Microbiology Commons^™

Genomic Degeneration And Reduction In The Fish Pathogen Mycobacterium Shottsi, David T. Gauthier, Janis H. Doss, M. Lagatta, T. Gupta, R.K. Karls, F. D. Quinn

Biological Sciences Faculty Publications

Mycobacterium shottsii is a dysgonic, nonpigmented mycobacterium originally isolated from diseased striped bass (Morone saxatilis) in the Chesapeake Bay, USA. Genomic analysis reveals that M. shottsii is a Mycobacterium ulcerans/Mycobacterium marinum clade (MuMC) member, but unlike the superficially similar M. pseudoshottsii, also isolated from striped bass, it is not an M. ulcerans ecovar, instead belonging to a transitional group of strains basal to proposed “Aronson” and “M” lineages. Although phylogenetically distinct from the human pathogen M. ulcerans, the M. shottsii genome shows parallel but nonhomologous genomic degeneration, including massive accumulation of pseudogenes accompanied by proliferation of …

Investigating Public Support For Biosecurity Measures To Mitigate Pathogen Transmission Through The Herpetological Trade, Elizabeth F. Pienaar, Diane J. Episcopio-Sturgeon, Zachary T. Steele

Biological Sciences Faculty Publications

The expanding global trade in herpetofauna has contributed to new infectious disease dynamics and pathways that allow for the rapid spread of pathogens geographically. Improved biosecurity is needed to mitigate adverse biodiversity, economic and human health impacts associated with pathogen transmission through the herpetological trade. However, general lack of knowledge of the pathogen transmission risks associated with the global trade in herpetofauna and public opposition to biosecurity measures are critical obstacles to successfully preventing pathogen transmission. In 2019 we administered a survey to 2,007 members of the public in the United States of America to ascertain their support for interventions …

Ozonized Biochar Filtrate Effects On The Growth Of Pseudomonas Putida And Cyanobacteria Synechococcus Elongatus Pcc 7942, Oumar Sacko, Nancy L. Engle, Timothy J. Tschaplinski, Sandeep Kumar, James Weifu Lee

Chemistry & Biochemistry Faculty Publications

Background

Biochar ozonization was previously shown to dramatically increase its cation exchange capacity, thus improving its nutrient retention capacity. The potential soil application of ozonized biochar warrants the need for a toxicity study that investigates its effects on microorganisms.

Results

In the study presented here, we found that the filtrates collected from ozonized pine 400 biochar and ozonized rogue biochar did not have any inhibitory effects on the soil environmental bacteria Pseudomonas putida, even at high dissolved organic carbon (DOC) concentrations of 300 ppm. However, the growth of Synechococcus elongatus PCC 7942 was inhibited by the ozonized biochar filtrates at …

Evidence For Metabolic Diversity In Meso-Neoproterozoic Stromatolites (Vazante Group, Brazil), Flavia Callefo, Fresia Ricardi-Branco, Mirian Mírian Liza Alves Forancelli Pacheco, Alexandre Ribeiro Cardoso, Nora Noffke, Verônica De Carvalho Teixeira, Itamar Tomio Neckel, Lara Maldanis, Emma Bullock, Dina Bower, Adalene Moreira Silva, Dario Ferreira Sanchez, Fabio Rodrigues, Douglas Galante

OES Faculty Publications

Deciphering the evolution of ecological interactions among the metabolic types during the early diversification of life on Earth is crucial for our understanding of the ancient biosphere. The stromatolites from the genus Conophyton cylindricus represent a datum for the Proterozoic (Meso to Neoproterozoic) on Earth. Their typical conical shape has been considered a result of a competition between microorganisms for space, light and nutrients. Well-preserved records of this genus from the "Paleontological Site of Cabeludo ", Vazante Group, São Francisco Craton (Southern Brazil) present in situ fossilized biofilms, containing preserved carbonaceous matter. Petrographic and geochemical analyses revealed an alternation between …

Defining The Realized Niche Of The Two Major Clades Of Trichodesmium: A Study On The West Florida Shelf, Kristina A. Confesor, Corday R. Selden, Kimberly E. Powell, Laura A. Donahue, Travis Mellett, Salvatore Caprara, Angela N. Knapp, Kristen N. Buck, P. Dreux Chappell

