Microbiology Commons^™

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 …

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, …

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 …

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 …

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 …

Partial Characterization Of The Extracellular Polysaccharides Produced By Mutants Of Rhizobium Leguminosarum And Rhizobium Trifolii, Lendell L. Cummins

Masters Theses

Extracellular polysaccharides (EPS) are suspected to be involved in some aspect of the Rhizobium-legume symbiotic process. Extracellular polysaccharides produced by R. leguminosarum and R. trifolii mutants were isolated and partially characterized by gas chromatography, column chromatography, double diffusion, polyacrylamide gel electrophoresis, and immunoelectrophoresis.

The R. leguminosarum mutants, derived from strain 128C53, all are deficient in EPS production (exo-). Strain ANU54 is missing the symbiotic plasmid, while plasmid-containing strains ANU54 pJB5JI and ANU54 pBR1AN nodulate (nod+) peas and clover respectively, but do not fix nitrogen (fix-).

R. leguminosarum mutants produce an EPS that is similar to parental LPS in composition. …

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 …

Raw Water Quality In The New Charleston, Illinois Pump Storage Reservoir, Kevin G. Lookis

Masters Theses

Charleston, Illinois, a city located in east-central Illinois, recently implemented a new concept in a raw water system. Due to storage capacity-sedimentation problems in the old Lake Charleston-Embarras River raw water supply, the Charleston Pump Storage Reservoir was constructed. The new system involved building a dike separating the Lake Charleston impoundment from the Embarras River. Water to increase the storage capacity of the reservoir can now be selectively pumped into the impoundment from the river.

Raw water quality in the reservoir was monitored on a weekly basis for a full year from August, 1981 to August, 1982. The levels of …

Inhibitors Affecting Bacterial Action In Reaerated Activated Sludge, Alice M. Dunn

Masters Theses

Inhibition of reaerated activated sludge was assessed using reduction of dissolved oxygen uptake by the microbial population as a criterion. Oxygen uptake is essential to biodegradation of the organic components in the activated sludge process; therefore, a decrease in dissolved oxygen uptake is an indication of reduced efficiency in wastewater treatment.

The irihibitors tested were cadmium, copper, lead, mercury, silver, 2,4-Dichlorophenoxyacetic acid (2,4-D), Terbacil and Zectran. The IIC₅₀, determined by extrapolation of plotted data, for each inhibitor in mg/l was Hg (20), Cu (50), Ag (43), Cd (115), Pb (550), and 2,4-D (430). The IIC₅₀ was determined …

A Study Of The Free-Floating Phytoplankton Of The Embarras River And Polecat Creek, Coles County, Il, Richard Gary Fried

Masters Theses

An investigation of 2 sites on the Embarras River and 1 site on Polecat Creek was undertaken for a one-year period. Special emphasis was given to phytoplankton species and to their seasonal variations. Approximately 170 species from 8 divisions of algae were encountered during the one-year period.

Temperature was found to have no direct correlation with algal periodicity.

Palmer's organic pollution genus index was applied to determine the approximate levels of organic pollution at all three sampling sites. Only one time during the year (10/1/79), did all three sites suggest high organic pollution.

Transeau's 6 ecological groups were used to …

Studies Of The Lysogenic Response As A Function Of Multiplicity Of Infection Of Lambdoid Bacteriophages, Mark Floyd Fedyk

Masters Theses

The lambdoid bacteriophages λimm^λ, λimm⁴³⁴ and λcI are ideal subjects for the study of gene product interactions at the molecular level due to their closely related genetic makeup. Previous work in this laboratory has indicated the possibility that phage λimm is sensitive to phage λimm⁴³⁴cI gene product and that phage λimm⁴³⁴ is sensitive to phage λimm^λcI gene product. The purpose of this research was to determine if the frequency of lysogeny for phage λimm^λ is affected by the multiplicity of coinfection with the λimm⁴³⁴ phage. By comparing the graph of …

The Biodegradation Of Crude Oil By Bacteria Isolated From Brine Pits, Larry Williams

Masters Theses

The biodegradation of crude oil by bacteria was studied. Hydrocarbon-utilizing bacteria were isolated from sixteen brine pits from an oil producing region of Clark County, Illinois. The bacteria were capable of growth from the vapors arising from crude oil as a sole carbon source. Some strains of bacteria were capable of emulsifying crude oil, apparently through the production of an emulsifying agent(s) that was found to be heat stable and cell bound but which can be partially stripped from the cell by autoclaving. The bacteria appear to belong to the genus Arthrobacter. The chemical environment of the pits was …

