AMERICAN SOCIETY FOR MICROBIOLOGY

VIRGINIA BRANCH 2012 ANNUAL MEETING

PROGRAM

Old Dominion University Higher Education Center

Hosts: Wayne Hynes, Holly Gaff, and David Gauthier November 2-3, 2012

Virginia Branch Officers 2012-2013

President: Laura Runyen-Janecky (University of Richmond, Lrunyenj@richmond.edu)

President-Elect: Janet Adams (J. Sargent Reynolds Community College, JAdams@reynolds.edu)

Treasurer: Lynn Lewis (University of Mary Washington, llewis@umw.edu)

Secretary: Robyn Puffenbarger (Bridgewater College, rpuffenb@bridgewater.edu)

Councilor: Wade Bell (Virginia Military Institute, BellWE@vmi.edu)

Alternate Councilor: Louise Temple (James Madison University, templelm@jmu.edu)

Immediate Past-President: Michaela Gazdik (Ferrum College)

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Welcome to the Virginia Branch ASM

2012 Annual Meeting

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VIRGINIA BRANCH ASM 2012 ANNUAL MEETING

FRIDAY, NOVEMBER 2, 2012

Old Dominion University Higher Education Center 12:00 – 4:00 pm Registration Desk (Check-in & On-site Registration) 1:30 – 1:45 pm Welcome

- Laura Runyen-Janecky, Department of Biology, University of Richmond, VA Branch ASM President

- Renee Olander, Associate Vice President for Regional Higher Education Centers

- Wayne Hynes, Professor & Chair, Department of Biological Sciences, Old Dominion University

1:45 – 2:00 pm The VA Branch - Celebrating 71 Years of Facilitating Scientific Exchange

in the Old Dominion: Paul Phibbs, Ph.D., Professor Emeritus of Microbiology & Immunology, The Brody School of Medicine, East Carolina University

All sessions held in Rooms 244A&B SESSION I Moderator: Barbara Mann (University of Virginia) 2:00 – 2:15 pm Spatial and Temporal Dynamics of Ehrlichia chaffeensis infected ticks in

Southeastern Virginia Chelsea L Wright, Robyn M Nadolny, Daniel E Sonenshine, Wayne L Hynes, and Holly D Gaff. Old Dominion University, Norfolk, VA.

2:15 – 2:30 pm Hyaluronate Lyase in Streptococcus pyogenes: The balance between self-

preservation and virulence Melanie Sloan and Wayne L. Hynes. Old Dominion University, Norfolk, VA.

2:30 – 2:45 pm Ticks of Southeastern Virginia and the Pathogens they Carry: Spatial and Temporal Trends Robyn Nadolny, Chelsea Wright, Sharon Vaturi, Daniel Sonenshine, David Gauthier, Wayne Hynes, and Holly Gaff. Old Dominion University, Norfolk, VA.

2:45 – 3:00 pm Investigating the Behavioral Response of Dermacentor variabilis and Amblyomma maculatum in the Presence of Attractants Pamela Kelman and Holly Gaff. Old Dominion University, Norfolk, VA.

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3:00 – 3:15 pm The Impact of Ammonium Nitrate Pollution to the Susceptibility of Rana catesbiena Tadpoles to the Pathogenic Fungi Batrachochytrium dendrobatidis

Alissa Sierra Harrell and Maureen Scott. Norfolk State University, Norfolk, VA.

3:15 – 3:30 pm Protein-protein interaction networks of bacteriophage using the yeast two-

hybrid system J. Harry Caufield and Peter Uetz. Virginia Commonwealth University, Richmond, VA.

3:30 – 4:00 pm BREAK: refreshments Note: Names in bold print denote competitors for “Best Student Presentation.” 4:00 – 5:00 pm ASM BRANCH LECTURE [Introduction of ASMBL speaker by Janet Adams,

J. Sargeant Reynolds Community College]

ASM BRANCH LECTURESHIP SPEAKER:

Dr. Joe Petrosino

Director, Alkek Center for Metagenomics and Microbiome Research Baylor College of Medicine, Houston TX

“The Human Microbiome: Impact on Health and Disease”

5:00 – 5:45 pm Poster Presentation Session 1 (presenters of odd numbered

posters will present) 5:45 – 6:30 pm Poster Presentation Session 2 (presenters of even numbered

posters will present) * Posters will be displayed for both sessions. 6:30 – 8:00 pm Buffet dinner

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VIRGINIA BRANCH ASM 2012 ANNUAL MEETING

SATURDAY, NOVEMBER 3, 2012 8:15 am Coffee and light breakfast SESSION II Moderator: Emily Hill (Virginia Commonwealth University) 9:15 – 9:30 am Francisella FopA plays a role in adherence to host cells

Brett Moreau and Barbara Mann. University of Virginia, Charlottesville, VA.

9:30 – 10:00 am Analysis of two Type IVb pili systems in Sinorhizobium meliloti

Hardik Zatakia, Cassandra Nelson, Ann Hirsch, and Birgit Scharf. Virginia Polytechnic Institute and State University, Blacksburg, VA.

10:00 – 10:15 am Sinorhizobium meliloti uses the chemoreceptor, McpU to sniff out host plants Benjamin A. Webb, Sherry Hildreth, Richard Helm, and Birgit E. Scharf Virginia Polytechnic Institute and State University, Blacksburg, VA.

10:15 – 10:30 am Expression of CRISPR3 Restores Exopolysaccharide Production to a pilA

Mutant in Myxococcus xanthus Regina Wallace, Wesley Black, and Zhaomin Yang. Virginia Polytechnic Institute and State University, Blacksburg, VA.

10:30 – 10:45 am The Potential for Replication and Transmission of Antibiotic Resistance

Plasmids in and to an E. coli Population in Agriculturally Impacted Stream Sediment Erika Gehr and James B. Herrick James Madison University, Harrisonburg, VA

10:45 – 11:00 am Isolation of Novel Bacillus Bacteriophages: Distinctive Morphologic and

Genomic Features 1McKenzie Quinn, 1Nicholas Minahan, 1Siara Rouzer, 1Steve Cresawn, 2Lynn Lewis, 2Kathy Loesser-Casey, and 1Louise Temple 1James Madison University, Harrisonburg, VA and 2University of Mary Washington

Note: Names in bold print denote competitors for “Best Student Presentation.” 11:00 – 11:15 pm Closing remarks

11:30 – 12:30 pm Lunch / Business Meeting

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Abstracts for Oral Presentations Spatial and Temporal Dynamics of Ehrlichia chaffeensis infected ticks in Southeastern Virginia Chelsea L Wright, Robyn M Nadolny, Daniel E Sonenshine, Wayne L Hynes, and Holly D Gaff. Old Dominion University, Norfolk, VA.

