Environmental Microbiology and Microbial Ecology Commons^™

The Human Oral Microbiome, Floyd E. Dewhirst, Tuste Chen, Jacques Izard, Bruce J. Paster, Anne C. R. Tanner, Wen-Han Yu, Abirami Lakshmanan, William G. Wade

Department of Food Science and Technology: Faculty Publications

The human oral cavity contains a number of different habitats, including the teeth, gingival sulcus, tongue, cheeks, hard and soft palates, and tonsils, which are colonized by bacteria. The oral microbiome comprises over 600 prevalent taxa at the species level, with distinct subsets predominating at different habitats. The oral microbiome has been extensively characterized by cultivation and culture-independent molecular methods such as 16S rRNA cloning. Unfortunately, the vast majority of unnamed oral taxa are referenced by clone numbers or 16S rRNA GenBank accession numbers, often without taxonomic anchors. The first aim of this research was to collect 16S rRNA gene …

Growth Of Lactic Acid Bacteria: Influence Of Protocooperation, Bacteriophage Infection, And Prebiotic Carbohydrates, Caitlin D. Goin

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

Lactic acid bacteria are a vital part of the fermented food industry and are the subject of much interest and research. Industry is especially interested in using modern molecular approaches to maintain and improve selected strains; several industrial uses could be improved by closer investigation, namely the protocooperation of S. thermophilus and L. bulgaricus, the role of CRISPRs in phage resistance, and the utilization of prebiotic carbohydrates. Questions such as what genes and pathways are shared during milk fermentation between S. thermophilus and L. bulgaricus, when and what genes are active during bacteriophage infection, and how and where …

Evolutionary Aspects Of Urea Utilization By Fungi, Dhammika H. M. L. P Navarathna, Steven D. Harris, David D. Roberts, Kenneth W. Nickerson

Kenneth Nickerson Papers

The higher fungi exhibit a dichotomy with regard to urea utilization. The hemiascomycetes use urea amidolyase (DUR1,2), whereas all other higher fungi use the nickel-containing urease. Urea amidolyase is an energy-dependent biotincontaining enzyme. It likely arose before the Euascomycete/Hemiascomycete divergence c. 350 million years ago by insertion of an unknown gene into one copy of a duplicated methylcrotonyl CoA carboxylase (MccA). The dichotomy between urease and urea amidolyase coincides precisely with that for the Ni/Co transporter (Nic1p), which is present in the higher fungi that use urease and is absent in those that do not. We suggest that …

Candida Albicans Cellwall Components And Farnesol Stimulate The Expression Of Both Inflammatory And Regulatory Cytokines In The Murine Raw264.7 Macrophage Cell Line, Suman Ghosh, Nina Howe, Katie Volk, Swetha Tati, Kenneth W. Nickerson, Thomas M. Petro

Kenneth Nickerson Papers

Candida albicans causes candidiasis, secretes farnesol, and switches from yeast to hyphae to escape from macrophages after phagocytosis. However, before escape, macrophages may respond to C. albicans’ pathogen-associated molecular patterns (PAMPs) through toll-like receptor 2 (TLR2) and dectin-1 receptors by expressing cytokines involved in adaptive immunity, inflammation, and immune regulation. Therefore, macrophages and the RAW264.7 macrophage line were challenged with C. albicans preparations of live wild-type cells, heat-killed cells, a live mutant defective in hyphae formation, a live mutant producing less farnesol, or an isolate producing farnesoic acid instead of farnesol. Interleukin-6 (IL-6), and IL-1b, IL- 10, and tumor …

Escherichia Coli O157:H7 And Other E. Coli Strains Share Physiological Properties Associated With Intestinal Colonization, Lisa Jacobsen, Lisa Durso, Tyrell Conway, Kenneth W. Nickerson

Kenneth Nickerson Papers

Escherichia coli isolates (72 commensal and 10 O157:H7 isolates) were compared with regard to physiological and growth parameters related to their ability to survive and persist in the gastrointestinal tract and found to be similar. We propose that nonhuman hosts in E. coli O157:H7 strains function similarly to other E. coli strains in regard to attributes relevant to gastrointestinal colonization.

