The objectives of this study were to determine whether feeding thermally processed peroxidized soybean oil (SO) induces markers of oxidative stress and alters antioxidant status in pig tissue, blood, and urine. Fifty-six barrows (25.3 ± 3.3 kg initial BW) were randomly assigned to dietary treatments containing 10% fresh SO (22.5 °C) or thermally processed SO (45 °C for 288 h, 90 °C for 72 h, or 180 °C for 6 h), each with constant air infusion rate of 15 liters/minute. Multiple indices of lipid peroxidation were measured in the SO including peroxide value (2.0, 96, 145, and 4.0 mEq/kg for 22.5, 45, 90, and 180 °C processed SO, respectively) and p-anisidine value (1.2, 8.4, 261, and 174 for 22.5, 45, 90, and 180 °C processed SO, respectively); along with a multitude of aldehydes. Pigs were individually housed and fed ad libitum for 49 d which included a 5 d period in metabolism crates for the collection of urine and serum for measures of oxidative stress. On day 49, pigs were euthanized to determine liver weight and analyze liver-based oxidative stress markers. Oxidative stress markers included serum, urinary, and liver thiobarbituric acid reactive substances (TBARS), and urinary F2-isoprostanes (ISP) as markers of lipid damage; serum and liver protein carbonyls (PC) as a marker of protein damage; and urinary and liver 8-hydroxy-2'-deoxyguanosine (8-OH-2dG) as a marker of DNA damage. Superoxide dismutase (SOD), and catalase activity (CAT) were measured in liver, glutathione peroxidase activity (GPx) was measured in serum and liver, and ferric reducing antioxidant power (FRAP) was measured in serum and urine as determinants of antioxidant status. Pigs fed 90 °C SO had greater urinary ISP (P = 0.02), while pigs fed the 45 °C SO had elevated urinary TBARS (P = 0.02) in comparison to other treatment groups. Pigs fed 45 °C and 90 °C SO had increased serum PC concentrations (P = 0.01) and pigs fed 90 °C SO had greater (P = 0.01) liver concentration of 8-OH-2dG compared to pigs fed the other SO treatments. Furthermore, pigs fed 90 °C SO had reduced serum GPx activity in comparison to pigs fed fresh SO (P = 0.01). In addition, pigs fed 180 °C SO had increased liver CAT activity (P = 0.01). Liver GPx and SOD or serum and urinary FRAP were not affected by dietary treatment. These results indicate that dietary peroxidized soybean oil induced oxidative stress by increasing serum PC while diminishing serum GPx, increasing urinary ISP and TBARS, and increasing 8-OH-2dG and CAT in liver.

While T cell contribution to IAV immunity is appreciated, data comparing methods to evaluate IFN-γ production by IAV-specific T cells elicited following vaccination is limited. To understand the differential immunogenicity between live-attenuated influenza virus (LAIV) and whole-inactivated virus (WIV) vaccines in relation to induction of peripheral T cell responses, ELISpot and ELISA were used to assess IFN-γ production by peripheral lymphocytes following antigen restimulation. Following restimulation, peripheral blood lymphocytes from WIV-vaccinated pigs had a greater quantity of IFN-γ secreting cells (SC) and IFN-γ secreted compared to LAIV vaccinated and non-vaccinated (NV) pigs. Pig age at time of WIV vaccination significantly impacted peripheral IAV-specific IFN-γ recall response, as did the inclusion of adjuvant in the WIV vaccine. Collectively, these data indicate that peripheral IAV-specific IFN-γ recall responses are not predictive of LAIV vaccination status, thus, are unlikely to be a useful surrogate for evaluating LAIV immunogenicity and predicting cross-protection. However, these data suggest that the evaluation of peripheral IFN-γ recall responses may be useful for identifying factors, such as animal age or vaccine formulation, that may impact parenterally-delivered WIV vaccine immunogenicity. Overall, results did not differ based upon the assay used to evaluate IFN-γ recall responses. Therefore, either ELISpot or ELISA could serve as a measure for evaluating IAV-specific IFN-γ cell-mediated immune responses in swine.

Vaccine associated enhanced respiratory disease (VAERD) has been described in pigs vaccinated with whole-inactivated influenza virus (WIV) following infection with heterologous influenza A virus (IAV). WIV vaccination elicits production of cross-reactive, non-neutralizing antibody to the challenge IAV strain suggesting circulating immune complexes and/or antibody dependent cell-mediated cytotoxicity could contribute to pathology. Moreover, pathological findings in VAERD lungs include significant lymphocytic infiltration and increased levels of proinflammatory cytokines. In order to further delineate the immune mechanism associated with VAERD versus partial-protection we defined the lymphocyte populations infiltrating the lungs following challenge, as well as level of complement activation. Post-challenge, IAV-specific IFNγ secreting cells in the lungs were increased in pigs with VAERD compared to non-vaccinated, challenged controls. CD8 T cells and NK cells were significantly higher in the lungs of pigs protected from homologous challenge, suggesting these cells may play a role in protection. Interestingly, pigs with VAERD had a significant increase in memory T cells that was not observed in pigs without VAERD. Additionally, complement activation was significantly higher in the lungs of pigs experiencing VAERD. These data further describe the lymphocytic pathology associated with VAERD and suggest a role for complement activation in the development of VAERD disease pathology.

This thesis is dedicated to the people who have changed my life in wondrous ways, without you, I would not be where I am today. To the memory of my grandfather, Gordon Olson, who not only taught me the value of education, but also taught me that it is okay to dance to the beat of my own drum. To the memory of a wonderful friend, Lois Lane, who was there for me when I needed someone the most; because of you I learned that my finest qualities are patience and perseverance. To my little rocks, Elyzah and Saphira, someday I hope you realize that the good times make the bad times better. Thank you for letting me read you journal articles in lieu of bed time stories, thank you for teaching me to make time for the simple things in life, and thank you for always believing in me when I doubted myself. To my doctors at the University of Iowa, Naurang Agrawal and Thomas Gellhaus, thank you for keeping me well so I could complete this journey. Lastly, I would like to thank those who told me to give up on my dreams, I am always up for a good challenge. iii