PCR screening for Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME), was conducted on Lone Star ticks (Amblyomma americanum) collected in 2010 and 2011 from multiple sites in southeastern Virginia. Spatial and temporal analyses were performed to determine if there was any clustering among geographical sites and times where and when E. chaffeensis was present. The temporal analyses indicate a significant temporal clustering of E. chaffeensis in both 2010 and 2011, with the risk of encountering E. chaffeensis-infected ticks being significantly higher during June (P<0.001). These results have public health implications suggesting a need for increased awareness of possible HME infections during times of increased presence of the organism in ticks. Hyaluronate Lyase in Streptococcus pyogenes: The balance between self-preservation and virulence Melanie Sloan and Wayne L. Hynes. Old Dominion University, Norfolk, VA. Group A streptococcus (GAS) avoids phagocytosis using a hyaluronic acid (HA) capsule. Hyaluronate lyase (HylA) is produced by some serotypes (4 and 22), paradoxically using HA as a substrate. Several investigators have hypothesized that HylA facilitates spreading in human tissue to promote bacterial spread. The current study involves the investigation of hyaluronate lyase as an alternative mechanism of obtaining carbon sources for growth. The gene coding for extracellular hyaluronate lyase; hylA, has been inactivated in a Hyl+ producer. The wild type and mutant enzyme producers were then subjected to desiccation, starvation, and intrapecific competition studies. Ticks of Southeastern Virginia and the Pathogens they Carry: Spatial and Temporal Trends Robyn Nadolny, Chelsea Wright, Sharon Vaturi, Daniel Sonenshine, David Gauthier, Wayne Hynes, and Holly Gaff. Old Dominion University, Norfolk, VA.

Tick-borne disease is on the rise in Virginia and nation-wide. Warming temperatures and changes in the ecology of plant and animals species are resulting in unpredictable outbreaks of tick-borne illness. In order to predict the risk of tick-borne disease, a detailed understanding of tick ecology and infectivity is required. The ODU Vector Ecology lab is conducting a long-term surveillance effort to quantify the ecology of ticks in southeastern Virginia and the diseases they carry. Tick populations and pathogen prevalence rates are proving to be spatially and temporally variable. Our data will parameterize models that will help predict tick-borne disease risk in Virginia and beyond.

Investigating the Behavioral Response of Dermacentor variabilis and Amblyomma maculatum in the Presence of Attractants Pamela Kelman and Holly Gaff. Old Dominion University, Norfolk, VA.

Amblyomma maculatum and Dermacentor variabilis are hard-bodied ticks that have been found in established populations in southeastern Virginia. In order to find whether or not it is possible to capture all life stages these two species of ticks in the field using sticky traps, a series of experiments were conducted using larval, adult and nymphal stages of A. maculatum and D. variabilis ticks. Using carbon dioxide and ammonia the movement of the ticks will be monitored recording progress toward or away from the source of the attractants over the course of ninety minutes. The Impact of Ammonium Nitrate Pollution to the Susceptibility of Rana catesbiena Tadpoles to the Pathogenic Fungi Batrachochytrium dendrobatidis Alissa Sierra Harrell and Maureen Scott. Norfolk State University, Norfolk, VA.

Ammonium Nitrate pollution is the most common form of agricultural pollution; the Environmental Protection Agency mandates that 10mg/L of ammonium nitrate is safe for human consumption. Rana catesbeiana pre-metamorphic tadpoles were exposed to 10mg/L of ammonium nitrate and 1 µg/L of

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thyroxine. This is a continuous study that has established all aquatic environments exposed to aquatic environments exposed to agricultural pollutant ammonium nitrate affect thyroxine by altering growth and developmental patterns in premetamorphic R. catesbiena tadpoles. It also exposed them the fungus Batrachochytrium dendrobatidis which causes Chitridiomycosis, the leading cause of mortality of frog populations worldwide.

Protein-protein interaction networks of bacteriophage using the yeast two-hybrid system J. Harry Caufield and Peter Uetz. Virginia Commonwealth University, Richmond, VA.

Protein-protein interactions can be a powerful way to study protein function as each interaction suggests functional connections to other proteins. We have used the yeast two-hybrid (Y2H) system to study protein interaction networks in several bacteriophage, including Cp-1 and Dp-1 from Streptococcus pneumoniae and phage lambda from E. coli. Initial results from our protein interaction screens of mycobacteriophage Giles are presented. We have also compared previous sets of interaction data to optimize Y2H results and present a strategy to optimize protein interaction screens to maximize the number of interactions detected while minimizing the number of false positives. Francisella FopA plays a role in adherence to host cells Brett Moreau and Barbara Mann. University of Virginia, Charlottesville, VA.

Francisella tularensis is an intracellular pathogen and the causative agent of tularemia. While much is known about Francisella survival once intracellular, little is known about pathogen molecules that promote adherence and internalization. A Francisella novicida transposon library and an in vitro competition assay were used to detect a mutant strain, FopA, which was deficient in attachment or internalization. Further characterization of FopA revealed that it was not attenuated in intracellular growth in epithelial cell lines, but was deficient in attachment. Further experiments are ongoing to further investigate this molecule�’s role in adhesion Analysis of two Type IVb pili systems in Sinorhizobium meliloti Hardik Zatakia, Cassandra Nelson, Ann Hirsch, and Birgit Scharf. Virginia Polytechnic Institute and State University, Blacksburg, VA.

The genome of the symbiotic bacterium Sinorhizobium meliloti encodes two Type IVb pili systems. In planta experiments proved that pili mutants have lower nodule occupancy and were deficient in attachment to plant roots. Using transcriptional fusions, we showed that expression of the structural pilin PilA1 peaked during late exponential phase and is regulated by ExpR, the global transcriptional regulator of EPS production. In the presence of AHL, ExpR binds directly to the pilA1 promoter region. Our work shows that the expression of Type IV pili in S. meliloti is regulated by ExpR, and is important for efficient nodulation of alfalfa.

