Environmental Microbiology and Microbial Ecology Commons^™

Project Alien, Rebecca Mccallin, Madelyn Hoying, Alex Evans, Matthew Nestler, Karli Rae Sutton, Garett Craig, Lucia Secaia Del Cid, Alexander Guy, Rachel Fernandez, Amanda Trusiak, Paige Aley, Ingabire Gakwerere, Nina Dorfner, Maria Mosbacher, Mary Flavin, Selvin Hernandez, Audrey Steen, Benjamin Kazimer

Undergraduate Research and Scholarship Symposium

Project ALIEN is a comprehensive plan to send humans to Mars to look for life on the Martian surface while exploring the viability and adaptability of terrestrial microbes in Martian atmospheric conditions. ALIEN will use a ballistic capture trajectory to get to Mars and stay in aerostationary orbit for a 30-day surface mission, during which two surface crewmembers will perform a variety of experiments to achieve the mission’s goals of Martian microbial discovery within brines of the Gale Crater and terrestrial microbe adaptability and viability to Martian conditions. Experimentation is based on the presumption that Martian microbes are metabolically similar …

Metabolic Network Analysis Of Filamentous Cyanobacteria, Daniel Alexis Norena-Caro

LSU Doctoral Dissertations

Cyanobacteria were the first organisms to use oxygenic photosynthesis, converting CO₂ into useful organic chemicals. However, the chemical industry has historically relied on fossil raw materials to produce organic precursors, which has contributed to global warming. Thus, cyanobacteria have emerged as sustainable stakeholders for biotechnological production. The filamentous cyanobacterium Anabaena sp. UTEX 2576 can metabolize multiple sources of Nitrogen and was studied as a platform for biotechnological production of high-value chemicals (i.e., pigments, antioxidants, vitamins and secondary metabolites). From a Chemical engineering perspective, the biomass generation in this organism was thoroughly studied by interpreting the cell as a microbial …

The Release, Transport, And Utilization Of Phosphorus From Bed-Sediments: A Study Of A Eutrophic Littoral Cove On Beaver Lake In Northwest Arkansas, James A. Mccarty

Graduate Theses and Dissertations

Eutrophication of surface waters not only impacts the environment but also water treatment processes, the most significant of which is from the effects of algae. During peak algal growth in many southern U.S. reservoirs, inflows that bring nutrients are at an annual minimum, and phosphorus released from bed-sediments is trapped in the hypolimnion. Littoral areas, described as the most productive zone of the lake, may be a possible source of phosphorus that fuels algal growth in the reservoir. I studied an isolated shallow cove in the War Eagle Creek arm of Beaver Lake in Northwest Arkansas to measure, quantify, and …

Improved Methods For The Quantification Of Viable Ascaris Suum And Application To Biosolids, Tanner Keyzers

Dissertations, Master's Theses and Master's Reports

The current method for detection of Ascaris ova in biosolids and wastewater has several drawbacks including being labor and time intensive as well as being difficult to quantify viable ova that have larvated during treatment. The goal of this thesis was to improve several methods for the detection and quantification of Ascaris ova during long term-storage of biosolids including improvement of quantifying viable ova using a heat and bleach treatment to promote movement of larvae inside of ova, and comparison of the traditional microscopic method with qPCR and PMA-qPCR to prevent the false-positive results. Improved methods to promote movement of …

Optimizing Genetic Manipulation Of Methanogens Through Faster Cloning Techniques, Merrisa Jennings

Biological and Agricultural Engineering Undergraduate Honors Theses

Methanogenesis is the biological production of methane. Only anaerobic archaea known as methanogens are capable of such a metabolic feat. They have strict living conditions and substrate sources which determine their rate of metabolism. This is of particular importance from a greenhouse gas reduction perspective or biogas capturing perspective. One of the best ways to optimize methanogen methane production is via genetic manipulation. The current procedures are timely though, therefore a faster cloning processes should be developed. The objective of this study was to optimize a premade genetic transformation kit known as the Gibson Kit. The Gibson Kit was supposed …

Chloride Salt Inhibition On Lipid Production In Wastewater-Grown Algae For Biofuel Production, Will Richardson

