Environmental Microbiology and Microbial Ecology Commons^™

Next Generation Sequencing Methods For Coastal Zone Water Quality Monitoring, Catherine Iskrenko

HCNSO Student Capstones

When analyzing the water quality of the coastal zone, culture-based techniques have been utilized most often to identify Fecal Indicator Bacteria in samples. Since the advent of the Sanger Method for DNA sequencing, other techniques have arisen that provide significantly more information on the microorganisms in sample, but they are still not the mainstream for water quality analysis. This capstone reviews and compares culture-based techniques, DNA sequencing, RNA sequencing, qPCR for biomarker, and 16S rDNA sequencing to highlight their merits and shortcomings for analyzing environmental water samples. The technique presented that provides the broadest range of information (including the identification …

Functions Of Ecosystems: Stream Metabolism As An Efficient And Effective Means To Gage The Health And Understand The Interworking Of Urban Streams In A Watershed Of Rock Island, Il, Ryan Johnson, Dr. Kevin Geedey

Celebration of Learning

Stream metabolism is a critical functional measure of stream health that integrates physical parameters like slope and discharge, with ecosystem functions like photosynthesis and respiration. Stream metabolism is widely studied; however, urban stream metabolism remains poorly understood. Stream metabolism was measured for five streams ranging from 1^st to 5^th orders from October 11^th to October 18^th 2017 and four streams ranging from 1^st to 4^th order from October 22^nd to 25^th 2017 located within an approximately 9.3 square kilometer watershed of Rock Island, IL that has an urban to suburban type of …

Microbiome And Bacterioplankton Rrna Gene Sequence Data Collected From Gulf Of Mexico Seawater Samples, Cruises Dp03 And Dp04 From April - August 2016, Cole Easson, Lindsey Freed

DEEPEND Datasets

Seawater was collected and filtered for microbiome and bacterioplankton sequencing and analyses at various depths during planned DEEPEND cruise expeditions to the GOM in 2016. Filters were stored and then processed for total environmental genomic DNA according to standard methods (see earthmicrobiome.org). 16S rRNA amplicon libraries covering the V4 hypervariable regions were generated with universal PCR primers and then sequenced on an Illumina MiSeq DNA sequencing platform. Raw paired-end sequences were joined and quality filtered in the bioinformatics program, QIIME. Vertical baseline characterizations will track alpha and beta diversity at different depths ranging from 0 – 1500 m, assess seasonal …

Nevada Desert Dust With Heavy Metals Suppresses Igm Antibody Production, Deborah E. Keil, Brenda J. Buck, Dirk Goossens, Brett Mclaurin, Lacey Murphy, Mallory Leetham-Spencer, Yuanxin Teng, James E. Pollard, Russell Gerads, Jamie C. Dewitt

Geoscience Faculty Research

Systemic health effects from exposure to a complex natural dust containing heavy metals from the Nellis Dunes Recreation Area (NDRA) near Las Vegas, NV, were evaluated. Several toxicological parameters were examined following lung exposure to emissive dust from three geologic sediment types heavily used for recreational off-road activities: yellow sand very rich in arsenic (termed CBN 5); a shallow cover of loose dune sand overlying a gravelly subsoil bordering dune fields (termed CBN 6); and brown claystone and siltstone (termed CBN 7). Adult female B6C3F1 mice were exposed by oropharyngeal administration to these three types of geogenic dusts at 0.01–100 …

Bioassessment Of A Drinking Water Reservoir Using Plankton: High Throughput Sequencing Vs. Traditional Morphological Method, Wanli Gao, Zhaojin Chen, Yuying Li, Yangdong Pan, Jingya Zhu, Shijun Guo, Lanqun Hu, Jin Huang

Environmental Science and Management Faculty Publications and Presentations

Drinking water safety is increasingly perceived as one of the top global environmental issues. Plankton has been commonly used as a bioindicator for water quality in lakes and reservoirs. Recently, DNA sequencing technology has been applied to bioassessment. In this study, we compared the effectiveness of the 16S and 18S rRNA high throughput sequencing method (HTS) and the traditional optical microscopy method (TOM) in the bioassessment of drinking water quality. Five stations reflecting different habitats and hydrological conditions in Danjiangkou Reservoir, one of the largest drinking water reservoirs in Asia, were sampled May 2016. Non-metric multi-dimensional scaling (NMDS) analysis showed …