Sinorhizobium meliloti uses the chemoreceptor, McpU to sniff out host plants Benjamin A. Webb, Sherry Hildreth, Richard Helm, and Birgit E. Scharf. Virginia Polytechnic Institute and State University, Blacksburg, VA. Sinorhizobium meliloti and alfalfa form an agriculturally important mutualism inside host plant roots that leads to drastic increase in plant yield. A prelude to this symbiosis is mediated by seed and root exudation of attractant compounds into the soil. Sinorhizobium meliloti uses chemotaxis to navigate through the soil toward host plants. Our behavioral studies and molecular techniques show that Sinorhizobium meliloti can discriminate between the seed exudates of select host plants and non-host plants and that the methyl accepting chemotaxis protein U (McpU) is an important part of chemotaxis toward host seed exudates.

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Expression of CRISPR3 Restores Exopolysaccharide Production to a pilA Mutant in Myxococcus xanthus Regina Wallace, Wesley Black, and Zhaomin Yang. Virginia Polytechnic Institute and State University, Blacksburg, VA.

Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR, can function as an adaptive defense system in prokaryotes. In Myxococcus xanthus, we uncovered a link between CRISPR3 and the regulation of exopolysaccharide production. A transposon insertion in CRISPR3 suppressed the EPS- phenotype of a pilA mutant. Since deletion of CRISPR3 failed to suppress and expression of CRISPR3 led to pilA suppression, we conclude it is a gain-of-function mutation. We suggest that one of the spacers may inhibit the expression of a negative regulator of EPS production, providing evidence that Type III-B systems may be involved in the regulation of host chromosomal genes.

The Potential for Replication and Transmission of Antibiotic Resistance Plasmids in and to an E. coli Population in Agriculturally Impacted Stream Sediment Erika Gehr and James B. Herrick. James Madison University, Harrisonburg, VA

Horizontal transfer of genetic information between transient and native bacterial populations may enhance the spread and recombination of resistance genes potentially playing a role in the formation of multi-resistant organisms in environmental reservoirs. Tetracycline resistance plasmids were compared using three isolation techniques -- traditional �“endogenous�” extraction from isolates, �“exogenous�” plasmid capture, and direct plasmid extraction from sediment --to determine the potential for plasmid born resistance in an E. coli population found in agriculturally-impacted stream sediment. Variation in plasmid extraction techniques eliminated some of the prejudice against the uncultivable environmental organisms and gave a more complete picture of the �“mobilome�” of tetracycline resistance plasmids.

Isolation of Novel Bacillus Bacteriophages: Distinctive Morphologic and Genomic Features 1McKenzie Quinn, 1Nicholas Minahan, 1Siara Rouzer, 1Steve Cresawn, 2Lewis, 2Kathy Loesser-Casey, and 1Louise Temple. 1James Madison University, Harrisonburg, VA and 2University of Mary Washington

Undergraduates at JMU and UMW isolated and characterized five bacteriophages of Bacillus thuringiensis Kurstaki as a part of the nationwide phage hunting program. Bacillus is a soil bacterium important in medicine and agriculture. All our sequenced BtK phages are Myoviridae (~100nm head diameter; ~200nm contractile tail) with circularly-permuted genomes of ~160Kb. Cluster BtA (3 phages) and Cluster BtB (2 phages) had 22% similarity between them. BtA phages had 8 tRNA genes and 1 pseudo-tRNA gene in the same locus and a self-splicing intron interrupting the DNA polymerase gene. Cluster BtB phages exhibited a broad host range among other Bacillus strains.

Abstracts for Poster Presentations

# 1 Self-Sterilizing Wooden Cutting Boards

Candice L. Bailey, Silvia A. Berland, Ricky D. Brockington, Norma J. Cavenas, Jessie-Sue A. Fickett, Angel L. Gray, Eric A. Hattoy, Tanya S. McAdoo, Leticia M. Mickels, Katrice S. Mobley, Jason A. Robertson, Amanda M. Rouse, Katina A. Sharpe, Carl W. Vermeulen. Paul D. Camp Community College, Suffolk, VA.

Wooden cutting boards are notorious for harboring pathogens within the end-grains and scorings because the boards are not dishwasher safe (they fall apart). However, boards can be made self-sterilizing by initially soaking newly constructed, hot boards with a special butcher's wax (73% linseed oil, 24% beeswax, 3% cetavlon). The boards killed applied E. coli, Vibrio spp, and Bacillus spp within 15 minutes. Ingested cetavlon (aka cetrimide) is non-toxic to eukaryotes, and for decades was used as the necessary non-volatile cationic detergent in shampoos. It is 80-times more antibacterial than is streptomycin (w/v).

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2 Wound Rinses and Bandages for Use in Major Disasters

Shari J. Artis, Kiara L. Barnes, Chelvia V. Biggs, Sandy L. Boone, Cornesia R. Council, Clarice P. Cypress, Adrian L. Daughtrey, Ashley L. Dobie, Deborah L. Elliott, Christopher M. Hall, Monique K. Helms, Ellen J. Hibbs, Cakia D. Jenkins, Cindy C. Jones, Leigh A. Jones, Timothy T. Jones, Coretta T. Manley, Varia F. McCloud, Jordyn B. McGrew, Millie, E. Mumford, Lynday M. Outland, Mattie F. Peterson, Melany R. Price, Trina S. Scott, Kecia L. Taliaferro, Jonalan L. Williams, Melissa N. Williams, Richard C.M. Wrenn, Judy K. Wyche, Carl W. Vermeulen. Paul D. Camp Community College, Suffolk, VA.

Under massive disaster conditions such as experienced in the Haitian quake, sanitized physiological rinses can be quickly made by boiling at 103c cotton-cloth-filtered, diluted seawater (3:1 fresh water to seawater) for only 3 minutes at sealevel. Bandages made from strips of cotton (best), linen or wool (worst) simultaneously boiled in this approximation of physiological saline are simultaneously sanitized. Sparks from batteries within vehicles caught in the debris can be used to ignite screening or other fine wires as kindling to produce the needed fires for boiling.

3 Bulk Potable Water for Under-Developed Areas

Megan A. Byrd, Janie T. D'Amico, Lindsay P. Dennys, Crystal L. Drake, Lakeshia V. Eley, Stephanie M. Fagan, Emily C. Flick, Chelsea M. Gephart, Kaitlin C. Gudger, Birgit Hileman, Miriam A. Hill-Brockington, Tami M. Holt, Ngbanda D. Mumbata, Ginger M. Nairn, Bryant Pruden, Shameaka N. Rawlings, Jennifer M. St. Clair, Melissa D. Stewart, Tamara L. Story, Rebecca B. Woodard, Carl W. Vermeulen. Paul D. Camp Community College, Suffolk, VA.

