Pathogenic Microbiology Commons^™

Cultivation Of Tropheryma Whipplei From Cerebrospinal Fluid, Matthias Maiwald, Axel Von Herbay, David N. Fredricks, Cleber C. Ouverney, Jon C. Kosek, David A. Relman

Faculty Publications, Biological Sciences

Whipple disease (WD) is a systemic disorder caused by the bacterium Tropheryma whipplei. Since the recognition of a bacterial etiology in 1961, many attempts have been made to cultivate this bacterium in vitro. It was eventually isolated, in 2000, from an infected heart valve, in coculture with human fibroblasts. Here we report the isolation of 2 new strains of T. whipplei from cerebrospinal fluid (CSF) of 2 patients with intestinal WD but no neurological signs or symptoms. One culture-positive specimen was obtained before treatment; the other was obtained 12 months after discontinuation of therapy, at a time of intestinal remission. …

Quorum Sensing In Candida Albicans: Probing Farnesol’S Mode Of Action With 40 Natural And Synthetic Farnesol Analogs, Roman Shchepin, Jacob M. Hornby, Erin Burger, Timothy Niessen, Patrick H. Dussault, Kenneth Nickerson

Kenneth Nickerson Papers

The dimorphic fungus Candida albicans produces extracellular farnesol (3,7, 11-trimethyl-2,6,10-dodecatriene- 1-ol) which acts as a quorum-sensing molecule (QSM) to suppress filamentation. Of four possible geometric isomers of farnesol, only the E,E isomer possesses QSM activity. We tested 40 natural and synthetic analogs of farnesol for their activity in an N-acetylglucosamine-induced differentiation assay for germ tube formation (GTF). Modified structural features include the head group, chain length, presence or absence of the three double bonds, substitution of a backbone carbon by S, O, N, and Se heteroatoms, presence or absence of a 3-methyl branch, and the bulkiness of the hydrophobic …

Prevalence Of Bacteria Of Division Tm7 In Human Subgingival Plaque And Their Association With Disease, Mary M. Brinig, Paul W. Lepp, Cleber C. Ouverney, Gary C. Armitage, David A. Relman

Faculty Publications, Biological Sciences

Members of the uncultivated bacterial division TM7 have been detected in the human mouth, but little information is available regarding their prevalence and diversity at this site. Human subgingival plaque samples from healthy sites and sites exhibiting various stages of periodontal disease were analyzed for the presence of TM7 bacteria. TM7 ribosomal DNA (rDNA) was found in 96% of the samples, and it accounted for approximately 0.3%, on average, of all bacterial rDNA in the samples as determined by real-time quantitative PCR. Two new phylotypes of this division were identified, and members of the division were found to exhibit filamentous …

Transcriptional Control Of The Opp Operon In Clostridium Difficile, S. Lindsey Davis

Inquiry: The University of Arkansas Undergraduate Research Journal

As a serious expense for both the hospital and the patient, nosocomial infections create a burden on the health care industry that is not easily overcome. Among the infections commonly contracted in the hospital environment, those associated with the Clostridium difficile bacterium account for millions of cases each year. Largely due to the nature of C. difficile infection as a response to the disruption of the normal flora of the colon caused by antibiotic activity, no completely effective treatment for this condition has been identified. It is this problem that forms the foundation for research devoted to the development of …

Eight Gram-Negative Bacteria Are 10,000 Times More Sensitive To Cationic Detergents Than To Anionic Detergents, Soumitra Rajagopal, Nicole Eis, Kenneth Nickerson

Kenneth Nickerson Papers

In liquid culture, eight typical Gram-negative bacteria were ca. 10,000-fold more sensitive to cationic detergents than to the anionic detergent sodium dodecyl sulfate. Cetyltrimethylammonium bromide (CTAB) was inhibitory at concentrations ranging from 0.0006% to 0.01%. Four pseudomonads able to form biofilms were ca. 1000-fold more resistant to CTAB on Luria–Bertani agar plates than they were in liquid culture. A lasI mutant of Pseudomonas aeruginosa was only able to tolerate 0.1% CTAB on Luria–Bertani agar plates but could tolerate 5% CTAB when supplemented with homoserine lactone containing culture supernatants.

