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      • HARVEST
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      Porcine Milk Fat Globule Membrane Proteins that Bind to F4ac Fimbriae and prevent attachment of Escherichia coli to enterocytes

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      NOVAKOVIC-THESIS.pdf (2.064Mb)
      Date
      2012-05-24
      Author
      Novakovic, Predrag
      Type
      Thesis
      Degree Level
      Masters
      Metadata
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      Abstract
      Post-weaning diarrhea (PWD) in pigs is caused by enterotoxigenic Escherichia coli (ETEC) which is often controlled by antibiotic feed supplements with the potential risk of generation of antibiotic resistant bacteria. Prevention of attachment of F4ac positive ETEC to intestinal mucosa may potentially be used as one of the antibiotic-free strategies for the control of this disease. F4ac-positive ETEC has been reported to bind to fat globule membranes in porcine milk. In this study, we used an affinity chromatography technique to identify individual milk fat globule membrane (MFGM) proteins that bind to F4ac fimbriae. An affinity column with covalently coupled F4ac fimbriae to activated Sepharose was created and the following F4ac-binding proteins were isolated from the porcine MFGM: lactadherin, butyrophilin, adipophilin, acyl-CoA synthetase 3, fatty acid binding protein 3, and xanthine dehydrogenase. Selected individual proteins of porcine MFGM, namely: xanthine dehydrogenase, butyrophilin, lactadherin, and fatty acid binding protein were isolated and tested for their inhibitory effects against attachment of F4ac positive ETEC and of F4ac fimbriae to either a small intestinal cell line (IPEC-J2) or primary porcine enterocytes by competitive ELISA. All of these proteins, except xanthine dehydrogenase, decreased attachment of F4ac positive E. coli and F4ac fimbriae to the intestinal cell line or primary enterocytes in a dose-dependent manner. Lactadherin demonstrated the strongest inhibitory effect on F4ac positive E. coli adherence to porcine enterocytes and the most potent interaction with F4ac fimbriae.
      Degree
      Master of Science (M.Sc.)
      Department
      Veterinary Pathology
      Program
      Veterinary Pathology
      Supervisor
      Simko, Elemir
      Committee
      Kidney, Beverly; Loewen, Matthew E.
      Copyright Date
      May 2012
      URI
      http://hdl.handle.net/10388/ETD-2012-05-448
      Subject
      Enterotoxigenic Escherichia coli (ETEC)
      Porcine milk fat globule membrane (MFGM) proteins
      F4ac-binding proteins
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