Microbiology Commons^™

Inoculum Size Effect In Dimorphic Fungi: Extracellular Control Of Yeast-Mycelium Dimorphism In Ceratocystis Ulmi, Jacob M. Hornby, Sarah M. Jacobitz-Kizzier, Donna J. Mcneel, Ellen C. Jensen, David S. Treves, Kenneth W. Nickerson

Papers in Microbiology

We studied the inoculum size effect in Ceratocystis ulmi, the dimorphic fungus that causes Dutch elm disease. In a defined glucose-proline-salts medium, cells develop as budding yeasts when inoculated at >106 spores per ml and as mycelia when inoculated at type, age of the spores, temperature, pH, oxygen availability, trace metals, sulfur source, phosphorous source, or the concentration of glucose or proline. Similarly, it was not influenced by added adenosine, reducing agents, methyl donors, amino sugars, fatty acids, or carbon dioxide. Instead, growing cells excreted an unknown quorum-sensing factor that caused a morphological shift from mycelia to budding yeasts. …

Detection Of Bacteria To Dental Tissue By Low-Light Imaging, G-Hong Robert Hsu

Loma Linda University Electronic Theses, Dissertations & Projects

Bacteria have been shown to play an important role in the development of pulp and periradicular disease. The source of the infective bacteria is most often the oral cavity. Previous studies have detected bacteria in teeth that are structurally intact that seemingly have no communication intraorally. Anachoresis has been proposed as a mechanism for localizing circulating bacteria to such teeth. In several previous bacteriologic anachoresis studies, bacteria have been found to localize in inflamed dental tissues. The criticism most commonly associated with these investigations has been the potential contribution of contaminant bacteria resulting in false positive findings.

Low-light image analysis …

The Biological Burden Of Antibiotic Resistance In Staphylococcus Aureus, Natasha Samantha Dean

Loma Linda University Electronic Theses, Dissertations & Projects

Antibiotic resistance is a major problem. Resistance is often mediated by genes on bacterial plasmids and dissemination of resistance is commonly achieved by plasmid transfer. The mechanisms of resistance that are presently understood suggest that resistance is energetically costly. If this is generally true, resistance would reduce the fitness of cells in the absence of antibiotics. We examined the hypothesis that resistance reduces the fitness of microorganism and requires continuous selection to be maintained. To test this hypothesis, we measured the effects of plasmid-borne resistance genes on cell growth by comparing the growth rates of Staphylococcus aureus with and without …

Oxalate Metabolism By The Acetogenic Bacterium Moorella Thermoacetica, Steven L. Daniel, Christine Pilsl, Harold L. Drake

Faculty Research & Creative Activity

Whole-cell and cell-extract experiments were performed to study the mechanism of oxalate metabolism in the acetogenic bacterium Moorella thermoacetica. In short-term, whole-cell assays, oxalate consumption was low unless cell suspensions were supplemented with CO2, KNO3, or Na2S2O3. Cell extracts catalyzed the oxalate-dependent reduction of benzyl viologen. Oxalate consumption occurred concomitant to benzyl viologen reduction; when benzyl viologen was omitted, oxalate was not appreciably consumed. Based on benzyl viologen reduction, specific activities of extracts averaged 0.6 μmol oxalate oxidized min−1 mg protein−1. Extracts also catalyzed the formate-dependent reduction of NADP+; however, oxalate-dependent reduction of NADP+ was negligible. Oxalate- or formate-dependent reduction …

Oxalate Metabolism By The Acetogenic Bacterium Moorella Thermoacetica, Steven Daniel, Christine Pilsl, Harold Drake

Faculty Research & Creative Activity

Whole-cell and cell-extract experiments were performed to study the mechanism of oxalate metabolism in the acetogenic bacterium Moorella thermoacetica. In short-term, whole-cell assays, oxalate consumption was low unless cell suspensions were supplemented with CO2, KNO3, or Na2S2O3. Cell extracts catalyzed the oxalate-dependent reduction of benzyl viologen. Oxalate consumption occurred concomitant to benzyl viologen reduction; when benzyl viologen was omitted, oxalate was not appreciably consumed. Based on benzyl viologen reduction, specific activities of extracts averaged 0.6 μmol oxalate oxidized min−1 mg protein−1. Extracts also catalyzed the formate-dependent reduction of NADP+; however, oxalate-dependent reduction of NADP+ was negligible. Oxalate- or formate-dependent reduction …

