbara Cavalazzi, Frances Westall, Sherry L. Cady, Roberto Barbieri, Frédéric Foucher

Geology Faculty Publications and Presentations

The chilled rinds of pillow basalt from the Ampere-Coral Patch Seamounts in the eastern North Atlantic were studied as a potential habitat of microbial life. A variety of putative biogenic structures, which include filamentous and spherical microfossil-like structures, were detected in K-phillipsite-filled amygdules within the chilled rinds. The filamentous structures (similar to 2.5 mu m in diameter) occur as K-phillipsite tubules surrounded by an Fe-oxyhydroxide (lepidocrocite) rich membranous structure, whereas the spherical structures (from 4 to 2 mu m in diameter) are associated with Ti oxide (anatase) and carbonaceous matter. Several lines of evidence indicate that the microfossil-like structures in …

Dur3 Is The Major Urea Transporter In Candida Albicans And Is Co-Regulated With The Urea Amidolyase Dur1,2, Dhammika H. M. L. P Navarathna, Aditi Das, Joachim Morschhauser, Kenneth W. Nickerson, David D. Roberts

Kenneth Nickerson Papers

Hemiascomycetes, including the pathogen Candida albicans, acquire nitrogen from urea using the urea amidolyase Dur1,2, whereas all other higher fungi use primarily the nickel-containing urease. Urea metabolism via Dur1,2 is important for resistance to innate host immunity in C. albicans infections. To further characterize urea metabolism in C. albicans we examined the function of seven putative urea transporters. Gene disruption established that Dur3, encoded by orf 19.781, is the predominant transporter. [¹⁴C]Urea uptake was energy-dependent and decreased approximately sevenfold in a dur3D mutant. DUR1,2 and DUR3 expression was strongly induced by urea, whereas the other putative transporter genes …

Iron Oxide And Calcite Associated With Leptothrix Sp. Biofilms Within An Estavelle In The Upper Floridan Aquifer, Lee J. Florea, Chasity L. Stinson, Josh Brewer, Rick Fowler, B Joe Kearns, Anthony M. Greco

International Journal of Speleology

In Thornton’s Cave, an estavelle in west-central Florida, SEM, EDS, and XRD data reveal biofilms that are predominantly comprised of FeOOH-encrusted hollow sheaths that are overgrown and intercalated with calcite. Fragments of this crystalline biofilm adhere to the walls and ceiling as water levels vary within the cave. Those on the wall have a ‘cornflake’ appearance and those affixed to the ceiling hang as fibrous membranes. PCR of DNA in the active biofilm, combined with morphologic data from the tubes in SEM micrographs, point to Leptothrix sp., a common Fe-oxidizing bacteria, as the primary organism in the biofilm. Recent discoveries …