Digital Commons Network^™

Enhancing Urban Water Quality Through Biological-Chemical Treatment: Aquatic Macroinvertebrate Community And Temporal Chlorophyll-A Response, Matthew Chaffee

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

With a growing human population, urbanization is impeding a plethora of natural waterways. Of these, urban ponds play a vital role in nutrient sequestration, flood prevention, and habitat sanctuaries. However, nutrient loading can reduce habitat effectiveness and promote harmful algae blooms. To reduce internal nutrient loads, a biological-chemical treatment strategy consisting of floating treatment wetlands (FTWs) and lanthanum were applied to two urban retention ponds, Densmore and Wilderness Ridge Ponds. To measure effectiveness, chlorophyll-a samples were collected and correlated with Sentinel-2. A novel band algorithm termed 3BR1 produced a strong correlation (R² = 0.72) to physical chlorophyll-a …

Connecting The Nebraska Water Quality Index To The Aquatic Microbial Community Of The North Platte River Basin, Nebraska, Paula R. Guastello

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

The Nebraska Water Quality Index, under development by the Nebraska Department of Environment and Energy, summarizes in a single value eight environmental parameters that have been monitored in Nebraska for nearly 20 years. Water quality parameters including those used in the Nebraska Water Quality Index have been shown in previous studies to impact bacterial growth. As such, this index has the potential to correlate with the freshwater microbial community. Here, I relate the Nebraska Water Quality Index to microbial community composition and structure using 16S rRNA gene amplicon sequence data collected from the North Platte River Basin, Nebraska. This index …

Gene Expression Changes Linked To Phenylpropanoid-Based Resistance To Fusarium Head Blight Of Wheat, Shiv Singla

Department of Plant Pathology: Dissertations, Theses, and Student Research

Fusarium graminearum is a devastating pathogen of wheat that causes Fusarium head blight (FHB) and contaminates the grain with the mycotoxin deoxynivalenol (DON). Resistance to FHB is quantitative and it is important to identify additional genes conferring resistance against it. The goal of this thesis was to examine if the constitutive expression of two sorghum phenylpropanoid pathway genes, SbCCoAOMT (encoding caffeoyl-CoA O-methyltransferase) and SbC3’H (encoding p-coumarate 3-hydroxylase), in the moderately-susceptible spring wheat CB037 can provide Type-I and Type-II resistance to F. graminearum and determine the underlying mechanisms of the enhanced resistance. The constitutive expression lines (CCoAOMT413, CCoAOMT421, C3H112, …

Assessing Functional Biodiversity For The Future Of Plants, Planet, And People, Ali Loker

Doctor of Plant Health Program: Dissertations and Student Research

Biodiversity plays a critical role in supporting life in global ecosystems and its links to ecosystem services and sustainability are recognized by scientific and non-scientific communities. Growing awareness of the importance of biodiversity is accelerated by discussions of its loss, and how to design interventions to conserve and mitigate a biodiversity crisis. Stakeholders are funding and implementing assessment strategies at various scales to help direct conservation efforts. There is also growing interest in measuring and communicating biodiversity outcomes.

Functional biodiversity characterizes the multiplicity of life forms into groups based on their diverse contributions to natural and agro-ecosystems. Assessing functional biodiversity …

Genome Sequence Of Sn1, A Bacteriophage That Infects Sphaerotilus Natans And Pseudomonas Aeruginosa, K.M. Damitha Gunathilake, Denise M. Tremblay, Pier-Luc Plante, Ellen Jensen, Kenneth Nickerson, Sylvain Moineau

Kenneth Nickerson Papers

Phage SN1 infects Sphaerotilus natans and Pseudomonas aeruginosa strains. Its genome consists of 61,858 bp (64.3% GC) and 89 genes, including 32 with predicted functions. SN1 genome is very similar to Pseudomonas phage M6, which contains hypermodified thymidines. Genome analyses revealed similar base-modifying genes as those found in M6.

