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Pages
- Title
- Repair of E. coli B130 DNA, damaged by mitomycin-c
- Creator
- Ginzburg, Irith
- Date
- 1968
- Collection
- Electronic Theses & Dissertations
- Title
- Conversion of the mitochondrial gene for mammalian cytochrome oxidase subunit II to a universal equivalent and expression in E. coli, in vitro, and in Xenopus oocytes
- Creator
- Cao, Jianli
- Date
- 1990
- Collection
- Electronic Theses & Dissertations
- Title
- Resistance of the newborn calf to oral challenge with a septicemia-producing Escherichia coli
- Creator
- Johnston, Noel Edgar
- Date
- 1975
- Collection
- Electronic Theses & Dissertations
- Title
- Microfiltration process for enhanced production of rDNA receptor cells of Escherichia coli
- Creator
- Anderson, Kevin Warren
- Date
- 1983
- Collection
- Electronic Theses & Dissertations
- Title
- Anti-late RNA and transcriptional control in T4 bacteriophage infected Escherichia coli
- Creator
- Frederick, Robert John, 1944-
- Date
- 1976
- Collection
- Electronic Theses & Dissertations
- Title
- Bioactivities of bovine anti-coliform antibodies elicited by J5 vaccinations
- Creator
- Chaiyotwittayakun, Anatachai
- Date
- 2003
- Collection
- Electronic Theses & Dissertations
- Title
- Physiological studies of recombinant protein expression in Escherichia coli
- Creator
- Wang, Kai
- Date
- 2006
- Collection
- Electronic Theses & Dissertations
- Title
- Strategies for improving synthesis of quinic acid and shikimic acid from D-glucose
- Creator
- Jancauskas, Justas
- Date
- 2008
- Collection
- Electronic Theses & Dissertations
- Title
- Isolation of salmonella and escherichia coli in feces of cull (market) dairy cows at slaughter
- Creator
- Akpinar, Özlem
- Date
- 2001
- Collection
- Electronic Theses & Dissertations
- Title
- Deconstructing the correlated nature of ancient and emergent traits : an evolutionary investigation of metabolism, morphology, and mortality
- Creator
- Grant, Nkrumah Alions
- Date
- 2020
- Collection
- Electronic Theses & Dissertations
- Description
-
Phenotypic correlations are products of genetic and environmental interactions, yet the nature of these correlations is obscured by the multitude of genes organisms possess. My dissertation work focused on using 12 populations of Escherichia coli from Richard Lenski's long-term evolution experiment (LTEE) to understand how genetic correlations facilitate or impede an organism's evolution. In chapter 1, I describe how ancient correlations between aerobic and anaerobic metabolism have...
Show morePhenotypic correlations are products of genetic and environmental interactions, yet the nature of these correlations is obscured by the multitude of genes organisms possess. My dissertation work focused on using 12 populations of Escherichia coli from Richard Lenski's long-term evolution experiment (LTEE) to understand how genetic correlations facilitate or impede an organism's evolution. In chapter 1, I describe how ancient correlations between aerobic and anaerobic metabolism have maintained - and even improved - the capacity of E. coli to grow in an anoxic environment despite 50,000 generations of relaxed selection for anaerobic growth. I present genomic evidence illustrating substantially more mutations have accumulated in anaerobic-specific genes and show parallel evolution at two genetic loci whose protein products regulate the aerobic-to-anaerobic metabolic switch. My findings reject the "if you don't use it, you lose it" notion underpinning relaxed selection and show modules with deep evolutionary roots can overlap more, hence making them harder to break. In chapter 2, I revisit previous work in the LTEE showing that the fitness increases measured for the 12 populations positively correlated with an increase in cell size. This finding was contrary to theory predicting smaller cells should have evolved. Sixty thousand generations have surpassed since that initial study, and new fitness data collected for the 12 populations show fitness has continued to increase over this period. Here, I asked whether cell size also continued to increase. To this end, I measured the size of cells for each of the 12 populations spanning 50,000 generations of evolution using a particle counter, microscopy, and machine learning. I show cell size has continued to increase and that it remains positively correlated with fitness. I also present several other observations including heterogeneity in cell shape and size, parallel mutations in cell-shape determining genes, and elevated cell death in the single LTEE population that evolved a novel metabolism - namely the ability to grow aerobically on citrate. This last observation formed the basis of my chapter 3 research where my collaborators and I fully examine the cell death finding and the associated genotypic and phenotypic consequences of the citrate metabolic innovation.
