Senior Honors Projects, 2010-current

Date of Award

Spring 2014

Document Type

Thesis

Degree Name

Bachelor of Science (BS)

Department

Department of Integrated Science and Technology

Advisor(s)

Louise Temple

Robert McKown

Jonathan Monroe

Abstract

The Shenandoah Valley of Virginia is a major poultry growing region in the United States, leading to a strong interest in controlling poultry zoonotic pathogens in order to prevent economic losses and transmission to humans. Bordetella avium, a Gram negative bacterium, is the causative agent of the upper respiratory illness, bordetellosis, in turkeys. An existing bordetellosis vaccine decreases severity of the disease but does not limit infection or transmission. An improved vaccine for bordetellosis is needed for better efficacy. Our research involves developing B. avium as a universal poultry live vaccine platform that will protect against bordetellosis and also serve to express foreign antigens. B. avium is hypothesized to have the ability to stably express foreign antigens, such as those from important poultry pathogens as Campylobacter jejuni, Clostridium perfringens, and Hemorrhagic enteritis virus (HEV), and produce a strong immune response to multiple pathogens following intranasal vaccination. The gene encoding Baa1, an autotransporter involved in host colonization in B. avium but shown to be non-essential, was selected as the location for heterologous genes, to produce an attenuated B. avium strain and express the foreign antigens at the cell surface. The potential vaccine delivery plasmid design consisted of the plasmid pBBR1-MCS, a broad host range plasmid, into which the Baa1 autotransporter was inserted with the foreign antigen of interest replacing the passenger domain of the autotransporter gene. Antigens from C. jejuni, such as a flagellin protein and an amino acid binding protein, known to elicit immune responses in poultry have been cloned into the passenger domain of the autotransporter gene to form the plasmid construct in the E. coli plasmid pCR2.1. After moving the construct into pBBR1-MCS, the expression will be tested in B. avium. Success of these efforts will allow this vaccine platform to be used for the delivery of any number of foreign antigens and response to pathogen threats.

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