Ethiopian Soils Harbor Natural Populations Of Rhizobia That Form Symbioses With Common Bean (Phaseolus Vulgaris L.), Serawit Kassa, Desta Beyene, Franklin Ampy, Amha Asseffa, Tadesse Gebremedhin, Peter Van Berkum

Department of Biology Faculty Publications

The diversity and taxonomic relationships of 83 bean-nodulating rhizobia indigenous to Ethiopian soils were characterized by PCR-RFLP of the internally transcribed spacer (ITS) region between the 16S and 23S rRNA genes, 16S rRNA gene sequence analysis, multilocus enzyme electrophoresis (MLEE), and amplified fragment-length polymorphism. The isolates fell into 13 distinct genotypes according to PCR-RFLP analysis of the ITS region. Based on MLEE, the majority of these genotypes (70%) was genetically related to the type strain of Rhizobium leguminosarum. However, from analysis of their 16S rRNA genes, the majority was placed with Rhizobium etli. Transfer and recombination of the 16S rRNA …

Phylogenetic Analyses Of Texas Isolates Indicate An Evolving Subtype Of The Clade B Feline Immunodeficiency Viruses, Eric A. Weaver, Ellen W. Collisson, Margaret Slater, Guan Zhu

Nebraska Center for Virology: Faculty Publications

Rigorous phylogenetic analyses were used to compare the nucleotide sequences of feline immunodeficiency virus strains isolated from Texas and throughout the world. The envelope V3-V4 sequences and capsid gene of the Texas isolates formed a cluster between subtypes B and E. Statistical comparisons with other published sequences confirmed that the Texas group is a unique cluster, possibly a new subtype, arising from subtype B.

Feline immunodeficiency virus (FIV) was initially isolated in 1987 from a cat in California with severe immunodeficiency and has been recognized as a common worldwide feline pathogen (11, 14, 19, 20, 33). FIV-infected cats exhibit a …

Q-Probes Study Of Replicate Specimens At The Clinical Laboratory, The Aga Khan University Hospital, Karachi, L Ali, I Siddiqui

Department of Pathology and Laboratory Medicine

Objective: The aim of this Q-probes study was to evaluate the precision of replicate specimens.Methods: This Q-probes study done at the Clinical Laboratory, The Aga Khan University Hospital Karachi is about routine chemistry analytes because of their importance in critical care patients. The analytes data was collected for six months from April to September 2002. There were total 358 samples, which were given for reanalysis during this period under the constant working conditions. After analysis, results of both the runs on different days, were compared to determine the percent difference between the results. After calculating the percent difference, the results …

Extracullular Atp Regulates Il-1beta Release From Microglial Cells Via Purinergic Receptor After In Vitro Trauma, Chengya Liang

Electronic Theses and Dissertations

Traumatic brain injury (TBI) induces a state of microglialactivation, which includes upregulation of macrophage functions and release inflammatory mediators such as certain inflammatory cytokines. Current literature suggests that interleukin-1Beta is an important cytokine mediator, which is dramatically increased after brain injury. Previous studies indicate that ATP is released by traumatically injured astrocytes and serves as a cell-to-cell mediator through purinergic receptors after in vitro injury. However, the mechanism of interleukin-1Beta release after traumatic brain injury remains poorly defined and is difficult to study using in vivo models. Using an in vitro model for traumatic brain injury (cell strain or stretch), …

Microbial Endemism And Biogeography, Brian P. Hedlund, James T. Staley

Life Sciences Faculty Research

The topic of microbial biogeography is almost 100 years old, however, when confronted with questions about the existence and extent of endemism in the microbial world, many microbiologists respond with opinions and theoretical arguments rather than examples of well-conducted studies. We begin this chapter with an overview of this debate as it applies to free-living prokayotes in part because there are relatively few good microbial biogeography studies. Furthermore, the arguments help to frame microbial biogeography in the larger context of biodiversity in that if endemism is common, then many more species exist.