En culture liquide, huit bactéries Gram négatif typiques étaient ca. 10,000 fois …

The Diversity And Abundance Of Parasites In Age Groups Of Yellow Perch In Lake Bemidji, Mn Applied To A Whittaker Curve, Kelly J. Main

Honors Capstones

Capstone submitted as a graduation requirement for the BSU Honors Program.

Comparison Of Tnf-A Levels Produced By Macrophages Treated With Brucella Abortus And Escherichia Coli Lps, Tajie Harris

Honors Capstones

Capstone submitted as a graduation requirement for the BSU Honors Program.

Effect Of Magnesium On The Production Of Scna And Scnr By Streptococcus Pyogenes, Strain Ff22, Laura E. Quinn

Biological Sciences Theses & Dissertations

The lantibiotic SA-FF22, produced by Streptococcus pyogenes strain FF22, was the first described antimicrobial peptide produced by a Streptococcus species. S. pyogenes is a group A Streptococcus responsible for such human illness as strep-throat, rheumatic fever, necrotizing fasciitis, endocarditis, and meningitis. Previous studies of SA-FF22 have shown that antimicrobial activity is lost in the presence of lmM magnesium. We hypothesize that the lack of SA-FF22 activity in the presence of magnesium is due to an absence of transcription of scnA, the gene encoding SA-FF22. The lack of transcriptional activation of scnA may be due to an absence of ScnR, a …

Escherichia Coli O157:H7 In Free-Ranging Deer In Nebraska, David G. Renter, Jan M. Sargeant, Scott E. Hygnstrom, Jeff D. Hoffmann, Jerry R. Gillespie

School of Natural Resources: Faculty Publications

In order to determine the prevalence and distribution of the human pathogen, Escherichia coli O157:H7, in free-ranging deer, hunters were asked to collect and submit fecal samples from deer harvested during a regular firearm season (14–22 November 1998). Prior to the season, 47% of the hunters with permits in the southeastern Nebraska (USA) study area indicated a willingness to participate in the study. Approximately 25% of successful hunters in the area submitted deer fecal samples. Escherichia coli O157:H7 was cultured from four (0.25%) of 1,608 total samples submitted. All of the fecal samples that were properly identified (1,426) and all …

Marine Planktonic Archaea Take Up Amino Acids, Cleber C. Ouverney, Jed A. Fuhrman

Faculty Publications, Biological Sciences

Archaea are traditionally thought of as “extremophiles,” but recent studies have shown that marine planktonic Archaea make up a surprisingly large percentage of ocean midwater microbial communities, up to 60% of the total prokaryotes. However, the basic physiology and contribution of Archaea to community microbial activity remain unknown. We have studied Archaea from 200-m depths of the northwest Mediterranean Sea and the Pacific Ocean near California, measuring the archaeal activity under simulated natural conditions (8 to 17°C, dark and anaerobic) by means of a method called substrate tracking autoradiography fluorescence in situ hybridization (STARFISH) that simultaneously detects specific cell types …

Integronlike Structures In Campylobacter Spp. Of Human And Animal Origin, Brigid Lucey, D. Crowley, P. Moloney, B. Cryan, M. Daly, F. O'Halloran, E. J. Threlfall, S. Fanning

Department of Biological Sciences Publications

Resistance to antimicrobial agents used to treat severe Campylobacter spp. gastroenteritis is increasing worldwide. We assessed the antimicrobial resistance patterns of Campylobacter spp. isolates of human and animal origin. More than half (n = 32) were resistant to sulphonamide, a feature known to be associated with the presence of integrons. Analysis of these integrons will further our understanding of Campylobacter spp. epidemiology.

The Molecular Characterisation Of Selected Epidemic Associated Gram Negative Pathogens In Southern Ireland, Mairéad Daly

Theses

This study was undertaken to evaluate various molecular techniques for the analysis of Gram negative bacterial associated epidemics. The first investigation dealt with bovine mastitis outbreaks, where the causative agent was identified as Pseudomonas aeruginosa. Epidemiological findings, suggested that all herds were infected from teat wipes that were contaminated with this organism. Initial investigations using the polymerase chain reaction (PCR), indicated a possible clonal relationship between all outbreak linked strains, with one exception. This finding was confirmed following pulsed-field gel electrophoresis (PFGE) and ribotype analysis. PCR was again employed to study a rather unusual epidemic, involving Salmonella tel-el-kebir over a …

