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

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

Evolution towards pathogenicity: Comparative analysis between Brucella and a recently isolated Pseudochrobactrum

Póster presentado Online en el XXVIII Congreso Nacional de Microbiología 28 de junio al 2 de julio, 2021 .- https://hal.inrae.fr/hal-03337244/documentLa brucelosis es una de las zoonosis bacterianas más ampliamente distribuidas en el mundo, contribuyendo de forma importante a la pobreza de muchos países en desarrollo. Ninguna de las vacunas actuales proporciona una protección completa, son virulentas para humanos e interfieren en el diagnóstico serológico. Por ello, para mejorar las vacunas actuales, es imprescindible seguirestudiando los mecanismos de virulencia de Brucella, el agente etiológico de la brucelosis. Ésta pertenece a las α-2 Proteobacteria, grupo que incluye desde bacterias ambientales hasta bacterias que han evolucionado a patógenos animales difícilmente detectados por el sistema inmune, como es el caso de Brucella. Para estudiar la evolución que se ha dado hacia la patogenicidad en esta familia y poder determinar mejor así los factores de virulencia de Brucella, es interesante comparar bacterias filogenéticamente próximas que son biológicamente diferentes. Por primera vez, se han aislado bacterias del género Pseudochrobactrum en un huésped natural de Brucella.Pseudochrobactrum es un género filogenéticamente cercano a Brucella que, hasta la fecha, sólo incluye bacterias ambientales. En este trabajo se han caracterizado estos nuevos aislamientos y teniendo en cuenta sus propiedades fisiológicas, su perfil de lípidos polares y ácidos grasos, y el análisis filogenético, se ha propuesto una nueva especie con el nombre de Pseudochrobactrum bovis. Los estudios comparativos con Brucella han revelado diferencias en la composición de la envoltura celular, el contenido genético y virulencia. Por ello, P. bovis puede representar un nuevo modelo para estudiar la evolución de Brucella hacia la virulenciaN

Serological survey of cattle toxoplasmosis in Medea, Algeria

International audienc

First genetic characterization of Toxoplasma gondii in stray cats from Algeria

International audienceToxoplasmosis is a parasitic disease with worldwide distribution and a major public health problem. In Algeria, no data are currently available about genotypes of Toxoplasma gondii isolated from animals or humans. The present study assesses for the first time the seroprevalence of toxoplasmosis in stray cats, and provides molecular characterization of T. gondii strains circulating in this feline population in Algiers, the capital city of Algeria. Sera from 96 stray cats were tested for the presence of antibodies against T. gondii using the modified agglutination test. The seroprevalence was 50% (48/96) using 1:6 as the positivity cut-off. Different organs samples from stray cats, including heart samples, were tested for the presence of Toxoplasma DNA using real-time PCR. T. Gondii DNA was detected in 90.6% (87/96) of hearts. Of these parasitic DNAs, 22 were submitted to genotyping through the analysis of 15 microsatellite markers. The identified genotypes (12 of 22) mainly belonged to the type II lineage

Evolution towards pathogenicity: Comparative analysis between Brucella and a recently isolated Pseudochrobactrum

International audienc

Pseudochrobactrum algeriensis sp. nov., isolated from lymph nodes of Algerian cattle

International audienceThree Gram-negative, rod-shaped, oxidase-positive, non-spore-forming, non-motile strains (C130915_07 T , C150915_16 and C150915_17) were isolated from lymph nodes of Algerian cows. On the basis of 16S rRNA gene and whole genome similarities, the isolates were almost identical and clearly grouped in the genus Pseudochrobactrum . This allocation was confirmed by the analysis of fatty acids (C 19:cyclo , C 18 : 1 , C 18 : 0 , C 16 : 1 and C 16 : 0 ) and of polar lipids (major components: phosphatidylethanolamine, ornithine-lipids, phosphatidylglycerol, cardiolipin and phosphatidylcholine, plus moderate amounts of phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine and other aminolipids). Genomic, physiological and biochemical data differentiated these isolates from previously described Pseudochrobactrum species in DNA relatedness, carbon assimilation pattern and growth temperature range. Thus, these organisms represent a novel species of the genus Pseudochrobactrum , for which the name Pseudochrobactrum algeriensis sp. nov. is proposed (type strain C130915_07 T =CECT30232 T =LMG 32378 T )