Fungi Of Forests: Examining The Diversity Of Root-Associated Fungi And Their Responses To Acid Deposition, Donald Jay Nelsen

Graduate Theses and Dissertations

Global importance of forests is difficult to overestimate, given their role in oxygen production, ecological roles in nutrient cycling and supporting numerous living species, and economic value for industry and as recreational zones. Fitness of the forest-forming trees strongly depends on microbial communities associated with tree roots. In particular, fungi impact tree fitness: mycorrhizal species provide water and nutrients for the trees in exchange for C, endophytic fungi play key roles in host defense against pathogenic organisms, and saprotrophic fungi decompose dead organic matter and facilitate nutrient cycling. In addition, pathogenic fungal species strongly affect forest fitness. Despite their importance, …

Range-Wide Prevalence And Impacts Of Pseudocercosporella Inconspicua On Lilium Grayi And An Assessment Of L. Superbum And L. Michauxii As Reservoirs, Cindy L. Barrett

Electronic Theses and Dissertations

Lilium grayi (Gray’s Lily), a southern Appalachian endemic species, is threatened by a Lilium-specific fungal pathogen, Pseudocercosporella inconspicua. The disease is characterized by tan lesions that can cause early senescence, while also lowering seed production and viability. This project tested for P. inconspicua conidia and accessed health at nine locations. The disease was present and ubiquitous across the range of L. grayi. Through identification of P. inconspicua conidia in the field, L. superbum (Turk’s Cap Lily) was identified as an additional host, while L. michauxii (Michaux’s Lily) was disease-free. However, infection was inducible in both species. With …

Do Novel Weapons That Degrade Mycorrhizal Mutualisms Explain Invasive Species Success?, Philip L. Pinzone Mr.

Biology Theses

Invasive plants often dominate novel habitats where they did not co-evolve with local species. Several hypotheses suggest mechanisms that explain increased exotic plant success, including 'novel weapons' and 'degraded mutualisms'. Japanese knotweed (Fallopia japonica) and European buckthorn (Rhamnus cathartica) are widespread plant invaders in North America that can dominate ecosystems. The goal of this study is to test whether these impacts are more consistent with novel weapons or degraded mutualism hypotheses. I examine tree seedling recruitment, (germination and initial survival) growth, (biomass) and mycorrhizal invasion (AMF content) as a function of F. japonica and R. cathartica …

Dissemination And Persistence Of Plasmid Located, Integron Associated Antibiotic Resistant Genes In Wastewater Treatment Plant Effluent And Stream Water Bacteria, _ Suhartono

Graduate Theses and Dissertations

Transmissible plasmid-mediated integrons play important role in the persistence and dissemination of antibiotic resistance throughout the environment. Plasmids from 139 multi-antibiotic resistant Escherichia coli recovered from wastewater treatment plant effluent and upstream and downstream receiving stream water in Northwest Arkansas were extracted and profiled. Genes of class 1 and class 2 integrase (intI), mobilization (mob), sulfamethoxazole resistance (sul), and trimethoprim resistance (dfr) were detected using PCR and confirmed through DNA sequencing. Plasmids from almost half of the isolates (47%) were transmissible with mobF12 gene as the most frequently detected mobilization gene. Plasmid-borne class 1 with and without class 2 integrons …

Global Worming: A Quantitative Study About Greenhouse Gas Flux In Surface Soils Facilitated By The Anecic Earthworm, Lumbricus Terrestris, Under Rising Global Temperature, Rachel Briden Frei, Jose A. Amador

Senior Honors Projects

Climate change is the long-term alteration in the Earth’s average weather conditions believed to be driven by greenhouse gases (GHG): carbon dioxide (CO₂), methane (CH₄) and nitrous oxide (N₂O). These alterations are expected to cause more extreme weather events, gradually warmer global temperatures and greater amounts of precipitation. Roughly 20% of the Earth’s CO₂, one-third of CH₄ and two-thirds of N₂O emissions, originate from soils, and earthworms are known to accelerate GHG. As climate change proceeds, there is expected to be an increase in global temperature of 2-6ºC. Temperature …

