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 ANTAGONISTIC MECHANISM OF METABOLITES FROM LACTOBACILLUS CASEI AGAINST FOODBORNE ENTEROHEMORRHAGIC ESCHERICHIA COLI(2022) Aditya, Arpita; Biswas, Debabrata; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Enterohemorrhagic Escherichia coli O157: H7 (EHEC), a foodborne enteropathogen, remains a significant public health concern since its discovery in 1982. With an incredibly low infectious dose (10-100 bacteria), this pathogen can cause self-limiting diarrhea, vomiting, and abdominal cramps. However, more complicated disease conditions such as bloody diarrhea or hemolytic colitis have been known to develop depending on the serotype involved in the infection, and on immune status and/or age of the patients. Due to its Shiga toxin (Stx) production ability, EHEC infection may lead to a kidney-related problem known as hemolytic uremic syndrome (HUS), which requires advanced medical care. Unlike other bacterial illnesses, therapeutic administration of antibiotics to treat EHEC infections is not recommended due to their controversial association with Stx production. As a result, only preventative/prophylactic and immune-supportive strategies are followed for EHEC infections. Using the antibacterial properties of probiotic bacteria and the metabolites they produce are promising alternative strategies for preventing EHEC infections. We have targeted the probiotic bacteria Lactobacillus casei to determine the mechanism of this alternative strategy. In our study, we have executed microbiological, molecular, chromatographic, and metagenomic approaches to determine the antagonistic mechanisms of action of their metabolites, specifically conjugated linoleic acid (CLA) produced by Lactobacillus casei, against the growth and metabolism of EHEC. The metabolites of wild-type L. casei (LCwt) were augmented by supplementing it with a prebiotic-like dietary component, namely peanut flour (PF) (LCwt+PF), while another LCwt was also genetically engineered (LCCLA) to over convert CLA from linoleic acid (LA). These modifications showed effective results in controlling EHEC both in vitro and in ex vivo conditions. Total metabolites present in cell-free culture supernatant (CFCS) of LCwt, LCwt+PF, and LCCLA were able to control the growth of EHEC without negatively hampering the relative abundance of Firmicutes and Bacteroidetes present in rumen fluid (RF). Among these CFCSs, CFCSCLA exerted the most desirable outcome by eliminating EHEC. In vitro studies demonstrated that, a lower concentration of purified CLA worked synergistically with other metabolites of LCwt and augmented their inhibitory activity against EHEC. The orchestrated effect of metabolites has been observed to downregulate the virulence genes, disrupt the cell membrane, interfere with cell division, and damage their genomic DNA. The probable effect of these metabolites, specifically CLA, on Stx production and neutralization was also investigated by assessing host cell cytotoxicity. Total metabolites of Lactobacillus spp. as well as CLA itself, showed improvement in cell survivability when exposed to Stx. Our findings established a ground to explore the effect of specific metabolites obtained from probiotic bacteria in control and prevention of EHEC. The findings also showed a promising association of purified CLA in neutralizing Stx which can be further explored to use it in therapeutic purposes.Item Effect of Air on Rumen Gas Production(2021) Rha, Rachel Youngah; Kohn, Richard A; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Ruminants may swallow air as they eat and ruminate throughout the day. However, it is unclear as to how the introduction of oxygen impacts fermentation pathways, bacteria, and yeast within this mostly anaerobic environment. Therefore, the focus of this thesis was to study air’s impact on rumen fermentation and to determine if probiotics could offset air’s impact on digestibility. An in vitro analysis of air and probiotics indicated the main effect of air decreased digestibility, the main effect of probiotics had variable effects, and probiotics had significant interactions with air. The interactions suggested yeast employing a potential alternative pathway with the introduction of oxygen. Utilizing published literature, a static and dynamic mathematical model was built to further analyze digestibility, gas composition, and uptake of oxygen within the rumen. Future studies will further develop this model with in vivo studies to further interpretation and understanding of rumen fermentation’s complex system.Item The Effects of Tryptophan and Probiotic Treatment on Behavior and Production Parameters of Laying Hens(2021) Bonilla Carrero, Paola Ivette; Dennis, Rachel L; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Dietary supplementation can impact behavior expression through microorganism’s activity in the gut microbiome and influence productivity in animal husbandry. Adding supplements to production animals’ diet can impact behaviors and productivity via gut-brain axis activity. We investigated the effects of either probiotic or antibiotic supplementation in addition to the effects of additional tryptophan (Trp) on overall behavior and production parameters. White Leghorn chickens (n=12) were supplemented with six dietary treatments in a 2 x 3 factorial design: probiotics (1 x 109 CFU/L) or the antibiotic erythromycin (125 mg/L) in combination with either normal (0.16%) or high (0.48%) Trp in drinking water. Results indicate that probiotics with tryptophan increased locomotion (P = 0.04), social (P = 0.04), and eating behavior (P = 0.02). Antibiotics with tryptophan increased fat pad (P = 0.04) and heart weight (P = 0.04). Supplementation affected behavioral expression of normal, comfort, and pecking behavior, potentially impacted by metabolic competition at the level of the gut microbiome.Item GENETICALLY ENGINEERED PROBIOTICS FOR DIAGNOSTICS AND DRUG DELIVERY: APPLICATIONS FOR CROHN’S DISEASE(2018) McKay, Ryan; Bentley, William E; Bioengineering; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)In the history of medicine, therapies have evolved while their mode of delivery has remained largely static. Generally, the active ingredient is formulated with an excipient to confer stability, and is ultimately delivered orally or intravenously in most applications. Crohn’s disease (CD), an illness with increasing global prevalence characterized by chronic inflammation of the intestines, is commonly treated with intravenously administered biologics. When these medicines spread throughout the body, only a small percentage acts at the desired site and side effects often arise. Thus, a targeted system is desired to localize treatment at sites of colonic inflammation. There is an entire field dedicated to localized delivery that typically employs drug-laden particles or capsules that can respond to local chemical or physical cues. We believe that bacteria can be “programmed” to respond analogously, and ultimately synthesize and deliver therapeutics. Nitric oxide (NO) levels are elevated at sites of intestinal inflammation, and thus serves as a targeting molecule that can attract programmed bacteria via a process called pseudotaxis. This is achieved by rewiring the native motility circuits of bacteria to respond to high NO levels. Additionally, localized treatment is attained by an NO- specific response whereby the designed bacteria produce and secrete a human protein reported to reduce inflammation in CD patients. This system may improve CD treatment via: 1) site-specific targeting to minimize side effects and increase efficacy, 2) in situ synthesis of the therapeutic avoids payload loss in the digestive tract and manufacturing obstacles associated with biologics, 3) probiotics are reported to provide innate benefits to CD patients, and 4) oral delivery is preferred by patients versus intravenous. We have also developed probiotics that fluoresce in response to NO which may serve as an ingestible biosensor for CD. We believe these reporter probiotics can assist in the diagnosis of CD by utilizing visualization of bacteria in a stool sample to reduce the need for invasive colonoscopies and biopsies. Overall, we have developed a platform of probiotic cells that respond to NO with applications for Crohn’s disease in mind, translating to noninvasive methods for both the diagnosis and treatment of CD.