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Infection and Immunity, December 2005, p. 8194-8203, Vol. 73, No. 12
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.12.8194-8203.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Novel Attenuated Salmonella enterica Serovar Choleraesuis Strains as Live Vaccine Candidates Generated by Signature-Tagged Mutagenesis

Yu-We Ku,1,2 Sean P. McDonough,1 Raghavan U. M. Palaniappan,1 Chao-Fu Chang,1,2* and Yung-Fu Chang1*

College of Veterinary Medicine, Cornell University, Ithaca, New York 14853,1 Institute of Medical Biotechnology, Central Taiwan University of Science and Technology, Taichung 40601, Taiwan2

Received 2 May 2005/ Returned for modification 10 June 2005/ Accepted 1 September 2005

Salmonella enterica serovar Choleraesuis is a host-adapted pathogen that causes swine paratyphoid. Signature-tagged mutagenesis (STM) was used to understand the pathogenicity of S. enterica serovar Choleraesuis in its natural host and also to develop novel attenuated live vaccine candidates against this disease. A library of 960 signature-tagged mutants of S. enterica serovar Choleraesuis was constructed and screened for attenuation in pigs. Thirty-three mutants were identified by the STM screening, and these mutants were further screened for attenuation by in vivo and in vitro competitive growth. Of these, 20 mutants targeting the outer membrane, type III secretion, transporter, lipopolysaccharide biosynthesis, and other unknown proteins were confirmed for attenuation. Five highly attenuated mutants (SC2D2 [ssaV], SC4A9 [gifsy-1], SC6F9 [dgoT], SC12B12 [ssaJ], and SC10B1[spiA]) were selected and evaluated for safety and protective efficacy in pigs by comparison with a commercially available vaccine strain. STM-attenuated live vaccine strains SC4A9 (gifsy-1) and SC2D2 (ssaV) were superior to commercially available live vaccine because they provided both safety and a protective immune response against challenge in pigs.

* Corresponding author. Mailing address for Chao-Fu Chang: 9206 Health Building, Institute of Biotechnology, Central Taiwan University of Science and Technology, Beitan District, Taichung City 40601, Taiwan. Phone: 886-22-363-2333. Fax: 886-22-363-2436. E-mail: cfchang{at}ntu.edu.tw. Mailing address for Yung-Fu Chang: Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, 14853. Phone: (607) 253-3675. Fax: (607) 253-3943. E mail: yc42@cornell.edu.

Editor: A. D. O'Brien

Infection and Immunity, December 2005, p. 8194-8203, Vol. 73, No. 12
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.12.8194-8203.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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