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Yi Han thesis 2020.pdf (2.35 MB)

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Development and Evaluation of Mucoadhesive Chitosan Nanoparticle-based Salmonella Vaccine for Oral Delivery in Broiler Birds

Han, Yi
http://rave.ohiolink.edu/etdc/view?acc_num=osu1587571015936815

Abstract Details

2020, Doctor of Philosophy, Ohio State University, Animal Sciences.
Salmonellosis causes around one million illness every year in the Unites States. The symptoms of the disease in human include diarrhea, fever and vomiting. Animal-origin foods especially Salmonella Enteritidis contaminated poultry-related produce, remain the most common source of Salmonella outbreaks, which is responsible for half of the total cases of food poisoning. With the huge consumer demand of chicken meat in the U.S., reducing Salmonella colonization in broilers can largely decrease the risk of foodborne salmonellosis. Current Salmonella vaccines in broiler have failed to provide protection and reduce food contamination. Therefore, our goal is to develop an innovative Salmonella oral vaccine which elicits robust immunity and reduces bacterial colonization and shedding in broilers. Our preliminary data showed gastric pH resistant biodegradable and mucoadhesive chitosan (CS) nanoparticles based vaccine containing entrapped immunogenic outer membrane proteins (OMP) and flagellin (FLA) of Salmonella Enteritidis (SE) with surface labeled flagella [CS(OMP+FLA)-F] was found targeted to intestinal immune sites and induced the expression of important Toll-like receptors in cecal tonsils of birds. In this study, we first prepared CS(OMP+FLA)-F and the temporal dynamic immune responses of the immunized broilers were examined after 1st dose to 3-day-old broilers and 2nd dose inoculated 2 weeks later. We observed increased mRNA expression level of Toll-like receptors (TLRs) 1, 4, 7 and 7 in the cecal tonsils of birds received CS(OMP+FLA)-F, especially at 7 days post 1st dose of vaccine delivery. The antigen specific splenocytes proliferation response was enhanced after 7 days post 1st and 2nd dose of vaccine. This study showed our candidate vaccine can induce both innate and adaptive immune responses. To determine the efficacy of candidate vaccine and investigate the ideal vaccination schedule for maximum protection, dose- and age-dependent study was carried out followed by SE challenge infection in broilers. Birds received vaccine at either 3-day or 3-week of age with one booster or not. Each dose of vaccine contained 10 or 50 μg OMP+FLA antigens in chitosan NPs. Our data showed that birds received 2 doses of the vaccine at 3-day and 3-week with 10 μg/dose or at 3-week and 4-week age with 50 μg/dose had the lowest challenge SE load in the cecal content by 0.6 log10 reduction. In those immunized birds, augmented antigen-specific antibody response, splenocytes proliferation and an increased frequency of IFNγ-producing T cells were observed. In addition, enhanced mRNA expression levels of TLR 2 and 4 and cytokines IL-4 and IL-10 were detected. We conclude from this study that 2 doses of the candidate vaccine has the capacity of inducing protective immune response against SE colonization and early vaccination is preferred. The third study was conducted with modified vaccine formulation by surface conjugation of nanoparticle with mannose for targeting dendritic cells. A commercial live Salmonella vaccine was included for vaccine efficacy comparison. Broilers were immunized orally at 3-day and boosted at 3-week of age, challenged with SE at 5-week of age and sacrificed 10 days post-challenge. Each dose of vaccine contained 10 or 50 μg OMP+FLA antigens, nanoparticle surface conjugated with FLA, mannose or both [CS(OMP+FLA)-F/M/F&M]. Birds received 10 μg/dose vaccine had a maximum reduction in SE load in the cecal content compared to birds received 50 μg/dose. CS(OMP+FLA)-F&M vaccination resulted in the highest SE reduction by over 1 log10 CFU in each gram of the cecal content, which was comparable to commercial live vaccine. Immunologically, it was associated with increased specific splenocyte proliferation response and recall memory T lymphocytes frequency, as well as specific IgG and IgA responses observed in 10 µg/dose CS(OMP+FLA) vaccinated birds. Enhanced expression of mRNA level of TLRs (TLR 1, 2, 3, 7) and cytokines (IL-1β, TNF-α, IL-10 and TGF-β1) were detected in CS(OMP+FLA) vaccinated birds’ cecal tonsils. Although the commercial vaccine elicited SE reduction effect, the related immune responses were not detected which needs further investigation. In conclusion, 10 μg/dose of antigens in mannose modified CS(OMP+FLA) vaccine administered twice orally to 3 day and 3 weeks old broilers provided protective immune response against SE colonization by inducing both innate and adaptive immune response in broilers.
Renukaradhya Gourapura (Advisor)
Ramesh Selvaraj (Advisor)
Michael Lilburn (Committee Member)
Sheila Jacobi (Committee Member)
Thaddeus Ezeji (Committee Member)
135 p.
Animal Sciences
Salmonella Enteritidis; Broiler; Chitosan nanoparticle vaccine; Antibody response; Innate immunity; Cell mediated immunity

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Citations

  • Han, Y. (2020). Development and Evaluation of Mucoadhesive Chitosan Nanoparticle-based Salmonella Vaccine for Oral Delivery in Broiler Birds [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1587571015936815

    APA Style (7th edition)

  • Han, Yi. Development and Evaluation of Mucoadhesive Chitosan Nanoparticle-based Salmonella Vaccine for Oral Delivery in Broiler Birds. 2020. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1587571015936815.

    MLA Style (8th edition)

  • Han, Yi. "Development and Evaluation of Mucoadhesive Chitosan Nanoparticle-based Salmonella Vaccine for Oral Delivery in Broiler Birds." Doctoral dissertation, Ohio State University, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=osu1587571015936815

    Chicago Manual of Style (17th edition)

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This open access ETD is published by The Ohio State University and OhioLINK.