Bacteriology Commons^™

Evaluation Of Intra-Species Diversity Of Oxalobacter Formigenes Strains Using Pulsed-Field Gel Electrophoresis (Pfge) And Multiplex Pcr, Nivedita Pareek

Masters Theses

Oxalobacter formigenes is a beneficial, strict anaerobic gut bacterium that plays an important role in the prevention of kidney stone disease with a unique characteristic of using only oxalate as a carbon and energy source. To date, twenty-one strains of O. formigenes have been isolated which are further divided into two groups based on differences in the patterns of cell membrane lipids, cellular proteins, and nucleic acid fragments. The objective of the study was to evaluate intra-species diversity of O. formigenes strains using pulsed-field gel electrophoresis (PFGE) and multiplex PCR.

Nine pure cultures of O. formigenes strains (5 strains group …

Xylan Metabolism By Caulobacter Crescentus, Viet Bui

Masters Theses

Bacteria present unique opportunities to explore the molecular mechanisms of nutrient acquisition processes at single cell level under various experimental conditions. Caulobacter crescentus is a gram-negative oligotrophic environmental bacterium with adaptive physiological responses, such as slow growth or complete arrest of cell cycle, presence of sessile and motile progenies with the sessile cells growing stalk to enhance nutrient uptake for growth in low-nutrient conditions. Details of plant polysaccharide-derived carbon compound, such as cellobiose, maltose, and xylose utilization by C. crescentus can be found but molecular details or mechanisms of polysaccharide utilization, such as xylan, has not been reported. Database analyses …

Characterization Of Pinitol Catabolism In Sinorhizobium Meliloti And Its Role In Nodule Occupancy, Angela Irene Kennedy-Mendez

Masters Theses

Sinorhizobium meliloti, which is the nitrogen fixing symbiont of Medicago sativa (alfalfa), was used to test the hypothesis that D-chiro-inositol is an intermediate of pinitol catabolism and that pinitol catabolism plays a role in S. melilot s ability to compete for nodule occupancy. An analytical method, involving gas chromatography and mass spectrometry (GCMS), was developed to detect and separate pinitol and D-chiro-inositol. The S. meliloti wild type, and three isogenic mutant strains (idhA, mocD, and mocE) were grown in minimal medium with pinitol and pinitol plus glycerol. The S. meliloti wild type can utilize pinitol as a sole carbon source, …

Resolution Of The Nutritional Requirements And Metabolic Profile Of Clostridium Scindens Atcc 35704, A Major Bile Acid-Dehydroxylating Anaerobe In The Human Gut, Rachana Shrestha

Masters Theses

Bile acids (cholate and chenodeoxycholate) are synthesized from cholesterol in the human liver, and secreted in small intestine for food digestion. However, a portion of these primary bile acids is converted in the colon to toxic secondary bile acids, deoxycholate and lithocholate. Secondary bile acid formation is the result of 7α-dehydroxylating anaerobes, and one of the key players in this conversion is Clostridium scindens. Interestingly, little is known about the basic physiology and nutrition of C. scindens. Therefore, the goal of this research was to understand the nutritional requirement, and determine the 7α-dehydroxylation activity and end products from …

Resolving The Nutritional Requirements Of The Bile Acid-Dehydroxylating Gut Bacterium Clostridium Scindens, Oindrila Paul

Masters Theses

Compared to the other parts of the human body, the gastrointestinal tract harbors a complex microbiota. The impact of the indigenous microbiota on host physiology is most pronounced in the colon, where the primary bile acids chenodeoxycholic acid and cholic acid are converted, via 7α-dehydroxylation, to the toxic secondary bile acids lithocholic acid and deoxycholic acid, respectively, by Clostridium scindens, an obligate anaerobe. Interestingly, other than bile acid dehydroxylation, little is known about the basic physiology of C. scindens. Understanding the metabolism of C. scindens will hopefully provide much-needed information as to how this gut bacterium impacts human health and …

The Isolation And Partial Characterization Of The Lipopolysaccharides From Rhizobium Leguminosarum Biovar Trifolii Strains Tb4, Tb104, And Tb112, Jill K. Miller

