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De Miguel Mj - Autor o Coautor
Muñoz Pm - Autor o Coautor
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Publicaciones > Artículo

The CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases.

Publicado en:Veterinary Research. 49 (1): 85-85 - 2018-09-05 49(1), doi: 10.1186/s13567-018-0583-1

De Miguel Mj; Muñoz Pm;

Afiliaciones

ACUNSA - Financiador
Aragon Government (Grupo de investigacion en desarrollo) [A13-17D] - Financiador
Artai - Financiador
Centro de Investigación y Tecnología Agroalimentaria de Aragón - Entidad de origen
Department of Biology, University of Medea, 26000, Medea, Algeria. - Autor o Coautor
Fundaciones Caja Navarra and Roviralta - Financiador
Inst Agroalimentario Aragon IA2, Ctr Invest & Tecnol Agroalimentaria Aragon CITA, Unidad Prod & Sanidad Anim, Zaragoza, Spain - Autor o Coautor
Institute for Tropical Health - Financiador
Instituto de Salud Tropical y Departamento de Microbiología y Parasitología-IDISNA, Universidad de Navarra, 31008, Pamplona, Spain. - Autor o Coautor
Instituto de Salud Tropical y Departamento de Microbiología y Parasitología-IDISNA, Universidad de Navarra, 31008, Pamplona, Spain. azuniga@unav.es. - Autor o Coautor
MINECOSpanish Ministry of Economy & Competitiveness [AGL2014-58795-C4-1-R, AGL2014-58795-C4-3-R] - Financiador
National Veterinary High School, Algiers, Algeria. - Autor o Coautor
Natl Vet High Sch, Algiers, Algeria - Autor o Coautor
Obra Social La CaixaLa Caixa Foundation - Financiador
PROFAND - Financiador
Ubesol - Financiador
Unidad de Producción y Sanidad Animal del Centro de Investigación y Tecnología Agroalimentaria de Aragón (CITA), Instituto Agroalimentario de Aragón (IA2), Zaragoza, Spain. - Autor o Coautor
Univ Medea, Dept Biol, Medea 26000, Spain - Autor o Coautor
Univ Navarra, Dept Microbiol & Parasitol IDISNA, E-31080 Pamplona, Spain - Autor o Coautor
Univ Navarra, Inst Salud Trop, E-31080 Pamplona, Spain - Autor o Coautor
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Resúmen

Resúmen: Brucella bacteria cause brucellosis, a major zoonosis whose control requires efficient diagnosis and vaccines. Identification of classical Brucella spp. has traditionally relied on phenotypic characterization, including surface antigens and 5-10% CO2 necessity for growth (CO2-dependence), a trait of Brucella ovis and most Brucella abortus biovars 1-4 strains. Although molecular tests are replacing phenotypic methods, CO2-dependence remains of interest as it conditions isolation and propagation and reflects Brucella metabolism, an area of active research. Here, we investigated the connection of CO2-dependence and carbonic anhydrases (CA), the enzymes catalyzing the hydration of CO2 to the bicarbonate used by anaplerotic and biosynthetic carboxylases. Based on the previous demonstration that B. suis carries two functional CAs (CAI and CAII), we analyzed the CA sequences of CO2-dependent and -independent brucellae and spontaneous mutants. The comparisons strongly suggested that CAII is not functional in CO2-dependent B. abortus and B. ovis, and that a modified CAII sequence explains the CO2-independent phenotype of spontaneous mutants. Then, by mutagenesis and heterologous plasmid complementation and chromosomal insertion we proved that CAI alone is enough to support CO2-independent growth of B. suis in rich media but not of B. abortus in rich media or B. suis in minimal media. Finally, we also found that insertion of a heterologous active CAII into B. ovis reverted the CO2-dependence but did not alter its virulence in the mouse model. These results allow a better understanding of central aspects of Brucella metabolism and, in the case of B. ovis, provide tools for large-scale production of diagnostic antigens and vaccines.

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