Phage SN1 was isolated in 1979 from activated sludge samples obtained from a wastewater treatment plant (Lincoln, Nebraska, USA) using S. natans ATCC 13338 as the host (1, 2). An early study showed that the siphophage SN1 has unusual bases in its genome as confirmed by cellulose thin-layer chromatography (1). Its …

Discovering Novel Polyextremotolerant Fungi, And Determining Their Ecological Role Within The Biological Soil Crust Consortium, Erin Carr

School of Biological Sciences: Dissertations, Theses, and Student Research

The ecological niche of polyextremotolerant fungi within oligotrophic ecosystems such as biological soil crusts has not yet been determined. These fungi persist in locations where nutrients are depleted while simultaneously surrounded by autotrophic microbes such as algae and cyanobacteria. Yet it has not been shown that they are engaging in any exchange of nutrients the way lichens do. However, there is seemingly no other way for these fungi to obtain vital nutrients, such as carbon or nitrogen, other than from these microbes. Here we have isolated polyextremotolerant fungi from cold desert biological soil crusts which are a microbial biofilm that …

Tiny Earth, Tinier Microbes: An Experiential Learning Approach To Antibiotic Discovery, Emily Kassing

Honors Theses

Antimicrobial resistance is one of the greatest global health challenges of the 21^st century as antibiotic discovery has slowed even as scientific knowledge about AMR has progressed. At the same time, science education has turned to active learning approaches like CUREs, or course-based undergraduate research experiences, to achieve educational objectives while engaging students in real-life research. The Tiny Earth Project is a global research initiative that seeks to crowdsource antibiotic discovery by recruiting undergraduate students to screen soil samples for antibiotic producers. The goal of this study was to determine the viability of translating the Tiny Earth programming to …

Genomic Analysis Of Acetoanaerobium Sp. Vlb-1, An Anaerobic Bacterium Isolated From Nebraska's Eastern Saline Wetlands, Claire Martin

Honors Theses

Through the analysis of genetic code, the genomic and physiological characteristics of an environmental anaerobic bacterial isolate, Acetoanaerobium sp. strain VLB-1, can be determined and significant pathways can be identified in its significance to the environment. This particular organism was isolated from the Eastern Saline Wetlands in Lincoln, NE. With this isolate, an investigation into elemental and amino acid cycling via the Stickland reaction and the Wood-Ljungdahl pathway was conducted to determine possible metabolic products. The Stickland reaction is a relatively newly discovered pathway, observed in the genus Clostridium. A. sticklandii is the main model for this method of …

Farnesol Secretion As A Possible Driving Force For Maintaining Candida Albicans As A Diploid, Kenneth Nickerson, Cory Boone, Kory Parker

Kenneth Nickerson Papers

Candida albicans is a pathogenic dimorphic fungus which is invariably found as a diploid in patients. C. albicans secretes the sesquiterpene farnesol both as a quorum sensing molecule which blocks the yeast to hypha conversion and as a virulence factor for pathogenicity. 20-25 μM farnesol kills other competing yeasts and fungi, often by triggering apoptosis, and yet wild type diploid C. albicans tolerates 300-500 μM farnesol. The recent availability of 10 haploid strains of C. albicans (5 mating type aand 5 mating type α) allowed us to compare their production of and sensitivity to farnesol. On average, the heterozygous diploid …

Soil Microbial Community Dynamics In Response To Prescribed Extreme Fires Following Juniperus Virginiana Invasion In The Loess Canyons Of Nebraska, Julie A. Fowler

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

In Nebraska and other regions of the Great Plains, the conifer Juniperus virginiana (eastern redcedar) is converting grasslands to dense woodlands. This is driven by the interacting drivers of fire suppression, altered grazing regimes, climate change and other anthropogenic factors, impacting the provisioning of ecosystem services. This vegetation state transition modifies water resource regulation and biogeochemical cycles leading to altered edaphic properties including soil microbial community composition. To restore these grasslands and control J. virginiana spread, prescribed extreme burns are implemented as a management tool through local prescribed burn associations. We hypothesized that the alternative state transition to dense J. …

Dietary Fiber Utilization In The Gut: The Role Of Human Gut Microbes In The Degradation And Consumption Of Xylose-Based Carbohydrates, Elizabeth Drey

Department of Food Science and Technology: Dissertations, Theses, and Student Research

Xylans are a family of xylose-based polysaccharides naturally present in fruits, vegetables, and cereal grains. Resistant to digestion by host enzymes, xylans reach the large intestine intact, where they are utilized by members of the gut microbiome. They are initially hydrolyzed by primary degraders that utilize extracellular xylanases to cleave xylan into smaller oligomers. These xylooligosaccharides (XOS) can then either be further hydrolyzed by primary degraders or can cross-feed secondary consumers, including Bifidobacterium. While several Bifidobacterium species have metabolic systems for XOS, studies have shown these species grow poorly on longer XOS and xylan substrates. In this thesis, …

Sterol Biosynthesis In Four Green Algae: A Bioinformatic Analysis Of The Ergosterol Versus Phytosterol Decision Point, Adam Voshall, Nakeirah T.M. Christie, Suzanne L. Rose, Maya Khasin, James L. Van Etten, Jennifer E. Markham, Wayne Riekhof, Kenneth Nickerson

