UMD Theses and Dissertations
Permanent URI for this collectionhttp://hdl.handle.net/1903/3
New submissions to the thesis/dissertation collections are added automatically as they are received from the Graduate School. Currently, the Graduate School deposits all theses and dissertations from a given semester after the official graduation date. This means that there may be up to a 4 month delay in the appearance of a given thesis/dissertation in DRUM.
More information is available at Theses and Dissertations at University of Maryland Libraries.
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Item AN INVESTIGATION ON A BACTERIOPHAGE ENDOLYSIN POSSESSING ANTIMICROBIAL ACTIVITY AGAINST ANTIBIOTIC-RESISTANT STAPHYLOCOCCUS AUREUS(2016) Linden, Sara Beth; Nelson, Daniel C; Molecular and Cell Biology; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Staphylococcus aureus is one of the most common causes of nosocomial (i.e. hospital-acquired) infection. Significantly, over 90% of S. aureus strains are resistant to penicillin, and since the mid-1980’s, methicillin-resistant S. aureus (MRSA) strains have become prevalent in hospitals worldwide, with resistance rates approaching 70%. In the U.S. alone, MRSA is responsible for over 100,000 invasive life threatening infections, such as necrotizing fasciitis, and causes 20,000 deaths annually. More worrisome, a variant known as community-acquired MRSA (CA-MRSA) is spreading in schools, gymnasiums, and even professional sports teams, where it infects otherwise healthy adolescents and young adults. Vancomycin is often considered the last antibiotic of choice against MRSA and other Gram-positive pathogens. However, rates of vancomycin-resistant enterococci (VRE) have already reached 30% and it is widely believed that emergence of vancomycin-resistant S. aureus (VRSA) is due to gene transfer during co-colonization of MRSA and VRE. Thus, alternative antimicrobial approaches are desperately needed. Endolysins, or peptidoglycan hydrolases, are phage-derived enzymes that actively lyse bacterial cells upon direct contact and may be considered such an alternative option. Moreover, the inability of bacteria to evolve resistance to endolysins is due to the specificity of the N-terminal catalytic domain, which cleaves a conserved peptidoglycan bond, and the C-terminal cell wall binding domain, which binds a cell surface moiety. This thesis represents an investigation into the endolysin PlyGRCS, which displays potent bacteriolytic activity against all antibiotic-resistant strains of S. aureus tested. This enzyme is active in physiologically relevant conditions (pH, NaCl, temperature), and its activity is greatly enhanced in the presence of calcium. PlyGRCS is the first endolysin with a single catalytic domain that cleaves two distinct sites in the peptidoglycan. Unlike antibiotics, PlyGRCS displays anti-biofilm activity, preventing, removing, and killing biofilms grown on abiotic and biotic surfaces. Engineering efforts were made to create an enzyme with a variable binding domain, which unfortunately displayed less activity than the wild type endolysin in the conditions tested. The antimicrobial efficacy of PlyGRCS was validated in a mouse model of S. aureus septicemia. The results from this study indicate that the endolysin PlyGRCS is a revolutionary therapeutic that should be further pursued for subsequent translational development.Item Low-Cost Paper-Based Assays for Multiplexed Genetic Analysis using Surface Enhanced Raman Spectroscopy(2013) Hoppmann, Eric Peter; White, Ian M; Bioengineering; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)In order to improve human health it is critical to develop low-cost sensors for chemical detection and healthcare applications. Low-cost chemical detectors can enable pervasive monitoring to identify health threats. Rapid yet accessible infectious disease diagnostics have the potential to improve patient quality of care, reduce healthcare costs and speed recovery. In both cases, when multiple targets can be detected with a single test (multiplexing), accessibility is improved through lowered costs and simplicity of operation. In this work we have investigated the practical considerations and applications of ink-jet printed paper surface enhanced Raman spectroscopy (SERS) devices. SERS enables specific simultaneous detection of numerous analytes using a single excitation source and detector. Sensitive detection is demonstrated in several real-world applications. We use a low-cost portable spectrometer for detection, further emphasizing the potential for on-site detection. These ink-jet printed devices are then used to develop a novel DNA detection assay, in which the multiplexing capabilities of SERS are combined with DNA amplification through polymerase chain reaction (PCR). In this assay, the chromatographic properties of paper are leveraged to perform discrimination within the substrate itself. As a test case, this assay is then used to perform duplex detection of the Methicillin-resistant Staphylococcus aureus (MRSA) genes mecA and femB, two genes which confer antibiotic resistance on MRSA. Finally, we explore statistical multiplexing methods to enable this assay to be applied to perform highly-multiplexed detection gene targets (5+), and demonstrate the differentiation of these samples using partial least-squares regression (PLS). By averaging the signal over a region of the SERS substrate, substrate variability was mitigated allowing effective identification and differentiation, even for the complex spectra from highly multiplexed samples which were impossible to visually analyze.Item Isolation and characterization of antimicrobial resistant Staphylococcus aureus in retail ground meats(2010) Li, Yi; Meng, Jianghong; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Staphylococcus aureus is commonly present in humans and animals. It can cause a variety of suppurative infections, food intoxication and toxic shock syndrome. Antimicrobial resistant S. aureus, especially methicillin-resistant S. aureus (MRSA), have emerged and are a major public health concern. There is an increasing risk of food production animals serving as a reservoir and transmitting S. aureus and MRSA in community environments. Due to the increased food safety risk posed by MRSA in addition to its multidrug resistance, we were interested in determining the prevalence of S. aureus and MRSA in retail meat and investing the multidrug resistance of the S. aureus isolates. A survey study was conducted, involving 480 retail ground meat samples (231 ground pork and 249 ground beef) collected in the Washington DC area from March 2009 to March 2010. Approximately 42.08% (n = 202) of the samples were identified as S. aureus positive and one MRSA isolate was recovered from a ground beef sample. Antimicrobial resistance testing showed 53.34% of recovered S. aureus isolates exhibited different levels of antimicrobial resistance to CLI, CHL, GEN, LEVO, CIP, SYN and TGC. The MRSA isolate was resistant to 8 of 22 antimicrobials tested. PFGE fingerprinting identified the MRSA isolate as USA300 subtype, which also carried genes of virulence factors PVL and protein A. Our findings indicated that antimicrobial resistant S. aureus strains were common in retail ground beef and port, and that MRSA could also be present in such products that could potentially serve as a reservoir.