Genetic And Structural Analyses Of Cytoplasmic Filaments Of Wild-Type Treponema Phagedenis And A Flagellar Filament-Deficient Mutant, Jacques Izard, William A. Samsonoff, Mary Beth Kinoshita, Ronald J. Limberger

Department of Food Science and Technology: Faculty Publications

Unique cytoplasmic filaments are found in the treponeme genus of spirochete bacteria. Their function is unknown, but their location underneath the periplasmic flagellar filaments (PFF) suggests a role in motility and/or cell structure. To better understand these unique structures, the gene coding for the cytoplasmic filaments, cfpA, was identified in various treponemal species. Treponema phagedenis cfpA was 2,037 nucleotides long, and the encoded polypeptide showed 78 to 100% amino acid sequence identity with the partial sequence of CfpA from T. denticola, T. vincentii, and T. pallidum subsp. pertenue. Wild-type T. phagedenis and a …

Insertional Inactivation Of Treponema Denticola Tap1 Results In A Nonmotile Mutant With Elongated Flagellar Hooks, Ronald J. Limberger, Linda L. Slivienski, Jacques Izard, William A. Samsonoff

Department of Food Science and Technology: Faculty Publications

The treponemal fla operon is comprised of numerous motility-related genes; however, the initial gene of this operon, tap1, has no known function. A recently developed system to generate specific mutants in Treponema denticola was utilized to determine if Tap1 was essential for motility. T. denticola tap1 and flanking DNA were identified, cloned, and sequenced, and a suicide plasmid that contained tap1 interrupted with an erythromycin resistance cassette (ermF and ermAM) was constructed. Because of potential polar effects from this cassette, a second plasmid that contained tap1 interrupted with a modified erythromycin resistance cassette that lacked the putative ermF transcription …

An Economic Analysis Of The Threat Of An Exotic Disease On Agriculture: The Case Of Sheep Pox In Western Australia, Nhu Che, Tom Kompas

Biosecurity published reports

Sheep pox is an acute disease that causes heavy losses due to high sheep mortality, costly eradication programs and ongoing problems with trade restrictions for many countries around the world. Australia until now has been unaffected, and although the risk of disease entry may be very slight, one case of sheep pox outbreak could cost the Western Australian economy as much as $50 million in damages, mostly from the trade disruption losses ( over the 21-month period of trading bans). The introduction of a Serology Test Program may reduce that amount by $35 million due to a shortening of the …

Chlorella Virus Pbcv-1 Encodes A Functional Homospermidine Synthase, Annette Kaiser, Michael Vollmert, Dorothea Tholl, Michael V. Graves, James Gurnon, Weimei Xing, Amber D. Lisec, Kenneth Nickerson, James L. Van Etten

Kenneth Nickerson Papers

Sequence analysis of the 330-kb genome of chlorella virus Paramecium bursaria chlorella virus 1 (PBCV-1) revealed an open reading frame, A237R, that encodes a protein with 34% amino acid identity to homospermidine synthase from Rhodopseudomonas viridis. Expression of the a237r gene product in Escherichia coli established that the recombinant enzyme catalyzes the NAD⁺-dependent formation of homospermidine from two molecules of putrescine. The a237r gene is expressed late in PBCV-1 infection. Both uninfected and PBCV-1-infected chlorella, as well as PBCV-1 virions, contain homospermidine, along with the more common polyamines putrescine, spermidine, and cadaverine. The total number of polyamine …

In Vitro Growth Characteristics Of Two Cryptococcus Neoformans Isolates, Kristy L. Jones, Juneann W. Murphy

Journal of the Arkansas Academy of Science

Cryptococcus neoformans is an opportunistic pathogen which attacks individuals with weakened immune systems. Two C. neoformans isolates, 184A and NU-2, were examined to determine characteristics that contribute to their difference in virulence. Both isolates were grown under tissue culture conditions, and different characteristics were tested at timed intervals. Isolate 184A was found to grow at a faster rate than isolate NU-2, with doubling times of 4 and 12 hours respectively. The polysaccharide capsule of isolate NU-2 doubled in size in four hours, while that of isolate 184A stayed approximately the same. The NU-2 yeast cell diameter without the polysaccharide capsule …

Recovery Of Rna Polymerase Ii Synthesis Following Dna Damage In Mutants Of Saccharomyces Cerevisiae Defective In Nucleotide Excision Repair, Michael S. Reagan, Errol C. Friedberg