In an effort to produce large amounts of potable water in underdeveloped areas, several international reports tell of using solar UV to sterilize in eight hours non-turbid surface waters in common 500 mL water bottles set on corrogated metal roofs. However, we found the bottles reportedly used were opaque to UV and the water more probably Pasteurized. By using UV-transparent ziploc bags, we found bacteria-laden water can be sterilized within two hours under bright sun if placed on reflective metal roofing even if placed slightly beneath a pond's surface so as to eliminate heating effects.

4 An shRNA-mediated Approach to Silence TGFbeta1 and VEGF-A Gene Expression in B16-F1

Melanoma Z. Charles Hand, Yoanthan T. Ararso, Osric A. Forrest, Pranay R. Reddy, and Kristian M. Hargadon. Hampden-Sydney College, Hampden Sydney, VA.

We have recently shown that the highly aggressive murine melanoma B16-F1 suppresses the maturation/activation of dendritic cells, an immune cell critical for induction of anti-tumor immunity. We have also recently demonstrated that B16-F1 melanoma secretes significantly higher levels of the immunosuppressive cytokines TGFbeta1 and VEGF-A than does the less tumorigenic B16 melanoma variant D5.1G4. Here, we utilized an shRNA-mediated gene silencing approach to interfere with expression of TGFbeta1 and VEGF-A in B16-F1 melanoma cells. These gene-silenced B16-F1 cell lines will serve as useful models for studying the role of TGFbeta1 and VEGF-A in melanoma-associated suppression of dendritic cells.

5 Quorum Sensing Based Actuation Regulation for Bacteria Powered Micro-robots (BacteriaBots)

Meghan Canter, Ali Sahari, Birgit Scharf, and Bahareh Behkam. Virginia Polytechnic Institute and State University, Blacksburg, VA.

A BacteriaBot is a bio-hybrid microrobot fabricated by interfacing an ensemble of live Escherichia coli with a micro-robot body. In this work, we explore a quorum sensing-based method to regulate motility in BacteriaBots. AI-2 was synthesized and introduced at known concentrations to quantify enhanced motility of E. coli MG1655 as a function of AI-2 concentration. Chemotaxis swim plate assay showed a halo diameter increase however, microscopy tracking and flagella staining did not show an increase in the swimming speed or the flagella count. This indicates that presence of AI-2 enhances chemotactic response but it does not enhance the motility speed.

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6 In Vitro Effects of Lindane on RAW 264.7 Macrophage Responses to LPS and IFNgamma

Mallory Stewart and David O. Freier. Lynchburg College, Lynchburg, VA. The dose response effect of lindane on RAW 264.7 murine macrophages will be evaluated. Lindane

is an organochlorine pesticide. Consumption, through ingestion or inhalation, has immunotoxic effects. RAW 264.7 cells seeded at 4 x 105 cells/well in a 24 well culture plate are exposed to 5, 50, or 200 µM lindane or vehicle controls. At 24 hours cells are washed four times and then stimulated with LPS (100 ng/ml) and IFN gamma (0.5 U/ml) for 24 hours. Effects of treatment are assessed by measurement of nitric oxide, expression of proteins (inducible nitric oxide synthase) and ELISA for IL-1beta.

7 Biochemical and Sequencing Procedures to Characterize and Confirm Possible Neisseria spp. from

Feline Oral Biofilm Cultures Julia R. Taylor and Dr. David O. Freier. Lynchburg College Lynchburg, VA.

In 2008, our lab tentatively identified an isolated strain from a male feline as a Neisseria spp. Cryogenically frozen samples (liquid nitrogen) are rapidly thawed and recultured in thioglycollate broth. Motility and aerotolerance are determined using motility agar. Differential gram staining confirms gram negative characterization and diplococci morphology. Colony morphology and hemolysis are assessed through culture on Chocolate Agar. Positive oxidase and catalase tests permit preliminary Neisseria confirmation followed by a negative DNase test to exclude B. catarrhalis. Additional biochemical tests and molecular identification using 16s rRNA sequencing will determine the specific phylogeny of the isolate.

8 Bacteriophage-mediated Antibiotic Resistance Kathleen Blevins and Lynn Lewis. University of Mary Washington, Fredericksburg, VA. In the 1940s, while the western world embraced antibiotics, the eastern world (specifically the Soviet Union) developed bacteriophages to treat bacterial infections. With the current shortage of antibiotics and the increase in antibiotic resistance, using bacteriophages as therapy is gaining acceptance. However, bacteriophages may cause antibiotic resistance, if a lysogenic phage contributes an antibiotic resistance gene to its host. Bacillus thuringiensis Kurstaki bacteriophages whose genomes contain a potential beta lactamase gene will be grown on the nonpathogenic Bacillus anthracis Delta Sterne, producing lysogens. The Bacillus anthracis Delta Sterne lysogens will be tested to see if they have acquired penicillin resistance.

9 Minimum Requirements for Flagellotropic Bacteriophage 7-7-1 Infection of Agrobacterium sp. H13-3 Katherine Broadway, Juin Yen, and Birgit Scharf. Virginia Polytechnic Institute and State University, Blacksburg, VA.

Bacteriophage 7-7-1 infects Agrobacterium sp. H13-3, using flagella as primary receptors. The analysis of fla and motA deletion strains showed, that rotating flagella are a requirement for the infection and propagation of 7-7-1. This phage is unique, in its specific host range and physical properties that differentiate it from other flagellotropic phages, such as PBS1 and phage infecting Bacillus subtilis and Escherichia coli, respectively. Not only does further research of phage 7-7-1 shed light on its infection cycle, but furthers the knowledge of the exact mechanism by which flagellotropic phages attach and infect their bacterial hosts.

10 Exploration of Sphingosine Kinase’s in RAW 264.7 Activation Stephanie DeRonde and David Freier . Lynchburg College, Lynchburg, VA. RAW 264.7 macrophages are murine derived cells used to evaluate innate responses and inflammation. Stimulus of pattern recognition receptors by lipopolysaccharide from E.coli (055:B5) and lipomannan from M. smegmatis will be compared in their ability to activate RAW 264.7 macrophages. Activation measures will include nitric oxide by Greiss reaction and isolation of protein�’s for western blot analysis of sphingosine kinase 1 and 2."

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11 Affects of trypsinization versus manual detachment of cells via scraping on RAW 264.7 murine macrophages Avery Eriksson and David Freier. Lynchburg College, Lynchburg, VA. Trypsinization and manual detachment of cells via scraping are two techniques used to remove RAW 264.7 macrophage cells from adherence to plastic surfaces for experimental use. This experiment was done to determine which method potentially affects cells more. Cells are treated dose responsively with lipopolysaccharide (from E. coli 055:B5) at 0 µl, 1 µl, 10 µl, and 100 ng/ml. Cells are incubated for 1, 4, or 16 hours after plating and then treated with LPS for 24 hours. RAW cell response is determined by measurement of nitric oxide production by the Greiss reaction.