Biological and Agricultural Engineering Undergraduate Honors Theses

Algae are increasingly being recognized as useful organisms for many applications in today’s world. Their ability to remove nitrogen, phosphorus, and trace metals from water while adding oxygen to water makes them an attractive tertiary treatment technology in municipal wastewater treatment facilities. At the same time, algae produce lipids and carbohydrates that are useful for biofuel production, and they are not a human food crop unlike many biofuel feedstocks. In this study the effect of increased chloride concentrations in wastewater was assessed on the ability of two species of algae, Chlorella vulgaris and Scenedesmus dimorphus, to function as a …

Bioaugmentation And Correlating Anaerobic Digester Microbial Community To Process Function, Kaushik Venkiteshwaran

Dissertations (1934 -)

This dissertation describes two research projects on anaerobic digestion (AD) that investigated the relationship between microbial community structure and digester function. Both archaeal and bacterial communities were characterized using high-throughput (Illumina) sequencing technology with universal 16S rRNA gene primers. In the first project, bioaugmentation using a methanogenic, aerotolerant propionate enrichment culture was investigated as a possible method to increase digester methane production. Nine anaerobic digesters, seeded with different biomass, were operated identically and their quasi steady state function was compared. Before bioaugmentation, different seed biomass resulted in different quasi steady state function, with digesters clustering into high, medium or low …

The Role Of Household Antimicrobials In The Proliferation Of Antibiotic Resistance During Anaerobic Digestion, Daniel Elliott Carey

Dissertations (1934 -)

Antimicrobial chemicals in consumer personal care products have been found to increase antibiotic resistance in pure culture studies. Although many studies focus on antibiotic resistance development pertinent to medical scenarios, resistance developed in natural and engineered environments might be significant and has become an emerging concern for human health. This dissertation focuses on the antimicrobial chemicals triclosan and triclocarban. These compounds are distinctly different from antibiotics and are used in products like soaps that are labelled as “antibacterial”. Municipal wastewater treatment plants receive triclocarban and triclosan loads higher than most contaminants of emerging concern because they are frequently used in …

Noninvasive Measurement Of Electrical Events Associated With A Single Chlorovirus Infection Of A Microalgal Cell, Seung-Woo Lee, Eun-Hee Lee, Gerhard Thiel, James L. Van Etten, Ravi Saraf

Department of Chemical and Biomolecular Engineering: Faculty Publications

Chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) contains a viral-encoded K⁺ channel imbedded in its internal membrane, which triggers host plasma membrane depolarization during virus infection. This early stage of infection was monitored at high resolution by recording the cell membrane depolarization of a single Chlorella cell during infection by a single PBCV-1 particle. The measurement was achieved by depositing the cells onto a network of one-dimensional necklaces of Au nanoparticles, which spanned two electrodes 70 μm apart. The nanoparticle necklace array has been shown to behave as a single-electron device at room temperature. The resulting electrochemical field-effect transistor …

Sulfate-Reducing Bacteria Community Analysis: Isco/Isb Coupled Remediation, Christopher K. Bartlett

Theses and Dissertations (Comprehensive)

Improved techniques for remediating groundwater systems are required for the more than 500,000 contaminated sites in North America. Many of these sites are the legacy of historical industrial operations, inappropriate disposal practices and accidental releases. The most widely observed contaminant at many of these sites is petroleum hydrocarbons (PHCs). Recently, remediation efforts that involve the sequential application of treatment technologies have gained widespread interest. One specific sequential technology application or treatment train employs the aggressive nature of a chemical oxidation followed by bioremediation for polishing. When persulfate is used as the chemical oxidant its natural degradation by-product is sulfate, an …

Production And Harvest Of Microalgae In Wastewater Raceways With Resource Recycling, Alexander Colin Roberts

Master's Theses

Microalgae can be grown on municipal wastewater media to both treat the wastewater and produce feedstock for algae biofuel production. However the reliability of treatment must be demonstrated, as well as high areal algae productivity on recycled wastewater media and efficient sedimentation harvesting. This processes was studied at pilot scale in the present research.