OES Faculty Publications

The cyanobacterium Trichodesmium plays an essential role supporting ocean productivity by relieving nitrogen limitation via dinitrogen (N-2) fixation. The two common Trichodesmium clades, T. erythraeum and T. thiebautii, are both observed in waters along the West Florida Shelf (WFS). We hypothesized that these taxa occupy distinct realized niches, where T. thiebautii is the more oceanic clade. Samples for DNA and water chemistry analyses were collected on three separate WFS expeditions (2015, 2018, and 2019) spanning multiple seasons; abundances of the single copy housekeeping gene rnpB from both clades were enumerated via quantitative PCR. We conducted a suite of statistical …

Mechanisms Of Stress Survival In Gram Positive Bacterial Pathogens, Asia Poudel

Chemistry & Biochemistry Theses & Dissertations

Anaerobic Gram-positive bacteria are not a well-characterized group but include many human pathogens that are resilient against stresses caused by the human immune system or by antibiotic treatment. This dissertation investigated the survival mechanisms of two clinically relevant Gram-positive organisms, Clostridioides difficile and Cutibacterium acnes under extracellular stresses. The response of the opportunistic skin pathogen Cutibacterium acnes to nanosecond electric pulses is characterized and found that growth in a biofilm, which usually protects bacteria from stress, renders this species more killable by this treatment. In addition, the stringent response (SR), a conserved bacterial stress survival mechanism, is studied in the …

Genetically Engineered Thermosynechococcus Elongatus Bp1: Assessment Of Potential Biorisks And Biofuel Production, Thu Ho Anh Nguyen-Jones

Chemistry & Biochemistry Theses & Dissertations

According to the International Energy Outlook 2019, released by the U.S. Energy Information Administration, it is projected that the energy consumption will increase up to 50% between 2018 and 2050 worldwide. As fossil fuel being a finite source of energy with the risk of depletion, many countries are now facing an energy security crisis. Therefore, it is important to develop other renewable and sustainable energy sources that will allow countries to shift away from depending on fossil fuels. Among several types of renewable energy, biofuel production using genetically engineered cyanobacteria is capturing much interest due to its many advantages. Different …

Evaluation Of Horizontal Gene Transfer Between Genetically Engineered Cyanobacteria And Gram-Negative Bacteria, Andriana Chrysovalanti Zourou

Chemistry & Biochemistry Theses & Dissertations

As the world population is increasing and societies become more technology driven, there is an imperative to develop ‘green energy’ sources to protect our planet. Cyanobacteria that have been genetically engineered to produce organic compounds that may be burnt as fuels show great potential, as they are an environmentally friendly and self-renewable, net carbon-neutral option. However, there are potential risks in the development and use of genetically modified organisms (GMOs). We need to understand in advance the risks that GMOs may pose to our environment and to animal and human health. This will enable experimental procedures, containment strategies and policies …

Evaluating The Role Of The Stringent Response Mechanism In Clostridioides Difficile Survival And Pathogenesis, Astha Pokhrel

Chemistry & Biochemistry Theses & Dissertations

The human pathogen Clostridioides difficile is increasingly tolerant of multiple antibiotics and causes infections with a high rate of recurrence, creating an urgent need for new preventive and therapeutic strategies. The stringent response, a universal bacterial response to extracellular stresses, governs antibiotic survival and pathogenesis in diverse organisms but has not previously been characterized in C. difficile. This dissertation explores the ability of C. difficile to mount the stringent response. The bacteria encode a full-length, canonical bifunctional Rel/Spo Homolog or RSH enzyme. C. difficile RSH is incapable of utilizing GTP as a substrate but readily synthesizes putative 5’-pGpp-3’ alarmones. …

Anaplasma Phagocytophilum Modulates Mammalian And Arthropod Signaling For Its Survival And Transmission, Supreet Khanal

Biomedical Sciences Theses & Dissertations

Vector-borne diseases (VBDs) are illnesses transmitted to humans and other animals by arthropods such as ticks, mosquitoes, and fleas. These arthropod vectors transmit infectious pathogens such as viruses, bacteria, and protozoa, to humans during blood-feeding. We have very few control strategies to treat or control these diseases. Human anaplasmosis, caused by the bacterium Anaplasma phagocytophilum, is the second most common tick-borne disease in the United States. This work defines three studies elucidating Anaplasma phagocytophilum-mediated modulation of cell signaling in mammalian cells and arthropod vector Ixodes scapularis ticks.

