Spores of thermophilic bacilli are a common concern during the manufacture of milk powder. Spores are believed to occur in high numbers in milk powder due to their ability to survive pasteurisation, attach to stainless steel surfaces, germinate, grow as biofilms and subsequently enter the product stream and thereby contaminate the final product. In this study, thirty one thermophilic bacilli isolates were obtained from a New Zealand milk powder production line and identified as either Anoxybacillus flavithermus or Geobacillus spp. using random amplified polymorphic DNA (RAPD) and species-specific PCR. Sporulation media and a polyethylene glycol two-phase separation system were modified to produce high yields of spores free from debris. The spores of four Geobacillus spp. isolates (CGT-8, D4, E7 and E11) were characterised in terms of structure (electron microscopy), surface charge (zeta potential), hydrophobicity (contact angle and microbial adhesion to hexadecane) and attenuated total reflectance infrared spectroscopy (ATR-IR). Spores from three of the four isolates possessed an exosporium while the fourth did not. However the integrity of the exosporium varied over time. The spores were negatively charged (-10 to -20 mV) at neutral pH and high ionic strength (0.1 M KC1). Both hydrophobicity assays revealed that the spores of the four isolates were relatively hydrophilic while ATR-IR revealed the spores' surfaces consisted of protein and polysaccharides. The influence of these spore characteristics on adhesion to a variety of substrata under high flow rates was examined using the extended Derjaguin, Landau, Verwey and Overbeek (XDLVO) theory. Spores generally attached in higher numbers to hydrophobic surfaces compared to hydrophilic surfaces, however this observation was more prevalent for isolate D4. This result indicated that a single mechanism could not describe the adhesion of spores from different strains. A series of glass surfaces with modified characteristics were produced in order to test the antifouling properties on the adhesion of D4 spores. Spores suspended in a high ionic strength medium (0.1 M KC1) attached in greater numbers (1 Log₁₀ CFU cm⁻�) to positively charged and hydrophobic surfaces compared with negatively charged and hydrophilic surfaces. A clean in place (CIP) procedure, reduced spore numbers on hydrophobic and hydrophilic surfaces by 1.5 and by 2.0 Log₁₀ CFU cm⁻�, respectively. When spores were suspended in milk, there was little difference in the number of spores attaching to the different surfaces (ie. 3.5 to 3.8 Log₁₀ CFU cm⁻�), and spore removal from surfaces via a CIP regime was unchanged (1.5 to 2.0 Log₁₀ CFU cm⁻� reduction) compared with spores that attached in simple 1:1 electrolyte media. The effects of a caustic wash on spore surface characteristics and adhesion was determined. There was a significant reduction in spore viability (2 Log₁₀ CFU mL⁻�) after a 30 min caustic wash at 65 �C in the current study, however surviving spores displayed a greater propensity to attach to stainless steel. Surface characterisation results revealed an increase in hydrophobicity and a greater negative charge on the spores' surface after treatment with NaOH. Surviving spores could potentially recontaminate sections of the plant which are cleaned with this recycled caustic wash solution, thereby seeding surfaces with spores at the beginning of the next processing run. In conclusion, while surfaces that reduce spore adhesion and enhance removal can be produced, exposure to complex solutions such as milk can reduce the anti-fouling effectiveness of such surfaces to spore adhesion.

Includes bibliographical references. / vi, 86, [56] leaves, [10] leaves of plates : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / The aim of this work is the further characterisation of the citrus exocortis viroid (CEV) replication complex in a tomato host system. The study employed three approaches. i. To raise polyclonal antiserum against a putative catalytic component of the CEV replication complex, the second largest subunit of tomato RNA polymerase II--ii. To develop a cell-free system for CEV RNA synthesis--iii. To use the polyclonal antiserum to the second largest subunit of RNA polymerase II, and a commercially available monoclonal antibody to the largest subunit, for antibody inhibition and immunoprecipitation experiments using the cell-free system. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 1997

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2005. / Härtill 5 uppsatser.

Thesis (M.S.)--Missouri University of Science and Technology, 2009. / Vita. The entire thesis text is included in file. Title from title screen of thesis/dissertation PDF file (viewed August 10, 2009) Includes bibliographical references (p. 58-64).

Due to increased plant resistance to the existing antibiotics produced, there is a need to develop alternatives. Nonribosomal peptides (NRPs) are important plant phytopathogens synthesized by nonribosomal peptide synthetases (NRPSs). In this study, a newly sequenced Bacillus strain Bacillus atrophaeus UCMB 5137 (63Z), found to have increased phytopathogenic activity, was investigated to gain insights to the possible reason behind this activity. NRPS modules were identified using a novel script that can act on unannotated, raw DNA sequences. The Structure Based Sequence Analysis Webserver was used to identify the amino acids incorporated into the final NRP, which were compared to the NRP database. Five NRPSs were found within the strain; fengycin/plipstatin, mycosubtilin, surfactin, bacillibactin and bacitracin. Some of the modules usually present for these NRPSs were not present in the test strain and only a few modules were found. A phylogenetic study was carried out and the topologies of the trees showed that genes were not transferred horizontally. It did, however, lead to the hypothesis that different NRPS genes are under different adaptive evolutionary pressures. Only slight conformational changes between L and D-conformation of amino acids were seen between the test and neighboring strains. All of the linker and terminal regions of synthetases were found to exhibit a large amount of conservation overall. Homology modeling was performed on the test strain on selected modules, TE and A-domains of fengycin and mycosubtilin synthetases. TE-domains between the different synthetases are different and specific for the NRP they facilitate release for. The NRPS from which the A-domain originates also influences substrate specificity as well as the module in which the A-domain occurs within the NRPS. Binding pockets of A-domains of differing substrate specificity were compared. Future work will include; refinement of the models and docking studies within the A-domain binding pocket. / Microsoft� Word 2010 / Adobe Acrobat 9.54 Paper Capture Plug-in