Kenneth Nickerson Papers

Animals and fungi produce cholesterol and ergosterol, respectively, while plants produce the phytosterols stigmasterol, campesterol, and β-sitosterol in various combinations. The recent sequencing of many algal genomes allows the detailed reconstruction of the sterol metabolic pathways. Here, we characterized sterol synthesis in two sequenced Chlorella spp., the free-living C. sorokiniana, and symbiotic C. variabilis NC64A. Chlamydomonas reinhardtii was included as an internal control and Coccomyxa subellipsoidea as a plant-like outlier. We found that ergosterol was the major sterol produced by Chlorella spp. and C. reinhardtii, while C. subellipsoidea produced the three phytosterols found in plants. In silico analysis of the …

Microbiota Accessible Carbohydrates And Susceptibility To Clostridioides Difficile Infection, Keegan Schuchart, Jennifer Auchtung, Thomas Auchtung

UCARE Research Products

This experiment looked into the effects of dietary microbiota accessible carbohydrates (MACs)on the composition of human GI microbe colonies and their susceptibility to pathogen infection. Three different concentrations of MAC ingrained media were tested in mini bioreactor arrays, treated with antibiotics, infected with Clostridioides difficile, then allowed to recover. Microbe community was examined for strain composition and diversity before and after the antibiotic treatment and Clostridioides difficile infection. After the initial mini bioreactor run, individual isolates were investigated in their ability to respond to specific MACs Arabinoglactina, Inulin, and Soluble starch at experimental concentrations.

Use Of Uav Imagery And Nutrient Analyses For Estimation Of The Spatial And Temporal Contributions Of Cattle Dung To Nutrient Cycling In Grazed Ecosystems, Amanda Shine

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

Nutrient inputs from cattle dung are crucial drivers of nutrient cycling processes in grazed ecosystems. These inputs are important both spatially and temporally and are affected by variables such as grazing strategy, water location, and the nutritional profile of forage being grazed. Past research has attempted to map dung deposition patterns in order to more accurately estimate nutrient input, but the large spatial extent of a typical pasture and the tedious nature of identifying and mapping individual dung pats has prohibited the development of a time- and cost-effective methodology. The first objective of this research was to develop and validate …

Triatomines: Trypanosomatids, Bacteria, And Viruses Potential Vectors?, Caroline Barreto Viera, Yanna Reis Praça, Kaio Luis Da Silva Bentes, Paula Beatriz Santiago, Sofia Marcelino Martins Silva, Gabriel Dos Santos Silva, Flávia Nader Motta, Izabela Marques Dourado Bastos, Jaime Martins De Santana, Carla Nunes De Araújo

Harold W. Manter Laboratory: Library Materials

Triatominae bugs are the vectors of Chagas disease, a major concern to public health especially in Latin America, where vector-borne Chagas disease has undergone resurgence due mainly to diminished triatomine control in many endemic municipalities. Although the majority of Triatominae species occurs in the Americas, species belonging to the genus Linshcosteus occur in India, and species belonging to the Triatoma rubrofasciata complex have been also identified in Africa, the Middle East, South-East Asia, and in the Western Pacific. Not all of Triatominae species have been found to be infected with Trypanosoma cruzi, but the possibility of establishing vector transmission …

Synthesis, Secretion, And Perception Of Abscisic Acid Regulates Stress Responses In Chlorella Sorokiniana, Maya Khasin, Rebecca E. Cahoon, Sophie Alvarez, Richard Beckeris, Seong-Il Eyun, Qidong Jia, Jean-Jack Riethoven, Kenneth Nickerson, Wayne R. Riekhof

Kenneth Nickerson Papers

Abscisic acid (ABA) is a phytohormone that has been extensively characterized in higher plants for its roles in seed and bud dormancy, leaf abscission, and stress responses. Genomic studies have identified orthologs for ABA-related genes throughout the Viridiplantae, including in unicellular algae; however, the role of ABA in algal physiology has not been characterized, and the existence of such a role has been a matter of dispute. In this study, we demonstrate that ABA is involved in regulating algal stress responses. Chlorella sorokiniana strain UTEX 1230 contains genes orthologous to those of higher plants which are essential for ABA …

How Science Makes America Great, Kenneth Nickerson

Kenneth Nickerson Papers

The proposed Federal Budget cuts funding for NIH, NSF, and CDC by 16% each and for the EPA by a whopping 31%. Ouch! The situation seems ironic because of President Trump’s slogan “Make America great again!”, which I agree with wholeheartedly. In addition to being a scientist, I also love history (especially military history) and science is at the top of my list of what has made the United States great. ... Why do we do it? I’m in science because it is rewarding. It is invigorating to discover how nature works. Scientists may act individually and in relative obscurity, …