12 Molecular typing of clinically relevant serovars of Salmonella from environmental samples

Timothy M. Smith, Jr. and David W. Buckalew. Longwood University, Farmville, VA. Salmonella enterica, includes seven groups or subspecies (ssp.) which are each separated into

serovars. The most significant pathogens for humans are found in group 1 which is Salmonella enterica ssp. Enterica, within which, are many serovars; the most clinically relevant being serovars Typhimurium, Enteritidis, Hadar, and Heidelberg. Using diagnostic media and multiplex PCR, clinically relevant Salmonella serovars were isolated and serotyped from tributaries and main channel of the upper Appomattox River watershed. Primers were designed to amplify alleles of the O, H1, and H2 antigens. Antigenic alleles were cross checked with catalogued antigenic formulas to identify isolate colonies to serovar.

13 Laboratory protocol for isolating, confirming, and molecular typing of serovars of Salmonella from

environmental samples Timothy M. Smith, Jr. and David W. Buckalew. Longwood University, Farmville, VA.

Our field and laboratory protocols used for the sampling, isolation, enumeration, and serological confirmation of species of Salmonella are provided in outline form. Included are the steps used for molecular determination of Salmonella enterica ssp. enterica to serovar.

14 Investigation of use of Heme as an Iron Source for Sodalis glossinidius via HemR

Gili Hrusa and Laura Runyen-Janecky. University of Richmond, Richmond, VA. Sodalis glossindius, a facultative intracellular symbiont of the tsetse fly, must acquire iron for

growth. High-affinity iron acquisition systems allow bacteria to fulfill their iron needs in environments where free iron is low. Previous Sodalis genomic investigations showed that Sodalis has putative genes for the HemRTUV heme transport system. In this study, we found that growth of Escherichia coli expressing the Sodalis hemR gene in low iron media is stimulated by heme and that heme stimulates growth of Sodalis in low iron media. We are currently testing the importance of HemR for heme growth stimulation using Sodalis mutants.

15 SEM Imaging of Neurospora crassa mutants

Jeff Illinik, Victor Townsend, Jr., and Philip Rock. Virginia Wesleyan College, Norfolk, VA. The growth and development of fungal mycelia are complex processes that are still rather poorly

understood. We have been using scanning electron microscopy to compare morphological and biochemical mutants of the model fungus, Neurospora crassa. The high resolution of scanning electron microscopy is useful for identifying structural changes in fungal hyphae associated with specific mutations, which are not readily apparent in the light microscope. An initial analysis has revealed changes in the ultrastructure of the crosswalls in microtubule motor protein (dynein and kinesin) mutants and may further elucidate the roles played by these proteins in hyphal growth.

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16 Histophilus somni survives in bovine monocytes by interfering with phagosome maturation Yu Pan and Thomas Inzana. Virginia Polytechnic Institute and State University, Blacksburg, VA. Virulent strains of Histophilus somni are resistant to killing by phagocytic cells. Intracellular growth of pathogenic strain 2336 declined during the first 24 h, but recovered after 48 h. However, commensal strain 129Pt was killed within 24h. These results were confirmed by fluorescence microscopy of bovine monocytes co-incubated with SYTO-9-stained bacteria. However, luxS and uspE mutants, which were compromised for growth in macrophages, were elongated within monocytes. Confocal microscopy indicated that EEA-1 co-localized with strains 2336 and 129Pt in monocytes, but LAMP-2 co-localized only with strain 129Pt. Therefore, virulent H. somni may survive intracellular killing by interfering with phagosome maturation.

17 A Study of Chloroquine’s Antiretroviral Characteristics Ryan Green and Lynn Lewis. University of Mary Washington, Fredericksburg, VA.

Murine Mammary Tumor Virus (MMTV) infected cells will be used to study the antiviral properties of the anti-malarial drug chloroquine. Studies have shown the activated progesterone receptor reduces chromatin density at the MMTV promoter increasing MMTV transcription and replication. In this study, the effectiveness of viral induction by progesterone and dexamethasone will be compared via TCID50, by treating infected tissue culture with progesterone or a dexamethasone control. Previous studies have also shown that chloroquine may decrease viral replication by preventing phagosomal acidification. The effects of chloroquine on MMTV will be studied via TCID50 using chloroquine treatments or an untreated control.

18 Studying the Oral Route of Transmission of Mycobacterium marinum in Danio rerio Ashley Moye, Brittany St. Jacques, Brandon Rowan, David Gauthier, Old Dominion University, Norfolk,

VA. The route of transmission of pathogenic Mycobacterium species to fishes is not well understood.

The suggested routes of transmission include: ingesting prey infected with Mycobacterium species; direct contact; or aqueous exposure via the skin, gills or intestine. The purpose of this study was to investigate the oral exposure route by inoculating a gelatin-based food with two doses of Mycobacterium marinum (1218R) and feeding to the model species, Danio rerio (zebrafish), over a period of four weeks. Mycobacterial presence in internal organs was determined by qPCR and bacteriology.

19 Analyzing the Importance of dnaK gene in the survival of Sodalis glossinidius during Heat Shock

Jose Santinni Roma and Laura Runyen-Janecky. University of Richmond, Richmond, VA. Sodalis glossinidius is bacterial symbiont that lives in the gut of the tsetse flies. Being able to tolerate changes in temperature is important for the survival of bacteria in organisms, like tsetse flies, that do not have a constant body temperature. Many bacteria, including Sodalis, contain DnaK, which functions as a chaperone protein that helps the bacteria survive during thermal stresses such as heat shock, a sudden change in temperature. This study aims to investigate the function and importance of the Sodalis dnaK gene in the survival of bacteria during heat shock.

20 Antibiotic Resistance of Enterococcus spp. in a Man-Made Drainage Lake Kristie Prtorich, M. Carpenter, H. Kurland, and J. Mott. James Madison University, Harrisonburg, VA. Newman Lake is a man-made lake on the campus of James Madison University formed through the damming of Sibert�’s Creek and serving as the collection point of storm-water drainage for the university. The lake has been found to contain elevated levels of fecal indicator bacteria. Potential sources of contamination include geese which aggregate around the lake. Water and geese feces samples were collected to isolate species of Enterococcus and determine their antibiotic resistance profiles. This data can be used to assess sources of contamination and potential risks to individuals and inform storm-water management planning to decrease impact on downstream waters.