Estimation Of Microbial Diversity In Poultry Litter Using Terminal Restriction Fragment Length Polymorphism And Isolation Of Phosphate Accumulating Bacteria From Poultry Litter, Yoganand Vadari

Masters Theses & Specialist Projects

The contamination of fresh water by phosphates in poultry litter results in substantial eutrophication of fresh water causing fish kills and other types of environmental damage. The poultry indus try in Kentucky is expanding rapidly. The number of broilers is increasing as more poultry farms are established in the state producing waste that needs disposal. Investigations were made to study the possibility of using microorganisms normally found in poultry litter to sequester phosphate, thereby delaying phosphate runoff after litter is applied to croplands. Little is known, however, about the microflora of poultry litter. Terminal restriction fragment length polymorphism of 16S …

Survival Of Mycobacterium Avium Subspecies Paratuberculosis In The Pol, John Rumsey

Electronic Theses and Dissertations

Mycobacterium avium subspecies paratuberculosis (map) is an intracellular pathogen that is known to parasitize macrophages and monocytes. Map infiltrates gastrointestinal tract host tissue where it is the known etiological agent of johne's disease in ruminants and implicated in the etiology of crohn's disease in humans. Map's ability to survive within macrophages enables it to disseminate throughout the rest of the host, possibly infecting other circulating blood leukocytes. In this study, the survival and fate of map strain atcc 43015 (human isolate) following phagocytosis was determined using in vitro murine macrophage cell line j774a.1 and polymorphonuclear cells (pmnc's) from five crohn's …

Evaluation Of The Host Range Of Polyvalent Staphylococcal Bacteriophages To Control Mrsa (Staphylococcus Aureus) From Irish Hospitals, Mohamed Farag M. Elbreki

Theses

This study describes firstly, the assembly of a collection of a varied spectrum of staphylococci which can be used to test the efficacy of bacteriophages for the biocontrol of problematic members of this genus. The collection assembled includes a variety of multi-antibiotic resistant MRSA (methicillin resistant S. aureus) strains, hVRSA (hetero vancomycin resistant S. aureus), VRSA and also teicoplenin resistant isolates. It also included a broad range of species of coagulase-negative {non-aureus) staphylococci including S. caprea, S. haemolyticus, S. saprophyticus, S. epidermidis, S. chromogenes, S. capitis, S. hominins and S. hyicus. All strains were characterized by morphology on Baird Parker, …

Genomes Of The Parapoxviruses Orf Virus And Bovine Papular Stomatitis Virus, Gustavo A. Delhon, E. R. Tuhlman, C. L. Afonso, Z. Lu, A. De La Concha-Bermejillo, H. D. Lehmkuhl, M. E. Piccone, G. F. Kutish, D. L. Rock

Nebraska Center for Virology: Faculty Publications

Bovine papular stomatitis virus (BPSV) and orf virus (ORFV), members of the genus Parapoxvirus of the Poxviridae, are etiologic agents of worldwide diseases affecting cattle and small ruminants, respectively. Here we report the genomic sequences and comparative analysis of BPSV strain BV-AR02 and ORFV strains OV-SA00, isolated from a goat, and OV-IA82, isolated from a sheep. Parapoxvirus (PPV) BV-AR02, OV-SA00, and OV-IA82 genomes range in size from 134 to 139 kbp, with an average nucleotide composition of 64% G+C. BPSV and ORFV genomes contain 131 and 130 putative genes, respectively, and share colinearity over 127 genes, 88 of which …

Identification And Characterization Of A Nontypeable Haemophilus Influenzae Putative Toxin-Antitoxin Locus, Dayle A. Daines, Justin Jarisch, Arnold L. Smith