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- Title
- The effects of genetic background on the evolution of antibiotic resistance and its fitness costs
- Creator
- Card, Kyle Joseph
- Date
- 2020
- Collection
- Electronic Theses & Dissertations
- Description
-
Antibiotic resistance is a growing public-health concern. Efforts to control the emergence and spread of resistance would benefit from an improved ability to forecast when and how it will evolve. To predict the evolution of resistance with accuracy, we must understand and integrate information about many factors, including a bacterium's evolutionary history. This dissertation centers on the effects of genetic background on the evolution of phenotypic resistance, its genetic basis, and its...
Show moreAntibiotic resistance is a growing public-health concern. Efforts to control the emergence and spread of resistance would benefit from an improved ability to forecast when and how it will evolve. To predict the evolution of resistance with accuracy, we must understand and integrate information about many factors, including a bacterium's evolutionary history. This dissertation centers on the effects of genetic background on the evolution of phenotypic resistance, its genetic basis, and its fitness costs. To address these issues, I used Escherichia coli strains from the long-term evolution experiment (LTEE) that independently evolved for multiple decades in an environment without antibiotics.First, I examined how readily these LTEE strains could overcome prior losses of intrinsic resistance through subsequent evolution when challenged with antibiotics. Second, I investigated whether lineages founded from different genotypes take parallel or divergent mutational paths to achieve increased resistance. Third, I tested whether fitness costs of resistance mutations are constant across different genetic backgrounds. In these studies, I focused attention on the interplay between repeatability and contingency in the evolutionary process. My findings demonstrate that genetic background can influence both the phenotypic and genotypic evolution of resistance and its associated fitness costs. I conclude this dissertation with a broader discussion about these and other factors that can influence the evolution of antibiotic resistance, and their clinical and public-health implications.
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- Title
- Physiologic and pathologic changes in calves given Escherichia colia endotoxin or Pasteurella multocida
- Creator
- Musa, Babiker El Hag
- Date
- 1971
- Collection
- Electronic Theses & Dissertations
- Title
- Coevolution of bacterial-phage interactions
- Creator
- Meyer, Justin R.
- Date
- 2012
- Collection
- Electronic Theses & Dissertations
- Description
-
Bacteria and their viruses, phage, are the most abundant and genetically diverse group of organisms on earth. Given their prevalence, it is no wonder that recent studies have found their interactions important for ecosystem function, as well as the health of humans. Unfortunately, because of technical challenges with studying microbes, some of the most basic questions on their interactions, such as who infects whom, and how their relationships evolved in the first place, remain unanswered....