The first study focused on mammalian PI3 kinases signaling in regulating …

Biology And Molecular Biology Of Ixodes Scapularis, Daniel E. Sonenshine, Ladislav Šimo, Justin D. Radolf (Ed.), D. Scott Samuels (Ed.)

Biological Sciences Faculty Publications

This chapter describes the biology of the tick Ixodes scapularis in relation to its role as the vector of the Lyme disease agent, Borrelia burgdorferi. Following a review of the internal anatomy of the tick, we review basic molecular processes that contribute to an understanding of the dynamics of the tick's specialized parasitic processes, including attachment behavior, salivation; silencing of host anti-inflammatory responses to enable blood ingestion at the dermal feeding site; hemoglobin digestion and reproduction. The chapter is divided into three parts: 1) systematic and anatomical characteristics of ticks; 2) host finding, attachment, salivary disruption of host defenses, blood …

Advancing Cyanobacteria Biomass Estimation From Hyperspectral Observations: Demonstrations With Hico And Prisma Imagery, Ryan E. O'Shea, Nima Pahlevan, Brandon Smith, Mariano Bresciani, Todd Egerton, Claudia Giardino, Lin Li, Tim Moore, Antonio Ruiz-Verdu, Steve Ruberg, Stefan G.H. Simis, Richard Stumpf, Diana Vaičiūtė

Biological Sciences Faculty Publications

Retrieval of the phycocyanin concentration (PC), a characteristic pigment of, and proxy for, cyanobacteria biomass, from hyperspectral satellite remote sensing measurements is challenging due to uncertainties in the remote sensing reflectance (∆R_rs) resulting from atmospheric correction and instrument radiometric noise. Although several individual algorithms have been proven to capture local variations in cyanobacteria biomass in specific regions, their performance has not been assessed on hyperspectral images from satellite sensors. Our work leverages a machine-learning model, Mixture Density Networks (MDNs), trained on a large (N = 939) dataset of collocated in situ chlorophyll-a concentrations (Chla), …

Exploring The Niche Of Rickettsia Montanensis (Rickettsiales: Rickettsiaceae) Infection Of The American Dog Tick (Acari: Ixodidae), Using Multiple Species Distribution Model Approaches, Catherine A. Lippi, Holly Gaff, Alexis L. White, Heidi K. St. John, Allen L. Richards, Sadie J. Ryan

Biological Sciences Faculty Publications

The American dog tick, Dermacentor variabilis (Say) (Acari: Ixodidae), is a vector for several human diseasecausing pathogens such as tularemia, Rocky Mountain spotted fever, and the understudied spotted fever group rickettsiae (SFGR) infection caused by Rickettsia montanensis. It is important for public health planning and intervention to understand the distribution of this tick and pathogen encounter risk. Risk is often described in terms of vector distribution, but greatest risk may be concentrated where more vectors are positive for a given pathogen. When assessing species distributions, the choice of modeling framework and spatial layers used to make predictions are important. …

In The Margins: Reconsidering The Range And Contribution Of Diazotrophs In Nearshore Environments, Corday R. Selden

OES Theses and Dissertations

Dinitrogen (N₂) fixation enables primary production and, consequently, carbon dioxide drawdown in nitrogen (N) limited marine systems, exerting a powerful influence over the coupled carbon and N cycles. Our understanding of the environmental factors regulating its distribution and magnitude are largely based on the range and sensitivity of one genus, Trichodesmium. However, recent work suggests that the niche preferences of distinct diazotrophic (N₂ fixing) clades differ due to their metabolic and ecological diversity, hampering efforts to close the N budget and model N₂ fixation accurately. Here, I explore the range of N₂ fixation …

Drivers Of Rickettsial Pathogen Transmission And Spillover In Local Tick Populations In Southeastern Virginia, Alexandra Cumbie

Biomedical Sciences Theses & Dissertations

Cases of spotted fever group rickettsiosis are becoming more prevalent in the United States. In Virginia, there are three human-biting ticks which are largely responsible for the spread of rickettsial pathogens and the increase in disease cases. These species include Dermacentor variabilis, Amblyomma americanum, and Amblyomma maculatum; all of which are vectors of rickettsial agents to vertebrate hosts. These species are sympatric as adults and have the potential to share large and small mammal hosts. Their interactions on and off host and their associated rickettsiae were the focus of this dissertation work. Amblyomma americanum is the vector …