A pilot facility was operated with nine CO₂-supplemented raceway ponds, each with a 33-m² surface area and a 0.3-m depth, continuously from March 6, 2013 through September 24, 2014. The ponds were operated as three sets of triplicates with two sets continuously fed …

Seeps And Springs At A Platteville “Observatory” On The River Bluffs, Bj Bonin, Greg Brick, Julia R. Steenberg

Sinkhole Conference 2015

Residential building construction along the Mississippi River bluffs in the 1970s created a unique enclosed outcrop of the Late Ordovician Platteville Formation at Lilydale, Minnesota. This outcrop was examined in early 2013 after a newly-formed spring flooded an elevator shaft the previous year, drawing attention to the foundation conditions. The Lexington Riverside property is a six story condominium complex constructed within the top of the bluff. A two-level underground parking garage was built into the bluff. Bedrock was mechanically excavated to accommodate the construction of the building, creating an unweathered rock surface. The space between the structure and the excavated …

The Contribution Of Syntrophic Fatty-Acid Degrading Microbial Communities To Anaerobic Digester Function And Stability, Prince Peter Mathai

Dissertations (1934 -)

Anaerobic digestion (AD), the conversion of complex organic matter to methane, occurs through a series of reactions mediated by different guilds of microorganisms. AD process imbalances, such as organic overload or high organic loading rates (OLR), can result in the accumulation of volatile fatty acids (VFA) e.g., propionate, which must be degraded to maintain stable reactor function. VFAs are metabolized by syntrophic fatty-acid degrading bacteria (SFAB) in association with methanogenic archaea (collectively, syntrophic microbial communities, SMC). Despite their indispensable role in AD, little is known about the ecology of SFAB, especially under stressed conditions. To facilitate ecological studies, four quantitative …

Concomitant Uptake Of Antimicrobials And Salmonella In Soil And Into Lettuce Following Wastewater Irrigation, J. Brett Sallach, Yuping Zhang, Laurie Hodges, Daniel D. Snow, Xu Li, Shannon L. Bartelt-Hunt

Department of Civil and Environmental Engineering: Faculty Publications

The use of wastewater for irrigation may introduce antimicrobials and human pathogens into the food supply through vegetative uptake. The objective of this study was to investigate the uptake of three antimicrobials and Salmonella in two lettuce cultivars. After repeated subirrigation with synthetic wastewater, lettuce leaves and soil were collected at three sequential harvests. The internalization frequency of Salmonella in lettuce was low. A soil horizon-influenced Salmonella concentration gradient was determined with concentrations in bottom soil 2 log CFU/g higher than in top soil. Lincomycin and sulfamethoxazole were recovered from lettuce leaves at concentrations as high as 822 ng/g and …

Methane Production By A Packed-Bed Anaerobic Digester Fed Dairy Barn Flush Water, Sean Richard Thomson

Master's Theses

Packed-bed digesters are an alternative to covered lagoon digesters for methane production and anaerobic treatment of dilute wastewaters such as dairy barn flush water. The physical media of packed-beds retain biofilms, often allowing increased treatment rates. Previous studies have evaluated several types of media for digestion of dilute wastewaters, but cost and media fouling have setback commercial development. A major operational cost has been effluent recirculation pumping.

In the present effort, a novel approach to anaerobic digestion of flush dairy water was developed at pilot-scale: broken walnut shells were used as a low-cost packed-bed medium and effluent recirculation was replaced …

Investigating Antibiotic Resistance Levels Of Salmonella Internalized In Lettuce Leaves, Jason B. Thomas

McNair Scholars Research Journal

Contamination of food crops by the human pathogen Salmonella is a food safety threat worldwide. Though using treated wastewater for irrigation is a sustainable practice, it may introduce trace levels of Salmonella that may contaminate food crops. Salmonella could develop resistance to antibiotics present in wastewater. The overall goal of the project is to increase the understanding of the public health risk associated with the use of treated wastewater to irrigate food crops. The objective of this particular study is to determine the antibiotic resistance level of Salmonella internalized in lettuce leaves. In this experiment, thirty-six plants of the lettuce …

Life Support: Long Term Storage Of Solid Waste In An Enclosed Membrane System, Emmy O. Trieu, Michael Flynn, Rocco Mancinelli

STAR Program Research Presentations

In deep space missions, maintaining life support is of the utmost priority. In such a closed system, human waste must be stored and treated. Simulated feces (ersatz) will be inoculated with microbes representing normal fecal flora, mixed with urine brine and shredded refuse of products typically used in space missions. Composting methods often use alternating layers of waste with scraps of carboniferous materials (finely shredded refuse). By preparing membrane bags with a homogenized ersatz and carboniferous refuse mixture and membrane bags with alternating layers of ersatz and carboniferous refuse, it may be possible to monitor anaerobic thermophillic digestion of the …