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 …

An Essay On Evolution: Evolution And The Origin Of Life Are Separate And Distinct Concepts, Kenneth Nickerson

Kenneth Nickerson Papers

Two years ago on a Saturday morning, I was asked quite pointedly by two friends how I could be both spiritual and a scientist. My questioning friends felt that these qualities were incompatible. It turned out their church taught that scientists were ‘the enemy’ because scientists believe in evolution. Since that time, Letters to the Editor regarding evolution have attracted my attention. Many of the published letters stated that the author did not believe in evolution, and argued in support of his/her position that God had created life. The authors thoroughly confuse two concepts which are separate and distinct: Evolution …

Microbiology Of Oil Fly Larvae, Kenneth Nickerson, Bradley A. Plantz

Kenneth Nickerson Papers

One animal beautifully adapted to the viscous asphalt of the La Brea tar pits is the oil fly Helaeomyia petrolei (Syn. Psilopa). As a normal part of its carnivorous existence the oil fly larval guts are filled with tar, with no adverse effects. Surface sterilized larvae 5 contained ca. 2 x 10 heterotrophic bacteria per larva. These bacteria have been identified as a mixture of enteric bacteria, most commonly Providencia rettgeri, and Acinetobacter spp. These bacteria were clearly growing because their numbers in the larval guts were 100 to 1,000 times greater than in free oil/asphalt. There is …

Candida Albicans Tup1 Is Involved In Farnesol-Mediated Inhibition Of Filamentous-Growth Induction, Bessie W. Kebaara, Melanie L. Langford, Dhammika H. M. L. P. Navarathna, Raluca Dumitru, Kenneth W. Nickerson, Audrey L. Atkin

Kenneth Nickerson Papers

Candida albicans is a dimorphic fungus that can interconvert between yeast and filamentous forms. Its ability to regulate morphogenesis is strongly correlated with virulence. Tup1, a transcriptional repressor, and the signaling molecule farnesol are both capable of negatively regulating the yeast to filamentous conversion. Based on this overlap in function, we tested the hypothesis that the cellular response to farnesol involves, in part, the activation of Tup1. Tup1 functions with the DNA binding proteins Nrg1 and Rfg1 as a transcription regulator to repress the expression of hypha-specific genes. The tup1/tup1 and nrg1/nrg1 mutants, but not the rfg1/rfg1 mutant, failed to …

The Effect Of Residue C:N Ratio On The Turnover Of N And C In Various Soil Organic Matter Fractions, Ana B. Wingeyer

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

Identifying soil organic matter (SOM) fractions that contribute to soil indigenous nitrogen (N) supply and understanding their turnover under different management constitute necessary tools toward an efficient N use. The objectives of this study were: i) trace the endpoint of carbon (C) flux from residue inputs into SOM; and ii) asses the role of the light fraction (LF), mobile humic acid (MHA) fraction and calcium humate (CaHA) fraction as N sources for heterotrophic decomposition of fresh plant residues with contrasting C:N ratio. A long-term aerobic soil incubation was carried out on ¹⁵N-labeled soil samples from Lincoln and Mead, NE. …

Farnesol Restores Wild-Type Colony Morphology To 96% Of Candida Albicans Colony Morphology Variants Recovered Following Treatment With Mutagens, Ellen C. Jensen, Jacob M. Hornby, Nicole E. Pagliaccetti, Chuleeon M. Wolter, Kenneth Nickerson, Audrey L. Atkin

Kenneth Nickerson Papers

Candida albicans is a diploid fungus that undergoes a morphological transition between budding yeast, hyphal, and pseudohyphal forms. The morphological transition is strongly correlated with virulence and is regulated in part by quorum sensing. Candida albicans produces and secretes farnesol that regulates the yeast to mycelia morphological transition. Mutants that fail to synthesize or respond to farnesol could be locked in the filamentous mode. To test this hypothesis, a collection of C. albicans mutants were isolated that have altered colony morphologies indicative of the presence of hyphal cells under environmental conditions where C. albicans normally grows only as yeasts. All …