Ixodes Scapularis Src Kinase Is Required For Rickettsial Pathogen Survival In Ticks, Jeremy W. Turck

Biological Sciences Theses & Dissertations

Anaplasma phagocytophilum is an obligate intracellular bacterium that causes disease in humans and animals. It is the causative agent for human anaplasmosis. A. phagocytophilum uses certain strategies to infect both vertebrates and invertebrates. It uses Ixodes scapularis ticks as a vector for spreading infection to other mammal species. This bacterium has a specific path for infection through the salivary glands of its vector host. It also suppresses certain functions such as the inhibition of apoptosis and ROS production in order to increase its survival in ticks. Src kinase, a non-receptor protein-tyrosine kinase, is a major player in cell signaling. Src …

Copper And Cobalt Metal Conjugation Enhances Flaviviral Replication, And Denv/Zikv Caused Thrombocytopenia Suppresses Integrin Signaling, Shovan Dutta

Biomedical Sciences Theses & Dissertations

Vector control is a global concern, and effective strategies have been on the way. Among the many different strategies, insect repellents to protect us from blood-feeding arthropods are very common. Food processing or preservatives in food expose us to several chemicals associated with insect repellents, insecticides, and pesticides. Using metallic conjugation with plant extracts or essential oils has become a widespread strategy to increase their efficacy. But it is worth it to be aware that some of those metallic compounds like Cobalt or Copper upon conjugation with thiosemicarbazone ligands derived from Citral and 2-acetylethiazole at the center coordination enhance both …

Varroa Destructor Mites Vector And Transmit Pathogenic Honey Bee Viruses Acquired From An Artificial Diet, Francisco Posada-Florez, Eugene V. Ryabov, Matthew C. Heerman, Yanping Chen, Jay D. Evans, Daniel E. Sonenshine, Steven C. Cook

Biological Sciences Faculty Publications

The ectoparasitic mite Varroa destructoris one of the most destructive pests of the honey bee (Apis mellifera) and the primary biotic cause of colony collapse in many regions of the world. These mites inflict physical injury on their honey bee hosts from feeding on host hemolymph and fat body cells/cellular components, and serve as the vector for deadly honey bee viruses, including Deformed wing virus (DWV) and the related Varroa destructor virus-1 (VDV-1) (i.e., DWV-like viruses). Studies focused on elucidating the dynamics of Varroa-mediated vectoring and transmission of DWV-like viruses may be confounded by viruses present in …

Potential Virus-Mediated Nitrogen Cycling In Oxygen-Depleted Oceanic Waters, M. Consuelo Gazitúa, Dean R. Vik, Simon Roux, Ann C. Gregory, Benjamin Bolduc, Brittany Widner, Margaret R. Mulholland, Steven J. Hallam, Osvaldo Ulloa, Matthew B. Sullivan

OES Faculty Publications

Viruses play an important role in the ecology and biogeochemistry of marine ecosystems. Beyond mortality and gene transfer, viruses can reprogram microbial metabolism during infection by expressing auxiliary metabolic genes (AMGs) involved in photosynthesis, central carbon metabolism, and nutrient cycling. While previous studies have focused on AMG diversity in the sunlit and dark ocean, less is known about the role of viruses in shaping metabolic networks along redox gradients associated with marine oxygen minimum zones (OMZs). Here, we analyzed relatively quantitative viral metagenomic datasets that profiled the oxygen gradient across Eastern Tropical South Pacific (ETSP) OMZ waters, assessing whether OMZ …

Innovative Approaches In The Discovery Of Aquatic Mycobacteriophages, Janis H. Doss

Biomedical Sciences Theses & Dissertations

Bacteriophages (phages), viruses that infect bacteria, have many applications in medicine, agriculture, molecular biology, and other fields. As antibiotic resistance becomes an increasing problem, interest in phages has grown. The traditional techniques of phage discovery are successful for some phages, but others require modified procedures to achieve detectable host infection.

