Microbiology Commons^™

Phytoplankton And Nutrient Dynamics In A Tidally Dominated Eutrophic Estuary: Daily Variability And Controls On Bloom Formation, Ryan E. Morse, Margaret R. Mulholland, Todd A. Egerton, Harold G. Marshall

OES Faculty Publications

To better understand nutrient dynamics and factors that promote the initiation of algal blooms, the Lafayette River, a tidal subestuary of Chesapeake Bay that experiences seasonal algal blooms, was sampled daily for a period of 54 d in the fall of 2005. Three phytoplankton blooms (chl a concentrations exceeding twice the average of monthly measurements from 2000 to 2009) occurred during this period: a mixed bloom of Akashiwo sanguinea and Gymnodinium sp., a monospecific Skeletonema costatum bloom, and a monospecific Gymnodinium sp. bloom. Over the sampling period, nutrient concentrations increased following precipitation events and were elevated between bloom periods but …

Electrotransfer Of Single-Stranded Or Double-Stranded Dna Induces Complete Regression Of Palpable B16.F10 Mouse Melanomas, Loree Heller, Vesba Todorovic, Maja Cemazar

Bioelectrics Publications

Enhanced tumor delivery of plasmid DNA with electric pulses in vivo has been confirmed in many preclinical models. Intratumor electrotransfer of plasmids encoding therapeutic molecules has reached Phase II clinical trials. In multiple preclinical studies, a reduction in tumor growth, increased survival or complete tumor regression have been observed in control groups in which vector or backbone plasmid DNA electrotransfer was performed. This study explores factors that could produce this antitumor effect. The specific electrotransfer pulse protocol employed significantly potentiated the regression. Tumor regression was observed after delivery of single-stranded or double-stranded DNA with or without CpG motifs in both …

Regulation Of Hyaluronate Lyase (Hyla) Expression By Regr A Transcriptional Repressor In Streptococcus Pyogenes, Alexis A. Kordis

Biological Sciences Theses & Dissertations

Streptococcus pyogenes, group A streptococci (GAS), causes diseases ranging from asymptomatic to life threatening. Some strains of S. pyogenes produce extracellular hyaluronate lyase (HylA), a potential virulence factor. HylA is an enzyme that degrades hyaluronic acid, which is found in the extracellular matrix of human tissues. The breakdown of the host tissue contributes to the spread of infection. The hylA gene is not constitutively expressed in vitro, which implies regulation.

A proposed regulator of hyaluronate lyase expression is RegR, a LacI/GalR like protein. A vector containing disrupted regR was electro-transformed into S. pyogenes M-type 22 strain 10403. Growth characteristics …

In Vitro Selection Of Chloroquine Tolerant Plasmodium Falciparum Parasites, Daniel A. Daley

Biological Sciences Theses & Dissertations

Malaria is among the most devastating human diseases, and a majority of lethal cases are caused by the protozoan parasite, Plasmodiumfalciparum. The emergence of multi-drug resistant P.falciparum is a major obstacle to malaria control and is highlighted by the abandonment of chloroquine (CQ) as a first-line treatment of P.falciparum infections worldwide. Chloroquine resistance (CQR) is associated primarily with mutations in the transmembrane digestive vacuole protein, PfCRT. However, CQR P.falciparum parasites harboring the same mutant pfcrt allele vary in their CQ response, suggesting the CQ response is multigenic in nature. No gene outside of pfcrt is completely associated with CQR. Thus, …

Section Abstracts: Biology With Microbiology And Molecular Biology

Virginia Journal of Science

Abstracts of the Biology with Microbiology and Molecular Biology Section for the 91st Annual Virginia Journal of Science Meeting, May 2013

Uncoupling Between Dinitrogen Fixation And Primary Productivity In The Eastern Mediterranean Sea, Eyal Rahav, Barak Herut, Noga Stambler, Edo Bar-Zeev, Margaret R. Mulholland

