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Infection and Immunity, April 2001, p. 2137-2143, Vol. 69, No. 4
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.4.2137-2143.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Reduced Virulence of a Bordetella bronchiseptica Siderophore Mutant in Neonatal Swine

Karen B. Register,1,* Thomas F. Ducey,2 Susan L. Brockmeier,1 and David W. Dyer2

Respiratory Diseases of Livestock Research Unit, USDA Agricultural Research Service National Animal Disease Center, Ames, Iowa 50010,1 and Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 731902

Received 18 September 2000/Returned for modification 9 November 2000/Accepted 4 January 2001

One means by which Bordetella bronchiseptica scavenges iron is through production of the siderophore alcaligin. A nonrevertible alcaligin mutant derived from the virulent strain 4609, designated DBB25, was constructed by insertion of a kanamycin resistance gene into alcA, one of the genes essential for alcaligin biosynthesis. The virulence of the alcA mutant in colostrum-deprived, caesarean-delivered piglets was compared with that of the parent strain in two experiments. At 1 week of age, piglets were inoculated with phosphate-buffered saline, 4609, or DBB25. Two piglets in each group were euthanatized on day 10 postinfection. The remainder were euthanatized at 21 days postinfection. Clinical signs, including fever, coughing, and sneezing, were present in both groups. Nasal washes performed 7, 14, and 21 days postinoculation demonstrated that strain DBB25 colonized the nasal cavity but did so at levels that were significantly less than those achieved by strain 4609. Analysis of colonization based on the number of CFU per gram of tissue recovered from the turbinate, trachea, and lung also demonstrated significant differences between DBB25 and 4609, at both day 10 and day 21 postinfection. Mild to moderate turbinate atrophy was apparent in pigs inoculated with strain 4609, while turbinates of those infected with strain DBB25 developed no or mild atrophy. We conclude from these results that siderophore production by B. bronchiseptica is not essential for colonization of swine but is required for maximal virulence. B. bronchiseptica mutants with nonrevertible defects in genes required for alcaligin synthesis may be candidates for evaluation as attenuated, live vaccine strains in conventionally reared pigs.

* Corresponding author. Mailing address: Swine Respiratory Diseases Project, USDA/ARS/National Animal Disease Center, P.O. Box 70, 2300 Dayton Rd., Ames, IA 50010. Phone: (515) 663-7700. Fax: (515) 239-8458. E-mail: kregiste{at}nadc.ars.usda.gov.

Infection and Immunity, April 2001, p. 2137-2143, Vol. 69, No. 4
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.4.2137-2143.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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