Fig. 2:
Neighbor-joining phylogram of the partial CAL (A), partial CHS (B), and partial β-tubulin (C) genes obtained from isolates A (18782B) and B (18782A), and the Sporothrix mexicana, S. pallida, S. brasiliensis, S. schenckii, and S. globosa reference strains constructed with MEGA version 4.0.2. Bootstrap values after 1,000 replicates are presented in the branch node. (D) Comparison of the partial ITS1 and 2 regions of all strains of S. brasiliensis demonstrated a high similarity between the type strain IPEC16490 and the patient isolates.
Fig. 3:
(A) Comparison of the T3B PCR fingerprinting profiles obtained for isolates A and B with the Sporothrix reference strains for the main species with clinical association. (1) Isolate A, (2) Isolate B, (3) S. brasiliensis (IPEC 16490), (4) S. globosa (IPEC 27135), (5) S. schenckii (IPEC27722), and (6) negative control. (M) Molecular marker DNA ladder, 100 bp (Invitrogen). (B) Primer-M13-driven DNA fingerprinting profiles of the two isolates included in this study (1) Isolate A and (2) Isolate B, and the reference strain (3) S. brasiliensis (IPEC 16490). The DNA molecular marker 1Kb (DNA ladder, Invitrogen) was loaded in the first and last wells. (C) (GACA)4-driven DNA fingerprinting profiles of the two isolates (1) Isolate A and (2) Isolate B, and the reference strain (3) S. brasiliensis (IPEC 16490). The DNA molecular marker 1Kb (DNA ladder, Invitrogen) was loaded in the first and last wells.
Fig. 5:
experimental infection. (A-B) Mice inoculated with the Sporothrix brasiliensis isolate B, presenting with nodules in the legs (circle) and enlarged testicles (arrow) on the 40th day of infection. (C) The survival of the mice following subcutaneous inoculation of 3.2 × 105 conidia of S. brasiliensis. The survival rate of the mice inoculated with isolate B was 90% by day 35 after the inoculation, whereas there was 100% survival in the other groups A, (A + B), and control. The data represent the survival rates of 10 animals per group. The control group was inoculated with phosphate-buffered saline (PBS). p < 0.05. (D) The number of S. brasiliensis colony forming units (CFU) isolated from the spleen. Groups of mice were subcutaneously inoculated with 3.2 × 105 conidia. Each point represents the mean number of CFU recovered from the spleens of three mice euthanized at the predetermined days 21, 35, and 49 after the inoculation with isolates A, B, or (A + B). p < .05. (E) The splenic index of the mice following the subcutaneous inoculation of 3.2 × 105 conidia of S. brasiliensis, and the control group inoculated with PBS. The spleen and body weight ratio of each infected and control mouse was determined. The ratios of the relative spleen weights of the infected mice were expressed as units in relation to the control. p < 0.5.
Fig. 6:
(A-J) Histological findings in the liver and lungs of mice inoculated with the Sporothrix brasiliensis isolates. HE. (A) Isolate A, liver, 21 days after the inoculation. Hepatitis, pyogranulomatous, necrotizing, focal, and moderate. (B) Isolate A, lung, 49 days after the inoculation. Pneumonia, pyogranulomatous (poorly organized), diffuse, and moderate. (C) Isolate (A + B), liver, 35 days after the inoculation. Hepatitis, pyogranulomatous, focal, and moderate. A perivascular well-organized granuloma was observed. (D) Isolate (A + B), lung, 49 days after the inoculation. Pneumonia, pyogranulomatous (poorly organized), diffuse, and moderate. (E) Isolate B, liver, 49 days after the inoculation. Hepatitis, pyogranulomatous, multifocal, and severe. Multiple well-organized granulomas were observed. (F) Isolate B, lung, 49 days after the inoculation. Pneumonia, pyogranulomatous, diffuse, and severe. A well-organized granuloma was observed. (G) Isolate B, liver, 49 days after the inoculation. Rare round-to-oval yeasts located in the centre of the granulomas. One of the yeasts shows a narrow-based budding (arrow). GMS. (H) Isolate B, lung, 49 days after the inoculation. Rare round-to-oval yeasts without budding (arrows) were observed in the centre of the granulomas. GMS. (I) Control mice, liver, 49 days after the inoculation, HE; (J) Control mice, lungs, 49 days after the inoculation, HE.
Fig. 7:
(A-E) Histological findings in the kidneys and heart of the mice inoculated with Sporothrix brasiliensis isolates. (A) Isolate B, kidney, 35 days after the inoculation. Pyogranulomatous inflammatory infiltrate in the perirenal adipose tissue. A well-organized granuloma was observed. HE. (B) Isolate (A + B), kidney, 35 days after inoculation. Interstitial nephritis, pyogranulomatous, focal, and mild. HE. (C) Isolate B, heart, 35 days after the inoculation. Suppurative valvular endocarditis, focal, and moderate. HE. (D) Isolate B, heart, 49 days after the inoculation. Pericardial mineralization (arrows) and fibrosis. HE. (E) Control mice, kidney, 35 days after the inoculation, HE.