Microbiology Commons^™

Response Of The Tick Dermacentor Variabilis (Acari: Ixodidae) To Hemocoelic Inoculation Of Borrelia Burgdorferi (Spirochetales), Robert Johns, Daniel E. Sonenshine, Wayne L. Hynes

Biological Sciences Faculty Publications

When Borrelia burgdorferi B31 low passage strain spirochetes were directly injected into the hemocoel of Dermacentor variabilis(Say) females, the bacteria were cleared from the hemocoel within < 24 h. Viable spirochetes were not found in hemolymph, salivary gland, or ovary tissues by subculturing or by IFA. The hemocyte population increased ≈6 times within the first 6 h after inoculation compared with the uninoculated controls. In contrast, the soluble total hemolymph protein content decreased inversely with the increase in hemocytes. Borreliacidal activity was demonstrated with cell-free hemolymph from D. variabilis. In vitro antimicrobial assays using hemolymph from borrelia-challenged and nonchallenged ticks resulted in 72% spirochete reductions compared with only 11.5%, respectively, within 1 h. Additional evidence of induced antimicrobial hemolymph protein activity was demonstrated by SDS-PAGE, which revealed upregulation of a lysozyme-like peptide (≈ 15 kDa) (22% increase) and the induction of a ≈ 5.8 kDa peptide in the B. burgdorferi-challenged ticks. In contrast with the nonvector borne Bacillus subtilis …

A Study On The Effects Of The N-Terminal Amino Acid Sequence On The Activation Of Human T-Cell Leukemia Virus Type 1 Protease, Hidayah Muhammad Kendall

Chemistry & Biochemistry Theses & Dissertations

Human T-cell leukemia virus type 1 (HTL V-1) is dependent upon the enzymatic activity of its protease for maturation. Maturation of the protease is facilitated by cleavage of specific amino acid residues, followed by dimerization. The effects of the amino acid sequence located N-terminally to the cleavage site on the ability of the protease to become active were the focus of the current study. These amino acid sequences were contributed by the plasmid vector into which the protease gene was inserted.

Surface probability analyses (SPAs) of the vectors, as well as for native sequences which produce the mature protease and …

An Extracellular Protein Produced By Staphylococcus Hominis Strain Αm With Antimicrobial Activity Against Mycobacterium Spp, Sandra May Jacobsen

Biological Sciences Theses & Dissertations

The inhibitory activity exhibited by Staphylococcus hominis strain αM toward Mycobacterium species was examined. These studies included determining the conditions that maximized inhibitory agent production, analyzing characteristics of the agent, determining the time of agent production, ascertaining plasmid possession and the role of plasmids in the production of this inhibitory agent, and ascertaining the antimicrobial activity of other strains of S. hominis. On solid and in liquid media, S. hominis strain αM exhibits a unique inhibitory activity toward Mycobacterium species that is not demonstrated by Staphylococcus epidermidis or other S. hominis strains. Maximum agent production and recovery was …

Nucleotide Sequence Of The Streptococcin A-Ff22 Lantibiotic Regulon: Model For Production Of The Lantibiotic Sa-Ff22 By Strains Of Streptococcus Pyogenes, Robert E. Mclaughlin, Joseph J. Ferretti, Wayne L. Hynes

Biological Sciences Faculty Publications

Streptococcin A-FF22 (SA-FF22) is a type AII linear lantibiotic produced by Streptococcus pyogenes strain FF22. Sequence analysis of an approximate 10 kb region of DNA showed it to contain nine open reading frames arranged in three operons responsible for regulation, biosynthesis and immunity of SA-FF22. This region is organized similarly to the Lactococcus lactis lacticin 481 region, however, unlike lacticin 481, a two-component regulatory system is essential for SA-FF22 production. located immediately downstream of the scn region is a putative transposase gene, the presence of which supports earlier data that indicated a mobile nature to this region. (©) 1999 Published …

Construction Of An Escherichia Coli Lac Repressor-Based System To Study Human Cytomegalovirus Gene Expression, Laura Fernanda Cageao-Luchetti

