Microbiology Commons^™

Proliferation And Differentiation Of Mast Cell Progenitors, Daniel Lee Jarboe

Theses and Dissertations

We have identified a late, committed stage in the differentiation of the mast cell progenitor just prior to granulation. This mast cell-committed progenitor (MCCP) differs from the more primitive bone marrow mast cell progenitor in that it is able to proliferate and differentiate in the absence of interleukin-3 (IL-3) when cultured on a monolayer of embryonic skin or 3T3 fibroblasts. The MCCP can be harvested from the mesenteric lymph nodes of mice in their fourteenth day of infection with the rodent hookworm Nippostrongylus brasiliensis and can be cloned in a methylcellulose culture system by supplementing the cultures with fibroblast-conditioned medium …

Quantitative Studies On Nodulation Competition Of Two Strains Of Bradyrhizobium Japonicum And Its Biological Control With A Bacteriophage, Jin Xiong

Masters Theses

Interstrain competition of Bradyrhizobium japonicum for nodulation of soybean (Glycine max) presents a serious problem to the field commercial inoculation practice. The more efficient, introduced strains are often outcompeted by the less efficient, indigenous strains. This study was conducted to determine if the competition could be effectively controlled by the massive introduction of a more efficient strain and/or the large scale reduction of a indigenous strain by the application of a bacteriophage specific to the indigenous one.

B. japonicum strain 61A123, common in Midwestern U.S. soils, and strain USDA110, representative of the strains used in commercial inocula, were chosen for …

Microbial Consumption Of Atmospheric Isoprene In A Temperate Forest Soil, Cory C. Cleveland, J. B. Yavitt

Ecosystem and Conservation Sciences Faculty Publications

Isoprene (2-methyl-1,3 butadiene) is a low-molecular-weight hydrocarbon emitted in large quantities to the atmosphere by vegetation and plays a large role in regulating atmospheric chemistry. Until now, the atmosphere has been considered the only significant sink for isoprene. However, in this study we performed both in situ and in vitro experiments with soil from a temperate forest near Ithaca, N.Y., that indicate that the soil provides a sink for atmospheric isoprene and that the consumption of isoprene is carried out by microorganisms. Consumption occurred rapidly in field chambers (672.60 +/- 30.12 to 2,718.36 +/- 86.40 pmol gdw day) (gdw is …

Reliable Detection Of Individuals Seropositive For The Human Immunodeficiency Virus (Hiv) By Competitive Immunoassays Using Escherichia Coli - Expressed Hiv Structural Proteins, G. J. Dawson, J. S. Heller, Charles Wood, R. A. Gutierrez, J. S. Webber, J. C. Hunt, S. A. Hojvat, D. Senn, S. G. Devare, R. H. Decker

Nebraska Center for Virology: Faculty Publications

We molecularly cloned the gag and env genes of the human immunodeficiency virus (HIV) and expressed fragments of these genes in Escherichia coli. Using the recombinant core and envelope proteins, we developed two competitive immunoassays (CIAs). Samples that recognized either the envelope or core proteins were considered positive for antibodies to HIV. This test system was comparable with western blot in detecting antibodies in patients with AIDS or AIDS-related complex that were repeatably reactive in the HIV screening test. All 360 individuals who were positive by western blot were positive by the CIA. A total of 844 samples repeatably …

Can We Guarantee The Safety Of Genetically Engineered Organisms In The Environment?, Kathleen H. Keeler, Frances E. Sharples

School of Biological Sciences: Faculty Publications

Can we guarantee the safety of genetically engineered organisms in the environment? To anticipate my remarks, the answer to the title question is “No, we cannot guarantee the safety of genetically engineered organisms released into the environment.” Indeed, it is a tenet of the scientific method that nothing can be proved, only disproved. Thus, we can never show that a release will be safe. We could only show that it would be unsafe, if that were the case. However, if the question is posed differently, for example, can we safely release genetically engineered organisms into the environment?, the answer is …

Partial Characterization Of Lpss From Rhizobium Fredii Usda 205 Before And After Nod Gene Induction, Bradley Lawrence Reuhs

Masters Theses

The lipopolysaccharides (LPS) from Rhizobium fredii USDA 205 and its nod- mutant, HC 205, which has been cured of the 192-MDa symbiotic plasmid, were isolated by the phenol-water extraction method. The LPS from the wild-type strain which was grown in association with host-plant root extract or a flavone analog to a constituent of the root extract, which is required for the induction of at least a portion of the nodulation genes, was also isolated. The conditions required for the induction of the nod genes were determined by work with a nod- mutant of R. fredii USDA 201 which …

The Effect Of Fluoride On Treponema Denticola, Carrie A. Norton Hughes

Dissertations

No abstract provided.

