Figure 1.
Growth of M. furfur NBRC 0656 in minimal media with (A) and without L-DOPA (B).
Figure 2.
ESR spectroscopy of melanin particles extracted from M. furfur NBRC 0656 grown on defined minimal medium with L-DOPA (A), mDixon medium (B), basal medium with L-Tryp (C) and basal medium with Arg (D) at 30°C for 7–10 days.
Figure 3.
Corresponding immunofluorescence and bright-field microscopy images demonstrating the labelling of yeast cells of M. furfur NBRC 0656 (A,B) and after (C,D) melanin extraction by the melanin-binding MAb 8D6. Bars represent 5 µm.
Figure 4.
Plate assay for demonstration of phenoloxidase activities for 10 days. M. furfur NBRC 0656 was inoculated into wells in laccase agar plate containing 1 mM of L-DOPA as the substrate.
C. neoformans H99 and S. cerevisiae MMCM 5211 were used as positive and negative controls, respectively. The development of an intense black color around the well indicated a positive test for phenoloxidase activity.
Figure 5.
Corresponding immunofluorescence and bright-field microscopy images demonstrating the labelling of yeast cells of M. furfur NBRC 0656 (A,B) and CBS 7019 (C,D) grown on MM with 1 mM L-DOPA and 1500 µg/ml kojic acid by the melanin-binding MAb 8D6. Bars represent 5 µm.
Figure 6.
The effect of L-DOPA and kojic acid on morphology of M. furfur NBRC 0656 when culturing in MM (A), MM with 1 mM L-DOPA (B), MM with 1 mM L-DOPA and kojic acid at 600 (C), 800 (D), 1,000 (E) 1,200 (F) 1,500 (G) µg/ml, and MM with 1,000 µg/ml kojic acid (H). Bars represent 5 µm.
Figure 7.
Effect of various concentrations of kojic acid on filament formation in M. furfur NBRC 0656 in microaerobic condition.
Data are means±SD (standard deviation of the means) based on at least three experiments. Statistically significant difference (two-tailed unpaired Student’s t-test) compared to control, MM and MM with L-DOPA, *p≤0.05.
Figure 8.
The mycelial productions of the various isolates in M. furfur.
M. furfur CBS 6046 (A), CBS 6000 (B), CBS 6001 (C), CBS 7019 (D) and two isolates from healthy person (E, F) were cultured on MM with 1 mM L-DOPA and kojic acid in 0.5% agar in microaerophilic environment at 30°C for 5–7 days. The concentrations of kojic acid were 1500 µg/ml for standard isolates (A-D) and 500 µg/ml for healthy isolates (E,F). Bars represent 5 µm.
Figure 9.
Corresponding immunofluorescence (A,C,E,G) and bright-field microscopy (B,D,F,H) images demonstrating the labelling of blastoconidia and short filaments from skin scrapings of pityriasis versicolor by the melanin-binding MAb 8D6.
Bars represent 5 µm.