Chronic microbial infections result from persistent host colonization that is not cleared via the immune response or therapeutics. Within the host, microbes can undergo adaptive evolution, whereby beneficial traits promoting persistence arise due to selection; these traits can therefore affect disease outcomes and treatment strategies. The Gram-negative opportunistic pathogen Pseudomonas aeruginosa is the primary cause of chronic, fatal respiratory infections in individuals with the heritable disease cystic fibrosis (CF). The goal of this dissertation is to identify adaptations that allow P. aeruginosa to persist in the host during chronic CF lung infection. To achieve this goal, P. aeruginosa was chronologically sampled from 3 CF patients, ranging from the first infecting bacterium (the ancestor) to ~40,000 generations post-infection. By comparing gene expression profiles of ancestral and evolved isolates sampled from multiple patients, I identified 24 parallel gene expression changes that occurred over time within each lineage, suggesting that these traits are beneficial to the bacterium. Because most of these traits had unknown physiological roles, I sought to characterize their biological significance. I used a gain-of-function genetic screen and discovered that a subset of these genes enhance biofilm formation, a sessile mode of growth proposed to be important during chronic CF lung infection. I showed that enhanced biofilm formation is due to increased production of the exopolysaccharide Psl, which is traditionally viewed as less critical for maintaining chronic infections than other virulence factors. Lastly, I demonstrated that a majority (~72%) of chronic P. aeruginosa isolates produce more Psl than their corresponding ancestor, suggesting that this exopolysaccharide is important during chronic infection and an adaptive trait. / text

In spite of abundant water resources, world is suffering from the scarcity of usable water. Seawater Reverse Osmosis (SWRO) desalination technology using polymeric membranes has been recognized as a key solution to water scarcity problem. However, economic sustainability of this advanced technology is adversely impacted by the membrane fouling problem. Fouling of RO membranes is a highly studied phenomenon. However, literature is found to be lacking a detailed study on kinetic and dynamic aspects of SWRO membrane fouling. The factors that impact the fouling dynamics, i.e., pretreatment and water quality were also not adequately studied at full–scale of operation. Our experimental protocol was designed to systematically explore these fouling aspects with the objective to improve the understanding of SWRO membrane fouling mechanisms. An approach with multiple analytical techniques was developed for fouling characterization. In addition to the fouling layer characterization, feed water quality was also analysed to assess its fouling potential. Study of SWRO membrane fouling dynamics and kinetics revealed variations in relative abundance of chemical and microbial constituents of the fouling layer, over operating time. Aromatic substances, most likely humic–like substances, were observed at relatively high abundance in the initial fouling layer, followed by progressive increase in relative abundances of proteins and polysaccharides. Microbial population grown on all membranes was dominated by specific groups/species belonging to different classes of Proteobacteria phylum; however, similar to abiotic foulant, their relative abundance also changed with the biofilm age and with the position of membrane element in RO vessel. Our results demonstrated that source water quality can significantly impact the RO membrane fouling scenarios. Moreover, the major role of chlorination in the SWRO membrane fouling was highlighted. It was found that intermittent mode of chlorination is better than continuous mode of chlorination of seawater, as anti–biofouling strategy. It was also confirmed that significant biofilm development was inevitable even with the use of chlorine to disinfect SWRO membranes. Our findings on the dynamic patterns of SWRO membrane fouling should help in further elaborating research projects focusing on the development of better strategies to minimize this troublesome phenomenon.

No description available.

This study examined some of the environmental, physiological, and genetic factors or mechanisms which contribute to L. monocytogenes’ desiccation survival under food processing conditions. Desiccation experiments were carried out on stainless steel coupons stored at 43% RH, 15°C. The level of initial contamination had no impact (p>0.05), whereas the presence of a mature biofilm, prior osmoadaptation, and the presence of salt (5%) and lard (20-60%) on the SS coupons significantly (p<0.05) increased the bacterium’s desiccation survival. An Lm568 transposon mutant library was constructed to screen for novel genes involved in desiccation survival. Fifteen tolerant and 16 sensitive desiccation mutants were sequenced. Interrupted genes involved in motility and FA membrane modification were the most common in tolerant mutants whereas energy and membrane transport related genes were the most recognized in sensitive mutants. Lastly, a spontaneous desiccation resistant Lm568 variant was isolated, emphasizing the importance of understanding desiccation tolerance for food safety.

Surgical site infections (SSIs) are an emerging cause of increased morbidity, mortality, and treatment cost, in veterinary medicine. Medical records were searched to evaluate for associations that could increase the risk of developing SSIs. Logistic regression was used to analyze the risk factors statistically, to determine their influence on SSI risk. An SSI incidence rate of 3.0% was found in this study for all small animal surgical procedures performed from September 2010 to July 2011, with implants, hypotension and surgical classification associated with increased likelihood of SSI. Active surveillance is crucial for the development of methods to prevent SSI’s. Biofilms contribute to the antimicrobial resistance properties commonly found in bacteria such as methicillin-resistant Staphylococcus pseudintermedius, which is found in canines. An enzyme known as DispersinB was studied to assess its effect on biofilm formation and degradation. DispersinB prevented the formation and eradicated biofilm in vitro. In vivo testing is required to further assess the effects of DispersinB. / Ontario Veterinary College Pet Trust, Canadian Institutes of Health Research, Kane Biotchech