Phenotypic And Functional Characterization Of Ia+ And La- Sublines Of The Human Monocyte-Like Cell Line, U937, Coles Meredith Squire

Theses and Dissertations

Sublines were derived from the U937 human monocytic cell line by limiting dilution techniques. Several of the sublines derived in this manner were found to constituitively express la, unlike most U937 cell lines previously examined. Thus the la+ and la- sublines were examined and characterized phenotypically and functionally. Morphologically the sublines, both la+ and la-, were found to be similar to the originally described U937 parent cell line. They were positive for production of non- specific esterase and expressed the HLA phenotype A(3,X), B(51,18), DR(2,X). The sublines were examined by immunofluorescence techniques with a large number of antibodies specific for …

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 …

Activation Of The Human Immunodeficiency Virus By Herpes Simplex Virus Type 1, Jeffrey M. Ostrove, John Leonard, Karen E. Weck, Arnold B. Rabson, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Herpes simplex virus type 1 (HSV-1) and some of its immediate-early genes stimulate expression of the human immunodeficiency virus (HIV) long terminal repeat (LTR) sequences and the replication of HIV itself. To demonstrate this, the HIV LTR was linked to the indicator gene chloramphenicol acetyltransferase (CAT) and transfected into Vero cells with or without the trans-activating gene (tat) of HIV. Infection of these cells with HSV-1 strain KOS or temperature-sensitive mutant tsB2l or tsE6 resulted in a large increase in CAT activity in the absence of tat and further augmentation in the presence of tat. This stimulation was seen at …

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 …

The Glycoprotein Toxin Of Bacillus Thuringiensis Subsp. Israelensis Indicates A Lectinlike Receptor In The Larval Mosquito Gut, Ganapathy Muthukumar, Kenneth Nickerson

Papers in Microbiology

The mosquito-active protein crystals produced by Bacillus thuringiensis subsp. israelensis contain covalently attached aminosugars which are critical for their larvicidal activity. The 50% lethal concentrations toward Aedes aegypti larvae were increased up to 10-fold by mild periodate treatment, up to 40-fold by forming the protein crystals in the presence of tunicamycin, and up to 7-fold by the presence during the mosquito bioassays of N-acetylglucosamine or its trimer, triacetylchitotriose. Periodate-treated crystals and crystals formed in the presence of tunicamycin had greatly reduced binding capacities for wheat germ agglutinin, an Nacetylglucosamine- specific lectin. These results suggest that the B. thuringiensis subsp. …

Temperature-Dependent Kinetic Variation Among Phosphoglucose Isomerase Allozymes From The Wing-Polymorphic Water Strider, Limnoporus Canaliculatusl, Anthony J. Zera

Anthony Zera Publications

Phosphoglucose isomerase (PGI) allozymes were isolated from the wing-polymorphic water strider, Lirnnoporus canaliculatus, and were characterized biochemically with respect to temperature-dependent kinetic and thermostability properties. At higher temperatures, the allozymes exhibited significant differences in Michaelis constant (K_m) values for substrates of both the forward and reverse reaction directions. Results were consistent with expectations of adaptive kinetic differentiation based on the latitudinal variation of PGI allele frequencies. PGI genotypes also differed with regard to maximal velocity (V_max)/K_m ratios at higher temperatures. These differences were due primarily, if not exclusively, to allozyme-dependent variation in K …

Microbial Degradation Of Oxalate In The Gastrointestinal Tracts Of Rats, Steven Daniel