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21 The innate immune response and melatonin: Interaction of responses of RAW 264.7 macrophages to TLR3 and TLR4 ligands with melatonin Alexandra Einstman and Dr. David O. Freier. Lynchburg College, Lynchburg, VA. Melatonin is a pleiotropic hormone mediated through interaction with G-protein coupled membrane bound receptors MT1 and MT2. These receptors are widely distributed in the body however remain many questions about the interaction of melatonin and how it affects particularly on the innate immune response. The direct effect of melatonin on RAW 264.7 cells responses to ligands for TLR3 and TLR4 will be compared in conjunction with melatonin using LPS from E.coli (0111:B4) [TLR4 ligand] and Polyinosinic:polycytidylic acid [TLR3 ligand].

22 Phylogenetics of Pilobolacea

Heaven Cerritos and Dale Beach. Longwood University, Farmville, VA. The family Pilobolaceae is a collection of fairly ubiquitous fungal species associated with the

Mucorales, and best known as �“Shotgun Fungi�” capable of shooting sporangia great distances. Traditional identification of isolates to species level requires measurement of asexual spores, and is often complicated by overlapping criteria, specifically in the dimensions of individual spores. Recent work describing the use of phylogenetic methods to characterize Pilobolaceae members via ribosomal RNA sequences proved sufficient to differentiate the genera within the family. We are extending this work by developing protein coding sequences as additional phylogenetic targets to increase sequence resolution for more specific classification.

23 Antimicrobial Analysis of a Novel Series of Amphiphiles

Kyle Bonifer and Crystal Manning. James Madison University, Harrisonburg, VA. Our research centers on the development of novel amphiphiles comprised of non-polar hydrocarbon tails, and polar head groups. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays tested the effectiveness of amphiphiles against pathogenic species of bacteria. MIC and MBC values were in the low micromolar range, specifically the compound M-P12,12 (meta-pyridine, 12 carbon chain, 12 carbon chain) was most effective for all strains tested, with MIC and MBC values of 2µM for S. aureus, 2µM for E. faecalis, 4µM for E. coli, and 8µM for P. aeruginosa. These values were less than control compounds DTAB, CTAB, and Lysol.

24 Seasonal variation in the persistence and viability of human adenovirus in estuarine water Wenda Quidort, Kimberly Reece, and Howard Kator. Virginia Institute of Marine Science, Gloucester Point, VA. Wastewater treatment plants can release enteric viruses resulting in environmental contamination with human health impacts. Determining the length of time that the virus persists and remains infective under various environmental conditions in estuarine waters is critical to assessing risk. Adenovirus is a good candidate viral indicator because it is stable in the environment, resistant to UV radiation, demonstrates little seasonal variation, and unlike norovirus, can be cultured to assess infectivity. Persistence and viability studies using human adenovirus were conducted during the summer, fall and winter. Viral DNA persistence was determined using qPCR and infectivity was assessed by integrated cell culture-qPCR.

25 Experimental Oral Transmission of Mycobacterium shottsii and M. pseudoshottsii to Chesapeake Bay Striped Bass (Morone saxatilis) Brittany St.Jacques, A Moye, B Rowan, and DT Gauthier. Old Dominion University, Norfolk, VA. The bacterial disease, mycobacteriosis, is present in Chesapeake Bay striped bass (Morone saxatilis) at high endemic levels. Mycobacteria isolated from these fish are predominantly two novel species: Mycobacterium pseudoshottsii and M. shottsii. Despite a century of research on finfish mycobacteriosis, only one paper has demonstrated transmission by biologically relevant exposure routes, while other accounts of natural transmission are anecdotal. This work examined experimental oral transmission of M. pseudoshottsii and M. shottsii to striped bass, via amended gel-based feed. M. pseudoshottsii was recovered

2012 VA-ASM

from spleens of 10% of exposed striped bass, indicating that ingestion is a viable natural transmission mode for this pathogen.

26 qPCR detection of human-specific viral and bacterial fecal markers in comparsion to standard fecal indicator bacteria N. Reid Mizelle, Kayla Mullins, and J. Brooks Crozier. Roanoke College, Roanoke, VA. The detection of human fecal pollution is important to human health and the environment. While the enumeration of fecal indicator bacteria (FIB) is the standard method for FIB detection , qPCR-based assays allow the detection and quantification of source-specific fecal bacteria and viruses with great precision. We report that there is no relationship between the number of FIBs and copy number of qPCR targets from sample sites along the Roanoke River and Tinker Creek (Roanoke, VA), even when the data were corrected for flow rate. More research will be needed to better establish appropriate qPCR water quality standards.

27 Site-Directed Mutagenesis and X-ray Crystallization of the ExsA Activator Protein of the Type III Secretion System in Pseudomonas aeruginosa Rika Judd, Cory Bernhards, and Florian Schubot. Virginia Polytechnic Institute and State University, Blacksburg, VA. Pseudomonas aeruginosa, a Gram-negative bacterium, causes disease in humans. P. aeruginosa utilizes a type III secretion system (T3SS) which injects toxic proteins into host cells. This T3SS is regulated by four interacting proteins (ExsA, ExsD, ExsC and ExsE), where ExsA activates transcription. In this study, site-directed mutagenesis was employed to change methionine 196 of ExsA to a cysteine in order to crosslink ExsA to a specially modified DNA promoter for crystallization. The ExsA M196C variant was successfully constructed, purified, and is ready for screening, the first steps of crystallization. Solving the structure of ExsA would give insight into how AraC-type transcriptional regulators work leading to the development of novel antibiotics for treating various bacterial infections.

28 Examination of cAMP levels in Mycobacteria

Michaela Gazdik and Kevin Reynolds. Ferrum College, Ferrum, VA. Mycobacterium species are unique because they contain many more annotated adenylate cyclases

(ACs), which are responsible for production of cyclic adenosine monophosphate (cAMP), compared to other bacteria. Evidence suggests that cAMP secreted from Mycobacterium tuberculosis (Mtb) plays a role in Mtb-host interactions during tuberculosis disease. Thus, characterization of mycobacterial cAMP secretion mechanisms will improve our understanding of how this bacterium establishes a successful infection. We are using a colorimetric cAMP assay to characterize the amount of cAMP produced and secreted under various environmental conditions, such as low pH, various fatty acids, and starvation. These levels will be compared between three different bacterial species, M. smegmatis, M. bovis BCG and Escherichia coli.

