PMC

SDS-PAGE under reducing conditions with Western blotting of different mycobacterial species. (A) The membrane containing the separated proteins was incubated with monospecific antiserum against AhpC. (B) The membrane containing the separated proteins was incubated with monospecific antiserum against AhpD. Lanes: 1, purified AhpC; 2, purified AhpD; 3, M. avium subsp. paratuberculosis 2E; 4, M. avium subsp. paratuberculosis ATCC 19698; 5, M. avium subsp. paratuberculosis 173p (goat clinical isolate); 6, M. avium subsp. paratuberculosis (goat clinical isolate); 7, M. avium subsp. avium D4; 8, M. intracellulare MNC 72; 9, Mycobacterium scrofulaceum MNC 95; 10, Mycobacterium kansasii MNC 861; 11, M. gordonae MNC 64; 12, Mycobacterium fortuitum MNC 103; 13, Mycobacterium phlei MNC 115; 14, M. bovis AN5; 15, M. tuberculosis H37Rv; 16, M. bovis BCG Moreau.

Dendrogram of sequence data for 27 nonpigmented M. szulgai, 12 pigmented M. szulgai, and some mycobacterial type strains in the MicroSeq database, with Mycobacterium fortuitum subsp. fortuitum as an out-group. GSA, gene sequence analysis. The dendrogram was generated by the neighbor joining (N Join) method, with the horizontal line at the top representing a 3.727% genetic difference.

RAW 264.7 cells infected with Mycobacterium fortuitum with a MOI 1:5 (cells:bacilli) were exposed to several concentrations of extract and rifampicin (0.5X, 1X, 2X and 4X MICs, 0 = untreated control) in triplicate. After 2, 4 and 6 days post infection, cells were lysed and plated on 7H10 agar to determine CFU/mL. Plots detail the bactericidal activity of extract of O. speciosus (a) and rifampicin (b) used as the reference positive control antibiotic. Values represent means ± SD

Relationship between SODs from different organisms. The dendrogram was constructed using the CLUSTAL_X program as described by Thompson et al. (). Relatedness is represented by the branch length. Bar, 0.1 amino acid substitution. GenBank accession numbers for the organisms represented are as follows: Gordonia sp. strain Kb2, CAC85367; Mycobacterium smegmatis MC² 155, YP_890640; Micrococcus luteus NCTC 2665, ZP_02944066; Brevibacterium linens BL2, ZP_00380098; Kineococcus radiotolerans SRS30216, YP_001363305; Nocardia farcinica IFM 10152, YP_116327; Mycobacterium abscessus, YP_001700872; Nocardia brasiliensis ATCC 700358 SodA, ABD64088; Rhodococcus sp. strain RHA1 SodA, YP_703964; Nocardia asteroides GUH2, P53651; Mycobacterium fortuitum, Q59519; Mycobacterium sp. strain MCS, YP_642204; Mycobacterium avium subsp. paratuberculosis K10, NP_959121; Mycobacterium avium 104, YP_879475; Kocuria rhizophila DC2201, YP_001853893; Mycobacterium lepraemurium, O86165; Mycobacterium smegmatis MC² 155 SodA, YP_890845; Mycobacterium avium subsp. paratuberculosis, ABZ81482; Renibacterium salmoninarum ATCC 33209, YP_001625417; Mycobacterium avium subsp. avium, ABZ81479.1; Arthrobacter sp. strain FB24, YP_831566.1; Arthrobacter pascens DMDC12, ABG76965.1; Mycobacterium avium subsp. paratuberculosis, ABZ81484.1; Mycobacterium avium subsp. hominissuis, ABZ81485.1; Mycobacterium leprae TN, NP_301180.1; Mycobacterium tuberculosis H37Rv, NP_218363.1; Arthrobacter aurescens TC1, YP_947831.1; Arthrobacter nitroguajacolicus, YP_001210461; Mycobacterium bovis AF 2122/97, NP_857513.1.