1. Glutamate decarboxylase-dependent acid resistance in Brucella spp.: distribution and contribution to fitness under extremely acidic conditions
- Author
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Maria Alessandra Damiano, Daniela Bastianelli, Daniela De Biase, Alessandra Occhialini, Axel Cloeckaert, Sascha Al Dahouk, Stephan Köhler, Centre d’études d’Agents Pathogènes et Biotechologies pour la Santé (CPBS), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP), Sapienza University [Rome], Bundesinstitut für Risikobewertung - Federal Institute for Risk Assessment (BfR), Institut de Recherche en Infectiologie de Montpellier (IRIM), Infectiologie Santé Publique (ISP-311), Université de Tours-Institut National de la Recherche Agronomique (INRA), This work was supported by grant 1329-485 from the Federal Institute for Risk Assessment, Germany (to A.O. and S.K.), and by funding from Sapienza University of Rome and Fondazione Roma (to D.D.B.). Travels and research stays of A.O., D.D.B., M.A.D., and D.B. were supported by the 2011-2012 Galilée program of Egide (Hubert Curien program), no. 25960UE, from the French and Italian Ministry of Foreign and Europeans Affairs. M.A.D. was a recipient was awarded a postdoctoral fellowship from the foundation Infectiopôle Sud, and A.O. was awarded a Congés pour Recherches ou Conversions Thématiques (CRCT) from Montpellier University, We thank E. Bianchini for her participation in genetic constructions of E. coli strains, B. Saadeh for preliminary GABA tests, C. Göllner for GAD tests, P. Bouhours for preparation of bacterial media, and I. Jacques and D. O'Callaghan for their generous gifts of several marine strains and of B. inopinata BO1 and B. inopinata-like BO2 strains, respectively., Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Infectiologie et Santé Publique (UMR ISP), Institut National de la Recherche Agronomique (INRA)-Université de Tours, and Institut National de la Recherche Agronomique (INRA)-Université de Tours (UT)
- Subjects
MESH: Acids/toxicity ,Glutamate decarboxylase ,MESH: Glutamic Acid/metabolism ,Gene Expression ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,MESH: Drug Tolerance ,Mice ,MESH: Animals ,Cloning, Molecular ,Brucella inopinata ,MESH: Glutamate Decarboxylase/metabolism ,MESH: Ochrobactrum/drug effects ,0303 health sciences ,Rana catesbeiana ,Ecology ,biology ,Glutamate Decarboxylase ,MESH: Brucella/enzymology ,Drug Tolerance ,Phenotype ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,MESH: Acids/metabolism ,Biotechnology ,MESH: Gene Expression ,Glutamic Acid ,Brucella pinnipedialis ,MESH: Escherichia coli/genetics ,Genetics and Molecular Biology ,Brucella ,Ochrobactrum ,Microbiology ,03 medical and health sciences ,medicine ,MESH: Brucella/drug effects ,Escherichia coli ,Animals ,Humans ,MESH: Cloning, Molecular ,extreme acid stress ,diagnostic phenotypic trait ,glutamate-dependent acid resistance ,MESH: Mice ,030304 developmental biology ,MESH: Humans ,MESH: Ochrobactrum/enzymology ,030306 microbiology ,biology.organism_classification ,bacterial infections and mycoses ,MESH: Rana catesbeiana ,Brucella ceti ,MESH: Brucella/genetics ,MESH: Escherichia coli/metabolism ,MESH: Brucella/isolation & purification ,Acids ,Food Science - Abstract
Brucella is an expanding genus of major zoonotic pathogens, including at least 10 genetically very close species occupying a wide range of niches from soil to wildlife, livestock, and humans. Recently, we have shown that in the new species Brucella microti , the glutamate decarboxylase (Gad)-dependent system (GAD system) contributes to survival at a pH of 2.5 and also to infection in mice by the oral route. In order to study the functionality of the GAD system in the genus Brucella , 47 isolates, representative of all known species and strains of this genus, and 16 strains of the closest neighbor genus, Ochrobactrum , were studied using microbiological, biochemical, and genetic approaches. In agreement with the genome sequences, the GAD system of classical species was not functional, unlike that of most strains of Brucella ceti , Brucella pinnipedialis , and newly described species ( B. microti , Brucella inopinata BO1, B. inopinata -like BO2, and Brucella sp. isolated from bullfrogs). In the presence of glutamate, these species were more acid resistant in vitro than classical terrestrial brucellae. Expression in trans of the gad locus from representative Brucella species in the Escherichia coli MG1655 mutant strain lacking the GAD system restored the acid-resistant phenotype. The highly conserved GAD system of the newly described or atypical Brucella species may play an important role in their adaptation to acidic external and host environments. Furthermore, the GAD phenotype was shown to be a useful diagnostic tool to distinguish these latter Brucella strains from Ochrobactrum and from classical terrestrial pathogenic Brucella species, which are GAD negative.
- Published
- 2014