Masters Theses

The lipopolysaccharides (LPSs) from Rhizobium leguminosarum biovar trifolii TB4 and two transposon (Tn5) symbiotic mutants, R. leguminosarum bv. trifolii TB104 and TB112, were isolated and partially characterized. Bv. trifolii TB4 is a derivative of R. leguminosarum bv. trifolii ANU843. The two mutant strains elicited incompletely developed clover nodules that exhibited low bacterial populations and very low nitrogenase activity. The LPSs were extracted using the hot phenol-water method and purified with gel filtration column chromatography using Sepharose 4B with an elution buffer consisting of EDTA and triethylamine. The hexose compositions were determined by gas chromatography of the alditol acetate derivatives. LPS …

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 …

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, …

The Effect Of A Bacteriophage On The Competition Of Rhizobium Japonicum Strains For Nodulation Of Glycine Max (L.) Merr, Lori Kelley

Masters Theses

The symbiotic relationship between Rhizobium japonicum (Kir.) Buch. and Glycine max (L.) Merr. was the concern of this thesis. This relationship occurs naturally in soybean fields. In the field there are many different strains of Rhizobium. Some of these strains fix atmospheric nitrogen more efficiently than others resulting in a higher crop yield. The problem is the more efficient nitrogen fixers are out-competed by less efficient nitrogen fixers. In order to solve this problem the more competitive strains have to be controlled. There are two main ways to do this. One is to increase the number of the desired …

Response Of A Nitrifying Biofilm To Copper, Denise Moldroski Mott

Masters Theses

In pure cultures of Nitrosomonas, copper is known to cause a stimulation of nitrification at low levels (≤ 0.03 mg Cu⁺²/1) and an inhibition affect at higher levels (≥ 0.05 mg Cu⁺²/1). Nitrifying activated sludges are much more tolerant of copper showing only 75% inhibition at 150mg Cu⁺²/1. The purpose of this paper was to determine the copper sensitivity of a nitrifying biofilm grown in a continuous flow trickling filter system.

A continuous flow trickling filter system was built using limestone chips for the biofilm substrate. The biofilm was established from secondary effluent. …

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 …

A Comparison Of The Acidic Extracellular Polysaccharides From Rhizobium Trifolii Mutants Affected In Trifolium Repens Root Hair Curling, Brian Andrew Hanley

Masters Theses

The compositions and sugar linkages of the acidic extracellular polysaccharide (EPS) from several Rhizobium trifolii mutants were compared. These mutants were derived from a wild type Rhizobium trifolii, ANU843, and are unable to cause normal root hair curling, Hac-, or nodulate clover, Nod-. Three of the mutants contain a Tn5 insertion in a 5.3 kb BglII fragment of the symbiotic plasmid (pSym). A fourth mutant is missing the pSym, which includes much of the 5.3 kb BglII fragment, both of these deletions result in Hac-Nod- phenotypes. When the 5.3 kb BglII fragment is cloned (pRT150) and reinserted into the …

Characterization Of The Lipopolysaccharides (Lps) From Four Transposon Generated Symbiotic Mutants Of Rhizobium Trifolii, Robert G. Shatters Jr.

Masters Theses

The LPSs from four symbiotic mutants of R. trifolii were isolated and characterized. The mutants were obtained from Dr. Barry Rolfe of The Australian National University. Three mutants have Tn5 insertions in the "nod" regions of the symbiotic plasmid. The fourth has a 30 kb deletion. This deletion includes the majority of the "nod" regions. All mutants fail to nodulate clover and fail to induce markedly curled root hairs, i.e. they are hac-nod-. The LPSs were purified as described by Carlson et al. 1978.⁷ The polysaccharide portion of the LPSs was purified by treating the LPSs in 1% HAc …

Comparison Of Surface Polysaccharides Of R. Leguminosarum Anu57 (Exo-1), R. Leg. Anu57(Pbr1an) (Nod+, Nif-), And R. Leg. Anu57(Pjb5j1) (Nod+, Nif-), Frank G. Hustmyer