Biology Faculty Publications

We have measured the kinetics of the recovery of mRNA synthesis in the inducible GAL10 and RNR3 genes after exposure of yeast cells to ultraviolet (UV) radiation. Such recovery is abolished in mutant strains defective in nucleotide excision repair (NER) of DNA, including a rad23 mutant. Mutants defective in the RAD7 or RAD16 genes, which are required for the repair of the non-transcribed strand but not the transcribed strand of transcriptionally active genes, show slightly faster recovery of RNA synthesis than wild-type strains. A strain deleted of the RAD26 gene, which is known to be required for strand-specific NER in …

Increase In Fluorescence Intensity Of 16s Rrna In Situ Hybridization In Natural Samples Treated With Chloramphenicol, Cleber C. Ouverney, Jed A. Fuhrman

Faculty Publications, Biological Sciences

Despite the numerous advantages of fluorescent in situ hybridization for the identification of single prokaryotic cells with 16S rRNA probes, use of the technique with natural samples, especially those from the marine environment, is still problematic. The low percentage of fluorescently labeled cells constitutes the primary problem for in situ hybridization of natural samples, probably due to low cellular rRNA content. This study represents an attempt to improve detection of marine prokaryotes by increasing cellular rRNA content without changing the species composition. Cells from three California coastal sites were treated with chloramphenicol, an inhibitor of protein synthesis and rRNA degradation, …

Communities Of Cloacal Bacteria In Tree Swallow Families, Michael P. Lombardo, Patrick A. Thorpe, R. Cichewicz, M. Henshaw, C. Millard, C. Steen, T. K. Zeller

Peer Reviewed Publications

Our aim in this study was to survey the communities of bacteria found in the cloacae of adult and nestling Tree Swallows (Tachycineta bicolor), determine if there were familial patterns of prevalence, and determine if there were relationships between bacteria loads and nestling size when 12 days old and fledging success.

Membrane Topology Of The Colicin A Pore-Forming Domain Analyzed By Disulfide Bond Engineering, Denis Duche, Jacques Izard, Juan M. Gonzalez-Manas, Michael W. Parker, Marcel Crest, Daniel Baty

Department of Food Science and Technology: Faculty Publications

Four colicin A double-cysteine mutants possessing a disulfide bond in their pore-forming domain were constructed to study the translocation and the pore formation of colicin A. The disulfide bonds connected a-helices 1 and 2, 2 and 10, 3 and 9, or 3 and 10 of the poreforming domain. The disulfide bonds did not prevent the colicin A translocation through the Escherichia coli envelope. However, the mutated colicins were able to exert their in vivo channel activity only after reduction of their disulfide bonds. In vitro studies with brominated phospholipid vesicles and planar lipid bilayers revealed that the disulfide bond that …

A Biosystematic Study Of Three Plant Pathogenic Fungi, Richard James Langham

Masters Theses

Isolates of Chalara australis, Eucalypt Ceratocystis, and Ceratocystis virescens were grown on Potato Dextrose Agar, Malt Extract Agar, and Czapek Agar and examined as part of a biosystematic study on these fungi. Basic micromorphological and macromorphological data on the cultures were recorded and the taxa were found to be morphologically similar. Tissue isolates and single spore isolates were crossed within these taxa and an interspecific cross performed. Chalara australis was found to be either asexual or have only one mating type present in the cultures studied. Eucalypt Ceratocystis was found to have a mating system characteristic of bipolar …

Direct Immunofluorescence Detection Of Giardia Spp. In Selected Mammals In Central Illinois, Using Anti-Giardia Polyclonal And Monoclonal Antibodies, Jeanette Aba Ackon Brown

Masters Theses

Giardia species occur in many kinds of mammals, and some of these hosts have been postulated as potential reservoirs for human infections. A study was initiated in fall 1990 to determine the distribution and frequency of giardiasis in populations of selected wild animals in 12 counties in Illinois. Fecal samples from 64 white-tailed deer, 13 coyotes, 9 muskrats, 6 raccoons, and 5 badgers were examined for the presence of Giardia spp. and specifically for G. lamblia. Anti-Giardia lamblia cysts polyclonal (PAb) and monoclonal (MAb), FITC-labeled antibody solutions were used for the direct immunofluorescence detection of Giardia cysts in …

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 …

Effects Of Corticosteroids And Of Cyclosporin A On Trypanosoma Musculi In Mice And In Culture, Philip Stephen Schuyler