Theses and Dissertations in Biomedical Sciences

An Escherichia coli lac repressor-based system was developed to study the roles of human cytomegalovirus (HCMV) genes during viral replication. To this end, a recombinant HCMV expressing the lac repressor was generated (RVlac), and an HCMV-specific promoter was targeted for conditional expression by inserting the lac operator sequence. The promoter of a nonessential gene was chosen in order to be able to assess parameters of repression and derepression of the operator-containing promoter in the endogenous locus, without having virus growth dependent on the specific inducer isopropylthiogalactoside (IPTG). The feasibility of this approach to conditionally express an HCMV promoter …

Vertical Profiles Of Virus-Like Particles And Bacteria In The Water Column And Sediments Of Chesapeake Bay, Usa, Lisa A. Drake, Keun-Hyung Choi, A. G. Edward Haskell, Fred C. Dobbs

OES Faculty Publications

Vertical profiles of virus-like particles (VLPs) and bacteria were determined by near-synoptic sampling through the water column and 15 to 25 cm into the sediment at 5 stations across the mouth of Chesapeake Bay, USA. VLPs were about 10 times more abundant in the pore water (grand mean = 3.6 x 10(8) VLPs ml(-1)) than in the water column (grand mean = 3.8 x 10(7) VLPs ml(-1)). Similarly, bacteria counts were about 3 times higher in the pore water (grand mean = 6.5 x 10(6) bacteria ml(-1)) than in the water column (grand mean = 2.4 x 10(6) bacteria ml(-1)). …

Thermophilic Bacterial Activity In A Deep-Sea Sediment From The Pacific Ocean, Fred C. Dobbs, Karen A. Selph

OES Faculty Publications

Thermophilic bacterial activity was detected in a deep-sea sediment sample from the South Pacific Ocean at 12 degrees S, 135 degrees W, an area of the seafloor distant from known hydrothermal venting. Incubation of sediments amended with C-14-glutamate indicated maximal respiration (evolution of (CO2)-C-14) and assimilation (incorporation of C-14 into acid-precipitated macromolecules) of substrate at 52 degrees C, relative to 4 and 22 degrees C. A parallel experiment at another site (2 degrees S, 140 degrees W) yielded no evidence of thermophily. Thermophilic bacteria may be deposited in deep-sea sediments following their long-distance dispersal from hydrothermal vents (e.g. the East …

Characterization Of P39, A Borrelia Burgdorferi Specific Protein, Jill M. Troyer

Theses and Dissertations in Biomedical Sciences

Borrelia burgdorferi, the causative agent of Lyme disease, expresses a species specific, 39 kiloDalton protein of unknown function called P39. This protein is highly immunogenic and the presence of anti-P39 antibodies in patient sera is used as an indicator of B. burgdorferi infection. P39 is also a candidate for vaccine developement due to conserved expression of P39 among B. burgdorferi isolates. Since little is presently known about this important molecule, the present study was designed to characterize P39 and was carried out in six separate investigations. First, infectious and noninfectious isolates of B. burgdorferi were established in vitro, and …

A Computer-Simulated Restriction Fragment Length Polymorphism Analysis Of Bacterial Small-Subunit Rrna Genes: Efficacy Of Selected Tetrameric Restriction Enzymes For Studies Of Microbial Diversity In Nature, Craig L. Moyer, James M. Tiedje, Fred C. Dobbs, David M. Karl

OES Faculty Publications

An assessment of 10 tetrameric restriction enzymes (TREs) was conducted by using a computer-simulated restriction fragment length polymorphism (RFLP) analysis for over 100 proximally and distally related bacterial small-subunit (SSU) rRNA gene sequences. Screening SSU rDNA clone libraries with TREs has become an effective strategy because of logistic simplicity, commercial availability, and economy. However, the rationale for selecting the type and number of TREs has not been systematically evaluated. Our objective nas to identify the optimal combination of TREs for RnP screening of cloned SSU rRNA genes from undefined bacterial clone libraries. After computer-simulated TRE digestion, the resultant fragments were …

Cascading Disturbances In Florida Bay, Usa: Cyanobacteria Blooms, Sponge Mortality, And Implications For Juvenile Spiny Lobsters Panulirus Argus, Mark J. Butler Iv, John H. Hunt, William F. Herrnkind, Michael J. Childress, Rodney Bertelsen, William Sharp, Thomas Matthews, Jennifer M. Field, Harold G. Marshall