Enhanced Hydrocarbon Biodegradation Using Bioaugmentation With Biowish™-Aqua Fog, Michael Robert Lehrer

Master's Theses

This study was done to determine the effectiveness of a commercially available bioaugmentation product, BiOWiSH^TM-Aqua FOG, for remediating petroleum-contaminated sandy soil. Biodegradation enhancement by BiOWiSH^TM-Aqua FOG was evaluated in laboratory microcosms by directly measuring total petroleum hydrocarbon (TPH) and indirectly using respirometry. Attempts were made to enrich hydrocarbon-degrading bacteria in BiOWiSH^TM-Aqua FOG, and the resulting enrichment cultures were screened using respirometry as well. Potential hydrocarbon-degrading bacteria in BiOWiSH^TM-Aqua FOG were isolated. Experiments were performed at bench-scale using microcosm bottles containing sand contaminated with either motor oil or No. 2 diesel fuel. The …

Investigation Of Bacillus Subtilis As A Biopesticide Against Botrytis Cinerea, Kenneth K. Ng

Master's Theses

The objective of this thesis was to investigate BiOWiSH^TM-Aqua, a commercial dry solid formulation containing a consortium of bacteria and yeast, as a biopesticide for treatment of Botrytis cinerea, a gray mold that affects strawberries. BiOWiSH^TM-Aqua was compared with another commercial product specifically used as a fungicide and bacteriocide, Serenade® Garden Disease Control Spray (concentrated Bacillus subtilis strain QST 713). Both laboratory tests as well as in vivo lab tests were conducted. BiOWiSH^TM-Aqua results varied widely from plate to plate, regardless of experimental conditions. In some of these plates, inhibition zones were observed …

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 …

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 …

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 …

Development Of Pyrf-Based Genetic System For Targeted Gene Deletion In Clostridium Thermocellum And Creation Of A Pta Mutant, Shital A. Tripathi, Daniel G. Olson, D. Aaron Argyros, Bethany B. Miller, Trisha F. Barrett, Daniel M. Murphy, Jesse D. Mccool, Anne K. Warner, Vineet B. Rajgarhia, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza

Dartmouth Scholarship

We report development of a genetic system for making targeted gene knockouts in Clostridium thermocellum, a thermophilic anaerobic bacterium that rapidly solubilizes cellulose. A toxic uracil analog, 5-fluoroorotic acid (5-FOA), was used to select for deletion of the pyrF gene. The ΔpyrF strain is a uracil auxotroph that could be restored to a prototroph via ectopic expression of pyrF from a plasmid, providing a positive genetic selection. Furthermore, 5-FOA was used to select against plasmid-expressed pyrF, creating a negative selection for plasmid loss. This technology was used to delete a gene involved in organic acid production, namely pta, which encodes …

Isolation And Characterization Of Bacteria From Extended Aeration Wastewater Treatment Plants., Randy Chin Kok Fei

Student Works (2010-2019)

In this study, three different wastewater treatment plants (STP1, STP2 and STP3) were sampled in Klang Valley. STP1 exhibited the best activated sludge health with healthy Sludge Volume (SV), Mixed Liquor Suspended Solids (MLSS) and Mixed Liquor Volatile Suspended Solids (MLVSS). STP2 were constantly disrupted by the mechanical failures which caused its dissolved oxygen (DO) to decrease whereas STP3 was troubled by the inadequate alkalinity in its mixed liquor causing acidic condition in aeration tank. The acidic condition resulted in pin-floc problem and contributed to high Total Suspended Solids (TSS) and low SV. Influent at STP3 was also found to …

Response Surface Optimization Of Crystal Violet Decolorization By Pellets Of Pyncnoporus Sanguineus., Siti Suhana Sulaiman

Student Works (2010-2019)

Ligninolytic enzymes of white-rot fungi have broad specificities and have been implicated in transformation and mineralization of organo-pollutant with chemical structure similar to lignin e.g. synthetic dyes. This study was carried out to optimize selected parameters for decolorization of Crystal violet by Pycnoporus sanguineus in shake flasks. The parameters studied were initial dye concentration (ppm), agitation speed (rpm) and process time. They were optimized using response surface methodology (RSM). Subsequent to the optimization, the decolorization process was scaled up in stirred tank reactor (STR). Effect of different geometry of impellers on the decolorization process was studied. Curved blade and angled …

