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Infection and Immunity, August 2007, p. 4050-4061, Vol. 75, No. 8
0019-9567/07/$08.00+0     doi:10.1128/IAI.00486-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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

Paola Caro-Hernández,1 Luis Fernández-Lago,1 María-Jesús de Miguel,2 Ana I. Martín-Martín,1 Axel Cloeckaert,3 María-Jesús Grilló,2,4 and Nieves Vizcaíno1*

Departamento de Microbiología y Genética, Edificio Departamental, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain,1 Centro de Investigación y Tecnología Agroalimentaria del Gobierno de Aragón, Unidad de Sanidad Animal, Carretera de Montañana 930, 50059 Zaragoza, Spain,2 INRA, UR1282, Infectiologie Animale et Santé Publique, IASP, Nouzilly F-37380, France,3 Instituto de Agrobiotecnología y Recursos Naturales, CSIC-UPNA, Ctra. Mutilva Baja, 31192 Pamplona, Spain4

Received 4 April 2007/ Returned for modification 10 May 2007/ Accepted 30 May 2007

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 genes 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) autoagglutination of bacterial suspensions, and (iii) assessment of susceptibility to polymyxin B, sodium deoxycholate, hydrogen peroxide, and nonimmune 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 between the OMs of B. ovis PA and B. abortus RB51 rough strains were observed. 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 the {Delta}omp22 mutant to nonimmune 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 {Delta}omp25d mutant were found, especially considering that the fully virulent {Delta}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 both of B. ovis in rams and of other Brucella species, should be thoroughly evaluated in future studies.

* Corresponding author. Mailing address: Departamento de Microbiología y Genética, Edificio Departamental, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain. Phone: 34-923-294 532. Fax: 34-923-224 876. E-mail: vizcaino{at}usal.es

{triangledown} Published ahead of print on 11 June 2007.

Editor: D. L. Burns

Infection and Immunity, August 2007, p. 4050-4061, Vol. 75, No. 8
0019-9567/07/$08.00+0     doi:10.1128/IAI.00486-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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