Mycobacterium is a diverse bacterial genus characterized by a unique cell wall containing mycolic acids, which aids in survival and pathogenesis. The aims of the present research were to isolate mycobacteriophages, use bioinformatics techniques to analyze mycobacterial prophages, and combine genetic analysis with multi-well plate host range studies to identify …

Repression Of Tick Microrna-133 Induces Organic Anion Transporting Polypeptide Expression Critical For Anaplasma Phagocytophilumsurvival In The Vector And Transmission To The Vertebrate Host, Ellango Ramasamy, Vikas Taank, John F. Anderson, Hameeda Sultana, Girish Neelakanta

Biological Sciences Faculty Publications

The microRNAs (miRNAs) are important regulators of gene expression. In this study, we provide evidence for the first time to show that rickettsial pathogen Anaplasma phagocytophilum infection results in the down-regulation of tick microRNA-133 (miR-133), to induce Ixodes scapularis organic anion transporting polypeptide (isoatp4056) gene expression critical for this bacterial survival in the vector and for its transmission to the vertebrate host. Transfection studies with recombinant constructs containing transcriptional fusions confirmed binding of miR-133 to isoatp4056 mRNA. Treatment with miR-133 inhibitor resulted in increased bacterial burden and isoatp4056 expression in ticks and tick cells. In contrast, treatment with …

Exosomes In Transmission And Blocking Of Arthropod-Borne Flaviviral Infection And Pathogenesis, Wenshuo Zhou

Biomedical Sciences Theses & Dissertations

Vector-borne diseases (VBDs) are human illnesses that are transmitted by vectors such as ticks, mosquitoes, and fleas. These arthropods transmit infectious pathogens such as viruses, protozoa, and bacteria, to humans during their blood-feeding. According to the estimation of the WHO, billions of people are affected by VBDs worldwide, and about 700,000 deaths are reported annually. Currently, very few reliable VBDs control approaches and vaccines available to treat various VBDs. Additionally, approaches to control arthropod’ proliferation and migration are difficult to develop. Furthermore, molecular determinants and mechanisms for the transmission are poorly understood. This work describes two studies identifying the novel …

Role Of Ixodes Scapularis Sphingomyelinase-Like Protein (Issmase) In Tick Pathogen Interactions, Pravesh Regmi

Biological Sciences Theses & Dissertations

Arthropod-borne diseases are one of the major concerns throughout the world. Ixodes scapularis (hard tick) is one of the major vectors that is involved in arthropod-borne disease transmission. Langat virus (LGTV) is a model pathogen that is very similar to other medically important flaviviruses such as Tick-Borne Encephalitis virus (TBEV) and Powassan virus (POWV). Sphingomyelinase-like protein (IsSMase, a Sphingomyelinase D or SMase D, a venomous protein ortholog of spiders) is an enzyme present in ticks that helps to catalyze the hydrolysis of the sphingomyelin (cell membrane lipid) into phosphocholine and ceramide. The objective of our study is to delineate the …

Diatom Community Composition Shifts Driven By Coherent Cyclonic Mesoscale Eddies In The California Current System, Zuzanna Maria Abdala

OES Theses and Dissertations

The California Current System (CCS) is characterized by an equatorward flowing eastern boundary current, as well as seasonal wind-driven coastal upwelling which supplies nutrient-rich waters to the surface and drives high coastal productivity. Cyclonic mesoscale eddies form off the coast in the CCS where they trap the highly productive upwelled coastal waters, along with their resident planktonic communities, and transport them offshore into the more oligotrophic California Current waters. The interaction between waters within and outside of the eddies is limited, and so the eddies act as natural mesocosms, where the resident phytoplankton population undergo ecological succession as the eddy …

Survival Of Enterococcus FaeciumIn Turkey Litter Under Different Temperature And Moisture Combinations, Steven G. Mcbride, Benjamin Holland, Pradeep Vasudevan, Joanna B. Mott

Virginia Journal of Science

Untreated poultry litter introduces a substantial load of fecal pathogens to the environment, impacting agriculture, public health and ecosystem function. There is substantial evidence that temperature and moisture are the primary drivers of fecal bacteria survival across ecosystems. However, both temperature and moisture effects have been shown to be modulated by the matrix in which the fecal bacteria are living. This context dependence highlights the importance of understanding fecal bacteria survival in a variety of matrices in order to implement effective waste management plans. In this study, we determined the survival patterns of Enterococcus faecium in post-use turkey litter under …