Biological Sciences Faculty Publications

Florida Bay, the shallow lagoon separating mainland Florida and the Florida Keys, USA, is experiencing an unprecedented series of ecological disturbances. In 1991, following reports of other ecosystem perturbations, we observed widespread and persistent blooms of cyanobacteria that coincided with the decimation of sponge communities over hundreds of square kilometers. Juvenile Caribbean spiny lobsters Panulirus argus, among other animals, rely on sponges for shelter; the impact of sponge loss on the abundance of lobsters and their use of shelter, in particular, has been dramatic. The loss of sponges on 27 experimental sites in hard bottom habitat in central Florida Bay …

Phylogenetic Diversity Of The Bacterial Community From A Microbial Mat At An Active, Hydrothermal Vent System, Loihi Seamount, Hawaii, Craig L. Moyer, Fred C. Dobbs, David M. Karl

OES Faculty Publications

The phylogenetic diversity of small-subunit rRNA genes associated,vith the domain Bacteria was examined (by using previously defined operational taxonomic units [C. L. Moyer, F. C. Dobbs, and D. M. Karl, Appl. Environ. Microbiol. 60:871-879, 1994]; those for Pele's Vents Bacteria are hereafter abbreviated PVB OTUs) with samples from a microbial mat at an active, deep-sea hydrothermal vent system. A cluster of phylogenetically related PVB OTUs (OTUs 2, 3, 6, and 8) was closely affiliated with Thiovulum sp. contained within the epsilon subclass of the class Proteobacteria and accounted for 60.5% of the small-subunit rRNA bacterial clone library from Pele's Vents. …

Pathogenicity Of Murine Cytomegalovirus Mutants, Victoria Jean Cavanaugh

Theses and Dissertations in Biomedical Sciences

The purpose of this study was to identified nonessential murine cytomegalovirus (MCMV) genes involved in pathogenesis in vivo. Our approach to identifyjng these genes consisted of constructing MCMV mutants, and then analyzing these mutants in vitro and in vivo. Recombinant viruses (RV) expressing the β-glucuronidase marker gene were constructed by site-directed insertion and deletion mutagenesis of the MCMV Hind III-J and -I regions of the viral genome. Mutations were targeted to this region of the MCMV genome because the corresponding region of the human CMV genome is nonessential and is involved in down-regulating major histocompatibility complex (MHC) class I expression …

The Kinetic Characterization Of Mutant L289p Of Recombinant Hiv-1 Reverse Transcriptase, Tryn Thomas Stimart

Chemistry & Biochemistry Theses & Dissertations

The Human Immunodeficiency Virus, type 1 (HIV-1) is the causative agent of the Acquired immunodeficiency Syndrome (AIDS). Currently, AIDS is the leading cause of death amongst young people in the United States. The large increase of HIV-1 infections over the past decade has spawned considerable research in fighting the spread of this disease. One area of intense HIV-1 research has focused on the enzyme Reverse Transcriptase (RT) which is an essential enzyme involved in the replication and life cycle of HIV-1. HIV-1 RT is composed of p66 and p51 subunits and is only active as a heterodimer in vivo. …

Structural Characterization Of A Novel Inhibitor Of Hiv Reverse Transcriptase (Hiv Rt), Greggory Jon Woitte

Chemistry & Biochemistry Theses & Dissertations

Human immunodeficiency virus (HIV) infections have become a leading cause of death among young people in the United States today. As the number of HIV infections increases, so too does the cost of treatment. Together, these numbers have prompted an increase in the development of pharmaceutical interventions. HIV reverse transcriptase (HIV RT) has become a suitable target for drug therapy because it is the sole enzyme responsible for HIV replication.