Show moreBacteria and their viruses, phage, are the most abundant and genetically diverse group of organisms on earth. Given their prevalence, it is no wonder that recent studies have found their interactions important for ecosystem function, as well as the health of humans. Unfortunately, because of technical challenges with studying microbes, some of the most basic questions on their interactions, such as who infects whom, and how their relationships evolved in the first place, remain unanswered. Here I report six studies on bacterial-phage interactions, each focused on understanding their pattern and the underlying biophysical, ecological, and evolutionary processes that shape them. To do this, I tested a number of hypotheses using laboratory experiments and analyses of natural microbial diversity. First, I tested whetherEsherichia coli cultured without phage would counter-intuitively evolve new interactions with phage. Typically bacterial traits responsible for phage resistance have pleiotropic consequences on growth, therefore as a side-effect of adapting to an abiotic environment, bacteria may also evolve to become more or less vulnerable to their parasites. After 45,000 generations of laboratory culturing without phage,E. coli gained resistance to lambda phage, gained sensitivity to a mutant T6 phage, and remained resistant to wild type T6. Each response was explained by understanding the pleiotropic costs or benefits of mutations that confer resistance. Because of pleiotropy, interactions may even evolve in the absence of one player.For the rest of my studies I examined how interactions evolve when host and parasite co-occur. First, I found that whenE. coli and phage lambda are cocultured,E. coli evolves resistance by reducing the number of phage genotypes that can infect it, whereas, lambda evolves to increase the number of bacterial genotypes it can infect. This antagonism produces a interaction matrix with a nested form where less derived host-ranges fall one within another. To determine whether this nested pattern is an artifact of the labratory environment, or if the pattern is general to natural communities, I performed a metaanalysis on already published phage-bacterial interaction matrices. The majority of networks were significantly nested (28 of 38). Lastly, I examined the molecular basis ofE. coli resistance to lambda and found that resistance often evolves through mutations inE. coli 'slamB , the gene for the phage receptor. Also, the strength of resistance is correlated with how the mutation perturbs the orientation specific features of the protein structure, primarily loop four which extends out of the cell membrane. For the final two chapters, I studied whether lambda could evolve to target a novel receptor and the evolutionary consequences of such an innovation. Under particular laboratory conditions,E. coli evolves resistance by down-regulating LamB, which sets the stage for lambda to evolve the necessary mutations to exploit a new protein receptor. When allowed to coevolve under this condition, lambda evolved to exploit another outer-membrane protein, OmpF. This new function is the result of a particular combination of four mutations in J, the gene for the protein ligand lambda uses to bind to its host. Once lambda evolves this novel interaction, an evolutionary arms-race begins that drives rapid diversification of the bacteria and phage. Overall, my studies show that coevolution between bacteria and phage, whether it be in the lab or in nature, produces nested interactions matrices. Secondly, antagonistic coevolution is a creative process able to generate new genotypes of host and parasite and promote the evolution of novel function. Lastly, costs for resistance have many important effects, from determining whether resistance will evolve or be lost, to the generation of diversity.
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- Title
- Comparison of Escherichia coli and Streptococcus faecalis as a test organism to determine the sanitary quality of food
- Creator
- Allen, Clarence Henry
- Date
- 1951
- Collection
- Electronic Theses & Dissertations
- Title
- Piliated Escherichia coli and porcine enteric disease in Michigan
- Creator
- Evans, Mark G.
- Date
- 1983
- Collection
- Electronic Theses & Dissertations
- Title
- Bioavailability of tetracycline in water and soil to Escherichia coli for expression of antibiotic resistance
- Creator
- Zhang, Yingjie
- Date
- 2013
- Collection
- Electronic Theses & Dissertations
- Description
-
Tetracyclines are a class of antimicrobials extensively used as human and veterinary medicine, and in livestock production since they were discovered in the 1940s. A large portion of tetracyclines administered to humans and animals are excreted and subsequently released into the environment, where they pose potential risks to ecosystem and human health. There is a growing concern that the presence of antibiotics such as tetracycline at trace levels in the environment is related to the...
Show moreTetracyclines are a class of antimicrobials extensively used as human and veterinary medicine, and in livestock production since they were discovered in the 1940s. A large portion of tetracyclines administered to humans and animals are excreted and subsequently released into the environment, where they pose potential risks to ecosystem and human health. There is a growing concern that the presence of antibiotics such as tetracycline at trace levels in the environment is related to the emergence and ever-increasing abundance of antibiotic resistance genes in natural and engineered microbial populations. However, basic knowledge at the molecular scale of bacterial access to tetracyclines present in environmental matrices and expression of antibiotic resistance genes remain nearly unknown. In this study, we used the E. coli MC4100/pTGM whole-cell bioreporter as an effective tool to investigate bioavailability of tetracycline in water and soil to bacteria for expression of antibiotic resistance genes. Our hypothesis was that the speciation of tetracycline dissolved in water and sorption by soil minerals controls the bioavailabilities for bacterial uptake and subsequent activation of antibiotic resistance genes. The results revealed that activation of antibiotic resistance in the E. coli bioreporter responded linearly to intracellular tetracycline concentration. The extent of tetracycline uptake by E. coli was modulated by tetracycline speciation. We have identified that zwitterionic tetracycline as the primary species favorable for bacterial uptake. Geochemical factors such as pH, salt composition and concentration influenced the fractional distributions of tetracycline species in aqueous solution and hence altered uptake by E. coli. In addition, the presence of organic ligands could also alter tetracycline speciation by releasing tetracycline from its metal complexes in aqueous solution. For tetracycline associated with Mg-smectite, desorption of tetracycline from clay to solution was the major exposure pathway for bacterial uptake and subsequent activation of antibiotic resistance in the diluted clay suspensions. In clay film cultivation, clay-sorbed tetracycline was still bioaccessible to E. coli evoking strong expression of antibiotic resistance. Direct contact of the E. coli bioreporters with clay surfaces and further formation of biofilms plausibly facilitated tetracycline transfer to bacteria. Overall, this study greatly advances the fundamental understanding of bioavailability of tetracycline in the environment to bacteria for expression of antibiotic resistance genes.
