Date of Award

11-2010

Degree Type

Dissertation

Degree Name

Doctor of Philosophy

Major

Animal Science

Major Professor

Jun Lin

Committee Members

Kelly R. Robbins; Arnold M. Saxton; Gina M. Pighetti; Chunlei Su

Abstract

Campylobacter is the major bacterial cause of human gastroenteritis in the United States and other developed countries. Poultry are considered a main source of human Campylobacter infections. Thus, reduction of Campylobacter load in poultry is significant in food safety and public health. However, no effective measure is commercially available to prevent Campylobacter colonization in poultry to date. Antimicrobial peptides (AMPs) are short and bactericidal peptides widely present in intestine to limit bacterial infections. Recently, AMPs have been increasingly recognized as a novel class of antibiotics (peptide antibiotics) to control foodborne pathogens. Notably, several potent anti-Campylobacter bacteriocins, a group of AMPs produced by commensal bacteria, dramatically reduced C. jejuni colonization in chickens and are being directed toward on-farm control of this pathogen to protect public health. As an important strategy to evade killing by potential peptide antibiotics and by host innate defense, AMP resistance mechanisms in C. jejuni are critical to understand, but are still unknown. In this dissertation, molecular basis of Campylobacter resistance to polymyxin B, the anti-Campylobacter bacteriocins (BCNs), and a chicken host defense AMP (fowlicidin-1) was comprehensively examined using both in vitro and in vivo systems. Although polymyxin B has been successfully used as a model peptide to study AMP resistance in other Gram-negative bacteria, functional genomics examination in this study suggested that polymyxin B is not a good surrogate to study Campylobacter resistance to physiologically relevant AMPs. Campylobacter only developed low-level BCN resistance with low frequency in vitro and in vivo; the acquired BCN resistance was not stable in Campylobacter. Genomic examination of two BCN resistant mutants using DNA microarray and random transposon mutagenesis revealed that the multidrug efflux pump CmeABC contributes to both intrinsic and acquired resistance of Campylobacter to the BCNs. Random transposon mutagenesis and targeted site-directed mutagenesis identified four genes (cbrR, tig, cjaB, and cj1583c) involved in Campylobacter resistance to fowlicidin-1. These genes were also required for optimal colonization of Campylobacter in chickens. Together, the findings from this dissertation revealed uniqueness and complexity of AMP resistance in Campylobacter and will enable us to develop more sustainable peptide antibiotics and novel intervention strategies to prevent and control Campylobacter infections in humans and animal reservoirs. Key words: Campylobacter, antimicrobial peptide resistance, polymyxin B, bacteriocins, fowlicidins

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