Humoral Immune Responses of White-Tailed Deer (Odocoileus virginianus) to Mycobacterium bovis BCG Vaccination and Experimental Challenge with M. bovis

Monitoring of the kinetics of production of serum antibodies to multiple mycobacterial antigens can be useful as a diagnostic tool for the detection of Mycobacterium bovis infection as well as for the characterization of disease progression and the efficacy of intervention strategies in several species. The humoral immune responses to multiple M. bovis antigens by white-tailed deer vaccinated with BCG orally via a lipid-formulated bait (n = 5), orally in liquid form (n = 5), and subcutaneously (n = 6) were evaluated over time after vaccination and after experimental challenge with virulent M. bovis and were compared to the responses by unvaccinated deer (n = 6). Antibody responses were evaluated by using a rapid test (RT), a multiantigen print immunoassay (MAPIA), a lipoarabinomannan enzyme-linked immunosorbent assay (LAM-ELISA), and immunoblotting to whole-cell sonicate and recombinant antigen MPB83. MAPIA and RT detected minimal to no antibody responses over those at the baseline to multiple M. bovis antigens in vaccinated white-tailed deer after challenge. This was in contrast to the presence of more readily detectable antibody responses in nonvaccinated deer with more advanced disease. The LAM-ELISA results indicated an overall decrease in the level of production of detectable antibodies against lipoarabinomannan-enriched mycobacterial antigen in vaccinated animals compared to that in nonvaccinated animals after challenge. Immunoblot data were inconsistent but did suggest the occurrence of unique antibody responses by certain vaccinated groups to Ag85 and HSP70. These findings support further research toward the improvement and potential use of antibody-based assays, such as MAPIA, RT, and LAM-ELISA, as tools for the antemortem assessment of disease progression in white-tailed deer in both experimental and field vaccine trials.

Essays on Industrial Organization and Environmental Economics

This dissertation studies environmental regulation issues in the hog production industry as well as forces behind the reorganization of the industry during the past two decades. Federal and State-level environmental regulations imposed on U.S. hog production during the year 2003 are examined in Chapter 1. Based on the number of regulations passed by the Federal government and states, the 2003 regulatory index is constructed. The regulatory stringency index suggests that state-level regulations vary across states and have increased over the years. In addition, state-level regulations are more stringent than federal regulations. Chapter 2 develops an empirically implementable theoretical model which allows us to investigate the long-run effects of environmental regulations on the U.S. hog industry. Hog feeding operations (HFOs) are divided into large feeding operations (LHFOs) and small feeding operations (SHFOs). The impact of the presence of a large number of LHFOs on the entry and exit of CHFOs is also examined. Results of this study suggest that: Increased state-level regulation stringency significantly lowers the output of SHFOs; increased state-level regulation stringency significantly lowers the output of LHFOs; increased state-level regulation stringency significantly lowers the number of SHFOs; SHFO output rises significantly in states that have a greater number of LHFOs; LHFO output rises significantly in states that have a greater number of LHFOs; the number of SHFOs significantly increases in states that have a greater numbers of LHFOs; regulation increases the average SHFO size; and regulation decreases the average LHFO size. Chapter 3 examines the importance of input availability, market attractiveness, agglomeration economies and environmental regulations on the reorganization of U.S. hog production for a panel of 22 U.S. hog producing states which include, Northern states, Southern states and Midwest states for the period 1994-2006. Results from this study suggest that: Hog production in a state is positively affected by hog production in a nearby state, confirming the presence of agglomeration economies; Environmental regulations and high corn price have negative effects on state-level U.S. hog production; High hog prices, and favorable labor cost, and land values attract hog production; and transportation cost has no effect on hog production. Advisors: Azzeddine Azzam and Karina Schoengold

Theoretical and Experimental Studies in Nuclear Magnetic Resonance

Nuclear magnetic resonance (NMR) is a tool used to probe the physical and chemical environments of specific atoms in molecules. This research explored small molecule analogues to biological materials to determine NMR parameters using ab initio computations, comparing the results with solid-state NMR measurements. Models, such as dimethyl phosphate (DMP) for oligonucleotides or CuCl for the active site of the protein azurin, represented computationally unwieldy macromolecules. 31P chemical shielding tensors were calculated for DMP as a function of torsion angles, as well as for the phosphate salts, ammonium dihydrogen phosphate (ADHP), diammonium hydrogen phosphate, and magnesium dihydrogen phosphate. The computational DMP work indicated a problem with the current standard 31P reference of 85% H3PO4(aq.). Comparison of the calculations and experimental spectra for the phosphate salts indicated ADHP might be a preferable alternative as a solid state NMR reference for 31P. Experimental work included magic angle spinning experiments on powder samples using the UNL chemistry department’s Bruker Avance 600 MHz NMR to collect data to determine chemical shielding anisotropies. For the quadrupolar nuclei of copper and scandium, the electric field gradient was calculated in diatomic univalent metal halides, allowing determination of the minimal level of theory necessary to compute NMR parameters for these nuclei.