29 Correlations between common biological indicators and Salmonella in environmental waters Timothy M. Smith, Jr., Sarah F. Lucento, and David W. Buckalew. Longwood University, Farmville, VA.

Fecal indicator bacteria (FIB) such as coliforms and and Escherichia coli have been used as a gauge for biological water quality since the 1920's. Recently, several studies suggest that FIB may not directly correlate with select pathogenic bacteria in natural waters. This study reports numbers of Salmonella spp. are higher than indicator bacteria in each of three streams assessed for both coliforms and E.coli and, in some cases, counts may be higher by an order of magnitude. Statistical analysis reveals there is a positive correlation of coliforms with Salmonella and of E. coli with Salmonella, but Salmonella counts are significantly higher than either FIB group.

2012 VA-ASM

30 Identification of a Francisella tularensis subsp. novicida capsule-like complex Kelly Freudenberger, Anna Champion, and Tom Inzana. Virginia Polytechnic Institute and State University, Blacksburg, VA.

We have identified a capsule-like-complex (CLC) of high molecular weight in Francisella novicida, which has previously been reported to be unencapsulated. This surface material appears similar to the reported CLC of the select agent, F. tularensis. A putative polysaccharide locus was identified in F. novicida that has significant homology to the locus involved in CLC production for F. tularensis. We hypothesize F. novicida could be making a similar CLC. With an F. novicida transposon library readily available, identification of the individual protein components of CLC could be more efficient than current techniques.

31 A Novel Application of Quantitative Mechanical Testing in Bacteria Abdullah Mamun, Claretta J. Sullivan, Donald Stone, and Abdelmageed Elmustafa. Eastern Virginia Medical School, Norfolk, VA. As bacteria navigate various environments, osmotic pressure can change dramatically due to changes in local solute concentration. The peptidoglycan, together with the cellular proteins, mitigates the osmotic stress that would otherwise result in lysis. We developed a methodology to determine nanomechanical properties of E. coli using a nanoindenter. The results from nanoindentation experiments on glutaraldehyde-fixed bacteria in air and water, and live bacteria in air are reported. In addition, a modeling and simulation of the bacterium was performed by combining the properties of the outer membrane, peptidoglycan and cytoplasmic membrane into a single effective layer using elasticity theory.

32 Assessing risk of tick-borne pathogens in the Hampton Roads Area James Lana and Holly Gaff. Old Dominion University, Norfolk, VA. Humans are affected by many tick-borne pathogens such as Borrelia burgdorferi, Rickettsia

rickettsii, and Ehrlichia chaffeensis. These pathogens are transmitted by Amblyomma americanum, Ixodes scapularis, and Dermacentor variabilis, which were the most common ticks found on humans through in the Hampton Roads area. As part of the larger tick surveillance project in the ODU Vector Ecology lab, ticks found on humans are submitted to the lab. In 2012, 359 ticks were submitted and analyzed to compare the frequency of each species in comparison with other collection techniques. This comparison allows more accurate interpretation of human disease risk than would be gained from other collection techniques alone. Future work will identify any differences in pathogens between ticks collected from humans and those collected through other techniques. Understanding the species of ticks present in our area and knowledge of their pathogens can better prepare people for treatment and preventative measures with future interactions of tick species.

33 Assessment of the Motility Test Technique Amanda Bristow and Emily Hill. Virginia Commonwealth University, Richmond, VA. Motility testing is often a required component of routine identification algorithms to properly speciate microorganisms isolated from patient specimens. Although motility tests are simple to perform, interpretation is difficult and can lead to misidentification. The goal of this research project is to compare motility test techniques available for use in the clinical microbiology laboratory in regard to accuracy, ease of performance, and turnaround time. American Type Culture Collection (ATCC) strains and organisms isolated from patient specimens will be tested. Results obtained will be used to recommend the motility test technique best suited for use in the clinical microbiology laboratory.

34 Methods for Capturing Escherichia coli Surface Dynamics Using AFM Natalie E. Lonergan and Claretta J. Sullivan. Eastern Virginia Medical School, Norfolk, VA. Atomic force microscopy (AFM), which permits high resolution, live cell imaging, has been under- utilized in research investigating bacterial surface dynamics. This is in part due to the difficulty of immobilizing bacteria to withstand AFM probe based forces in nutrient media. The current study outlines a

2012 VA-ASM

reliable method of sample preparation for AFM imaging of live Escherichia coli in Luria Bertani (LB) broth media. E. coli cells were stably immobilized on poly-L-lysine coated glass in diluted PBS, and were recovered in minimal media prior to imaging in LB broth.

35 Structure/function analysis of a dual function receptor protein, FegA Christopher Romeo and Stephanie Stockwell. James Madison University, Harrisonburg, VA. The Gram-negative bacterium, Bradyrhizobium japonicum, performs symbiotic nitrogen fixation for soybean. FegA is a Bj strain 61A152 outer membrane receptor required for ferrichrome utilization and symbiosis. These functions are distinct, as Bj USDA110 ferrichrome receptor, FhuA, is dispensable for symbiosis. We hypothesize FegA�’s unique role in planta is the result of structural differences between the N-terminal domains (NTDs) of FegA and FhuA. Recombinant plasmids containing fegA NTD deletion and chimeric alleles were introduced into the fegAB mutant for complementation of iron uptake. Constructs with altered NTDs fail to complement fegAB-, suggesting this domain is critical for iron uptake and/or stability.

36 Role of two large, novel surface proteins in Bordetella avium virulence Nathaniel Tate Burkholder, Stephanie Stockwell, and Louise Temple-Rosebrook. James Madison University, Harrisonburg, VA.

Bordetella avium is a gram negative bacterium that causes bordetellosis in turkeys, but lacks common virulence factors found in other members of the Bordetella genus. Two large, novel genes in B. avium, bav1944-5, are adjacent to a type I secretion system and encode putative adhesins. A knock out mutant in this locus is being tested for tracheal binding affinity, serum resistance, toxicity to cultured cilia, and biofilm formation. The mutant was observed to exhibit wild-type attachment levels, but showed less resistance to serum. A potential toxic domain in BAV1945 was identified, suggesting these proteins may play multiple roles in virulence.

37 Host range infectivity and sensitivity of Bacillus pumilus bacteriophages Anne E. Schindler, Skyelar Decker, Andrew Montgomery, and Louise Temple. James Madison University, Harrisonburg, VA.