Effects Of Epichloë Coenophiala−Tall Fescue Symbiosis On Plant-Microbe-Soil Interactions In A Temperate Pasture, Lindsey C. Slaughter

Theses and Dissertations--Plant and Soil Sciences

Plants interact in myriad ways with microorganisms to influence ecosystem processes such as nutrient cycling, which can regulate ecosystem response to global change. One important plant-microbe symbiosis occurs between cool-season grasses and asexual fungal Epichloë endophytes, such as tall fescue (Schedonorus arundinaceus) and Epichloë coenophiala. Because the common toxic strain of the endophyte (CTE) harms grazing livestock, non-livestock toxic endophyte (NTE) strains have been developed and are increasingly deployed in pastures. Little is known about how these symbioses impact other plant-microbe interactions and microbe-mediated soil processes in grassland ecosystems. I conducted three studies to determine how E. …

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 …

Potential For Biocontrol Of Environmental Vibrio Spp. By Predatory Bacteria Bacteriovorax, Patrick Tennis

OES Theses and Dissertations

The scale of predator-prey interactions varies from the iconic (e.g., lions and gazelles) to the microscopic (e.g., protists and bacteria). Bdellovibrio-and-like-organisms (BALOs) are bacteria that prey upon other Gram-negative bacteria, including pathogens. In particular, the halophilic genus Vibrio, many species of which are disease agents, has been shown to be susceptible to attack by BALOs. One motivation of the present research is to assess the potential for biocontrol of Vibrio by Bacteriovorax. This investigation is the first to test the susceptibility of Vibrio strains (n = 26) originating from a variety of geographic locations to predation by …

Effects Of Treated Wastewater Effluent On Microbial Community Structure In A Natural Receiving Aquatic System, Matthew D. Hladilek

Masters Theses

Despite our dependency on treatment facilities to condition wastewater for eventual release to the environment, our knowledge regarding the effects of treated water on the local watershed is extremely limited. Responses of these lotic systems to the treated wastewater effluent have been traditionally investigated by examining the benthic macroinvertebrate assemblages and community structure; however, these studies do not address the microbial diversity of the water systems. In the present study, planktonic and benthic bacterial community structure were examined at fourteen sites (from 60 m upstream to 12,100 m downstream) and at two time points along an aquatic system receiving treated …

Effects Of Sex And Mycorrhizal Fungi On Gas Exchange In The Dioecious Salt Marsh Grass Distichlis Spicata, Kassandra Reuss-Schmidt, Todd N. Rosenstiel, Sally R. Rogers, Allie G. Simpson, Sarah M. Eppley

Biology Faculty Publications and Presentations

Premise of research. In dioecious plant species, males and females often differ in physiology, and mycorrhizal fungal relationships are likely to influence these differences. However, few data are available on the potential role of mycorrhizal fungi in altering sex-specific physiology and population sex ratios of dioecious plant species.

Methodology. In this study, we measured leaf gas exchange in a multifactorial greenhouse experiment with and without mycorrhizal fungal additions and under field conditions in Distichlis spicata, a dioecious C4 salt marsh grass, displaying extreme spatial sex ratio variation.

Pivotal results. We found a significant interaction between gas exchange, plant sex, 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 …

Interactions Of Various Bacterial Populations With Chemical And Physical Factors From Seasonal Inputs And Outputs Of Retention Ponds, Felicia A. Krelwitz

All Student Theses and Dissertations

Retention ponds at Governors State University play an important role in collecting and treating storm water runoff before leaving campus and entering Thorn Creek. Many chemical and physical factors influence the diversity of bacterial populations in freshwater ecosystems. The main objective of this study was to compare the interactions of various bacterial populations with chemical and physical factors from seasonal inputs and outputs of Governors State University retention ponds before being discharged into Thorn Creek. The retention ponds studied include the Café Settling pond, Café pond and Beaver pond. Water and sediment were collected from inputs and outputs every other …