Biological Sciences Faculty Publications

Background: Certain strains of an obligate parasite of the human upper respiratory tract, nontypeable Haemophilus influenzae (NTHi), can cause invasive diseases such as septicemia and meningitis, as well as chronic mucosal infections such as otitis media. To do this, the organism must invade and survive within both epithelial and endothelial cells. We have identified a facilitator of NT(Hi) survival inside human cells, virulence-associated protein D (vapD_Hi, encoded by gene H10450). Both vapD_Hi and a flanking gene, H10451, exhibit the genetic and physical characteristics of a toxin/antitoxin ( TA) locus, with VapD_Hi serving as the …

Endocrine-Disrupting Effects Of Cattle Feedlot Effluent On An Aquatic Sentinel Species, The Fathead Minnow, Edward Orlando, Alan Kolok, Gerry Binzcick, Jennifer Gates, Megan Horton, Christy Lambright, L. Earl Gray Jr., Ana M. Soto, Louis J. Guilette

Biology Faculty Publications

Over the last decade, research has examined the endocrine-disrupting action of various environmental pollutants, including hormones, pharmaceuticals, and surfactants, in sewage treatment plant effluent. Responding to the growth of concentrated animal feeding operations (CAFOs) and the pollutants present in their wastewater (e.g., nutrients, pharmaceuticals, and hormones), the U.S. Environmental Protection Agency developed a new rule that tightens the regulation of CAFOs. In this study, we collected wild fathead minnows (Pimephales promelas) exposed to feedlot effluent (FLE) and observed significant alterations in their reproductive biology. Male fish were demasculinized (having lower testicular testosterone synthesis, altered head morphometrics, and smaller …

Morphological Characteristics And Species Separation Of Hawaiian Postlarval Amphidromous Fishes, M. Eric Benbow, Albert J. Burky, Carl M. Way

Biology Faculty Publications

There are five Hawaiian amphidromous fishes (Gobiidae: Lentipes concolor, Awaous guamensis, Sicyopterus stimpsoni, Stenogobius hawaiiensis; Eleotridae: Eleotris sandwicensis). Amphidromous adults deposit eggs on the stream bottom. After hatching, larvae drift to the ocean for growth followed by postlarval migration back into the streams. Postlarvae were collected to construct a dichotomous identification key based on 12 morphological measures and ray counts from four fins. Overall, the presense of fused pelvic fins, standard length (SL), and fin ray numbers were the most useful in species separation. Gobies were separated from the eleotrid by the former having fused …

Ultrastructure Of Mycobacterium Marinum Granuloma In Striped Bass Morone Saxatilis, D. T. Gauthier, W. K. Vogelbein, C. A. Ottinger

Biological Sciences Faculty Publications

An emerging epizootic of mycobacteriosis currently threatens striped bass Morone saxatilis populations in Chesapeake Bay, USA. Several species of mycobacteria, including Mycobacterium marinum, species resembling M. avium, M. gordonae, M. peregrinum, M. scrofulaceum and M. terrae, and the new species M. shottsii have been isolated from diseased and healthy bass. In this study, we describe the ultrastructure of developing M. marinum granulomas in experimentally infected bass over a period of 45 wk. The primary host response to injected mycobacteria was formation of large macrophage aggregations containing phagocytosed bacilli. M. marinum were always contained within phagosomes. Close …

Cd40-Associated Traf 6 Signaling Is Required For Disease Induction In A Retrovirus-Induced Murine Immunodeficiency, Kathy A. Green, Cory L. Ahonen, W. James Cook, William R. Green

Dartmouth Scholarship

LP-BM5 retrovirus-infected C57BL/6 mice develop splenomegaly, lymphadenopathy, hypergammaglobulinemia, and immunodeficiency; thus, this disease has been named mouse AIDS. In this syndrome, CD154/CD40 interactions are required for but do not mediate disease by upregulation of CD80 or CD86. We report here that there is nonetheless a necessity for CD40 signaling competence, specifically an intact tumor necrosis factor receptor-associated factor 6 (TRAF 6) binding site.