Isothermal Environmental Heat Energy Utilization By Transmembrane Electrostatically Localized Protons At The Liquid-Membrane Interface, James Weifu Lee

Chemistry & Biochemistry Faculty Publications

This study employing the latest theory on transmembrane electrostatic proton localization has now, for the first time, consistently elucidated a decades-longstanding bioenergetic conundrum in alkalophilic bacteria and more importantly discovered an entirely new feature: isothermal environmental heat utilization by electrostatically localized protons at the liquid-membrane interface. It was surprisingly revealed that the protonic motive force (equivalent to Gibbs free energy) from the isothermal environmental heat energy utilization through the electrostatically localized protons is not constrained by the overall energetics of the redox-driven proton pump system because of the following: (a) the transmembrane electrostatically localized protons are not free to move …

Factors Affecting The Microbiome Of Ixodes Scapularis And Amblyomma Americanum, R. Jory Brinkerhoff, Chris Clark, Kelly Ocasio, David T. Gauthier, Wayne L. Hynes

Biological Sciences Faculty Publications

The microbial community composition of disease vectors can impact pathogen establishment and transmission as well as on vector behavior and fitness. While data on vector microbiota are accumulating quickly, determinants of the variation in disease vector microbial communities are incompletely understood. We explored the microbiome of two human-biting tick species abundant in eastern North America (Amblyomma americanum and Ixodes scapularis) to identify the relative contribution of tick species, tick life stage, tick sex, environmental context and vertical transmission to the richness, diversity, and species composition of the tick microbiome. We sampled 89 adult and nymphal Ixodes scapularis (N …

Growth Phase Proteomics Of The Heterotrophic Marine Bacterium Ruegeria Pomeroyi, Dasha Krayushkina, Emma Timmins-Schiffman, Jessica Faux, Damon H. May, Michael Riffle, H. Rodger Harvey, Brook L. Nunn

OES Faculty Publications

The heterotrophic marine bacterium, Ruegeria pomeroyi, was experimentally cultured under environmentally realistic carbon conditions and with a tracer-level addition of ¹³C-labeled leucine to track bacterial protein biosynthesis through growth phases. A combination of methods allowed observation of real-time bacterial protein production to understand metabolic priorities through the different growth phases. Over 2000 proteins were identified in each experimental culture from exponential and stationary growth phases. Within two hours of the ¹³C-labeled leucine addition, R. pomeroyi significantly assimilated the newly encountered substrate into new proteins. This dataset provides a fundamental baseline for understanding growth phase differences in molecular …

Deepep: A Deep Learning Framework For Identifying Essential Proteins, Min Zeng, Min Li, Fang-Xiang Wu, Yaohang Li, Yi Pan

Computer Science Faculty Publications

Background: Essential proteins are crucial for cellular life and thus, identification of essential proteins is an important topic and a challenging problem for researchers. Recently lots of computational approaches have been proposed to handle this problem. However, traditional centrality methods cannot fully represent the topological features of biological networks. In addition, identifying essential proteins is an imbalanced learning problem; but few current shallow machine learning-based methods are designed to handle the imbalanced characteristics. Results: We develop DeepEP based on a deep learning framework that uses the node2vec technique, multi-scale convolutional neural networks and a sampling technique to identify essential proteins. …

Acute Systemic Inflammatory Response To Lipopolysaccharide Stimulation In Pigs Divergently Selected For Residual Feed Intake, Haibo Liu, Kristina M. Feye, Yet T. Nguyen, Anoosh Rakhshandeh, Crystal L. Loving, Jack C. M. Sekkers, Nicholas K. Gabler, Christopher K. Tuggle

Mathematics & Statistics Faculty Publications

Background: It is unclear whether improving feed efficiency by selection for low residual feed intake (RFI) compromises pigs’ immunocompetence. Here, we aimed at investigating whether pig lines divergently selected for RFI had different inflammatory responses to lipopolysaccharide (LPS) exposure, regarding to clinical presentations and transcriptomic changes in peripheral blood cells.

Results: LPS injection induced acute systemic inflammation in both the low-RFI and high-RFI line (n = 8 per line). At 4 h post injection (hpi), the low-RFI line had a significantly lower (p= 0.0075) mean rectal temperature compared to the high-RFI line. However, no significant differences in complete blood count …