Prevalence And Serovars Of Salmonella In The Feces Of Free-Ranging White-Tailed Deer (Odocoileus Virginianus) In Nebraska, David G. Renter, David P. Gnad, Jan M. Sargeant, Scott E. Hygnstrom

School of Natural Resources: Faculty Publications

To determine the prevalence and serovars of Salmonella in free-ranging deer, we cultured feces from white-tailed deer (Odocoileus virginianus) harvested by hunters during a regular firearm season in southeastern Nebraska (USA). We recovered Salmonella from 5 (1%; 95% confidence interval: 0.37– 2.20%) of 500 samples and identified four different Salmonella enterica serovars [Litchfield (1), Dessau (1), Infantis (2), and Enteritidis (1)]. Although the prevalence of Salmonella in free-ranging deer appears to be low, the serovars recovered are known to be pathogenic to humans and animals.

Determination Of Mrna Half-Lives In Candida Albicans Using Thiolutin As A Transcription Inhibitor, Bessie W. Kebaara, Lindsey E. Nielsen, Kenneth Nickerson, Audrey L. Atkin

Kenneth Nickerson Papers

A method for determining mRNA half-lives in the polymorphic fungus Candida albicans is described. It employs growth in a defined medium, the inhibition of transcription with thiolutin (10–20 μg/mL), and quantitative Northern blotting. The method is effective for the A72, SC5314, and CAI-4 strains of C. albicans, and for mRNAs that have a wide variety of decay rates and steady-state abundances. The range of half-lives detected (from 4–168 min) shows that this method is effective for mRNAs with widely varying half-lives. The mRNA decay rates obtained are compared with those for orthologous mRNAs from Saccharomyces cerevisiae. This procedure …

High Phosphate (Up To 600 Mm) Induces Pseudohyphal Development In Five Wild Type Candida Albicans, Jacob M. Hornby, Raluca Dumitru, Kenneth Nickerson

Kenneth Nickerson Papers

A method is described for the formation of nearly 100% pseudohyphae populations of wild-type Candida albicans A72. The method employs fungal growth at 37 °C (ca. 5×10⁶ cells/ml) in a glucose–proline–N-acetyl-glucosamine medium supplemented with up to 600 mM phosphate (KH₂PO₄/K₂HPO₄ 1:1) at pH 6.5. Four other strains of C. albicans (MEN, 10261, SG5314 and CAI-4) also formed pseudohyphae under these conditions, although the phosphate response profiles differed in the concentration required for each strain to form pseudohyphae.

Quorum Sensing In Candida Albicans: Probing Farnesol’S Mode Of Action With 40 Natural And Synthetic Farnesol Analogs, Roman Shchepin, Jacob M. Hornby, Erin Burger, Timothy Niessen, Patrick H. Dussault, Kenneth Nickerson

Kenneth Nickerson Papers

The dimorphic fungus Candida albicans produces extracellular farnesol (3,7, 11-trimethyl-2,6,10-dodecatriene- 1-ol) which acts as a quorum-sensing molecule (QSM) to suppress filamentation. Of four possible geometric isomers of farnesol, only the E,E isomer possesses QSM activity. We tested 40 natural and synthetic analogs of farnesol for their activity in an N-acetylglucosamine-induced differentiation assay for germ tube formation (GTF). Modified structural features include the head group, chain length, presence or absence of the three double bonds, substitution of a backbone carbon by S, O, N, and Se heteroatoms, presence or absence of a 3-methyl branch, and the bulkiness of the hydrophobic …

Eight Gram-Negative Bacteria Are 10,000 Times More Sensitive To Cationic Detergents Than To Anionic Detergents, Soumitra Rajagopal, Nicole Eis, Kenneth Nickerson