OES Faculty Publications

In the nitrogen (N)-impoverished photic zones of many oceanic regions, prokaryotic organisms fixing atmospheric dinitrogen (N₂; diazotrophs) supply an essential source of new nitrogen and fuel primary production. We measured dinitrogen fixation and primary productivity (PP) during the thermally stratified summer period in different water regimes of the oligotrophic eastern Mediterranean Sea, including the Cyprus Eddy and the Rhodes Gyre. Low N₂ fixation rates were measured (0.8-3.2μmol N m-2 d-1) excluding 10-fold higher rates in the Rhodes Gyre and Cyprus Eddy (~20μmol N m^-2 d^-1). The corresponding PP increased from east to west (200-2500μmol …

Cpg And Interleukin-15 Synergize To Enhance Ifn-Gamma Production By Activated Cd8(+) T Cells, Dustin Cobb, Sigi Guo, Ronald B. Smeltz

Bioelectrics Publications

Interleukin-15 (IL-15) regulates the development and maintenance of memory CD8(+) T cells. Paradoxically, we previously reported that IL-15 could enhance CD8(+) T-cell responses to IL-12, a proinflammatory cytokine required for optimal priming of effector CD8(+) T cells. To expand the physiological relevance of these findings, we tested IL-15 for its ability to enhance T-cell responses to bacterial CpG. Expectedly, CpG enhanced the production of IFN-gamma by CD8(+) T cells polyclonally activated with anti-CD3. However, addition of IL-15 to CpG-stimulated cultures led to a striking increase in IFN-gamma production. The effect of CpG and IL-15 was also evident with CD8(+) T …

Combined Effects Of Co2 And Light On Large And Small Isolates Of The Unicellular N2-Fixing Cyanobacterium Crocosphaera Watsonii From The Western Tropical Atlantic Ocean, Nathan S. Garcia, Fei-Xue Fu, Cynthia L. Breene, Elizabeth K. Yu, Peter W. Bernhardt, Margaret R. Mulholland, David A. Hutchins

OES Faculty Publications

We examined the combined effects of light and pCO₂ on growth, CO₂-fixation and N₂-fixation rates by strains of the unicellular marine N₂-fixing cyanobacterium Crocosphaera watsonii with small (WH0401) and large (WH0402) cells that were isolated from the western tropical Atlantic Ocean. In low-pCO₂-acclimated cultures (190ppm) of WH0401, growth, CO₂-fixation and N₂-fixation rates were significantly lower than those in cultures acclimated to higher (present-day approximate to 385ppm, or future approximate to 750ppm) pCO² treatments. Growth rates were not significantly different, however, in low-pCO₂-acclimated cultures of …

The Relative Contribution Of Methanotrophs To Microbial Communities And Carbon Cycling In Soil Overlying A Coal-Bed Methane Seep, Christopher T. Mills, Gregpry F. Slater, Robert F. Dias, Stephanie A. Carr, Christopher M. Reddy, Raleigh Schmidt, Kevin W. Mandernack

Chemistry & Biochemistry Faculty Publications

Seepage of coal-bed methane (CBM) through soils is a potential source of atmospheric CH₄ and also a likely source of ancient (i.e. ¹⁴C-dead) carbon to soil microbial communities. Natural abundance ¹³C and ¹⁴C compositions of bacterial membrane phospholipid fatty acids (PLFAs) and soil gas CO₂ and CH₄ were used to assess the incorporation of CBM-derived carbon into methanotrophs and other members of the soil microbial community. Concentrations of type I and type II methanotroph PLFA biomarkers (16:1ω8c and 18:1ω8c, respectively) were elevated in CBM-impacted soils compared with a control site. Comparison …

The Use Of Metagenomic Approaches To Analyze Changes In Microbial Communities, Girish Neelakanta, Hameeda Sultana

