The Three Flagellar Loci ofPA Are Dispensable for Virulence in Cellular Models and Mice.

Abstract

is a facultative intracellular bacterium that causes a non-zoonotic ovine brucellosis mainly characterized by male genital lesions and is responsible for important economic losses in sheep farming areas. Studies about the virulence mechanisms ofhave been mostly performed with smooth (bearing O-polysaccharide in lipopolysaccharide) zoonotic species, and those performed withhave revealed similarities but also relevant differences. Except for few strains recently isolated from unconventional hosts,species are non-motile but contain the genes required to assemble a flagellum, which are organized in three main loci of about 18.5, 6.4, and 7.8 kb. Although these loci contain different pseudogenes depending on the non-motilespecies, smooth16M builds a sheathed flagellum under particular culture conditions and requires flagellar genes for virulence. However, nothing is known in this respect regarding otherstrains. In this work, we have constructed a panel ofPA mutants defective in one, two or the three flagellar loci in order to assess their role in virulence of this rough (lacking O-polysaccharide)species. No relevant differences in growth, outer membrane-related properties or intracellular behavior in cellular models were observed between flagellar mutants and the parental strain, which is in accordance with previous results with16M single-gene mutants. However, contrary to thesemutants, unable to establish a chronic infection in mice, removal of the three flagellar loci indid not affect virulence in the mouse model. These results evidence new relevant differences betweenand, two species highly homologous at the DNA level and that cause ovine brucellosis, but that exhibit differences in the zoonotic potential, pathogenicity and tissue tropism.