Cd46-Mediated Transduction Of A Species D Adenovirus Vaccine Improves Mucosal Vaccine Efficacy, Zenaido T. Camacho, Mallory A. Turner, Michael A. Barry, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

The high levels of preexisting immunity against Adenovirus type 5 (Ad5) have deemed Ad5 unusable for translation as a human vaccine vector. Low seroprevalent alternative viral vectors may be less impacted by preexisting immunity, but they may also have significantly different phenotypes from that of Ad5. In this study we compare species D Ads (26, 28, and 48) to the species C Ad5. In vitro transduction studies show striking differences between the species C and D viruses. Most notably, Ad26 transduced human dendritic cells much more effectively than Ad5. In vivo imaging studies showed strikingly different transgene expression profiles. The …

Phylogenetic Analyses Of Texas Isolates Indicate An Evolving Subtype Of The Clade B Feline Immunodeficiency Viruses, Eric A. Weaver, Ellen W. Collisson, Margaret Slater, Guan Zhu

Nebraska Center for Virology: Faculty Publications

Rigorous phylogenetic analyses were used to compare the nucleotide sequences of feline immunodeficiency virus strains isolated from Texas and throughout the world. The envelope V3-V4 sequences and capsid gene of the Texas isolates formed a cluster between subtypes B and E. Statistical comparisons with other published sequences confirmed that the Texas group is a unique cluster, possibly a new subtype, arising from subtype B.

The Hemolymph Jh Titer Exhibits A Large-Amplitude, Morph-Dependent, Diurnal Cycle In The Wing-Polymorphic Cricket, Gryllus Firmus, Zhangwu Zhao, Anthony J. Zera

Anthony Zera Publications

The hemolymph juvenile hormone (JH) titer was measured in over 500 flight-capable and flightless, adult female Gryllus firmus at 3–6 h intervals during each of days 2–8 of adulthood. The flight-capable morph exhibited a large-amplitude daily cycle in the hemolymph JH titer, while the flightless morph exhibited a barely perceptible cycle. The JH titer cycle was observed on all days in the flight-capable morph, but the large amplitude cycle (>15–20 fold increase in mean titer; >100-fold increase in some individuals), began on day 5. For both the large and small amplitude cycles, the JH titer peaked near the end …

High Phosphate (Up To 600 Mm) Induces Pseudohyphal Development In Five Wild Type Candida Albicans, Jacob M. Hornby, Raluca Dumitru, Kenneth Nickerson

Kenneth Nickerson Papers

A method is described for the formation of nearly 100% pseudohyphae populations of wild-type Candida albicans A72. The method employs fungal growth at 37 °C (ca. 5×10⁶ cells/ml) in a glucose–proline–N-acetyl-glucosamine medium supplemented with up to 600 mM phosphate (KH₂PO₄/K₂HPO₄ 1:1) at pH 6.5. Four other strains of C. albicans (MEN, 10261, SG5314 and CAI-4) also formed pseudohyphae under these conditions, although the phosphate response profiles differed in the concentration required for each strain to form pseudohyphae.

Isolation And Characterization Of Mycobacteria From Striped Bass Morone Saxatilis From The Chesapeake Bay, Martha W. Rhodes, Howard Kator, Ilsa Kaattari, David Gauthier, Wolfgang Vogelbein, Christopher A. Ottinger

Biological Sciences Faculty Publications

Mycobacteriosis in striped bass Morone saxatilisof Chesapeake Bay, USA, was first diagnosed in 1997 based on the presence of granulomatous inflammation and acid-fast bacteria in skin and spleen. To confirm histopathology, bacteriological detection and identification of mycobacteria were begun using splenic tissue from fish with and without skin ulcerations. On the basis of initial studies using a variety of selective and nonselective media, decontamination, homogenization and incubation conditions, a simple and quantitative recovery method using aseptic necropsy of splenic tissue was developed. Optimal recovery was obtained by spread-plating homogenates on Middlebrook 7H10 agar with incubation for 3 mo at …

Dissecting The Loci Of Low-Level Quinine Resistance In Malaria Parasites, Michael T. Ferdig, Roland A. Cooper, Jianbing Mu, Bingbing Deng, Deirdre A. Joy, Xin-Zhuan Su, Thomas E. Wellems