Biological Sciences Faculty Publications

Microbes are the most abundant biological entities found in the biosphere. Identification and measurement of microorganisms (including viruses, bacteria, archaea, fungi, and protists) in the biosphere cannot be readily achieved due to limitations in culturing methods. A non-culture based approach, called “metagenomics”, was developed that enabled researchers to comprehensively analyse microbial communities in different ecosystems. In this study, we highlight recent advances in the field of metagenomics for analyzing microbial communities in different ecosystems ranging from oceans to the human microbiome. Developments in several bioinformatics approaches are also discussed in context of microbial metagenomics that include taxonomic systems, sequence databases, …

Thalassiosira Spp. Community Composition Shifts In Response To Chemical And Physical Forcing In The Northeast Pacific Ocean, P. Dreux Chappell, Leann P. Whitney, Traci L. Haddock, Susanne Menden-Deuer, Eric G. Roy, Mark L. Wells, Bethany D. Jenkins

Biological Sciences Faculty Publications

Diatoms are genetically diverse unicellular photosynthetic eukaryotes that are key primary producers in the ocean. Many of the over 100 extant diatom species in the cosmopolitan genus Thalassiosira are difficult to distinguish in mixed populations using light microscopy. Here, we examine shifts in Thalassiosira spp. composition along a coastal to open ocean transect that encountered a 3-month-old Haida eddy in the northeast Pacific Ocean. To quantify shifts in Thalassiosira species composition, we developed a targeted automated ribosomal intergenic spacer analysis (ARISA) method to identify Thalassiosira spp. in environmental samples. As many specific fragment lengths are indicative of individual Thalassiosira spp., …

Next-Peak: A Normal-Exponential Two-Peak Model For Peak-Calling In Chip-Seq Data, Nak-Kyeong Kim, Rasika V. Jayatillake, John L. Spouge

Mathematics & Statistics Faculty Publications

Background: Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) can locate transcription factor binding sites on genomic scale. Although many models and programs are available to call peaks, none has dominated its competition in comparison studies.

Results: We propose a rigorous statistical model, the normal-exponential two-peak (NEXT-peak) model, which parallels the physical processes generating the empirical data, and which can naturally incorporate mappability information. The model therefore estimates total strength of binding (even if some binding locations do not map uniquely into a reference genome, effectively censoring them); it also assigns an error to an estimated binding location. The comparison study …

Stromatolites And Miss—Differences Between Relatives, N. Noffke, S. M. Awramik

OES Faculty Publications

Benthic microorganisms form highly organized communities called “biofilms.” A biofilm consists of the individual cells plus their extracellular polymeric substances (EPS). In marine and non-marine environments, benthic microbial communities interact with the physical sediment dynamics and other factors in the environment in order to survive. This interaction can produce distinctive sedimentary structures called microbialites. Binding, biostabilization, baffling, and trapping of sediment particles by microorganisms result in the formation of microbially induced sedimentary structures (MISS); however, if carbonate precipitation occurs in EPS, and these processes happen in a repetitive manner, a multilayered build-up can form—stromatolites. Stromatolites and MISS are first found …

Dinitrogen Fixation In Aphotic Oxygenated Marine Environments, Eyal Rahav, Edo Bar-Zeev, Sarah Ohayon, Hila Elifantz, Natalia Belkin, Barak Herut, Margaret R. Mulholland, Iiana Berman-Frank

OES Faculty Publications

We measured N₂ fixation rates from oceanic zones that have traditionally been ignored as sources of biological N₂fixation; the aphotic, fully oxygenated, nitrate (NO¯₃)-rich, waters of the oligotrophic Levantine Basin (LB) and the Gulf of Aqaba (GA). N₂ fixation rates measured from pelagic aphotic waters to depths up to 720 m, during the mixed and stratified periods, ranged from 0.01 nmol N L^-1 d^-1 to 0.38 nmol N L^-1 d^-1. N₂ fixation rates correlated significantly with bacterial productivity and heterotrophic diazotrophs were identified from aphotic as well as …

Inactivation Of Bacterial Opportunistic Skin Pathogens By Nonthermal Dc-Operated Afterglow Atmospheric Plasma, L. C. Heller, C. M. Edelblute, A. M. Mattson, X. Hao, J. F. Kolb

Bioelectrics Publications

AIMS: Multidrug-resistant opportunistic pathogens are clinically significant and require the development of new antimicrobial methods. In this study, Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus aureus cells were exposed to atmospheric plasma on agar plates and in vitro on porcine skin for the purpose of testing bacterial inactivation.

