No description available.

No description available.

No description available.

No description available.

Thesis (Ph.D.)--University of Delaware, 2007. / Principal faculty advisor: David L. Kirchman, College of Marine and Earth Studies. Includes bibliographical references.

Call number: LD2668 .T4 1963 R57 / Master of Science

A seliberia-like bacterium (SLO), isolated from reverse osmosis membranes was characterized by morphological, physiological and DNA studies. The helically twisted cells of this organism were often observed in star-shaped clusters. Depending on nutritional conditions, cells ranged from 0.5 to 21 um in length and possessed prosthecae. Small motile cells were produced by asymmetric fission or by a budding process. Ovoid "generative" cells were observed in mixed culture conditions or when the pure culture isolate was grown in the presence of humic acid. The SLO oxidatively utilized glucose, maltose, xylose, cellobiose, and several amino acids as sole carbon and energy sources. The organism is a strict aerobe and does not anaerobically respire. The moles percent guanine plus cytosine (mol% G + C) of the SLO DNA was 38% as compared with 63-67% for Seliberia stellata. Although the cellular morphology and physiology of the SLO closely resembles that of S. stellata, the SLO is considered to be a new species of Seliberia based on the presence of prosthecae and the mol% G + C.

Oxidative assimilation of glucose-U-C¹⁴ by several aerobic bacteria was found to involve the assimilation of radioactivity into nitrogenous cell components, principally proteinaceous, in conjunction with the reincorporation of endogenously produced ammonia. In one of these bacteria, Pseudomonas aeruginosa, if the cells were starved or treated with chloramphenicol/ prior to glucose-C¹⁴ the amount of assimilation, especially into protein, was decreased. The incorporation into nucleic acids and lipids was increased by the antibiotic, but was only slightly affected by starvation. A determination of the cytological sites of the assimilated material showed that, in control cell extracts, the soluble proteins of the cytoplasm contained most of the C¹⁴. Starved or antibiotic treated cell fractions had substantially less of the label in these proteins, whereas the radioactivity incorporated into the ribosomal ribonucleic acid and the "membrane" lipids was greater. A study of the aminoacyl-soluble ribonucleic acid synthetases in P. aeruginosa revealed that these enzymes were present only in the cytoplasm. Starving the cells resulted in decreased activity of the synthetases, but they were rapidly reactivated during oxidative assimilation. The large amount of heterologous reactions between bacterial soluble ribonucleic acids and synthetases indicated that little species specificity existed. However, cross reactions between the systems in bakers' yeast and the bacteria were poor, showing that some degree of species specificity was present in these instances. Preliminary experiments on the route of assimilation of ammonia in P. aeruginosa and in P. fluorescens gave no evidence for the direct amination of pyruvate by alanine dehydrogenase, but did demonstrate a requirement for concurrent substrate oxidation while ammonia was being incorporated. In contrast, several lines of evidence indicated that ammonia was assimilated via ∝-ketoglutarate in P. aeruginosa. / Land and Food Systems, Faculty of / Graduate

AbstractThe bactericidal effect of ozone at concentrations of 1 and 2 p.p.m. by volume, was tested on separately atmized resting cell suspensions of Aerobacter aerogenes, Bacillus megathreium, and Escherichia coli B impinged on Millipore filters. The exposure of each species required the use of 18 randomly selected filters containing impinged organisms. Of the 18 filters, 6 were used in the control. The temperature and the relative humidity of the ozone flow were approximately 25ºC and 20 per cent, respectively, while the flow-rate across each filter was 3 liters per minute. It was found that all of the organisms tested were completely destroyed by both 2 and 2 p.p.m. ozone within 2 minutes of exposure.Ball State UniversityMuncie, IN 47306