Bioremediation Of Crude Oil Spills Impacts Microbial Communities And The Magnetic Susceptibility Of Sediments, Carol L. Beaver, Anja Williams, Estella Atekwana, Gamal Abdel Aal, Farag Mewafy, Lee Slater, Silvia Rossbach

Research and Creative Activities Poster Day

A worldwide increase in the production, shipment, and storage of crude oil has led to an increase in oil spills, and efforts to physically remove this oil from the environment are often insufficient. As a result, hydrocarbon contamination that is not eliminated by suction, excavation, or by dispersants remains in the water and sediments to be degraded by microbes. Initially, the bioremediation of oil occurs aerobically, but as oxygen is depleted, microbes must switch to alternate electron acceptors. Gradients of terminal electron acceptors form over time, which in conjunction with the toxic properties of oil, have an effect on the …

Managing Soil Microbial Communities With Organic Amendments To Promote Soil Aggregate Formation And Plant Health, Shawn T. Lucas

Theses and Dissertations--Plant and Soil Sciences

The effects of managing soil with organic amendments were examined with respect to soil microbial community dynamics, macroaggregate formation, and plant physio-genetic responses. The objective was to examine the possibility of managing soil microbial communities via soil management, such that the microbial community would provide agronomic benefits. In part one of this research, effects of three amendments (hairy vetch residue, manure, compost) on soil chemical and microbial properties were examined relative to formation of large macroaggregates in three different soils. Vetch and manure promoted fungal proliferation (measured via two biomarkers: fatty acid methyl ester 18:2ω6c and ergosterol) and also stimulated …

Interactions Of Zooplankton And Phytoplankton With Cyanobacteria, Rebecca Alexander

School of Natural Resources: Dissertations, Theses, and Student Research

Cyanobacteria are a major concern in Nebraska reservoirs and are capable of producing toxins that can cause skin irritations and gastrointestinal problems, as well as affect the nervous system. It is important to determine the mechanisms that can cause cyanobacteria blooms due to the effect they can have on human health. The interaction of zooplankton and other phytoplankton groups with cyanobacteria is important because there is a biological component in surface waters that should be taken into consideration along with the physical and chemical parameters that have been noted to promote cyanobacteria. For example, zooplankton have the ability to alter …

Fecal Indicator Bacteria In Knitting Mill Creek With A Comparison Of Methods Used To Enumerate E. Coli, Manju Nagarajan

OES Theses and Dissertations

This research seeks to evaluate the impact of an urban, multi-use neighborhood on the bacteriological quality of water in Knitting Mill Creek, a blind arm of the Lafayette River, a sub-estuary of the lower Chesapeake Bay. A principal objective was to determine the effects of rainfall, a surrogate for run-off, on water-column concentrations of fecal-indicator bacteria (FIB). I collected water samples weekly (September 2009-December 2010) and biweekly (January 2010-December 2011) at a storm-sewer outfall and a nearby marina and quantified their E. coli and enterococci concentrations using Colilert-18 and Enterolert, most-probable-number assays. In addition, I took monthly samples for these …

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 …

Biogeochemical Behavior Of Dissolved Arsenic And Uranium Concentrations In Public Water Supply Wells, Kevin J. Mcvey

School of Natural Resources: Dissertations, Theses, and Student Research

Public water supply (PWS) wells currently contain dissolved uranium concentrations above the federally mandated maximum contaminant level (MCL) of 30 ppb (parts per billion) and dissolved arsenic concentrations above the 10 ppb MCL. Both uranium and arsenic are known to cause various forms of cancer in humans. Variations in total uranium concentrations in PWS wells in Nebraska indicate a relationship to the duration and rate of pumping in specific wells. Although total arsenic concentrations show some variability over time in specific wells, the relationship to pumping is not as clear. Previous studies show that iron and sulfur bacteria present in …

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.