METHODS AND RESULTS: Microbial inactivation at varying exposure durations was tested using a nonthermal plasma jet generated with a DC voltage from ambient air. The observed reduction in colony forming units was quantified as log₁₀ reductions.

CONCLUSIONS: Direct plasma exposure significantly inactivated seeded bacterial cells by approx. 6 log_{10 …}

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

OES Theses and Dissertations

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

Identification Of Persistent Long Range Interactions In GA95 And GB95 Through Thermal Unfolding Simulations, Milen Redai Tesfamariam

Chemistry & Biochemistry Theses & Dissertations

For over five decades, different experiments have been performed to research how proteins attain their native three dimensional structures. However, the folding problem continues to be a puzzle in modern science. The design of two proteins that have maximal sequence identity but different folds and functions is one method that is being used to study the relationship between protein structure and amino acid sequence. In particular, mutant proteins of Streptococcus protein G, G_A and G_B, have 95% sequence identity and a 3a helix fold and β4/a fold, respectively. Molecular dynamics simulations of G_A95 …

Section Abstracts: Biology With Microbiology And Molecular Biology

Virginia Journal of Science

Abstracts of the Biology with Microbiology and Molecular Biology Section for the 90th Annual Meeting of the Virginia Academy of Science, May 23-25, 2015, Norfolk State University, Norfolk, Virginia.

Effects Of The Parasitic Dinoflagellate Hematodinium Sp. On Blue Crab (Callinectes Sapidus) Activity Predation And Habitat Selection, John M. Tiggelaar Ii

Biological Sciences Theses & Dissertations

The blue crab Callinectes sapidus occurs along the Atlantic and Gulf coasts of the United States, and in high salinity regions, they are subject to lethal infection by the parasitic dinoflagellate Hematodinium. In Virginia's seaside estuaries, the prevalence of Hematodinium infection of C. sapidus can sometimes exceed 50%, threatening the commercial fishery for this species. Indeed, other commercially important crustacean fisheries have approached collapse due to Hematodinium infections. Most studies of this host-parasite interaction have focused on epidemiology, host-pathogen dynamics, and pathogen transmission, and little is known about the impact of the parasite on host behavior and population dynamics. …

Regulation Of The Vapbc-1 Toxin-Antitoxin Locus In Nontypeable Haemophilus Influenzae, Susan D. Cline, Sehresh Saleem, Dayle A. Daines

Biological Sciences Faculty Publications

Nontypeable Haemophilus influenzae (NTHi) are human-adapted commensal bacteria that can cause a number of chronic mucosal infections, including otitis media and bronchitis. One way for these organisms to survive antibiotic therapy and cause recurrent disease is to stop replicating, as most antimicrobials target essential biosynthetic pathways. Toxin-antitoxin (TA) gene pairs have been shown to facilitate entry into a reversible bacteriostatic state. Characteristically, these operons encode a protein toxin and an antitoxin that associate following translation to form a nontoxic complex, which then binds to and regulates the cognate TA promoter. Under stressful conditions, the labile antitoxin is degraded and the …

Toxin-Antitoxic Loci Vapbc-1 And Vapxd Contribute To Survival And Virulence In Nontypeable Haemophilus Influenzae, Dabin Ren, Anna N. Walker, Dayle A. Daines

Biological Sciences Faculty Publications

Background: Nontypeable Haemophilus influenzae (NTHi) is a significant human pathogen responsible for respiratory tract infections and the most common cause of recurrent otitis media. Type II toxin-antitoxin (TA) systems are genetic elements that code for a stable protein toxin and a labile antitoxin that are thought to be involved in metabolic regulation of bacteria by enabling a switch to a dormant state under stress conditions. The contribution to infection persistence of the NTHi TA loci vapBC-1 and vapXD was examined in this study.