Phytohormone Signaling In Chlorella Sorokiniana: Perspectives On The Evolution Of Plant Cell-To-Cell Signaling, Maya Khasin

School of Biological Sciences: Dissertations, Theses, and Student Research

Cell-to-cell communication is a key aspect of microbial physiology and population dynamics, and a cornerstone in understanding the evolution of multicellularity. Quorum sensing in bacteria is a canonical example of microbial cell-to-cell signaling, in which bacteria use small molecule signals in order to monitor their population size and modulate their physiology accordingly. We propose that the evolution of plant hormone signaling arose in unicellular green algae, analogously to quorum sensing in bacteria, and that the complexity of these pathways required the recruitment of increasingly specific enzymes to increasingly sophisticated gene networks throughout the course of phytohormone signaling evolution. Using Chlorella …

Quorum Sensing In Candida Albicans: Farnesol Versus Farnesoic Acid, Wayne R. Riekhof, Kenneth Nickerson

Kenneth Nickerson Papers

Candida albicans is a clinically important dimorphic fungus that exhibits either a budding yeast or a mycelial-hyphal or pseudohyphal growth, depending on environmental conditions. The yeast-to-mycelia morphologic transition, which is generally regarded as an important virulence determinant, depends on the inoculum size of liquid cultures. The yeast form is favored when cultures are inoculated at > 10⁶ cells∙mL^–1, whereas the mycelial form is favored at inoculum densities < 10⁶∙mL^–1. Farnesoic acid (FA) and farnesol (FOH) are two related sesquiterpene quorum- sensing molecules that, upon accumulation, prevent the yeast-to-mycelial conversion. Oh et al. showed that C. albicans …

Deciphering Fungal Dimorphism: Farnesol’S Unanswered Questions, Kenneth Nickerson, Audrey L. Atkin

Kenneth Nickerson Papers

Candida albicans excretes E,E-farnesol as a virulence factor and quorum sensing molecule that prevents the yeast to hyphal conversion. Polke et al. (2016) identified eed1Δ/Δ as the first farnesol hypersensitive mutant of C. albicans. eed1Δ/Δ also excretes 10X more farnesol and while able to form hyphae, it cannot maintain hyphae. This mutant enables new research into unanswered questions, including the existence of potential farnesol receptors and transporters, regulation of farnesol synthesis, and relationships among farnesol, germ tube formation and hyphal maintenance. The eed1 farnesol hypersensitivity can be explained by higher internal concentrations of farnesol or lower thresholds for response. One …

Fungi In The Healthy Human Gastrointestinal Tract, Heather E. Hallen-Adams, Mallory J. Suhr

Department of Food Science and Technology: Faculty Publications

Many species of fungi have been detected in the healthy human gut; however, nearly half of all taxa reported have only been found in one sample or one study. Fungi capable of growing in and colonizing the gut are limited to a small number of species, mostly Candida yeasts and yeasts in the family Dipodascaceae (Galactomyces, Geotrichum, Saprochaete). Malassezia and the filamentous fungus Cladosporium are potential colonizers; more work is needed to clarify their role. Other commonly-detected fungi come from the diet or environment but either cannot or do not colonize (Penicillium and Debaryomyces species, which are …

Comparative Genomics, Transcriptomics, And Physiology Distinguish Symbiotic From Free-Living Chlorella Strains, Cristian F. Quispe, Olivia Sonderman, Maya Khasin, Wayne R. Riekhof, James L. Van Etten, Kenneth Nickerson

Kenneth Nickerson Papers

Most animal–microbe symbiotic interactions must be advantageous to the host and provide nutritional benefits to the endosymbiont. When the host provides nutrients, it can gain the capacity to control the interaction, promote self-growth, and increase its fitness. Chlorella-like green algae engage in symbiotic relationships with certain protozoans, a partnership that significantly impacts the physiology of both organisms. Consequently, it is often challenging to grow axenic Chlorella cultures after isolation from the host because they are nutrient fastidious and often susceptible to virus infection. We hypothesize that the establishment of a symbiotic relationship resulted in natural selection for nutritional and metabolic …

From Milpas To The Market: A Study On The Use Of Metal Silos For Safer And Better Storage Of Guatemalan Maize, José Rodrigo Mendoza

Department of Food Science and Technology: Dissertations, Theses, and Student Research