Faculty Research & Creative Activity

Rates of oxalate degradation by mixed bacterial populations in cecal contents from wlld rats ranged from 2.5 to 20.6 µmol/g (dry weight) per h. The oxalate-degrading activity in cecal contents from three strains of laboratory rats (Long-Evans, Wistar, and Sprague-Dawley) from four commercial breeders was generally lower, ranging from 1.8 to 3.5 µmollg (dry weight) of cecal contents per h. This activity did not increase when diets were supplemented with oxalate. Wben Sprague-Pawley rats from a fifth commercial breeder were fed an oxalate diet, rates of oxalate degradation in cecal contents increased from 2.0 to 23.1 µmollg (dry weight) per …

Carbon Monoxide-Dependent Chemolithotrophic Growth Of Clostridium Thermoautotrophicum, M. Savage, Zhongren Wu, Steven Daniel, Leon Lundie, Harold Drake

Faculty Research & Creative Activity

The acetogen Clostridium thermoautotrophicum was cultivated under CO-dependent chemolithotrophic conditions. CO-dependent growth profiles and energetics indicated that supplemental CO2 was fundamental to efficient growth at the expense of CO. Overall product stoichiometry approximated 6.5CO --> CH3CO2H + 3.5CO2 + 0.6 cell C + 0.5 unrecovered C. Initial CO/CO2 ratios of 2 to 4 yielded optimal doubling times and cell yields. Maximal YCO values approximated 2.5 g of cell dry weight per mol of CO consumed; Y H2 , was considerably lower than Y CO Cross-transfer growth experiments and protein profiles indicated differential expression of genes between CO and methanol cultures.

Carbon Monoxide-Dependent Chemolithotrophic Growth Of Clostridium Thermoautotrophicum, M. Dean Savage, Zhongren Wu, Steven L. Daniel, Leon L. Lundie Jr, Harold L. Drake

Faculty Research & Creative Activity

The acetogen Clostridium thermoautotrophicum was cultivated under CO-dependent chemolithotrophic conditions. CO-dependent growth profiles and energetics indicated that supplemental CO2 was fundamental to efficient growth at the expense of CO. Overall product stoichiometry approximated 6.5CO --> CH3CO2H + 3.5CO2 + 0.6 cell C + 0.5 unrecovered C. Initial CO/CO2 ratios of 2 to 4 yielded optimal doubling times and cell yields. Maximal YCO values approximated 2.5 g of cell dry weight per mol of CO consumed; Y H2 , was considerably lower than Y CO Cross-transfer growth experiments and protein profiles indicated differential expression of genes between CO and methanol cultures.

Microbial Degradation Of Oxalate In The Gastrointestinal Tracts Of Rats, Steven L. Daniel

Faculty Research & Creative Activity

Rates of oxalate degradation by mixed bacterial populations in cecal contents from wlld rats ranged from 2.5 to 20.6 µmol/g (dry weight) per h. The oxalate-degrading activity in cecal contents from three strains of laboratory rats (Long-Evans, Wistar, and Sprague-Dawley) from four commercial breeders was generally lower, ranging from 1.8 to 3.5 µmollg (dry weight) of cecal contents per h. This activity did not increase when diets were supplemented with oxalate. Wben Sprague-Pawley rats from a fifth commercial breeder were fed an oxalate diet, rates of oxalate degradation in cecal contents increased from 2.0 to 23.1 µmollg (dry weight) per …

Estimation Of Food Limitation In Daphnia Pulex From Boulder Basin, Lake Mead, Thomas Mark Bartanen

Publications (WR)

In February, 1982 I began a year-long study to determine if growth and reproduction in Daphnia Pulex were limited by the amount of food available in Boulder Basin, Lake Mead. To determine this, I made monthly collections of Daphnia Pulex and natural lake seston from an already established station in Boulder Basin. I cultured the Daphnia Pulex under simulated field conditions in a flow-through feeding apparatus using four different food regimes; 1-natural lake seston filtered through 80 um mesh to remove other zooplankton, 2-lake seston (as above) with an enrichment of 10³ cells-ml^-1 of Chlamydomonas reinhardti, 3-lake …

Structural Disulfide Bonds In The Bacillus Thuringiensis Subsp. Israelensis Protein Crystal, Graham A. Couche, Mary A. Pfannenstiel, Kenneth W. Nickerson