Fucoidan, a sulfated polysaccharide isolated from the brown algae Fucus vesiculosus, has been shown to block a variety of cell adhesion related events including metastasis. In addition, fucoidan has also …

Characterization Of An Antiviral Agent Based On Nonionic Surfactants And It's Effects On Human Dermal Fibroblasts, Ji Young Li

Biological Sciences Theses & Dissertations

The AIDS ·pandemic has directed various research endeavors towards finding an appropriate method for eliminating all potentially infectious material within bone allografts before implantation into a recipient. To that effect, Panavirocide was conceived by Medicine and Applied Science·s, Inc. This compound, in which three nonionic surfactants serve as the active agents, has been shown to inactivate HIV-1 particles within blood and blood products. Because Panavirocide has never been used with allografts, the purpose of the present research was to characterize the nonionic surfactants for the putative alteration of the formulation by determining the critical micelle concentration values and to determine …

Translational Regulation Of The C-Jun Proto-Oncogene, Anil Sehgal

Theses and Dissertations in Biomedical Sciences

The v-jun oncogene was originally isolated from the ASV17 virus in 1987. Ever since its isolation, extensive work has been done to understand the role of the v-jun oncogene in cell transformation. The c-Jun protein is a transcription factor which binds to the DNA target TGACTCA. The c-Jun protein binds to DNA in the form of dimers. It can form homodimers with itself and heterodimers with Jun family (JunB and JunD), Fos family (FosB, Fra1 and Fra2), or with CREB family members through the leucine zipper motif. Because the c-jun proto-oncogene plays an important role in cell transformation, extensive work …

Estimation Of Diversity And Community Structure Through Restriction-Fragment-Length-Polymorphism Distribution Analysis Of Bacterial 16s Ribosomal-Rna Genes From A Microbial Mat At An Active, Hydrothermal Vent System, Loihi Seamount, Hawaii, Craig L. Moyer, Fred C. Dobbs, David M. Karl

OES Faculty Publications

PCR was used to amplify (eu)bacterial small-subunit (16S) rRNA genes from total-community genomic DNA. The source of total-community genomic DNA used for this culture-independent analysis was the microbial mats from a deep-sea, hydrothermal vent system, Pele's Vents, located at Loihi Seamount, Hawaii. Oligonucleotides complementary to conserved regions in the 16S rRNA-encoding DNA (rDNA) of bacteria were used to direct the synthesis of PCR products, which were then subcloned by blunt-end ligation into phagemid vector pBluescript II. Restriction fragment length polymorphism patterns, created by using tandem tetrameric restriction endonucleases, revealed the presence of 12 groups of 16S rRNA genes representing discrete …

A Comparison Of No And N20 Production By The Autophic Nitrifier Nitrosomonas Europaea And The Heterotrophic Nitrifier Alcaligenes Faecalis, Iris C. Anderson, Mark Poth, Julie Homstead, David J. Burdige

OES Faculty Publications

Soil microorganisms are important sources of the nitrogen trace gases NO and N₂O for the atmosphere. Present evidence suggests that autotrophic nitrifiers such as Nitrosomonas europaea are the primary producers of NO and N₂O in aerobic soils, whereas denitrifiers such as Pseudomonas spp. or Alcaligenes spp. are responsible for most of the NO and N₂O emissions from anaerobic soils. It has been shown that Alcaligenes faecalis, a bacterium common in both soil and water, is capable of concomitant heterotrophic nitrification and denitrification. This study was undertaken to determine whether heterotrophic nitrification might be …

The Distribution And Occurrence Of Aerobic Heterotrophic Antimicrobic-Producing Bacteria Isolated From The Lower Chesapeake Bay, Linda Lea Gilmer

Biological Sciences Theses & Dissertations

Three different estuarine microenvironments in the lower Chesapeake Bay were investigated for the occurrence and geographical distribution of antimicrobic-producing bacteria. Samples were taken from the water column, fish skin, and algal surfaces. A total of 1472 bacterial isolates were assayed for antimicrobial activity against fungal, Gram-negative and Gram-positive bacterial test organisms. Of the total bacterial isolates assayed, 5. 2% were active antimicrobic producers. Algal surf aces were shown to have a higher incidence of antimicrobic-producing bacteria than the water column or fish skin. No antimicrobic activity was demonstrated by bacteria isolated from the flounder fish skin. The majority of the …

Kinetic Characterization Of A Recombinant C-Terminal Mutant Of Reverse Transcriptase From The Human Immunodeficiency Virus, Thomas S. Heard