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- Title
- Development of a nanoparticle DNA based biosensor for the Shiga-like toxin 1 detection using co-polymerization hybridization readout amplification
- Creator
- Anderson, Michael Jeffrey
- Date
- 2012
- Collection
- Electronic Theses & Dissertations
- Description
-
Approximately 3.4 million deaths occur each year due to lack of clean water. The Shigatoxigenic group of Escherichia coli (STEC) bacteria causes approximately 2.2 million of these deaths and nearly a billion cases of illness. Current approved testing methods for Escherichia coli require a 12-24 hours growth of the sample in order to test for an indicator species. This indicator species only suggests possible E. coli O157:H7 presence, but does not confirm it. Direct culture testing cannot...
Show moreApproximately 3.4 million deaths occur each year due to lack of clean water. The Shigatoxigenic group of Escherichia coli (STEC) bacteria causes approximately 2.2 million of these deaths and nearly a billion cases of illness. Current approved testing methods for Escherichia coli require a 12-24 hours growth of the sample in order to test for an indicator species. This indicator species only suggests possible E. coli O157:H7 presence, but does not confirm it. Direct culture testing cannot identify the pathogenic STEC bacteria either. Confirmation of toxicity requires molecular methods of identification leading to specialized equipment needs and higher testing costs. Typical molecular methods use polymerase chain reaction (PCR) to amplify a very specific target DNA sequence, indicating the presence of a specific microorganism. PCR requires a thermocycler and a clean laboratory environment. Recent work has been successfully explored using nanomaterial based systems to identify the same genes used in a PCR reaction in a simpler detection system. The nanomaterials are used for both signal amplification and as the detection molecule. In this dissertation work a biosensor utilizing nanoparticles has been developed for the detection of Shiga-like toxin 1 gene that is present in Escherichia coli O157:H7. The system has been successfully constructed using a novel carbohydrate coated gold nanoparticle for delivery of the self assembling co-polymerization DNA oligonucleotide reporters. A set of self-assembled single-stranded DNA (ssDNA) molecules has been developed to amplify a target sequence without the use of enzymes. The full detection system incorporates the gold nanoparticle and a magnetic microparticle into a DNA recognition step. The gold nanoparticle is used for both co-polymerization detection and electrochemical detection. Input material (genomic DNA) is extracted using a modified commercial extraction method to produce PCR quality DNA from whole bacterial cells. When extraction and recognition elements are combined, a limit of detection for E. coli O157:H7 of 105 cells/mL using co-polymerization and 101 cells/mL with electrochemical detection has been achieved. Sample preparation from spiked culture samples until final detection takes a total of 7 hours. Electrochemical detection provides only a presence or absence indicator, where tethered co-polymerization is able to provide quantitative values of the input bacterial concentration. So a combination of co-polymerization amplification and electrochemical detection can provide the potential for more sensitive and quantitative measurement without the need for enzymes as in PCR applications.
