Supplementary MaterialsS1 Fig: GI colonization is dependent upon the addition of

Supplementary MaterialsS1 Fig: GI colonization is dependent upon the addition of Sm to the drinking water. individual groups of mice at levels comparable to those of the parental strain. (TIFF) ppat.1005061.s004.tiff (349K) GUID:?99DDC75A-CB81-4759-A9B8-333990BE84C6 S5 Fig: Newman and showed impaired GI colonization following inoculation by oral gavage with 109 CFU. Fecal pellets were cultured quantitatively at the indicated time points. Each dot indicates the CFU double mutant of RN4220, which lacks alpha- and beta-strain, and fecal pellets were cultured quantitatively at indicated time points. Each dot indicates the CFU strain Newman and the mutant were killed under low pH conditions (glycine buffer, pH 3.0C3.6). (TIF) ppat.1005061.s007.tif (205K) GUID:?A118C6DB-9299-41FA-B2F9-DBD7AD8CFCB8 S8 Fig: Newman was more sensitive than the parental strain to overnight treatments with the proteolytic enzymes pepsin and trypsin. (TIFF) ppat.1005061.s008.tiff (443K) GUID:?35ABE305-1319-48BE-B9E0-E521CB7E6B51 S9 Fig: Compared to the control strain V8, strain Newman was only weakly proteolytic, and no significant differences were observed between the protease activity of culture supernatants derived from the WT or the mutant. (TIFF) ppat.1005061.s009.tiff (958K) GUID:?10720EE7-3DE1-4C2E-8E3F-791BBCB8762D S10 Fig: Autolysin extracts from strain Newman and Newman showed comparable lytic activity toward peptidoglycan prepared from the wild type strain Lafferty. (TIFF) ppat.1005061.s010.tiff (309K) GUID:?601754F0-3347-4DAF-90FB-E812CA7B44BD S11 Fig: The Newman WT and mutant both colonized the mouse GI tract, whereas the and the double mutant were not recovered from the mouse stools 7 and 14 days after inoculation. Each dot indicates the CFU 0.05; ** 0.01.(TIFF) ppat.1005061.s011.tiff (108K) GUID:?CA73ECD9-FC2D-4E93-B0F8-D98362D728AC CH5424802 distributor Data Availability StatementAll relevant data are within the paper and its Supporting Information files. Abstract colonizes the nose, throat, skin, and gastrointestinal (GI) tract of humans. GI carriage of is usually difficult to CH5424802 distributor eradicate and has been shown to facilitate the transmission of the bacterium among individuals. Although staphylococcal colonization of the GI tract is asymptomatic, it increases the likelihood of CH5424802 distributor contamination, particularly skin and soft tissue infections caused by USA300 isolates. We established a mouse model of persistent GI colonization and characterized the impact of selected surface antigens on colonization. In competition experiments, an acapsular mutant colonized better than the parental strain Newman, whereas mutants defective in sortase A and clumping factor A showed impaired ability to colonize the GI tract. Mutants lacking protein A, clumping factor B, poly-N-acetyl glucosamine, or SdrCDE showed no defect in colonization. An wall teichoic acid (WTA) mutant (and mutants showed reduced adherence in vitro to intestinal epithelial cells. The mutant was recovered in lower numbers than the wild type strain in the murine stomach and duodenum 1 h after inoculation. This reduced fitness correlated with the in vitro susceptibility of the mutant to bile salts, proteases, and a gut-associated defensin. Newman showed enhanced susceptibility to autolysis, and an autolysin double mutant abrogated this phenotype. However, the mutant did not survive better in the mouse GI tract than the mutant. Our results indicate that this failure of the mutant to colonize the GI tract correlates with its poor adherence and susceptibility to bactericidal factors within the mouse gut, but not to enhanced activity of its major autolysin. Author Summary persistently colonizes ~20% of the human population, and 40C60% of humans are intermittently colonized by this Adam23 bacterium. The most common reservoir for is the anterior nares, and the incidence of staphylococcal disease in higher in individuals who are colonized. Rectal colonization by isolates, reflecting gastrointestinal (GI) carriage, has recently been recognized as an important reservoir from which person to person transmission occurs. We developed a murine model of GI colonization to investigate bacterial factors that promote staphylococcal colonization of the gut. We identified several surface-associated antigens that modulate colonization of the GI tract and identified a surface glycopolymer (cell wall teichoic acid) as critical for the early actions in colonization. The failure of the teichoic acid.