Papers in Microbiology

We examined disulfide bonds in mosquito larvicidal crystals produced by Bacillus thuringiensis subsp. israelensis. Intact crystals contained 2.01 x 10^-8 mol of free sulfhydryls and 3.24 x 10-^-8 mol of disulfides per mg of protein. Reduced samples of alkali-solubilized crystals resolved into several proteins, the most prominent having apparent molecular sizes of 28, 70, 135, and 140 kilodaltons (kDa). Non-reduced samples contained two new proteins of 52 and 26 kDa. When reduced, both the 52- and 26-kDa proteins were converted to 28-kDa proteins. Furthermore, both bands reacted with antiserum prepared against reduced 28-kDa protein. Approximately 50% of …

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 …

The Determination Of Antibody Levels In Endodontic Patients Against Eleven Microorganisms Using Ifa And Elisa Techniques, William C. Payne

Loma Linda University Electronic Theses, Dissertations & Projects

Endodontic (root canal) infections are generally considered to be caused by the invasion of microorganisms which destroy pulp tissue and extend to the periapical region. No definitive study has identified a specific bacterium or group of bacteria as etiological agents. We quantitively measured antibodies present in the serum of 30 normal individuals and patients with either chronic (n = 30) or acute (n = 30) periapical lesions with the indirect fluorescent antibody (IFA) and enzyme-linked immunosorbent assay (ELISA) for IgG and IgM against 11 oral microorganisms routinely isolated from such lesions. The results were analyzed statistically. Analysis of variance of …

Amino Sugars In The Glycoprotein Toxin From Bacillus Thuringiensis Subsp. Israelensis, Mary Ann Pfannenstiel, Ganapathy Muthukumar, Graham A. Couche, Kenneth W. Nickerson

Papers in Microbiology

The carbohydrate content of purified Bacillus thuringiensis subsp. israelensis crystal toxin was determined by six biochemical tests, column chromatography on an amino acid analyzer, and the binding of 11 fluorescent lectins. The crystals contained approximately 1.0% neutral sugars and 1.7% amino sugars. The anmino sugars consisted of 70% glucosamine and 30% galactosamine. No N-acetylneuraminic acid (sialic acid) was detected. The presence of amino sugars was confirmed by the strong binding of fluorescent wheat germ agglutinin and the weak binding of fluorescent soybean agglutinin. These lectins recognize N-acetyl-D-glucosamine and N-acetyl-D-galactosamine, respectively. The lectin-binding sites appeared evenly distributed among …

Cyanonews (Vol. 3, No. 1, April 1987), Jeff Elhai

CyanoNews

CyanoNews was a newsletter that served the cyanobacteriological community from 1985 to 2003, with content provided by readers (sort of a blog before there were blogs). The newsletter reported new findings from the lab, summaries of recent meetings (often provided by graduate students and post-docs entering the field), positions sought or available, life transitions, a compendium of recent cyanobacteria-related articles, and other items of interest to those who study cyanobacteria.

Cyanonews (Vol. 3, No. 2, September 1987), Jeff Elhai

CyanoNews

CyanoNews was a newsletter that served the cyanobacteriological community from 1985 to 2003, with content provided by readers (sort of a blog before there were blogs). The newsletter reported new findings from the lab, summaries of recent meetings (often provided by graduate students and post-docs entering the field), positions sought or available, life transitions, a compendium of recent cyanobacteria-related articles, and other items of interest to those who study cyanobacteria.

Nucleotdie Sequence Of An Anti-Fluorescyl Hapten Antibody Heavy Chain Variable Region Gene From A Balb/C Mouse Hybridoma Cell Line, Zhen-Qian Liu, Charles Wood, Tai Te Wu

Nebraska Center for Virology: Faculty Publications

We report here the nucleotide sequence of an anti-fluorescyl hapten antibody heavy chain variable region gene from a BALB/c mouse hybridoma cell line 3-13 (1). This gene segment was contained in a 5.0 kb EcoRI fragment isolated by a 700 bp probe 3' to the mouse heavy chain joining minigene region between the XbaI and EcoRI sites. Mature protein starts at *. The first 21 amino acid residues have also been confirmed by direct amino acid sequencing and they are identical to those of immunoglobulin MOPC104E heavy chain (2). Complementarity determining regions (CDRs) are underlined. The third CDR is unusually …