Chemistry & Biochemistry Theses & Dissertations

The human immunodeficiency virus (HIV) reverse transcriptase (RT) (EC 2.7.7.49) is the central replication enzyme for HIV. In general, the kinetic mechanism for this and all other polymerases involves the ordered binding of two substrates: a primer-template (PT) followed by a deoxyribonucleoside triphosphate (dNTP). Previous investigations prompted this research when it was discovered that the substrate dNTP, in absence of PT, could protect a recombinant c-terminal mutant HIV-1 RT from inhibition by pyridoxal-5'-monophosphate (PLP), an active-site dNTP inhibitor. In contrast, the non-mutant recombinant HIV-1 RT required both substrates for protection from PLP inhibition. This investigation sought to determine if this …

A Genetic And Physiological Study Of The Role Of Extracellular Copper-Binding Proteins In Copper Detoxification By The Marine Bacterium Vibrio Alginolyticus, Valerie J. Harwood

Theses and Dissertations in Biomedical Sciences

Supernatant proteins in Vibrio alginolyticus batch cultures were analyzed by SDS-PAGE before copper was added, 24 and 48 hours after the addition of copper, and in 24 hour control (no Cu) cultures. Two proteins, one 21 kilodalton (kDa) and one 19 kDa, were found to be copper-induced, and were designated copper-binding protein 1 (CuBP1) and CuBP2. CuBP1 and CuBP2 became detectable in supernatants during the Cu-induced lag phase, and increased in concentration over the following 48 hours. Chloramphenicol inhibited production of these proteins. Gel-to-gel variability was implicated as the dominant factor determining whether one or two Cu-induced proteins were detected …

The Study Of Natural Product(S) From Opsanus Tau (Linnaeus), The Oyster Toadfish, Miki Taira

Chemistry & Biochemistry Theses & Dissertations

Opsanus tau (Linnaeus), the Oyster toadfish, commonly inhabits the Chesapeake Bay. The Oyster toadfish has been categorized as a venomous and/or ichthyotoxic fish; however, chemical studies to substantiate these claims have not been forthcoming.

The oyster toadfish has dorsal and opercular spines which are believed to secrete a venom(s). Based on our findings, the oyster toadfish does not secrete venom(s) from these spines.

The oyster toadfish secretes large amounts of a mucus when it is irritated. 3-Octanone was believed to be the major ichthyotoxic component of the mucus from the oyster toadfish. Based on our investigation, 3-octanone is present only …

Electrostatic Effects On Airborne Asbestos Monitoring, Roxanne Francis

Biological Sciences Theses & Dissertations

The affects of electrostatic forces on the Membrane Filter Method (MFM) used for air-borne asbestos monitoring have been investigated for many years. Several studies have indicated that these forces interfere with the collection of asbestos fibers on the membrane filter, which results in an underestimated analysis of the airborne concentration. By varying the electroconductivity of the extension cowls on the filter cassettes, it was speculated that the significance of the electrostatic forces could be statistically analyzed. Four sets of filter cassettes with grounded conductive (GC) extension cowls, ungrounded conductive (UC) extension cowls, grounded nonconductive (GN) extension cowls, and ungrounded nonconductive …

The Composition And Concentrations Of Cyanobacteria In The North Landing River, Virginia And Upper Currituck Sound, North Carolina, Sonja E. Midgette

Biological Sciences Theses & Dissertations

A 12 month study of cyanobacteria composition and concentrations in the North Landing River and upper Currituck Sound was conducted from March 1986 to February 1987. Epifluorescence microscopy and the fluorochrome proflavin were used to determine cell concentrations and vertical, station to station and seasonal distribution of these cells. Cyanobacterial assemblage patterns were coupled with pH, temperature, salinity and water transparency readings.

Cyanobacteria were found to be a persistent and prolific phytoplankton component of both the river and upper sound. For a comparison of vertical distribution for each station, an analysis of variance for. two groups of equal size was …

Vertical Distribution Of Phytoplankton Populations Along The Northeastern Continental Shelf Margin Of The United States, Bruce Burwell Wagoner

Biological Sciences Theses & Dissertations

A study of phytoplankton vertical distribution at six stations along the northeast continental shelf of the United States was conducted in April 1984.