Kenneth Nickerson Papers

In liquid culture, eight typical Gram-negative bacteria were ca. 10,000-fold more sensitive to cationic detergents than to the anionic detergent sodium dodecyl sulfate. Cetyltrimethylammonium bromide (CTAB) was inhibitory at concentrations ranging from 0.0006% to 0.01%. Four pseudomonads able to form biofilms were ca. 1000-fold more resistant to CTAB on Luria–Bertani agar plates than they were in liquid culture. A lasI mutant of Pseudomonas aeruginosa was only able to tolerate 0.1% CTAB on Luria–Bertani agar plates but could tolerate 5% CTAB when supplemented with homoserine lactone containing culture supernatants.

En culture liquide, huit bactéries Gram négatif typiques étaient ca. 10,000 fois …

Escherichia Coli O157:H7 In Free-Ranging Deer In Nebraska, David G. Renter, Jan M. Sargeant, Scott E. Hygnstrom, Jeff D. Hoffmann, Jerry R. Gillespie

School of Natural Resources: Faculty Publications

In order to determine the prevalence and distribution of the human pathogen, Escherichia coli O157:H7, in free-ranging deer, hunters were asked to collect and submit fecal samples from deer harvested during a regular firearm season (14–22 November 1998). Prior to the season, 47% of the hunters with permits in the southeastern Nebraska (USA) study area indicated a willingness to participate in the study. Approximately 25% of successful hunters in the area submitted deer fecal samples. Escherichia coli O157:H7 was cultured from four (0.25%) of 1,608 total samples submitted. All of the fecal samples that were properly identified (1,426) and all …

Pyoveridine Production By Pseudomonas Aeruginosa Exposed To Metals Or An Oxidative Stress Agent, Kim-Hien Thi Dao, Katherine E. Hamer, Christine L. Clark, Lawrence G. Harshman

School of Biological Sciences: Faculty Publications

Siderophores are low molecular mass compounds used by many microorganisms to scavenge dissolved iron, which is typically rare in environments. Pseudomonas aeruginosa (PAO1) was exposed to metals and methyl viologen in low-iron medium in order to monitor the abundance of a specific siderophore (pyoverdine) and monitor growth over time. In this study it was discovered that cadmium can stimulate pyoverdine abundance. Cadmium may interact with and sequester the Fur regulatory protein, which represses siderophore synthesis under conditions of iron sufficiency. Mercury did not stimulate pyoverdine production at concentrations that strongly inhibited bacterial growth, suggesting that pyoverdine production is not governed …

Chlorella Virus Pbcv-1 Encodes A Functional Homospermidine Synthase, Annette Kaiser, Michael Vollmert, Dorothea Tholl, Michael V. Graves, James Gurnon, Weimei Xing, Amber D. Lisec, Kenneth Nickerson, James L. Van Etten

Kenneth Nickerson Papers

Sequence analysis of the 330-kb genome of chlorella virus Paramecium bursaria chlorella virus 1 (PBCV-1) revealed an open reading frame, A237R, that encodes a protein with 34% amino acid identity to homospermidine synthase from Rhodopseudomonas viridis. Expression of the a237r gene product in Escherichia coli established that the recombinant enzyme catalyzes the NAD⁺-dependent formation of homospermidine from two molecules of putrescine. The a237r gene is expressed late in PBCV-1 infection. Both uninfected and PBCV-1-infected chlorella, as well as PBCV-1 virions, contain homospermidine, along with the more common polyamines putrescine, spermidine, and cadaverine. The total number of polyamine …

Can We Guarantee The Safety Of Genetically Engineered Organisms In The Environment?, Kathleen H. Keeler, Frances E. Sharples

School of Biological Sciences: Faculty Publications

Can we guarantee the safety of genetically engineered organisms in the environment? To anticipate my remarks, the answer to the title question is “No, we cannot guarantee the safety of genetically engineered organisms released into the environment.” Indeed, it is a tenet of the scientific method that nothing can be proved, only disproved. Thus, we can never show that a release will be safe. We could only show that it would be unsafe, if that were the case. However, if the question is posed differently, for example, can we safely release genetically engineered organisms into the environment?, the answer is …

