Characterization of the intracellular Campylobacter-like organism of porcine proliferative enteritis
Abstract
The first objective of this research was to localize the intracellular Campylobacter-like organism (CLO) of porcine proliferative enteritis (PPE) from fresh PPE(+) intestines using differential centrifugation (DC). Conventionally reared pigs affected with the intestinal adenomatosis form of PPE were euthanitized and necropsied; sections of intestines with typical PPE lesions were flushed with physiologic saline to remove surface debris. Surface epithelial cells were harvested by scraping the cells free from the underlying lamina propria. The cells were washed and lysed. Subcellular organelles and the CLO were separated by DC. The second objective was to purify the CLO from the sedimentation fraction obtained by DC by immunoaffinity chromatography and flow cytometry. Flow cytometric separation was attempted by specifically labelling the CLO with a fluorescent dye to allow separation from other bacteria and subcellular components within the sedimentation fraction. Immunoaffinity chromatography was performed by coupling of CLO antibody to a gel bed. A suspension of the sedimentation fraction was applied to the column. The antigen-antibody bonds were broken by applying an eluant to collect purified CLO. The third objective was to characterize the CLO by bacteriologic procedures, gas chromatography and transmission electron microscopy. The DC resulted in concentration of the CLO in the mitochondrial fraction (MF). Purification techniques failed to yield purified suspensions of CLO. Characterization studies revealed that the CLO would not grow using routine bacteriologic campylobacter isolation techniques. However, a coryneform, Oerskovia xanthineolytica groups A-4, was isolated from a microsomal fraction. Antibodies against the coryneform failed to react with the CLO. Gas chromatographic fatty acid analyses on known Campylobacter spp., the coryneform, CLO (+) MF's and normal MF's also failed to identify the CLO. Ultrastructural studies revealed that some morphological characteristics of the CLO were different than those of other Campylobacter spp, isolated from swine.
Degree
Ph.D.
Advisors
Thacker, Purdue University.
Subject Area
Veterinary services|Microbiology
Off-Campus Purdue Users:
To access this dissertation, please log in to our
proxy server.