Abstract

The biota of black fungi in humid indoor environments was established using a protocol that consisted of non-selective and selective isolation procedures. In total, 113 samples were taken from bathrooms of residences in The Netherlands, Germany and Austria. Samples were processed either (i) directly by culturing on agar media, or (ii) by pre-incubating samples for enrichment in mineral solutions with perlite granules under constant toluene atmosphere for three months. Dilutions from the latter were then cultured and incubated as were those directly plated to agar media. Black colonies were selected and identified by sequencing the rDNA Internal Transcribed Spacer (ITS) region. Twenty-eight strains of black fungi were found in 26 positive samples without enrichment, and 42 strains were isolated from 38 positive samples after enrichment in toluene. The great majority of black fungal species were members of the order Chaetothyriales, which is the main order of melanized human opportunistic pathogens. Cladosporium species (Capnodiales) were the most frequent isolates when no enrichment was applied, as opposed to Exophiala species (Chaetothyriales) with enrichment. The enrichment method provides insight into a fungal biota commonly occurring in homes which has previously been overlooked. Several species have been previously known only from cutaneous infections and could suggest that bathrooms are a likely reservoir of these fungi.

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