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- Creators: Arizona State University
- Creators: Haydel, Shelley
Description
Little is known about the diversity and role of bacteriophages in carbon (C) rich ecosystems such as peatlands in tropical and temperate regions. In fact, there is no currently published assessment of phage abundance on diversity in a key tropical ecosystem such as Amazon peatlands. To better understand phage assemblages in terrestrial ecosystems and how bacteriophages influence organic C cycling to final products like CO2 and CH4, phage communities and phage-like particles were recovered, quantified, and viable phage particles were enriched from pore water from contrasting Amazon peatlands. Here we present the first results on assessing Amazon bacteriophages on native heterotrophic bacteria. Several steps to test for methodological suitability were taken. First, the efficiency of iron flocculation method was determined using fluorescent microscopy counts of phage TLS, a TolC-specific and LPS-specific bacteriophage, and Escherichia coli host pre- and post-extraction method. One-hundred percent efficiency and 0.15% infectivity was evidenced. Infectivity effects were determined by calculating plaque forming units pre and post extraction method. After testing these methods, fieldwork in the Amazon peatlands ensued, where phages were enriched from pore water samples. Phages were extracted and concentrated by in tandem filtering rounds to remove organic matter and bacteria, and then iron flocculation to bind the phages and allow for precipitation onto a filter. Phage concentrates were then used for overall counts, with fluorescent microscopy, as well as phage isolation attempts. Phage isolations were performed by first testing for lysis of host cells in liquid media using OD600 absorbance of cultures with and without phage concentrate as well as attempts with the cross-streaking methods. Forty-five heterotrophic bacterial isolates obtained from the same Amazon peatland were challenged with phage concentrates. Once a putative host was found, steps were taken to further propagate and isolate the phage. Several putative phages were enriched from Amazon peatland pore water and require further characterization. TEM imaging was taken of two phages isolated from two plaques. Genomes of selected phages will be sequenced for identification. These results provide the groundwork for further characterizing the role bacteriophage play in C cycling and greenhouse gas production from Amazon peatland soils.
ContributorsSpring, Jessica Lynette (Author) / Cadillo-Quiroz, Hinsby (Thesis director) / Haydel, Shelley (Committee member) / Misra, Rajeev (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
Groundwater contamination is of environmental and human health concern. Bioremediation is a nature-based method for contaminant treatment. Bioremediation, which relies on the ability of microorganisms to destroy or transform contaminants, must be reliable and cost-competitive in comparison to more traditional treatment methods. Two hurdles must be overcome to enhance bioremediation’s effectiveness and competitiveness: i) being able to degrade recalcitrant compounds, and ii) being able to control the growth rate and location of microorganisms involved in bioremediation in the subsurface. My dissertation adds foundational knowledge and engineering application on how to biodegrade recalcitrant emerging and legacy halogenated compounds. Generating biotransformation knowledge on the recalcitrant emerging contaminants called per- and polyfluoroalkyl substances (PFAS) may lead to solutions for protecting both people and the planet. In my dissertation, I analyzed PFAS biotransformation and microbial defluorination literature via meta-analytical and bibliometric methods to identify unexplored topics and experimental conditions. The metanalytical work identified trends in PFAS microbial biotransformation science to inform future experimental design.
The second hurdle which must be overcome is being able to control bacterial growth in the subsurface. During bioremediation implementation microbial overgrowth may clog injection wells and the subsurface, leading to reduced porosity and treatment efficacy. Contaminant treatment schemes based on aerobic cometabolism frequently exhibit overgrowth at subsurface injection points for O2 (the electron acceptor) and a labile hydrocarbon (e.g., propane). My dissertation work experimentally evaluated acetylene as a microbial inhibitor for use in controlling microbial overgrowth during trichloroethene (TCE) aerobic cometabolism. I demonstrated that acetylene reduces the likelihood of microbial overgrowth of TCE-degrading microorganisms in soil-free microcosms and aquifer soil columns while retaining TCE degradation capacity. Cumulatively, my dissertation provides foundational knowledge for academics and bioremediation practitioners to develop robust and reliable bioremediation technologies.
ContributorsSkinner, Justin Paul (Author) / Delgado, Anca G. (Thesis advisor) / Rittmann, Bruce E (Committee member) / Chu, Min Ying Jacob (Committee member) / Arizona State University (Publisher)
Created2023