Members of the genus Bacillus are mainly Gram positive, aerobic rod shaped, endospore-forming bacteria that are increasingly being recognised for their ability to promote plant growth and antagonise fungal pathogens. From a biological control perspective, Bacillus spp. strains that produce antifungal compounds are of particular interest. In this study, aerobic endospore-formers were isolated from an undisturbed indigenous grassland soil and screened for antifungal activity and other plant growth promoting traits. Endospore-formers were also isolated from rhizosphere soil associated with the roots of maize, wheat and kale grown in pots containing soil from the same grassland site. Microbial diversity amongst isolates showing antifungal activity was investigated using different molecular fingerprinting methods, namely, intergenic transcribed spacer–PCR (ITS-PCR), random amplified polymorphic DNA-PCR (RAPD-PCR) and 16S rRNA gene amplification and sequencing. Characterization of the active antimicrobial compound(s) associated with selected isolates was also attempted. Prior to isolating from bulk and rhizosphere soils, samples were pre-heated to eliminate heat sensitive vegetative cells. Mean endospore counts were; wheat rhizosphere, Log 6.03 c.f.u g-1 soil; maize rhizosphere, Log 5.88 c.f.u g-1 soil; kale rhizosphere Log 5.90 c.f.u g-1 soil; and bulk soil Log 5.67 c.f.u g-1soil. A total of three hundred and eighty-four isolates were screened for antagonism towards Rhizoctonia solani using dual-culture plate bioassays. Thirty four of the isolates (~9%) mostly isolated from the bulk soil inhibited R. solani at varying degrees. Differences in antimicrobial interactions were apparent in in vitro bioassay; supposedly due to different concentrations and/or types of antimicrobial compounds. Biochemical tests for amylase, cellulase, chitinase, and proteinase activity, siderophore production and inorganic phosphate solubilisation were conducted. None of the isolates possessed all of these attributes and only a few showed multiple traits. Ninety-one percent of the isolates exhibited proteinase activity, 76% were able to hydrolyze starch whereas only four displayed cellulase activity. Only four isolates from the bulk-soil were capable of solubilising inorganic phosphate. ITS-PCR and 16S rRNA gene sequence analysis showed high levels of genetic homology amongst isolates and the majority were closely associated with representatives of the B. cereus group. Isolate C76 was the exception, being closely matched with B. subtilis. ITS-PCR banding profile was useful for distinguishing between species but did not distinguish within species. RAPD-PCR distinguished finer levels of genetic diversity between and within sample sets, with primer OPG-11 showing the greatest levels of heterogeneity. DNA extraction methods and the influence of template DNA dilution were investigated to determine their influence on RAPD-PCR analysis reproducibility. Prominent bands were comparable for crude template- and kit-extracted DNA but slight changes in band intensity and in some instances, additional faint bands were observed. At the highest DNA concentrations tested (7 μg/ml), further bands with molecular weights above 2.5 kbp were apparent. Strict standardization of PCR conditions greatly reduced variability of the RAPD-PCR analysis. Isolates from the different sample sets were screened for the presence of genetic markers associated with the biosynthesis of zwittermicin A, an aminopolyol antibiotic produced by some members of the B. cereus group. In an initial screen only one isolate, W96, yielded PCR amplicons consistent with those previously reported in the literature for the zwittermicin A genes. Later a further sixteen isolates grouped with W96 on the basis of the RAPD-PCR fingerprinting profiles, were screened for the presence of these genes. Of these, only six showed PCR amplification products similar to W96. Sequence homology testing against the GenBank database confirmed the presence of the zwittermicin A genes in these isolates. Isolate W96 was selected for further extraction and characterization of its antifungal compound(s). However, after culturing in various broth media cell free supernatants of W96 failed to show antifungal activity in vitro even when the supernatants were concentrated 20-fold. These findings provide a general overview of the diversity of aerobic endospore-forming bacteria present in an undisturbed indigenous grassland soil that exhibited antifungal activity in vitro and the limited influence tested crop rhizospheres have on this diversity. Combined use of ITS-PCR, 16S rRNA sequencing and RAPD-PCR techniques served as a rapid and effective means of grouping isolates for further investigations of their potential use as biocontrol agents and plant growth promoting rhizobacteria. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.