Over 60 bacteriophages infecting Bacillus pumilus BL8 were isolated from the environment by freshman at JMU, of which eight have been completely sequenced. Twenty-two bacteriophage were examined through plaque assays for host range sensitivity on four strains of Bacillus pumilus: BL8, ATCC7061, F036B, and SAFR. Two strains, BL8 and ATCC7061, contain a prophage that can be induced by infection with certain other phages. Podoviridae, myoviridae, and siphoviridae morphologies were observed by electron microscopy. Titer and plaque morphology varied by host strain. Further research into the genomics and biology of this collection is currently being conducted by an undergraduate research team.

38 Discovering Methicillin Resistant Mobile Genetic Element in Viral Fraction of Environmental Samples

Christophe Langouët-Astrie, Monica Chowdhury, Rachel Cross, Nicholas Minahan, Kim Okafor, Laurel Owens, Morgan Pate, Emily Pelto, McKenzie Quinn, Brooke Sauder, and Dr. Louis Temple. James Madison University, Harrisonburg, VA.

Methicillin resistant Staphylococcus aureus (MRSA) is a growing problem in clinical settings. MRSA is caused by a mutation in the mecA gene, which causes a misfolding of a penicillin binding protein on S. aureus membrane, and prevents the antibiotic methicillin from binding to the membrane. Recent studies in Spain have found the mecA gene in viral genomes in environmental samples. Our research has been to develop a protocol for collecting environmental samples and isolating viral fractions from the samples, detect the presence of the mecA gene via PCR, therefore confirming the findings from previous work.

2012 VA-ASM

39 Direct capture of plasmids conferring resistance to late-generation cephalosporins from agriculturally-impacted stream sediments Seana Sears, Elena Balkanska, and James Herrick. James Madison University, Harrisonburg, VA. Streams harbor large numbers of native and introduced bacteria, which can be both recipients and donors of antibiotic resistance genes on mobile elements. Plasmid-borne resistance to late-generation expanded-spectrum beta lactams is of increasing concern in clinical settings but has not been detected in natural environments. Using a method developed in our laboratory, a rifampicin-resistant E. coli was used to capture, without cultivation, resistance plasmids directly from stream sediment impacted by agricultural runoff. Transconjugants were resistant to the third-generation cephalosporins cefotaxime and ceftazidime. To our knowledge, this is the first evidence of transmissible expanded-spectrum beta-lactam resistance in a stream environment. Attendees and participants Adams, Janet jadams@reynolds.edu Balkanska, Elena balkanee@jmu.edu Beach, Dale beachdl@longwood.edu Bell, Wade bellwe@vmi.edu Blevins, Kathleen kblevins@mail.umw.edu Bonifer, Kyle Bonifeks@gmail.com Bristow, Amanda bristowak@mymail.vcu.edu Broadway, Katherine bkatie5@vt.edu Buckalew, David buckalewdw@longwood.edu Burket, Jessica burketja@evms.edu Burkholder, Nathaniel Tate burkhont@dukes.jmu.edu Cabral, Guy gacabral@vcu.edu Canter, Meghan Carpenter, Marc Caufield, Harry J. caufieldjh@vcu.edu Cerritos, Heaven heaven.cerritos@live.longwood.edu Champion, Anna annac@vt.edu DeRonde, Stephanie Einstman, Alexandra einstman_a@students.lynchburg.edu Eriksson, Avery eriksson_a@students.lynchburg.edu Fisher, Deborah Neely dneely-fisher@reynolds.edu Freier, David freier@lynchburg.edu Freudenberger, Kelly Gauthier, David dgauthie@odu.edu Gazdik,Michaela mgazdik@ferrum.edu Gehr, Erika erika.gehr@gmail.com Hand, Charlie handc14@hsc.edu Herrick, James herricjb@jmu.edu Hill, Emily s2emcrab@vcu.edu Hrusa, Gili Marie gili.hrusa@richmond.edu Hynes, Wayne whynes@odu.edu Illinik, Jeff jjillinik@vwc.edu Judd, Rika Kator, Howard kator@vims.edu

2012 VA-ASM

Kelman, Pamela pkelm001@odu.edu Kurland, Hilary Lana, James jlana001@odu.edu Langouet-Astrie, Christophe langoucj@dukes.jmu.edu Lewis, Lynn llewis@umw.edu Lonergan, Natalie lonergne@evms.edu Mann, Barbara bjm2r@virginia.edu Manning, Crystal mannincf@dukes.jmu.edu Marciano-Cabral, Francine fmcabral@vcu.edu McBride, Steven mcbridsg@dukes.jmu.edu McCallister, Shannon McKenna, Kristin mckennkr@dukes.jmu.edu Meng, Bowen bmeng@vt.edu Minahan, Nicholas minahant@dukes.jmu.edu Moreau, Brett gbm5pn@virginia.edu Morris, Dennis dennis.morris@millercoors.com Moye, Ashley amoye005@odu.edu Nadolny, Robyn rnado002@odu.edu Okafor, Kimberly Owens, Laurel bass_ball@cox.net Pan, Yu pyu08@vt.edu Partin, Michael partinmt@dukes.jmu.edu Petrosino, Joe Phibbs, Paul Prtorich, Kristie prtorikn@dukes.jmu.edu Puffenbarger, Robyn rpuffenb@bridgewater.edu Quidort, Wenda wendi@vims.edu Quinn, McKenzie quinnmr@dukes.jmu.edu Railling, Patricia raillip@evms.edu Reynolds, Kevin Daniel kreynolds@ferrum.edu Rock, Philip prock@vwc.edu Roma, Jose Santinni josesantinni.roma@richmond.edu Runyen-Janecky, Laura Lrunyenj@richmond.edu Scharf, Birgit bscharf@vt.edu Schindler, Anne Sears, Seana searssp09@gmail.com Sharp, Julia sharpja@evms.edu Smith, Timothy timothy.smith@live.longwood.edu Spencer, Jacqueline spencerj@tncc.edu Sloan, Melanie St. Jacques, Brittany bstja001@odu.edu Stewart, Mallory stewart_m@students.lynchburg.edu Sullivan, Claretta sullivcj@evms.edu Taylor, Julia ` Taylor_JR@students.lynchburg.edu Temple, Louise templelm@jmu.edu Uetz, Peter uetz@vcu.edu Vermeulen, Carl ecoligist@yahoo.com Wallace, Regina Reginaw@vt.edu Webb, Benjamin mercury1@vt.edu Wright,Chelsea cwrig024@odu.edu Zatakia, Hardik hzatakia@vt.edu

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2012 VA-ASM