Biofilms are complex and highly adapted communities of microorganisms found attached to surfaces. Among the best characterized infectious multi-cellular biofilms is the oral community known as dental plaque. Streptococcus mutans resides in the oral biofilm, and is one of the main causative agents of dental caries. Streptococci are known to monitor their population density using a peptide pheromone (CSP)/two component signalling system (ComDE) in a process classically known as quorum sensing (QS). Previous work in S. mutans has implicated the QS system in genetic competence, the stress response, bacteriocin production and biofilm formation. Our objective in this work was to thoroughly characterize the transcriptional and phenotypic response to CSP in S. mutans, and determine its role in biofilm formation. We have shown that the CSP pheromone is more than simply a QS signal, and is also an inducible ‘alarmone’ capable of communicating stress in the population. We have demonstrated that elevated concentrations of CSP such as those that occur during stress trigger autolysis in a fraction of the population. Importantly, we have shown that autolysis in S. mutans occurs via a novel mechanism of action: intracellular accumulation of a self-acting bacteriocin. We have also identified and characterized the autolysis immunity protein, which is differentially regulated from the bacteriocin to allow survival at low cell density. A second regulatory system was shown to govern expression of autolysis immunity in the absence of CSP signaling, and also contribute to the oxidative stress response in the biofilm. Finally, we present evidence that autolysis is involved in the release of DNA in the biofilm, which contributes to the architecture of the extracellular matrix and may provide a mechanism for the dissemination of fitness-enhancing genes under stress. Together, our data provides a mechanistic link between phenotypes previously ascribed to the CSP pheromone in S. mutans.

Remediation of DNAPL contaminated sites in fractured rock has proven to be very difficult. No current technology can be used to remediate such sites in a timely and economic manner due to the inherent heterogeneity of fractured rock and back diffusion of contaminants stored in the rock matrix. This study was conducted in order to evaluate the viability of biostimulation of native biofilm as a means to control flow and back diffusion at fractured rock sites. A field trial was conducted at an uncontaminated site in southern Ontario. The site is underlain by dolomites of the Lockport formation. Three major fracture zones have been identified in the study area. Two closely spaced (5.04 m) boreholes were used to isolate a fracture zone at a depth of 17 m with straddle packers. These boreholes were used to create an injection-withdrawal system with recirculation, which was used for tracer injection in order to load the rock matrix with a conservative dye tracer and to inject nutrients for 21 days in order to stimulate the growth of biofilm in the fracture. Evaluation of the ability of the biofilm to control flow through the fracture was conducted through pulse interference tests. Pulse interference tests were conducted before and after the injection of nutrients. The results from the pulse interference tests showed a maximum 65% reduction in transmissivity, which is equivalent to a 28% reduction in fracture aperture shortly after the cessation of biostimulation. In order to investigate the effect of the biofilm stimulation on matrix diffusion the rock matrix was loaded with Lissamine, a conservative fluorescent dye tracer prior to biostimulation and its concentration was monitored at injection and withdrawal wells. The effect that biostimulation had on matrix diffusion was determined by comparing field concentration measurements with a model that simulates a system unaffected by biofilm stimulation. The biostimulation lowered the concentration of tracer attributable to back diffusion at the withdrawal well by about 20% for approximately 30 days following the cessation of biostimulation. It is also thought that large amounts of tracer might have been trapped in the biofilm as it formed and was then released back into the fracture as the biofilm deteriorated. / Thesis (Master, Civil Engineering) -- Queen's University, 2009-08-11 19:27:44.232

No description available.

No description available.

Membrane biofilm reactors (MBfR) utilize membrane fibers for bubble-less transfer of gas by diffusion and provide a surface for biofilm development. Nitrogen removal was attempted using MBfR in various configurations - nitrification, denitrification and consecutive nitrification and denitrification. Effects of loading rate and dissolved oxygen on nitrification performance were primarily investigated in a stand-alone nitrifying MBfR. Specific nitrification rate increased linearly with specific loading rate, up to the load of 3.5 g N/m²d. Beyond that load, substrate diffusion limitation inhibited further increase of specific nitrification rate. 100% oxygen utilization was achievable under limited oxygen supply condition. Effects of mineral precipitation, dissolved oxygen and temperature on hydrogenotrophic denitrification were investigated in a stand-alone denitrifying MBfR. Mineral precipitation, caused by intended pH control, caused the deterioration of denitrification performance by inhibiting the diffusion of hydrogen and nitrate. Operating reactor in various dissolved oxygen conditions showed that the denitrification performance was not affected by dissolved oxygen in MBfR. Optimum temperature of the hydrogenotrophic denitrification system was around 28°C. Total nitrogen removal in a two-step MBfR system incorporating sequential nitrification and hydrogen-driven autotrophic denitrification was investigated in order to achieve nitrogen removal by autotrophic bacteria alone. Long-term stable operation, which proved difficult in previous studies due to excessive biofilm accumulation in autotrophic denitrification systems, was attempted by biofilm control. Nitrification performance was very stable throughout the experimental periods over 200 days. Performance of autotrophic denitrification was maintained stably throughout the experimental periods, however biofilm control by nitrogen sparging was required for process stability. Biofilm thickness was also stably maintained at an average of 270 µm by the gas sparging biofilm control. According to the cost analysis of denitrifying MBfR, hydrogenotrophic denitrification can be an economical tertiary treatment option compared to conventional denitrifying filter although its economic feasibility highly depends on the cost of hydrogen gas. Although this study was conducted in a lab-scale, the findings from this study can be a valuable stepping stone for larger scale application and open the door for system modifications in future.