Masters Theses

Extracellular polysaccharides (EPS) and lipopolysaccharides (LPS) are suspected to be involved in the legume/Rhizobium symbiotic process. Polysaccharide compositions from the EPS, LPS, capsular polysaccharide (CPS), and small polysaccharide (SmPS) were compared between Rhizobium leguminosarum ANU57 (parent, Exo^-1) and two mutants R. leg. ANU57(pBR1AN) (nod⁺, nif^-) and R. leg. ANU57(pJB5J1) (nod⁺, nif^-) both containing a Sym (symbiotic) plasmid insertion. R. leg. ANU57(pBR1AN) and R. leg. ANU57(pJB5J1) are nod⁺ on clover and peas, respectively. The nod⁺ mutants produce larger amounts of EPS and decreased amounts of LPS compared to …

Siderocapsa Major-- Fact Or Artifact?, Ray W. Swanson

Masters Theses

The validity of the genus Siderocapsa, a group of unicellular iron-depositing bacteria, has been a subject of debate for many years. Lack of photomicroscopic and cultural studies has kept this group in taxonomic obscurity. Bacteria were collected from the Campus pond, Eastern Illinois University, Charleston, Illinois, by a submerged slide/coverslip technique. These bacteria were identified as Siderocapsa major according to the description of Molisch (1909). Scanning electron microscopy of Siderocapsa major microcolonies has shown cells (0.5 by 1.15 μm), in a clear central well (2-6 μm), surrounded by an amorphous material which accumulates with the age of the colony. …

Pesticide Effects In A Simulated Soil Ecosystem, Raymond J. Samp

Masters Theses

The screening of pesticides to determine which were most inhibitory to bacteria was accomplished by testing the effects of 10 pesticides on 9 different organisms using the disk assay method. Results showed that Gram positive bacteria were more sensitive to all pesticides tested than were the Gram negative bacteria. In addition, the hormone herbicides were found to be the most inhibitory to these bacteria and 2,4-D (2,4-dichlorophenoxyacetic acid), a hormone herbicide, was chosen for subsequent in vivo studies.

A soil perfusion apparatus was used to determine the effects of 2,4-D in a typical soil ecosystem. Bacterial plate counts and nutrient …

Studies On Muropeptide Amino Acids Of Rhizobium Japonicum, Mohammed Alauddin

Masters Theses

The isolation of Rhizobium japonicum cell walls was performed by ultrasonication and the enzymatic digestion with DNase, RNase and pronase separately. Crude cell walls were washed with M NaCl and 5% SDS and were isolated by 20,000 g centrifugation. Muropeptide amino acids were freed by hydrolysis at 121C for 10 minutes in 6N HCl.

Detection of Rhizobium japonicum murein layer amino acids was done by 2-dimensional instant thin layer chromatography on silica gel plates. The solvents found most favorable, were n-butanol: acetic acid: water and n-propanol: 34% ammonium hydroxide. Amino acid spots were developed by spraying ninhydrin on the chromatograms, …

Morphological And Physiological Effects Of 2,4-D On Gram-Negative And Gram-Positive Bacteria, John H. Rupnow

Masters Theses

Manometric, spectrophotometric, photomicrographic and quantitative biochemical techniques were employed to determine the morphological and physiological effects of 2,4-D on Gram-positive and Gram-negative bacteria.

The growth and motility of Gram-positive bacteria appeared to be more adversely affected than the growth and motility of Gram-negative species. The terminal oxidation, lysis, and chain length of both Gram-positive and Gram-negative bacteria did not appear to be affected by the concentrations used. The average cell size of both Gram-positive and Gram-negative bacteria was observed to increase after exposure to 1000 ppm 2,4-D. The normal leakage of amino acid and ultraviolet light absorbing cell constituents appeared …

Lysis Of The Rhizobiaceae, Craig Richard Baumrucker

Masters Theses

The following members of the Rhizobiaceae were tested for their lytic response to various treatments: Agrobacterium tumefaciens, Chromobacterium violaceum, Rhizobium phaseoli, Rhizobium trifolii, Rhizobium lupini, and Rhizobium japonicum. Lysis was observed in spent growth media and as a function of saline concentration, pH, buffer composition, disodium (ethylenedinitrilo) tetraacetate (EDTA) concentration, and lysozyme.

Cells were harvested by centrifugation, washed, and resuspended in various treatment solutions. The lytic response was followed spectrophotometrically.

The different responses of the six cultures was applied to a summation of reclassification proposals evolved through modern taxonomic techniques such as DNA homology …