Diversity Of Estrogen Degrading Microorganisms In Las Vegas Wash And Lake Mead, Nevada, Usa, Susanna Blunt, Jim Bruckner, Jenny C. Fisher, Duane P. Moser

Lake Mead Science Symposium

Endocrine disrupting chemicals (EDCs) are a subject of intense research as more studies reveal their persistence in the environment and detrimental effects on wildlife. Steroid hormones, including the natural and synthetic estrogens estrone (E1), 17-beta-estradiol (E2) and 17- alpha-ethinyl estradiol (EE2), are among the most bioactive and have been detected at low concentrations in waterways downstream from wastewater treatment plants. Las Vegas Wash, a stream flowing into Lake Mead and fed primarily by treated wastewater, provides a unique experimental system in which to study the role microorganisms play in the fate and dispersal of these compounds in surface waters.

Spatial And Temporal Patterns In The Microbial Diversity Of A Meromictic Soda Lake In Washington State, Pedro A. Dimitriu, Holly C. Pinkart, Brent M. Peyton, Melanie R. Mormile

All Faculty Scholarship for the College of the Sciences

The microbial community diversity and composition of meromictic Soap Lake were studied using culture-dependent and culture-independent approaches. The water column and sediments were sampled monthly for a year. Denaturing gradient gel electrophoresis of bacterial and archaeal 16S rRNA genes showed an increase in diversity with depth for both groups. Late-summer samples harbored the highest prokaryotic diversity, and the bacteria exhibited less seasonal variability than the archaea. Most-probable-number assays targeting anaerobic microbial guilds were performed to compare summer and fall samples. In both seasons, the anoxic samples appeared to be dominated by lactate-oxidizing sulfate-reducing prokaryotes. High numbers of lactate- and acetate-oxidizing …

Environmental Controls On The Landscape-Scale Biogeography Of Stream Bacterial Communities, Noah Fierer, Jennifer L. Morse, Sean T. Berthrong, Emily S. Bernhardt, Robert B. Jackson

Environmental Science and Management Faculty Publications and Presentations

We determined the biogeographical distributions of stream bacteria and the biogeochemical factors that best explained heterogeneity for 23 locations within the Hubbard Brook watershed, a 3000-ha forested watershed in New Hampshire, USA. Our goal was to assess the factor, or set of factors, responsible for generating the biogeographical patterns exhibited by microorganisms at the landscape scale. We used DNA fingerprinting to characterize bacteria inhabiting fine benthic organic matter (FBOM) because of their important influence on stream nutrient dynamics. Across the watershed, streams of similar pH had similar FBOM bacterial communities. Streamwater pH was the single variable most strongly correlated with …

Microbial Nitrogen Limitation Increases Decomposition, Joseph M. Craine, Carl Morrow, Noah Fierer

Dartmouth Scholarship

With anthropogenic nutrient inputs to ecosystems increasing globally, there are long-standing, fundamental questions about the role of nutrients in the decomposition of organic matter. We tested the effects of exogenous nitrogen and phosphorus inputs on litter decomposition across a broad suite of litter and soil types. In one experiment, C mineralization was compared across a wide array of plants individually added to a single soil, while in the second, C mineralization from a single substrate was compared across 50 soils. Counter to basic stoichiometric decomposition theory, low N availability can increase litter decomposition as microbes use labile substrates to acquire …

Sulfur-Oxidizing Bacteria In Soap Lake (Washington State), A Meromictic, Haloalkaline Lake With An Unprecedented High Sulfide Content, Dimitry Y. Sorokin, Mirjam Foti, Holly C. Pinkart, Gerard Muyzer

All Faculty Scholarship for the College of the Sciences

Culture-dependent and -independent techniques were used to study the diversity of chemolithoautotrophic sulfur-oxidizing bacteria in Soap Lake (Washington State), a meromictic, haloalkaline lake containing an unprecedentedly high sulfide concentration in the anoxic monimolimnion. Both approaches revealed the dominance of bacteria belonging to the genus Thioalkalimicrobium, which are common inhabitants of soda lakes. A dense population of Thioalkalimicrobium (up to 10⁷ cells/ml) was found at the chemocline, which is characterized by a steep oxygen-sulfide gradient. Twelve Thioalkalimicrobium strains exhibiting three different phenotypes were isolated in pure culture from various locations in Soap Lake. The isolates fell into two groups …

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 …