Results: Deletions in vapBC-1, vapXD and vapBC-1 vapXD significantly decreased the survival of NTHi co-cultured with …

In Vitro Antimicrobial Effect Of A Cold Plasma Jet Against Enterococcus Faecalis Biofilms, Chunqi Jiang, Christoph Schaudinn, David E. Jaramillo, Paul Webster, J. William Costerton

Bioelectrics Publications

The hypothesis that a cold plasma jet has the antimicrobial effect against Enterococcus faecalis biofilms was tested in vitro. 27 hydroxyapatite discs were incubated with E. faecalis for six days to form a monoculture biofilm on the disc surface. The prepared substrata were divided into three groups: the negative control, the positive control (5.25% NaOCl solution), and the plasma treatment group. Resultant colony-forming unit counts were associated with observations of bacterial cell morphology changes using scanning electron microscopy (SEM). Treatment of E. faecalis biofilm with the plasma and 5.25% NaOCl for 5min resulted in 93.1% and 90.0% kill (P < 0.0001), respectively. SEM detected that nearly no intact bacteria were discernible for the plasmaexposed HA disc surfaces. The demonstrated bactericidal effect of the plasma with direct surface contact may be due to the enhanced oxidation by the locally produced reactive plasma species.

Mutation In The Plasmodium Falciparum Crt Protein Determines The Stereospecific Activity Of Antimalarial Cinchona Alkaloids, Carol E. Griffin, Jonathan M. Hoke, Upeka Samarakoon, Junhui Duan, Jianbing Mu, Michael T. Ferdig, David C. Warhurst, Roland A. Cooper

Biological Sciences Faculty Publications

The Cinchona alkaloids are quinoline aminoalcohols that occur as diastereomer pairs, typified by (-)-quinine and (+)-quinidine. The potency of (+)-isomers is greater than the (-)-isomers in vitro and in vivo against Plasmodium falciparum malaria parasites. They may act by the inhibition of heme crystallization within the parasite digestive vacuole in a manner similar to chloroquine. Earlier studies showed that a K76I mutation in the digestive vacuole-associated protein, PfCRT (P. falciparum chloroquine resistance transporter), reversed the normal potency order of quinine and quinidine toward P. falciparum. To further explore PfCRT-alkaloid interactions in the malaria parasite, we measured the in …

Novel Lineages Of Prochlorococcus And Synechococcus In The Global Oceans, Sijun Huang, Steven W. Wilhelm, H. Rodger Harvey, Karen Taylor, Nianzhi Jiao, Feng Chen

OES Faculty Publications

Picocyanobacteria represented by Prochlorococcus and Synechococcus have an important role in oceanic carbon fixation and nutrient cycling. In this study, we compared the community composition of picocyanobacteria from diverse marine ecosystems ranging from estuary to open oceans, tropical to polar oceans and surface to deep water, based on the sequences of 16S-23S rRNA internal transcribed spacer (ITS). A total of 1339 ITS sequences recovered from 20 samples unveiled diverse and several previously unknown clades of Prochlorococcus and Synechococcus. Six high-light (HL)-adapted Prochlorococcus clades were identified, among which clade HLVI had not been described previously. Prochlorococcus clades HLIII, HLIV and …

Diversity, Distribution, And Expression Of Diazotroph Nifh Genes In Oxygen-Deficient Waters Of The Arabian Sea, Amal Jayakumar, Mamoon M. D. Al-Rshadat, Bess B. Ward, Margaret R. Mulholland