The Influence Of Carbon Source Types And Nitrate On The Performance Of The Enhanced Biological Phosphorus Removal Systems, Shamim Ara Begum

UNLV Theses, Dissertations, Professional Papers, and Capstones

This research focuses on two issues in enhanced biological phosphorus removal (EBPR). The first issue encompasses the effect of the type of carbon sources (i.e. volatile fatty acids and non volatile fatty acids) on the microbiology of EBPR. The second issue deals with the influence of denitrification on EBPR. Few studies have been performed to investigate these two issues. The recent findings on these areas are summarized below to establish the knowledge gaps this dissertation attempted to address. Details on the specific findings can be found in the literature review portion of this proposal.

Temperature Influence And Heat Management Requirements Of Microalgae Cultivation In Photobioreactors, Thomas Hagen Mehlitz

Master's Theses

Microalgae are considered one of the most promising feedstocks for biofuel production for the future. The most efficient way to produce vast amounts of algal biomass is the use of closed tubular photobioreactors (PBR). The heat requirement for a given system is a major concern since the best algae growth rates are obtained between 25-30 °C, depending on the specific strain. A procedure to determine temperature influence on algal growth rates was developed for a lab-scale PBR system using the species Chlorella. A maximum growth rate of 1.44 doublings per day at 29 °C (optimal temperature) was determined. In addition, …

N-Glycan Modification In Aspergillus Species, Elke Kainz, Andreas Gallmetzer, Christian Hatzl, Juergen H. Nett, Huijuan Li, Thorsten Schinko, Robert Pachlinger, Harald Berger, Yazmid Reyes-Dominguez, Andreas Bernreiter, Tillmann Gerngross, Stefan Wildt, Joseph Strauss

Dartmouth Scholarship

The production by filamentous fungi of therapeutic glycoproteins intended for use in mammals is held back by the inherent difference in protein N-glycosylation and by the inability of the fungal cell to modify proteins with mammalian glycosylation structures. Here, we report protein N-glycan engineering in two Aspergillus species. We functionally expressed in the fungal hosts heterologous chimeric fusion proteins containing different localization peptides and catalytic domains. . This strategy allowed the isolation of a strain with a functional -1,2-mannosidase producing increased amounts of N-glycans of the Man 5 GlcNAc 2 type. This strain was further engineered by the introduction of …

Analysis Of Bacterial Population And Distribution In The Developing Strata Of A Constructed Wetland Used For Chlorinated Ethene Bioremediation, Milton J. Clausen Jr.

Theses and Dissertations

Chlorinated hydrocarbons and their degradation products are among of the most common organic groundwater contaminates in the United States. These compounds attack the central nervous system in animals and can affect the photosynthesis of plants. These compounds are also resistant to degradation in the environment and, because of this, pose a risk to any ecosystem in which they are present. This study identified the dominant microbial species in a constructed treatment wetland at Wright-Patterson AFB, Dayton, Ohio using 16S rRNA gene sequence analysis. Samples were taken from three different depths and during each of the four seasons. These samples were …

Cellulose Utilization By Clostridium Thermocellum: Bioenergetics And Hydrolysis Product Assimilation, Yi-Heng P. Zhang, Lee R. Lynd

Dartmouth Scholarship

The bioenergetics of cellulose utilization by Clostridium thermocellum was investigated. Cell yield and maintenance parameters, Y(X/ATP)True = 16.44 g cell/mol ATP and m = 3.27 mmol ATP/g cell per hour, were obtained from cellobiose-grown chemostats, and it was shown that one ATP is required per glucan transported. Experimentally determined values for G(ATP)P-T (ATP from phosphorolytic beta-glucan cleavage minus ATP for substrate transport, mol ATP/mol hexose) from chemostats fed beta-glucans with degree of polymerization (DP) 2-6 agreed well with the predicted value of (n-2)/n [corrected] (n = mean cellodextrin DP assimilated). A mean G(ATP)(P-T) value of 0.52 +/- 0.06 was calculated …