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- Title
- Enzymatic characteristics of a-galactosidases from Aspergillus niger, coffee beans and Escherichia coli
- Creator
- Gavriel, Penelope
- Date
- 1984
- Collection
- Electronic Theses & Dissertations
- Title
- Breaking biofilms : regulation of Type II secretion system in V. cholerae and the formation of the hyper-pseudopilus
- Creator
- Sloup, Rudolph E.
- Date
- 2016
- Collection
- Electronic Theses & Dissertations
- Description
-
Vibrio cholerae is the causative agent of the human disease cholera, it resides in aquatic resevoirs and forms biofilms, which are closely associated communities of bacteria embedded in polysaccharides, DNA, and proteins. In V. cholerae biofilm formation is regulated by the second messenger molecule cyclic di-GMP (c-di-GMP). A genetic screen for promoters regulated by the c-di-GMP revealed a novel promoter (PepsG) in the eps operon encoding the V. cholerae Type 2 secretion system (T2SS). The...
Show moreVibrio cholerae is the causative agent of the human disease cholera, it resides in aquatic resevoirs and forms biofilms, which are closely associated communities of bacteria embedded in polysaccharides, DNA, and proteins. In V. cholerae biofilm formation is regulated by the second messenger molecule cyclic di-GMP (c-di-GMP). A genetic screen for promoters regulated by the c-di-GMP revealed a novel promoter (PepsG) in the eps operon encoding the V. cholerae Type 2 secretion system (T2SS). The T2SS, which exports proteins from the periplasm to the extracellular space, is phylogenetically related to Type 4 pili. The major pseudopilin is encoded by epsG which forms a short piston like structure necessary for secretion. I hypothesized that differential regulation of the eps operon extends the pseudopilin forming a structure called a hyper-pseudopilus outside the cell where it promotes biofilm development. In Chapter 2, I determined that the promoter upstream of the operon (PepsC1) is induced four fold by c-di-GMP and this induction is mediated by the c-di-GMP binding transcription factor VpsR directly. High levels of c-di-GMP were found to decrease the activity of extra cellular proteases secreted by the T2SS, however this effect was not a direct result of regulation of the T2SS as determined by mutation of the VpsR binding site in PepsC1. I was unable to establish a phenotype for the transcriptional control of the eps operon. This work establishes T2S as a new phenotype which is transcriptionally controlled by c-di-GMP and the biofilm associated transcription factor VpsR. In Chapter 3, I show that overexpression of epsG in a continuous flow cell system increased V. cholerae biofilms while a ΔepsG strain showed no biofilm formation. However, there was no change in activity of T2S dependent serine proteases while epsG was over expressed indicating increased biofilms is not likely due to increased secretion. Polyclonal antibody stained EpsG was also detectable on the surface of WT cells and long pseudopili were visualized with over expression of epsG. This evidence suggests the T2SS forms a hyper-pseudopilus important for biofilm formation. In Chapter 4, I present my work identifying novel anti-biofilm compounds. In 2011 Escherichia coli O104:H4 caused the deadliest E. coli outbreak in modern times resulting in 54 deaths and the highest rate of hemolytic uremic syndrome ever recorded. Subsequently, we showed a correlation between biofilm gene expression and virulence factor expression. I sought to identify small molecule compounds effective at inhibiting O104:H4 biofilms. I discovered at a concentration of 0.01% the nonionic surfactants polysorbate 80 (PS80) and polysorbate 20 (PS20) were found to inhibit biofilm formation by 90% and 91% respectively. These compounds were able to disperse preformed biofilms. Treatment of mice infected with E. coli O104:H4 resulted in high bacterial loads and inflammation. While addition of PS80 in the drinking water of the mice did not reduce bacterial loads, it completely abolished inflammation symptoms. PS80 is an FDA approved compound, well studied and effective at low nanomolar concentrations that reduces symptoms of infection in mice. which establishes it as an excellent candidate for further study as an anti-infective agent with anti-biofilm capabilities
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- Title
- Cell division in cis-platinum(II)diamminodichloride-induced filaments of Escherichia coli : dependence upon completion of deoxyribonucleic acid repair and a protease activity
- Creator
- Markham, Bruce Edward
- Date
- 1980
- Collection
- Electronic Theses & Dissertations