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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 …

Intrinsic Challenges In Ancient Microbiome Reconstruction Using 16s Rrna Gene Amplification, Kirsten Ziesemer, Allison Mann, Krithivasan Sankaranarayanan, Hannes Schroeder, Andrew T. Ozga, Bernd W. Brandt, Egija Zaura, Andrea Waters-Rist, Menno Hoogland, Domingo C. Salazar-Garcia, Mark Aldenderfer, Camilla Speller, Jessica Hendy, Darlene A. Weston, Sandy J. Macdonald, Gavin H. Thomas, Matthew J. Collins, Cecil M. Lewis Jr., Corinne Hofman, Christina Warinner

Biology Faculty Articles

To date, characterization of ancient oral (dental calculus) and gut (coprolite) microbiota has been primarily accomplished through a metataxonomic approach involving targeted amplification of one or more variable regions in the 16S rRNA gene. Specifically, the V3 region (E. coli341–534) of this gene has been suggested as an excellent candidate for ancient DNA amplification and microbial community reconstruction. However, in practice this metataxonomic approach often produces highly skewed taxonomic frequency data. In this study, we use non-targeted (shotgun metagenomics) sequencing methods to better understand skewed microbial profiles observed in four ancient dental calculus specimens previously analyzed by amplicon …

Bacteriophages Isolated From Lake Michigan Demonstrate Broad Host-Range Across Several Bacterial Phyla, Kema Malki, Alex Kula, Katherine Bruder, Emily Sible, Thomas Hatzopoulos, Stephanie Steidel, Siobhan C. Watkins, Catherine Putonti

Biology: Faculty Publications and Other Works

BACKGROUND:

The study of bacteriophages continues to generate key information about microbial interactions in the environment. Many phenotypic characteristics of bacteriophages cannot be examined by sequencing alone, further highlighting the necessity for isolation and examination of phages from environmental samples. While much of our current knowledge base has been generated by the study of marine phages, freshwater viruses are understudied in comparison. Our group has previously conducted metagenomics-based studies samples collected from Lake Michigan - the data presented in this study relate to four phages that were extracted from the same samples.

FINDINGS:

Four phages were extracted from Lake Michigan …

A Tail Of Two Phages: Genomic And Functional Analysis Of Listeria Monocytogenes Phages Vb_Lmos_188 And Vb_Lmos_293 Reveal The Receptor-Binding Proteins Involved In Host Specificity, Aidan Casey, Kieran Jordan, Horst Neve, Aidan Coffey, Olivia Mcauliffe

Department of Biological Sciences Publications

The physical characteristics of bacteriophages establish them as viable candidates for downstream development of pathogen detection assays and biocontrol measures. To utilize phages for such purposes, a detailed knowledge of their host interaction mechanisms is a prerequisite. There is currently a wealth of knowledge available concerning Gram-negative phage-host interaction, but little by comparison for Gram-positive phages and Listeria phages in particular. In this research, the lytic spectrum of two recently isolated Listeria monocytogenes phages (vB_LmoS_188 and vB_LmoS_293) was determined, and the genomic basis for their observed serotype 4b/4e host-specificity was investigated using comparative genomics. The late tail genes of these …

Label-Free Surface-Enhanced Raman Spectroscopy-Linked Immunosensor Assay (Slisa) For Environmental Surveillance, Vinay Bhardwaj

FIU Electronic Theses and Dissertations

The contamination of the environment, accidental or intentional, in particular with chemical toxins such as industrial chemicals and chemical warfare agents has increased public fear. There is a critical requirement for the continuous detection of toxins present at very low levels in the environment. Indeed, some ultra-sensitive analytical techniques already exist, for example chromatography and mass spectroscopy, which are approved by the US Environmental Protection Agency for the detection of toxins. However, these techniques are limited to the detection of known toxins. Cellular expression of genomic and proteomic biomarkers in response to toxins allows monitoring of known as well as …

An Exploration Of The Phylogenetic Placement Of Recently Discovered Ultrasmall Archaeal Lineages, Jeffrey M. O'Brien