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. …

Bacteriophage Populations In Wastewater Effluent, Guy William Lawrence

Masters Theses

Effluent samples were taken at various stages of sewage treatment at a local wastewater treatment plant. Primary, secondary, tertiary and post-chlorinated effluents were sampled. The agar overlay method was used to determine the numbers of plaque-forming bacteriophage particles on thirty-six species of enteric bacteria from the Eastern Illinois University stock culture collection. Twenty-three pure culture phage isolates were prepared from selected plaques and their host range specificity was determined. For every host organism tested, a fifty percent or greater reduction of bacteriophage was evident through the sewage treatment process. Results obtained from investigation of host range specificity indicate that potential …

Interactions In Lambdoid Bacteriophage Populations, Bruce Edward Mitchell

Masters Theses

The lambdoid bacteriophages are ideal subjects for the study of populational interactions at the molecular level because they do not mutually exclude one another in mixed infection of the same host cell. Previous work done in this laboratory using the lambdoid phage pairs λ-434 hy mi and λ-∅80 have demonstrated a density-dependent reduction in progeny yield per infected cell (burst size) as a function of increasing multiplicity of infection (m.o.i.). The λ cI gene which codes for the repress or molecule has been demonstrated to be the cause of the interaction in the λ-434 hy mi phage pair. …

The Effects Of Residual Chlorine On Selected Microflora Of Kickapoo And Cassell Creek, James A. Mcgaughey

Masters Theses

The survival of fecal coliforms, fecal streptococci, and heterotrophs after exposure to chlorine were compared in receiving waters of the Mattoon and Charleston, Illinois waste treatment facilities employing membrane filter techniques and Standard Plate Counts. Water samples were taken from receiving waters upstream from the treatment plant, from secondary effluent and chlorinated secondary effluent within the plant, and from downstream receiving waters. Stream flow contribution and plant effluent contribution required the calculation of dilution factors to determine expected numbers of the various microflora found downstream assuming no residual chlorine effect. Total chlorine concentration was measured by amperometric titration at the …

The Effect Of Ergosterol On The Desaturation Of 14c-Labeled Oleic Acid In Tetrahymena, Maynard E. Neville Jr.

Masters Theses

The ciliated protozoan Tetrahymena pyriformis-Strain W desaturated ¹⁴C-labeled fatty acid substrate in both sterol supplemented (ergosterol) and nonsupplemented cultures. Cis-octadecenoic acid (¹⁴C-9-18:1), when added to the medium during the log phase of growth, was incorporated into the cells and also desaturated at the Δ⁶ and Δ¹² positions. The ¹⁴C labeled substrate and products, linoleate (9,12-18:2) or linolenate (6,9,12-18:3) were recovered and separated by silver-nitrate Unisil column chromatography. Initially, recovery of ¹⁴C compounds varied widely when expressed as the percent of total counts per minute recovered from column chromotography, and a variety of …

A Statistical Approach To The Study Of The Ecosystems Of Seven Ponds In East-Central Illinois, Gregory Lee Orr

Masters Theses

Gross primary productivity, heterotrophic bacterial numbers, and net phytoplankton densities of seven ponds in Coles County, Illinois, were studied in relation to physical, chemical, and biological habitat variables (light intensity and duration, turbidity, water temperature, pH, dissolved oxygen, sulfur, nitrogen, phosphorus, production, bacteria, and phytoplankton). Ten observations were made for each pond (except where otherwise noted) from 17 June through 25 August 1974. Stepwise multiple linear regression analyses of the data were used in order to determine those environmental factors which were important in predicting (i.e., significantly correlated with) bacterial and phytoplankton densities, and production. A multiple linear regression equation …

A Photomicrographic Guide To The Saprolegniaceae Of Illinois, Ronald L. Butler

Masters Theses

There is a paucity of information concerning the occurrence and distribution of the Saprolegniaceae of Illinois. These fungi, commonly called "water molds", are of particular interest as many are potential parasites of fish, crustaceans, and other aquatic plant and animal life. It is of considerable interest to students of the fungi and of practical value to the fisheries biologists and environmentalists to have available a means of identification for these organisms.

This paper provides a descriptive and photomicrographic account of the Saprolegniaceae of Illinois. Included herein are dichotomous keys to the genera of this family as well as generic descriptions …

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 …

A Study Of Soil Amoebae Infective To Mice, Pamela Mcdaniel

Masters Theses

On January 1, 1973, 10 soil samples were collected from areas in Coles County, Illinois, in an effort to isolate soil amoebae. All 10 soil samples were cultured on bacto-nutrient agar and colonies of amoebae genera isolated were Acanthamoeba and Hartmannella. Amoebabacteria cultures prepared from soil isolates were injected into each of 10 adult mice which were sacrificed at the end of the nine days and examined for evidence of infection and pathogenicity. Lesions were observed in two mice and amoebae cysts and bacterial colonies were present in nine mice. Amoeba-bacteria cultures were purified using streptomycin antibiotic discs and …