Loma Linda University Electronic Theses, Dissertations & Projects

The in vivo and in vitro effects of cyclosporin A and corticosteroids on Trypanosoma musculi were examined. Mice that received continuous administration of 6 mg/liter/day of dexamethasone in drinking water and 200 mg/kg hydrocortisone acetate as a single dose 24 hrs prior to injection with T. musculi had significantly higher and longer parasitemias, than controls. Body and spleen weights and B cell responsiveness were lower in test animals and there were some deaths. Mice injected with T. musculi that received 100 mg/kg/day cyclosporin A had longer, but not higher, parasitemias than controls. Corticosteroids appear to partially inactive antibody production (i.e. …

Analysis Of The Charleston, Illinois Water Supply For The Presence Of Organic Mutagens Utilizing The Ames Salmonella/Microsome Assay, Steven Lee Washburn

Masters Theses

Water samples from the Charleston, Illinois water supply and selected agricultural run-off areas were tested for the presence of organic mutagenic chemicals. The Ames Salmonella/microsome mutagenicity assay utilizing strains TA98 and TA100 was used to analyze concentrated water samples. A special set of samples was exposed to the S9 activation system coupled with the TA98 strain to simulate the mammalian metabolic system.

No mutagens were detected in any of the samples tested. Several positive and negative control systems were included in the testing system. Negative control systems were used to determine the average spontaneous reversion rates for each strain for …

Fecal Coliform Membrane Filtration Recovery Techniques, Pamela Rose Rinebold

Masters Theses

Researchers over the years have been trying to improve recovery techniques for fecal coliform bacteria. Improved techniques would provide better estimates of the number of fecal coliform bacteria as well as better estimates of fecal contamination with potential pathogenicity of food and water.

Stressed, but viable, fecal coliform bacteria if placed under the appropiate conditions, can recover, thus presenting the possibility of inadequate disinfection. Chlorine sanitization, as done in potable and wastewater treatment, is one such method of stressing bacteria.

Techniques of recovery were studied using presumably unstressed and chlorine stressed samples. Phase one of this study was the comparison …

Prevalence Of Extreme Detergent Resistance Among The Enterobacteriaceae, Vance Kramer, Kenneth Nickerson, Nancy V. Hamlett, Caroline O'Hara

Kenneth Nickerson Papers

The detergent-resistance properties of 208 independent isolates of the Enterobacteriaceae have been examined. Of these bacterial strains, 200 were able to grow in the presence of ≥5% sodium dodecyl sulfate, including all members of the Klebsielleae tribe. This resistance does not appear to be plasmid encoded. It is proposed that detergent-resistant organisms he termed saponotolerant or saponophilic, by analogy with other microorganisms occupying harsh ecological niches. In contrast to their prevalent resistance to anionic detergents, not one of the 208 strains tested was found to grow in the presence of three different cationic detergents. This sensitivity to cationic detergents may …

Distribution Of Naegleria Fowleri In Selected Northeast Arkansas Lakes, Robert Dean Evans, Lawrence W. Hinck

Journal of the Arkansas Academy of Science

Seven northeast Arkansas recreational lakes were examined for the presence of pathogenic and nonpathogenic Naegleria fowleri. Cultural differentiation and microscopic morphology were used as species determining tests, while mouse pathogenicity tests were conducted to determine virulence. Only one isolate met all criteria utilized for definite identification of Naegleria fowleri, although Naegleria type organisms were found in all of the lakes. None of the isolates were pathogenic in mice.

The Development Of Techniques For The Enumeration Of Selected Pathogenic Bacteria In Lynnhaven Estuarine Sediments, Mark T. Frank

Biological Sciences Theses & Dissertations

Methodologies for the improved recovery of selected pathogenic bacteria from estuarine sediments were investigated. Laboratory and field studies were conducted to document the sensitivity of the techniques for isolating Salmonella species, Pseudomonas aeruginosa, and Vibrio parahaemolyticus. Vibrio parahaemolyticus-like organisms were enriched in a seawater yeast extract broth and isolated from Thiosulfate Citrate Bile Salts Sucrose plated media. Enrichment procedures with and without elevated temperatures and subsequent transfers to selective and differential plated media have been examined for the isolation of Salmonella species and P. aeruginosa. These pathogens were initially assayed for utilizing primary enrichments of asparagine …