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J Med Microbiol 53 (2004), 175-181; DOI: 10.1099/jmm.0.05421-0
© 2004 Society for General Microbiology
ISSN 0022-2615

Production of melanin by Aspergillus fumigatus

Sirida Youngchim1,2, Rachael Morris-Jones1, Roderick J. Hay3 and Andrew J. Hamilton1

1Dermatology Department, St Johns Institute of Dermatology, Guy's Hospital, Kings and St Thomas’ Medical Schools, London, UK 2Microbiology Department, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand 3Faculty of Medicine and Health Sciences, Queen University, Belfast, Northern Ireland, UK

Correspondence Sirida Youngchim syoungch{at}mail.med.cmu.ac.th

Received August 11, 2003
Accepted November 29, 2003

Melanins, or melanin-like compounds, may play a role in the pathogenesis of a number of human fungal infections. This study investigated the production of melanin by the important opportunistic pathogen Aspergillus fumigatus. Conidia from A. fumigatus were harvested and treated with proteolytic enzymes, denaturant and hot, concentrated acid; this yielded dark particles which were similar in size and shape to the original propagules. Electron spin resonance spectroscopy revealed that the conidial-derived particles were stable free radicals consistent with an identification as melanin. Melanin particles were used to immunize BALB/c mice in order to produce a total of five anti-melanin monoclonal antibodies (mAbs). The latter mAbs were strongly reactive both with intact conidia and with extracted melanin particles by ELISA and immunofluorescence reactivity. Immunofluorescence labelling with the novel mAbs was used to examine the temporal expression of melanin during in vitro culture of A. fumigatus –melanization was confined to conidial structures and was absent from hyphae. SDS-PAGE L-3,4-dihydroxyphenylalanine (L-DOPA) substrate analysis confirmed the presence of a laccase-type activity in conidial extracts, but not in hyphae. Melanin-binding mAbs were used to detect the presence of melanized conidia in three patients with nasal aspergilloma, indicating that in vivo melanization may occur during infection.

Abbreviations: DHN, dihydroxynaphthalene; DOPA, 3,4-dihydroxyphenylalanine; ESR, electron spin resonance; IF, immunofluorescence; SEM, scanning electron microscopy; TEM, transmission electron microscopy.




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