Honors Scholar Theses

In recent years, several new clades within the domain Achaea have been discovered. This is due in part to microbiological sampling of novel environments, and the increasing ability to detect and sequence uncultivable organisms through metagenomic analysis. These organisms share certain features, such as small cell size and streamlined genomes. Reduction in genome size can present difficulties to phylogenetic reconstruction programs. Since there is less genetic data to work with, these organisms often have missing genes in concatenated multiple sequence alignments. Evolutionary Biologists have not reached a consensus on the placement of these lineages in the archaeal evolutionary tree. There …

How Many Particles Are Present In The Air? Bioaerosol Detection Using An Air Particle Counter, Angie Pamela Rivera, Parag Vaishampayan

STAR Program Research Presentations

Relative cleanliness in terms of particle abundance in spacecraft assembly facilities is determined by particle counts carried out in clean rooms during resting conditions. Particle counters assess total particles and particle size distribution, but do not distinguish inert particles from biological particles, which may include bacterial spores that are resistant to standard cleanroom sterilization procedures. Current cleanroom certifications do not fully assess the effects of human presence on spacecraft contamination since humans are known symbionts to enumerate microorganisms and assessments are performed at rest when there is no human presence. In this study, contamination risks and bioburden in spacecraft assembly …

Collecting Diverse Microorganisms From Rover Spacecraft, Jennifer I. Jacobs, Arianna Jefferson, Heidi Aronson, James Tan, Wayne Schubert, Parag Vaishampayan

STAR Program Research Presentations

. The Planetary Protection discipline at NASA’s Jet Propulsion Laboratory develops and implements procedures to prevent both forward and backward contamination between the Earth and solar system bodies. However, there will always be some microorganisms that will be resistant to the strictest of sterilization methods. In order understand the microorganisms found on spacecraft during assembly, and to rapidly identify them, a mass spectrometry approach was developed. As an experimental approach, a custom database was created for a subset of microorganisms in the Planetary Protection Archive. In order to make the database as accurate and efficient as possible, several different procedures …

Isolation Of Possible Biocontrol Endophytic Bacteria From Solanum Tuberosum Effective Against Streptomyces Scabies., Annie Flatley, Luke Ogle, Adam Noel, Erica Fraley, Alaxandra Goodman, Donna Becker

Poster Sessions

Use of biological control offers a cost effective and environmentally safe method for controlling plant diseases. Biocontrol agents that can colonize roots and live endophytically within plant tissue should allow for effective disease control. The goal of this research was to develop protocols to isolate putative Streptomyces species from potato stem and tuber tissue due to their ability to produce inhibitory compounds which could potentially reduce diseases caused by Streptomyces scabies. Endophyte isolation from Solanum tuberosum (potato) plants (stem and tuber tissues) that were grown in a biocontrol field trial were the focus of this study. Several surface …

Use Of Two Pathogen-Inhibiting Streptomyces Isolates For Biocontrol Of Scab Disease Of Raphanus Sativus, Tyler Park, Nichole Klingler, Adam Noel, Luke Ogle, Annie Flatley, Donna Becker

Poster Sessions

Streptomyces species are ubiquitous soil bacteria that are promising as biological control agents due to their prolific antibiotic production that can inhibit soil-borne plant pathogens. This includes Streptomyces scabies, which causes scab disease on underground storage organs. The goal of this research was to test two Streptomyces isolates that have known inhibitory and biocontrol abilities against S. scabies in a potato (Solanum tuberosum) agricultural system for their effectiveness in biocontrol of this pathogen in a radish (Raphanus sativus) greenhouse assay. Pathogenic S. scabies were mixed into either pasteurized or unpasteurized soil prior to planting. Radish …

Prokaryotic Diversity In The Rhizosphere Of Organic, Intensive, And Transitional Coffee Farms In Brazil, Adam Caldwell, Livia Silva, Cynthia Da Silva, Cleber Ouverney

Faculty Publications, Biological Sciences

Despite a continuous rise in consumption of coffee over the past 60 years and recent studies showing positive benefits linked to human health, intensive coffee farming practices have been associated with environmental damage, risks to human health, and reductions in biodiversity. In contrast, organic farming has become an increasingly popular alternative, with both environmental and health benefits. This study aimed to characterize and determine the differences in the prokaryotic soil microbiology of three Brazilian coffee farms: one practicing intensive farming, one practicing organic farming, and one undergoing a transition from intensive to organic practices. Soil samples were collected from 20 …