Biological Sciences Faculty Publications

Quinine (QN) remains effective against Plasmodium falciparum, but its decreasing efficacy is documented from different continents. Multiple genes are likely to contribute to the evolution of QN resistance. To locate genes contributing to QN response variation, we have searched a P. falciparum genetic cross for quantitative trait loci (QTL). Results identify additive QTL in segments of chromosomes (Chrs) 13, 7 and 5, and pairwise effects from two additional loci of Chrs 9 and 6 that interact, respectively, with the QTL of Chrs 13 and 7. The mapped segments of Chrs 7 and 5 contain pfcrt, the determinant of …

Virulence Factors Of The Group A Streptococci And Genes That Regulate Their Expression, Wayne Hines

Biological Sciences Faculty Publications

Group A streptococci produce a wide variety of extracellular (cell-associated and released) virulence factors. The function of these factors varies and includes roles in adhesion, spreading, tissue destruction, immune system evasion, and cell toxicity. How these factors are regulated with regard to one another is important for this organism's ability to bring about the variety of diseases this microbe is capable of causing. Therefore, along with the multitude of virulence factors, there are a number of regulatory systems that regulate, either directly or indirectly, the production of these factors and therefore influence the pathogenesis of group A streptococcal infections.

A Survey Of Antibiotic Resistance Among Coliform Bacteria Isolated From The Missouri River, Sara E. Mcdonnell, Amy M. Treonis

Biology Faculty Publications

The prevalence of antibiotic resistant microorganisms in the environment is not well known but could represent a challenge to maintaining public health in the future. Fecal waste from livestock facilities, where antibiotics are routinely used, is recognized as a significant source of pollution to surface waters in the United States. We collected water from the Missouri River across a winter to spring seasonal change in order to survey the density of coliform bacteria. We tested bacterial isolates for resistance to penicillin, tetracycline, ampicillin, erythromycin, and ciprofloxacin. Coliform density in Missouri River water generally was high throughout the study (0-405 cfu …

Evaluation Of Immunogenicity Of Transgenic Chloroplast Derived Protect, Vijay Koya

Electronic Theses and Dissertations

Anthrax, a fatal bacterial infection is caused by Bacillus anthracis, a gram-positive, spore forming, capsulated, rod shaped organism. Centers for Disease Control (CDC) lists anthrax as Category A biological agent due to its severity of impact on human health, high mortality rate, acuteness of the disease and potential for delivery as a biological weapon. The currently available human vaccine in the United States (AVA anthrax vaccine adsorbed) is prepared from Alum adsorbed formalin treated supernatant culture of toxigenic, non-encapsulated strain of Bacillus anthracis with the principle component being protective antigen (PA83). Evaluation of anthrax vaccine given to nearly 400,000 US …

Cellular Immune Response And Gene Expression Profiling In Crohn's Dise, Claudia Romero

Electronic Theses and Dissertations

Despite the chronic debate in the etiology of crohn's disease (cd), a debilitating inflammatory bowel disease (ibd) closely related to ulcerative colitis (uc), an emerging interest in a possible mycobacterial role has been marked. Granuloma and pathologic manifestations in cd resemble aspects found in tuberculosis, leprosy and paratuberculosis. The latter, a chronic enteritis in cattle, goat, sheep and primates, which is similar to human enteritis, also known as cd, is caused by a fastidious, slow growing mycobacterium avium subspecies paratuberculosis (map). Due to the similarities between cd and paratuberculosis, a mycobacterial cause in cd has been proposed. Recent discovery of …

Dinitrogen Fixation And Release Of Ammonium And Dissolved Organic Nitrogen By Trichodesmium Ims101, Margaret R. Mulholland, Deborah A. Bronk, Douglas G. Capone

OES Faculty Publications

Two methods used to measure dinitrogen (N₂) fixation (acetylene reduction and ¹⁵N₂ uptake) often result in different N₂ fixation rates. Part of the discrepancy may arise from the observation that Trichodesmium can release a fraction of their recently fixed N₂ as dissolved organic nitrogen (DON) and/or ammonium (NH4 ⁺). To resolve outstanding issues regarding N₂ fixation and the production of dissolved combined nitrogen (N) by Trichodesmium, we conducted a comprehensive analysis of N₂ fixation and the production of DON and NH₄⁺ in cultures of Trichodesmium IMS101. We performed …