This project aimed to implement the use of metal silos to improve quality and safety of maize consumed by inhabitants of the highlands of Guatemala. This manuscript includes a literature review of the maize production chain in Guatemala, a survey about agricultural practices used in the region of study, as well as a characterization of the analyzed maize regarding its mycoflora, nutritional composition, and insect infestation. To better understand the current situation regarding agricultural practices and maize consumption, a survey was carried out. Sample consisted of 280 families representing 14 rural communities distributed in the townships of Todos Santos and …

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 …

Killer Toxin From Several Food-Derived Debaryomyces Hansenii Strains Effective Against Pathogenic Candida Yeasts, Nabaraj Banjara, Kenneth Nickerson, Mallory J. Suhr, Heather E. Hallen-Adams

Kenneth Nickerson Papers

Candida yeasts are the dominant fungi in the healthy human microbiome, but are well-known for causing disease following a variety of perturbations. Evaluation of fungal populations from the healthy human gut revealed a significant negative correlation between the foodborne yeast, Debaryomyces hansenii, and Candida species. D. hansenii is reported to produce killer toxins (mycocins) effective against other yeast species. In order to better understand this phenomenon, a collection of 42 D. hansenii isolates was obtained from 22 cheeses and evaluated for killer activity against Candida albicans and Candida tropicalis over a range of temperatures and pH values. Twenty three …

Sequence-Based Methods For Detecting And Evaluating The Human Gut Mycobiome, Mallory J. Suhr, Nabaraj Banjara, Heather E. Hallen-Adams

Department of Food Science and Technology: Faculty Publications

We surveyed the fungal microbiota in 16 fecal samples from healthy humans with a vegetarian diet. Fungi were identified using molecular cloning, 454 pyrosequencing and a Luminex analyte specific reagent (ASR) assay, all targeting the ITS region of the rRNA genes. Fungi were detected in each fecal sample and at least 46 distinct fungal operational taxonomic units (OTUs) were detected, from two phyla — Ascomycota and Basidiomycota. Fusarium was the most abundant genus, followed by Malassezia, Penicillium, Aspergillus, and Candida. Commonly detected fungi such as Aspergillus and Penicillium, as well as known dietary fungi Agaricus bisporus and …

The Human Gut Mycobiome: Pitfalls And Potentials — A Mycologist's Perspective, Mallory J. Suhr, Heather E. Hallen-Adams

Department of Food Science and Technology: Faculty Publications

We have entered the Age of the Microbiome, with new studies appearing constantly and whole journals devoted to the human microbiome. While bacteria outnumber other gut microbes by orders of magnitude, eukaryotes are consistently found in the human gut, and are represented primarily by the fungi. Compiling 36 studies spanning from 1917 to 2015, we found at least 267 distinct fungal taxa have been reported from the human gut, and seemingly every new study includes one or more fungi not previously described from this niche. This diversity, while impressive, is illusory. If we examine gut fungi, we will quickly observe …

Fungi Inhabiting The Healthy Human Gastrointestinal Tract: A Diverse And Dynamic Community, Heather E. Hallen-Adams, Stephen D. Kachman, Jaehyoung Kim, Ryan Legge, Inés Martínez

Department of Food Science and Technology: Faculty Publications

Fungal DNA was selectively amplified, and the ITS region sequenced, from fecal samples taken from 45 healthy human volunteers at one (21 volunteers) or two (24 volunteers) time points. Seventy-two operational taxonomic units, representing two phyla and ten classes of fungi, were recovered. Candida yeasts, notably C. tropicalis (present in 51 samples), and yeasts in the Dipodascaceae (39 samples), dominated, while 38 OTUs were detected in a single sample each. Fungi included known human symbionts (Candida, Cryptococcus, Malassezia and Trichosporon spp.), common airborne fungi (Cladosporium sp.) and fungi known to be associated with food ( …

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 …

Candida Albicans Quorum Sensing Molecules Stimulate Mouse Macrophage Migration, Jessica C. Hargarten, Tyler C. Moore, Thomas M. Petro, Kenneth W. Nickerson, Audrey L. Atkin

Kenneth Nickerson Papers

The polymorphic commensal fungus Candida albicans causes life-threatening disease via bloodstream and intra-abdominal infections in immunocompromised and transplant patients. Although host immune evasion is a common strategy used by successful human fungal pathogens, C. albicans provokes recognition by host immune cells less capable of destroying it. To accomplish this, C. albicans white cells secrete a low-molecular-weight chemoattractive stimulant(s) of macrophages, a phagocyte that they are able to survive within and eventually escape from. C. albicans opaque cells do not secrete this chemoattractive stimulant( s). We report here a physiological mechanism that contributes to the differences in the interaction of C. …