Genomic Signatures Of Cooperation And Conflict In The Social Amoeba, Elizabeth A. Ostrowski, Yufeng Shen, Xiangjun Tian, Richard Sucgang, Huaiyang Jiang, Jiaxin Qu, Mariko Katoh-Kurasawa, Debra A. Brock, Christopher Dinh, Fremiet Lara-Garduno, Sandra L. Lee, Christie L. Kovar, Huyen H. Dinh, Viktoriya Korchina, Laronda Jackson, Shobha Patil, Yi Han, Lesley Chaboub, Gad Shaulsky, Donna M. Muzny, Kim C. Worley, Richard A. Gibbs, Stephen Richards, Adam Kuspa, Joan E. Strassmann, David C. Queller

Biology Faculty Publications & Presentations

• Molecular evolution analyses reveal the history of social conflict
• Genes that mediate social conflict show signatures of frequency-dependent selection
• Balanced polymorphisms suggest that cheating may be stable and endemic

Cooperative systems are susceptible to invasion by selfish individuals that profit from receiving the social benefits but fail to contribute. These so-called "cheaters" can have a fitness advantage in the laboratory, but it is unclear whether cheating provides an important selective advantage in nature. We used a population genomic approach to examine the history of genes involved in cheating behaviors in the social amoeba Dictyostelium discoideum, testing whether these genes experience …

Investigating The Driving Mechanisms Behind Differences In Bleaching And Disease Susceptibility Between Two Scleractinian Corals, Pseudodiploria Strigosa And Diploria Labyrinthiformis, Zoe A. Pratte

FIU Electronic Theses and Dissertations

Disease and bleaching are two conditions which commonly lead to coral death. Among coral species, susceptibility to disease and bleaching is variable, and Pseudodiploria strigosa tends to be diseased more than Diploria labyrinthiformis, while D. labyrinthiformis bleaches more readily. The focus of this dissertation was to investigate and compare multiple components of these two coral species, and identify how they may relate to disease and bleaching resistance. Compenetnts examined included the surface mucopolysacharide layer (SML) thickness, gene expression, microbial associates, and a white plague aquarium study. The SML thickness decresased with increasing temperature regardless of coral species, indicating that SML …

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 ( …

Epiphytic Diatom Community Structure In A Karst Riverine System, Gregory John Barren

Masters Theses & Specialist Projects

The goal of this study was to assess the epiphytic diatom community structure of two host species along a karst gradient in the upper Green River, Kentucky to a gain a better understanding of the role of diatoms in the food web. The host species studied were Podostemum ceratophyllum and Cladophora. Percent cover of P. ceratophyllum and Cladophora were quantified in the four study reaches. The host species were sampled near-shore and mid-channel in each reach in September and October of 2013. After diatoms were extracted from the host and enumerated the density and diversity were quantified. Twelve genera were …

Is There Specificity In A Defensive Mutualism Against Soil Versus Lab Nematodes, Dictyostelium Discoideum Farmers And Their Bacteria?, Boahemaa Adu-Oppong,, David C. Queller, Joan E. Strassmann

Biology Faculty Publications & Presentations

Background: The social amoeba Dictyostelium discoideum is a soil-dwelling microbe, which lives most of its life cycle in the vegetative stage as a predator of bacteria and as prey for nematodes. When bacteria are sparse, amoebae aggregate into a multicellular fruiting body. Some clones of D. discoideum have agriculture (Brock et al., 2011). They carry bacteria through the social stage, eat them prudently, and use some bacteria as defence against non-farming D. discoideum competitors. Caenorhabditis elegans preys on D. discoideum in the laboratory but does not encounter it in nature because C. elegans lives on rotten fruit. The nematode …

Concurrent Coevolution Of Intra-Organismal Cheaters And Resisters, S R. Levin, D A. Brock, David C. Queller, Joan E. Strassmann

Biology Faculty Publications & Presentations

The evolution of multicellularity is a major transition that is not yet fully understood. Specifically, we do not know whether there are any mechanisms by which multicellularity can be maintained without a single-cell bottleneck or other relatedness-enhancing mechanisms. Under low relatedness, cheaters can evolve that benefit from the altruistic behaviour of others without themselves sacrificing. If these are obligate cheaters, incapable of cooperating, their spread can lead to the demise of multicellularity. One possibility, however, is that cooperators can evolve resistance to cheaters. We tested this idea in a facultatively multicellular social amoeba, Dictyostelium discoideum. This amoeba usually exists as …