A total of 168 taxa were identified from nine phylogenetic groups collected at depths up to 150 m. Highest cell concentrations and cell volumes were in the upper 30 m. Greatest cell concentrations were produced by diatoms and a pico-nanoplankton component, with cell volumes mainly the product of diatoms and dinoflagellates. A high degree of variability was observed between phylogenetic groups within each station, but less variability was noted for phylogenetic categories at different stations.

Little difference …

Interactions Between Aquatic Chromogenic Bacteria And Pathogenic Vibrio Spp., John Louis Harris

Biological Sciences Theses & Dissertations

Interactions between populations inhabiting an ecosys­tem are responsible, in large part, for determining which species coexist in that ecosystem. In this study, interac­tions between selected Vibrio species, V. alginolyticus and V. anguillarum, and marine heterotrophic, chromogenic bac­teria in aquatic microcosms were investigated. The ultimate goal of this project was to evaluate the feasibility of ex­ ploiting competitive interactions as a means to biologically control growth of the pathogenic marine bacteria, V. alginolyticus and V. anguillarum. In the majority of ex­periments (14/16), if chromogenic species were allowed to develop stable populations before a Vibrio spp. was intro­duced into a …

Seasonal Phytoplankton Assemblages In The Nansemond River, Virginia, Cindy Eleanor Shomers

Biological Sciences Theses & Dissertations

A monthly phytoplankton study was conducted on the Nansemond River from November 1984 to October 1985. Light microscopy was utilized to characterize the resident populations, their cell concentrations and to provide a general comparison to other regional studies of river systems considered more polluted. The results divided the flora into two major categories. The first group was a pico-nanoplankton component composed of Cyanobacteria and Chlorophycean cells 1-10 um in diameter. The second group was a microplankton assemblage of Bacillariophyceae, Dinophyceae, Cryptophyceae, Chlorophyceae, Cyanobacteria and Euglenophyceae. The pico-nanoplankton had a unimodal abundance pattern with peak concentrations occurring in spring. The microplankton …

A Simple Solid-Phase Electrophoretic Procedure For The Separation Of Plasmid Dna, Linda Ann Simurra

Biological Sciences Theses & Dissertations

A method was developed for extraction of plasmid DNA from bacterial cells embedded in agarose blocks. Cell - containing blocks were treated with various lysing reagents and inserted into the wells of an agarose gel. Upon electrophoresis the plasmid DNA migrated out of the embedding block and into the gel leaving intact chromosomal DNA in the well. This method was tested with various organisms and found to be effective for plasmid isolation. In comparison to "traditional" procedures, this new method is less tedious since chemical separation of plasmids is not required prior to electrophoresis. Also, a higher yield of plasmid …

Preservation Of Atp In Hypersaline Environments, Bruce J. Tuovila, Fred C. Dobbs, Paul A. Larock, B. Z. Siegel

OES Faculty Publications

High concentrations of particulate ATP were found in the anoxic brines of the Orca Basin and East Flower Garden, Gulf of Mexico. Other measurements indicative of growth and respiration suggested that the microbial community in the brines was inactive, but somehow the ATP associated with the cells persisted. Conceivably, when cells growing just above the interface sank into the brine, the increased osmotic stress could elicit an osmoregulatory response resulting in increased ATP. It was also possible that hydrolytic enzymes were inactivated, resulting in the preservation of ATP. Experiments in which a culture of marine bacteria was suspended in menstrua …

An Annual Study Of Phytoplankton Composition And Associated Environmental Conditions Of Lake Trashmore, Virginia Beach, Virginia, Seba B. Sheavly

Biological Sciences Theses & Dissertations

A 12 month study of Lake Trashmore was conducted on phytoplankton composition, inorganic nutrients and their seasonal associations. The dominant phytoplankton groups observed were diatoms, chlorophyceans and cyanobacteria. Phytoplankton assemblage patterns were coupled with pH, temperature, dissolved oxygen, nitrites and silicates. Diatoms dominated during periods of high nutrient concentrations, cooler temperatures and an unstable water column. Cyanobacteria forms dominated during periods of warmer temperatures, higher pH and decreased nitrogen and dissolved oxygen concentrations. A seasonal community shift occurred from cyanobacterial dominance to chlorophycean dominance when pH levels decreased. The lake is eutrophic based on indices of chlorophyll a concentrations, transparency …