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Infect. Immun. doi:10.1128/IAI.00486-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Role of the Omp25/Omp31 Family in the Outer Membrane Properties and Virulence of Brucella ovis

Departamento de Microbiología y Genética, Edificio Departamental, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain, Centro de Investigación y Tecnología Agroalimentaria to del Gobierno de Aragón, Unidad de Sanidad Animal, Carretera de Montañana, 930, 50059 Zaragoza, Spain, INRA, UR1282, Infectiologie Animale et Santé Publique, IASP, Nouzilly, F-37380, France, and Instituto de Agrobiotecnología y Recursos Naturales, CSIC-UPNA, Ctra. Mutilva Baja, 31192, Pamplona, Spain

^* To whom correspondence should be addressed. Email: vizcaino{at}usal.es.

	Abstract

The genes coding for the five outer membrane proteins (OMPs) of the Omp25/Omp31 family expected to be located in the outer membrane (OM) of rough virulent Brucella ovis PA were inactivated to evaluate their role in virulence and OM properties. The OM properties of the mutant strains and of the mutants complemented with the corresponding wild-type gene were analyzed, in comparison with the parental strain and rough B. abortus RB51, in several tests: (i) binding of anti-Omp25 and anti-Omp31 monoclonal antibodies (ii) auto-agglutination of bacterial suspensions and (iii) susceptibility to polymyxin B, sodium deoxycholate, hydrogen peroxide, and non-immune ram serum. A tight balance of the members of the Omp25/Omp31 family was seen to be essential for the stability of the B. ovis OM and important differences in the OM were observed between B. ovis PA and B. abortus RB51 rough strains. Regarding virulence, the absence of Omp25d and Omp22 from the OM of B. ovis PA led to a drastic reduction in spleen colonization in mice. While the greater susceptibility of mutant omp22 to non-immune serum and its difficulty in surviving in the stationary phase might be on the basis of its dramatic attenuation, no defects in the OM able to explain the attenuation of the omp25d mutant were found, mainly considering that the fully virulent omp25c mutant displayed more important OM defects. Accordingly, Omp25d, and perhaps Omp22, could be directly involved in the penetration and/or survival of B. ovis inside host cells. This aspect, together with the role of Omp25d and Omp22 in the virulence of both B. ovis in rams and other Brucella species, should be thoroughly evaluated in future studies.

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