OES Faculty Publications

The Arabian Sea oxygen minimum zone (OMZ), the largest suboxic region in the world's oceans, is responsible for up to half of the global mesopelagic fixed nitrogen ( N ) loss from the ocean via denitrification and anammox. Dinitrogen (N₂) fixation is usually attributed to cyanobacteria in the surface ocean. Model prediction and physiological inhibition of N₂ fixation by oxygen, however, suggest that N₂ fixation should be enhanced near the oxygen-deficient zone (ODZ) of the Arabian Sea. N₂ fixation and cyanobacterial nifH genes (the gene encoding dinitrogenase reductase) have been reported in surface waters overlying …

Eukaryotic Microbes In The Deep Sea: Abundance, Diversity, And The Effect Of Pressure, Danielle Morgan-Smith

OES Theses and Dissertations

The dark ocean is vast, high in pressure, cold, and scarce in resources, but has been shown to support a diverse and active microbial community wherever it is studied. Such studies, however, are scarce due to the difficulty of sampling at such depths, and are difficult to interpret due to compounding effects of pressure and temperature on physiology. Protists, functionally defined as the microbial portion of the domain Eukarya, are particularly neglected in studies of deep-sea microbiology. Here, I present three studies on microbial eukaryotes in the deep sea: first, a study of the abundance of microbial eukaryotes in the …

Grazing On Synechococcus Spp. By The Red-Tide Dinoflagellate Karenia Brevis: Implications For Bloom Dynamics In The Gulf Of Mexico, Leo A. Procise

OES Theses and Dissertations

Karenia brevis, the toxic dinoflagellate responsible for massive red tides in the Gulf of Mexico (GOM), causes fish kills, shellfish poisoning, and acute respiratory irritation in humans. Bloom initiation and maintenance have been linked to the physical environment as well as various nutrient input mechanisms. To date, efforts to quantify nitrogen (N) sources fueling K. brevis blooms in the GOM have not included mixotrophic grazing although many dinoflagellates, including K. brevis, are known to be capable of mixotrophy. This dissertation reports field and laboratory results demonstrating that natural bloom populations and K. brevisisolates from the West Florida …

The Fecal Viral Flora Of Wild Rodents, Tung G. Phan, Beatrix Kapusinszky, Chunlin Wang, Robert K. Rose, Howard L. Lipton, Eric L. Delwart

Biological Sciences Faculty Publications

The frequent interactions of rodents with humans make them a common source of zoonotic infections. To obtain an initial unbiased measure of the viral diversity in the enteric tract of wild rodents we sequenced partially purified, randomly amplified viral RNA and DNA in the feces of 105 wild rodents (mouse, vole, and rat) collected in California and Virginia. We identified in decreasing frequency sequences related to the mammalian viruses families Circoviridae, Picobirnaviridae, Picornaviridae, Astroviridae, Parvoviridae, Papillomaviridae, Adenoviridae, and Coronaviridae. Seventeen small circular DNA genomes containing one or two replicase genes distantly related to the Circoviridae representing several potentially …

Iron Limitation Of A Springtime Bacterial And Phytoplankton Community In The Ross Sea: Implications For Vitamin B12 Nutrition, Erin M. Bertrand, Mak A. Saito, Peter A. Lee, Robert B. Dunbar, Peter N. Sedwick, Giacomo R. Ditullio

OES Faculty Publications

The Ross Sea is home to some of the largest phytoplankton blooms in the Southern Ocean. Primary production in this system has previously been shown to be iron limited in the summer and periodically iron and vitamin B-12 colimited. In this study, we examined trace metal limitation of biological activity in the Ross Sea in the austral spring and considered possible implications for vitamin B-12 nutrition. Bottle incubation experiments demonstrated that iron limited phytoplankton growth in the austral spring while B-12, cobalt, and zinc did not. This is the first demonstration of iron limitation in a Phaeocystis antarctica-dominated, early …

Section Abstracts: Biology With Microbiology And Molecular Biology

Virginia Journal of Science

Abstracts of the Biology with Microbiology and Molecular Biology Section for the 89th Annual Meeting of the Virginia Academy of Science, May 25-27, 2011, University of Richmond, Richmond VA.