Marine Sequestration Of Carbon In Bacterial Metabolites, Oliver J. Lechtenfeld, Norbert Hertkorn, Yuan Shen, Matthias Witt, Ronald Benner

Faculty Publications

Linking microbial metabolomics and carbon sequestration in the ocean via refractory organic molecules has been hampered by the chemical complexity of dissolved organic matter (DOM). Here, using bioassay experiments and ultra-high resolution metabolic profiling, we demonstrate that marine bacteria rapidly utilize simple organic molecules and produce exometabolites of remarkable molecular and structural diversity. Bacterial DOM is similar in chemical composition and structural complexity to naturally occurring DOM in sea water. An appreciable fraction of bacterial DOM has molecular and structural properties that are consistent with those of refractory molecules in the ocean, indicating a dominant role for bacteria in shaping …

Changing Bacterial Growth Efficiencies Across A Natural Nutrient Gradient In An Oligotrophic Estuary, Amber A. Kiger

FIU Electronic Theses and Dissertations

Recent studies have characterized coastal estuarine systems as important components of the global carbon cycle. This study investigated carbon cycling through the microbial loop of Florida Bay by use of bacterial growth efficiency calculations. Bacterial production, bacterial respiration, and other environmental parameters were measured at three sites located along a historic phosphorus-limitation gradient in Florida Bay and compared to a relatively nutrient enriched site in Biscayne Bay. A new method for measuring bacterial respiration in oligotrophic waters involving tracing respiration of ¹³C-glucose was developed. The results of the study indicate that ¹³C tracer assays may provide a better …

Urbanization Changes The Composition And Bioavailability Of Dissolved Organic Matter In Headwater Streams, Thomas B. Parr, Christopher Cronan, Tsutomu Ohno, Stuart Findlay, Sean Smith, Kevin Simon

Publications

Population growth in cities has resulted in the rapid expansion of urbanized land. Most research and management of stream ecosystems affected by urban expansion has focused on the maintenance and restoration of biotic communities rather than their basal resources. We examined the potential for urbanization to induce bottom-up ecosystem effects by looking at its influence on dissolved organic matter (DOM) composition and bioavailability and microbial enzyme activity. We selected 113 headwater streams across a gradient of urbanization in central and southern Maine and used elemental and optical analyses, including parallel factor analysis of excitation-emission matrices, to characterize DOM composition. Results …

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. …

Examining Egg Surface Morphology And Microbial Content Of Sceloporus Virgatus Eggshells, Linnaea Arnett, Stacey Weiss Dr., Mark Martin Dr.

Summer Research

Eggs laid by the Sceloporus virgatus lizard are exposed to a variety of potentially protective microorganisms when passed through the cloaca, the opening at the end of both the reproductive and digestive tracts. Past data have indicated that eggs dissected out of the female before passing through the cloaca often have decreased hatching success, potentially due to the lack of protective bacteria on the dissected eggshell surface. This study examined the microorganisms found on both laid and dissected eggs at 0 weeks of incubation, as well as microbial and fungal content on the eggshells after 3 weeks of incubation in …

Proteomic Adaptations To Starvation Prepare Escherichia Coli For Disinfection Tolerance, Zhe Du, Renu Nandakumar, Kenneth W. Nickerson, Xu Li

Kenneth Nickerson Papers

Despite the low nutrient level and constant presence of secondary disinfectants, bacterial re-growth still occurs in drinking water distribution systems. The molecular mechanisms that starved bacteria use to survive low-level chlorine-based disinfectants are not well understood. The objective of this study is to investigate these molecular mechanisms at the protein level that prepare starved cells for disinfection tolerance. Two commonly used secondary disinfectants chlorine and monochloramine, both at 1 mg/L, were used in this study. The proteomes of normal and starved Escherichia coli (K12 MG1655) cells were studied using quantitative proteomics. Over 60-min disinfection, starved cells showed significantly higher disinfection …

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 …