The Strain Improvement Of Aspergillus Ficuum Nrrl 3135, Eric A. Utt

Retrospective Theses and Dissertations

A strain improvement program was initiated to increase the yield of extracellular phytase (EC 3.1.3.8) in Aspergillus ficuum NRRL 3135. A minimal plating medium containing 2.99 g liter^-1 a-ketoglutaric acid was developed to decrease and stabilized conidiogenesis time and facilitate the isolation of single colonies. Exposure of A. ficuum conidia to 0.5 mg ml^-1 N-methyl-N’-nitro-N-nitrosoguanidine (NTG) for 15 min at pH 6.5 caused a one log kill and an increase in the mutation rate of 800-fold. Treatment of A. ficuum conidia with 46 ergs mm^-2 sec ^-1 UV radiation for 30 sec resulted in a one …

The Effects Of Frozen Storage And Repeated Freeze-Thawing On Salmonella In Varied Concentrations Of Waste Activated Sludge, Joseph Toman Jr.

Masters Theses

Commonly, waste activated sludge is applied to farmland as a fertilizer supplement and soil conditioner. The use of waste activated sludge is not only an efficient way of sludge disposal, but also an economical one. Much concern has been raised about the safety of such a practice. Harmful pathogens, like Salmonella, are commonly present in waste material. Treatment at wastewater plants is successful in eliminating these organisms from final effluent, but the solids generated are full of indigenous microorganisms, including Salmonella.

Work of this paper attempts to determine the safety of the sludge product after simulated winter conditions …

Antibiotic Effects On Coliform Enumeration, Keith Leonard Wozniak

Masters Theses

The inhibitory effects of Bacillus polymyxa on Escherichia coli were studied. Significant coliform inhibition was observed. The inhibitory agent (presumably polymyxin) was active against coliforms after a one hour holding time. Different suspending menstrua were analyzed for their effects on coliform inhibition. It was observed that Peptone water provided the highest recovery of coliforms while phosphate buffer provided less protection. Two methods of enumeration were also studied as to their ability to quantify coliforms inhibited by B. polymyxa. Comparable inhibition was shown using both methods. No significant differences exist between the Standard Plate Count and Membrane Filter techniques. Overall, …

Productive, Persistent Infection Of Human Colorectal Cell Lines With Human Immunodeficiency Virus, Akio Adachi, Scott Koenig, Howard Gendelman, Daryl Daugherty, Sebastiano Gattoni-Celli, Anthony Fauci, Malcom A. Martin

Nebraska Center for Virology: Faculty Publications

Thirteen adherent human non-lymphocyte cell lines were tested for their susceptibility to infection by human immunodeficiency virus. Productive infection could be demonstrated in three of five colorectal carcinoma cell lines examined; the other eight human non-lymphocyte cell lines were uninfectible. A susceptible colon carcinoma cell line (HT29), as well as normal colonic mucosa, was shown to contain a 3.0-kilobase species of poly(A)⁺ CD4 RNA, whereas uninfectible colon carcinoma and rhabdomyosarcoma cell lines synthesized no detectable T4 RNA. A persistently infected colon carcinoma cell line was established that continued to produce progeny human immunodeficiency virus for more than 10 weeks …

Analysis Of Immune Enhancing Effects Of Interleukin-2, Sharareh Sazesh

Retrospective Theses and Dissertations

The effectiveness of IL-2 in the enhancement of immune responses in B6-AKR and normal B6 mice was investigated. In vivo treatment of chimeras with murine IL-2 partially restored PFC responses of their splenocytes to SRBC. This treatment suppressed splenocytes IL-2 production in chimeras. In vivo treatment of chimeras with rhIL-2 resulted in a slight enhancement of mitogenic responses to Con-A. This treatment had no significant effect on mitogenic responses to PHA. The rhIL-2 treatment did not result in suppression of IL-2 production by chimeric splenocytes. Recombinant human IL-2 enhanced mitogenic responses of normal B6 and chimeric mice to PHA in …