Stenotrophomonas maltophilia is increasingly emerging as an opportunistic pathogen of global concern. Due to its inherent resistance to several classes of antibiotics including carbapenems and its ability to acquire mobile resistance elements, treatment of infections caused by S. maltophilia is a constant challenge for clinicians. Trimethoprim-sulphamethoxazole (TMP-SMX) is the generally accepted antibiotic of choice for the treatment of infections caused by this organism, but resistance to the drug is increasingly being reported; hence, the need for alternative therapeutic options. In this study, the antimicrobial susceptibility profile of 110 commensal S. maltophilia isolates obtained from Nkonkobe municipality, Eastern Cape Province, Republic of South Africa was investigated. Twenty-one antibiotics including TMP-SMX and the newer fluoroquinolones; levofloxacin, gatifloxacin and moxifloxacin were included in the antibiotic panel. About 63.4 percent of the isolates were susceptible to TMP-SMX with a resistance rate of 28.2 percent. The fluoroquinolones were more effective with susceptibilities ranging from 76 percent to 94.7 percent. Resistance to the fluoroquinolones ranged from 1.3 percent to 2.7 percent. Levofloxacin was the most effective fluoroquinolone tested. Phenotypic dectection of extended spectrum β-lactamases (ESBLs) showed double disc synergy test (DDST) positivity in 59.5 percent of the isolates. Cefepime was the most sensitive indicator cephalosporin in the DDST with 77.3 percent of suspected ESBL-producing isolates showing cefepime-clavulanic acid synergy. Isolates exhibited nine different ESBL phenotypes, however, PCR amplification of the bla genes revealed four isolates that possessed genes belonging to the CTX-M group (CTX-M-1 and CTX-M-8 groups). ESBL genes are usually carried on mobile elements such as plasmids and transposons which may also bear genes that mediate resistance to aminoglycosides, tetracyclines, TMP-SMX and fluoroquinolones. ESBL positive isolates appeared more susceptible to the fluoroquinolones compared to TMP-SMX but there was no significant relationship between ESBL production and susceptibility to these drugs (p > 0.05). The newer fluoroquinolones are a possible alternative treatment option for S. maltophilia infections in this environment but further studies and clinical investigations are needed to determine the in vivo efficacy of these drugs.

No description available.

The interplay between geology and biology has shaped the Earth during billions of years. Microbe-mineral interactions are prime examples of this interplay and underscore the importance of microorganisms in making Earth a suitable environment for all forms of life. The present thesis takes an interdisciplinary approach to obtain an integrated understanding of microbe-mineral interactions. More specifically it addresses how the composition and distribution of biogenic weathering agents (siderophores) differ with regard to soil horizon and mineral type in situ, what siderophore type soil microorganisms produces under laboratory conditions, what role microbial surface attachment plays in mineral weathering reactions and what central roles and applications siderophores have in the environment. Podzol, the third most abundant soil in Europe, and most abundant in Scandinavia, was chosen for a field experiment, where three minerals (apatite, biotite and oligoclase) were inserted in the organic, eluvial and upper illuvial soil horizons. The study started with an investigation of the siderophore composition in the bulk soil profile and on the mineral surfaces (paper I), which was followed by a study of the siderophore producing capabilities of microorganisms isolated from the soil profile under laboratory conditions (paper II). Subsequently, a study was done on the impact of microbial surface attachment on biotite dissolution (paper III). Finally, the roles of siderophores in nature and their potential applications were reviewed (paper IV). The major findings were that the concentration of hydroxamate siderophores in the soil attached to the mineral surfaces was greater than those in the surrounding bulk soil, indicating that the minerals stimulate the microbial communities attached to their surfaces to produce more siderophores than the microorganisms in the bulk soil. Each mineral had a unique assemblage of hydroxamate siderophores, that makes the mineral type one of the main factors affecting siderophore composition in the natural environment. Siderophore production varied between the microbial species originating from different soil horizons, suggesting that the metabolic properties of microbes in deep soil horizons function differently from those at upper soil horizons. Microbial surface attachment enhanced the biotite dissolution, showing that attached microbes has a greater influence on weathering reactions in soil than planktonic populations. In conclusion, our findings reflected that the complicated relationship between microorganisms and mineral surfaces reinforces the central theme of biogeochemistry that the mineral controls the biological activity in the natural environments. However, the importance of these relationships to the biogeochemical systems requires further investigation. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: In press. Paper 3: In press.</p><p> </p>

Call number: LD2668 .T4 1952 J61 / Master of Science

Call number: LD2668 .T4 1985 C348 / Master of Science