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Antimicrobial Agents and Chemotherapy, March 2007, p. 912-922, Vol. 51, No. 3
0066-4804/07/$08.00+0     doi:10.1128/AAC.00568-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The ß-Defensin Gallinacin-6 Is Expressed in the Chicken Digestive Tract and Has Antimicrobial Activity against Food-Borne Pathogens

Albert van Dijk,¹ Edwin J. A. Veldhuizen,¹ Stefanie I. C. Kalkhove,^1, Johanna L. M. Tjeerdsma-van Bokhoven,¹ Roland A. Romijn,² and Henk P. Haagsman^1*

Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, 3584 CM The Netherlands,¹ ABC Protein Expression Center, Utrecht University, 3584 CH Utrecht, The Netherlands²

Received 28 April 2006/ Returned for modification 10 August 2006/ Accepted 18 December 2006

Food-borne pathogens are responsible for most cases of food poisoning in developed countries and are often associated with poultry products, including chicken. Little is known about the role of ß-defensins in the chicken digestive tract and their efficacy. In this study, the expression of chicken ß-defensin gallinacin-6 (Gal-6) and its antimicrobial activity against food-borne pathogens were investigated. Reverse transcription-PCR analysis showed high expression of Gal-6 mRNA in the esophagus and crop, moderate expression in the glandular stomach, and low expression throughout the intestinal tract. Putative transcription factor binding sites for nuclear factor kappa beta, activator protein 1, and nuclear factor interleukin-6 were found in the Gal-6 gene upstream region, which suggests a possible inducible nature of the Gal-6 gene. In colony-counting assays, strong bactericidal and fungicidal activity was observed, including bactericidal activity against food-borne pathogens Campylobacter jejuni, Salmonella enterica serovar Typhimurium, Clostridium perfringens, and Escherichia coli. Treatment with 16 µg/ml synthetic Gal-6 resulted in a 3 log unit reduction in Clostridium perfringens survival within 60 min, indicating fast killing kinetics. Transmission electron microscopy examination of synthetic-Gal-6-treated Clostridium perfringens cells showed dose-dependent changes in morphology after 30 min, including intracellular granulation, cytoplasm retraction, irregular septum formation in dividing cells, and cell lysis. The high expression in the proximal digestive tract and broad antimicrobial activity suggest that chicken ß-defensin gallinacin-6 plays an important role in chicken innate host defense.

Published ahead of print on 28 December 2006.

Present address: PodiCeps B.V., Yalelaan 46, 3584 CM Utrecht, The Netherlands.

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