Prevalence Of Extreme Detergent Resistance Among The Enterobacteriaceae, Vance Kramer, Kenneth Nickerson, Nancy V. Hamlett, Caroline O'Hara

Kenneth Nickerson Papers

The detergent-resistance properties of 208 independent isolates of the Enterobacteriaceae have been examined. Of these bacterial strains, 200 were able to grow in the presence of ≥5% sodium dodecyl sulfate, including all members of the Klebsielleae tribe. This resistance does not appear to be plasmid encoded. It is proposed that detergent-resistant organisms he termed saponotolerant or saponophilic, by analogy with other microorganisms occupying harsh ecological niches. In contrast to their prevalent resistance to anionic detergents, not one of the 208 strains tested was found to grow in the presence of three different cationic detergents. This sensitivity to cationic detergents may …

Microorganisms And Soil Structure, T. M. Mccalla

Department of Agronomy and Horticulture: Faculty Publications

SUMMARY

Laboratory tests were made to determine the effectiveness of different compounds and microbial groups in increasing the stability of Peorian loess lumps against the action of falling water drops. The influence of these on percolation tests in the laboratory was also determined.

Many organic substances-dextrose, sucrose, starch, peptone, cullulose, and gum arabic-did not themselves contribute directly to soil-structure stability, though these substances do furnish energy material for soil microorganisms, which can convert them readily into either microbial tissue or decomposition products that increase soil-structure stability. Lignin, proteins, oils, fats, waxes, resin, and paraffin increased the stability of lumps of …

Distribution Of Microorganisms In The Soil As Affected By Plowing And Subtilling Crop Residues, Roy C. Dawson, Virginia T. Dawson, T. M. Mccalla

Nebraska Agricultural Experiment Station: Historical Research Bulletins

The purpose of this investigation was primarily to determine what effects rate and method of application of residues have on the distribution of microorganisms within the plow-depth layer of soil.

Studies On Restaurant Sanitation And The Effect Of Sodium Azide On The Physiology Of Streptococcus Salivarius, Kenneth D. Rose

Open Access Master's Theses (through 2010)

Whether restaurant eating utensils serve as vectors in the transmission of respiratory and related diseases is still a matter for question, in spite of the overwhelming evidence in support of this contention.This is due to largely to the lack of epidemiological evidence, the tracing of an outbreak of a disease directly and unquestionably to contaminated eating utensils.The very nature of the vectors concerned obviates any possibility of obtaining this epidemiological evidence, since the contaminating agent exists as such only momentarily at the best. This research problem was undertaken with the hope that a technique designed to demonstrate the appearance of …

The Geographic Distribution Of Azotobacter And Rhizobium Meliloti In Nebraska Soils In Relation To Certain Environmental Factors, H. B. Peterson, T. H. Goodding

Nebraska Agricultural Experiment Station: Historical Research Bulletins

In this investigation a survey of Nebraska soils has been made in order to determine the distribution of the aerobic nonsymbiotic and symbiotic nitrogen fixers of the genera Azotobacter and Rhizobium respectively. In connection with this survey, some of the characteristics of the soils which may bring about this distribution were studied. Up to this time practically no research has been reported on the microflora of the soils of Nebraska. Hence there is little basis for predicting the activity of these organisms under environmental conditions as they exist here. It is hoped that this work will not only supply some …

Two Lincoln (Nebraska) Typhoid Fever Epidemics Of 1911 And 1912., Herbert H. Waite

School of Biological Sciences: Faculty Publications

On August 29, 1912, I was requested by the Board of Health of the city of Lincoln to make an investigation to determine if possible the origin and cause of an outbreak of a disease reported as typhoid fever which was then prevailing. This outbreak was said to be most prevalent in one particular portion of the city and consequently attention was turned to that portion. Of the cases reported an investigation was made of over 60 individuals and after considering all the probable causes for the majority of those affected the conclusion was reached that all could be logically …