Theses and Dissertations at Montana State University (MSU)
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Item Microbial ecology of nitrifying simulated premises plumbing(Montana State University - Bozeman, College of Letters & Science, 2012) Encarnacion, Gem Deangkinay; Chairperson, Graduate Committee: Anne Camper; Mark D. Burr and Anne K. Camper were co-authors of the article, 'Bacterial communities in nitrifying simulated premises plumbing' in the journal 'Water research' which is contained within this thesis.; Anne K. Camper was a co-author of the article, 'Detection and enrichment of a nitrifying community containing a novel archaeon from simulated premises plumbing' in the journal 'PLoS One' which is contained within this thesis.Because of the Stage 2 Disinfectants and Disinfection Rule limiting then concentration of disinfection by products in drinking water, the use of chloramine as an alternative to chlorine has been increasing. However, the ammonia introduced by chloramination can lead to nitrification which results in the production of nitrite and nitrate, leading to regulatory violations. Nitrification in reactors with copper and polyvinyl chloride (PVC) surfaces was established by indigenous organisms from Bozeman tap water and has been stably maintained for more than 6 years. Statistical analyses of polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) profiles determined that the active bacterial populations were different in the two systems. The assemblage of the organisms was also different from the starting population (BAC influent) suggesting both material and ammonia/carbon source affect the population. No known ammonia oxidizing bacteria were detected suggesting the role of different group for ammonia oxidation. Fluorescence in situ hybridization (FISH) detected archaea in the biofilm from both reactors. Archaeal 16S rRNA gene sequences were found to be phylogenetically affiliated with known archaeal ammonia oxidizers. Two archaeal amoA sequences were amplified from the system as determined by DGGE. We propose to provisionally classify a detected archaeon as Candidatus Nitrosotenuis bozemanii, based on its affinity to Nitrosotenuis uzonensis (Hatzenpichler et al., in preparation). Bacterial abundances were comparable in the two systems but archaeal abundances were higher in the PVC reactor suggesting material effect on the overall microbial population composition and density. Enrichment in modified synthetic Crenarchaeota medium yielded a culture of archaea and bacteria that consistently oxidizes ammonia to nitrate. Attempts to isolate the archaeal component using antibiotics failed, suggesting the disruption of a possible beneficial relationship between the archaea and bacteria. Genes involved in the transformation of nitrogen within the system were also investigated and hao distantly related to that of ammonia oxidizing bacteria was detected but its potential role remains unknown. This study provides evidence of archaea associated with biofilms in drinking water and while further analysis is needed to definitively elucidate their role, results of this study prompts the reevaluation of the current concept of nitrification in drinking water.Item Analysis of microbial biofilm community composition within constructed wetlands(Montana State University - Bozeman, College of Letters & Science, 2010) Faulwetter, Jennifer Lynn; Chairperson, Graduate Committee: Anne Camper; Vincent Gagnon, Carina Sundberg, Florent Chazarenc, Mark D. Burr, Jacques Brisson, Anne K. Camper and Otto R. Stein were co-authors of the article, 'Microbial processes influencing performance of treatment wetlands: a review' in the journal 'Ecological engineering' which is contained within this thesis.; Mark D. Burr, Otto R. Stein and Anne K. Camper were co-authors of the article, 'Characterization of sulfate reducing bacteria in constructed wetlands' in the journal 'Proceedings of the 11th international conference on wetland systems for water pollution control, Indore, India' which is contained within this thesis.; Mark D. Burr, Albert E. Parker, Otto R. Stein and Anne K. Camper were co-authors of the article, 'The effect of plant species and sample location on bacterial biofilm communities associated with constructed wetland microcosms' in the journal 'The international society for microbial ecology journal' which is contained within this thesis.; Mark D. Burr, Albert E. Parker, Otto R. Stein and Anne K. Camper were co-authors of the article, 'The influence of sulfate reducing bacteria and ammonia oxidizing bacteria on nutrient cycling in constructed wetland microcosms' in the journal 'Microbial ecology' which is contained within this thesis.; Mark D. Burr, Alfred B. Cunningham, Frank M. Stewart, Anne K. Camper and Otto R. Stein were co-authors of the article, 'Floating treatment wetlands for domestic wastewater treatment' in the journal 'Water science and technology' which is contained within this thesis.Constructed wetlands (CWs) are ecologically-based water treatment systems that provide cost-effective amelioration of waterborne pollutants. Fundamental understanding of removal mechanisms, especially microbial processes, limits greater usage of constructed wetlands as a wastewater treatment system. The influence of plant species selection, season, and organic load rate on pollutant removal was previously linked to the redox condition of the sub-surface wetland environment. The goal of this research was to determine which of these environmental variables (including spatial location within the CW) influenced the dominant microbial populations and/or the activity of various sub-populations. Once identified, a constructed wetland might be optimized for growth of microorganisms involved in removal of a specific pollutant. To assess environmental factors, microbial population samples were taken in six locations (effluent, 3 root and 2 gravel areas) within replicate unplanted microcosms and wetland microcosms planted with Deschampsia cespitosa or Leymus cinereus during the summer (24°C) and winter (4°C) seasons. Microcosms were fed a synthetic domestic wastewater in 20-day batches for at least 12 months prior to sampling. The most recent techniques in molecular biology including denaturing gradient gel electrophoresis (DGGE) and quantitative PCR were utilized and included treatment with and without propidium monoazide (PMA) to distinguish between "live" and "dead" microbial communities. Primer sets targeted the entire bacterial community (16S rDNA) and two functional groups, nitrifying bacteria (amoA gene) and sulfate reducing bacteria (dsrB gene). Results indicated that overall microbial community structure (16S rDNA) was affected by general location within the microcosm (effluent, root, gravel) as well the plant species present. Specific microbial groups appeared to be affected differently with relative gene quantities of sulfate reducing bacteria and nitrifying bacteria being influenced by a combined effect of plant species and season. For dsrB, D. cespitosa had the lowest relative gene quantities overall. Both genes were more abundant in the summer season, indicating seasonal importance. Location within the microcosms was also important, with anoxic environments (column bottom) being more important for dsrB presence and a diverse population of cultivated sulfate reducers. The roots were an important location for both microbial diversity and activity for all genes investigated.