[Sampling and detection methods of bioaerosols for the risk assessment of microorganisms in work environments]

Bioaerosols including bacteria and fungi have been almost unrecognized as pollutants of work environments in Japan. The combination of filter sampling and DNA staining by ethidium bromide (EtBr) was examined for the detection and evaluation of total numbers of bioaerosols, including viable and dead microorganisms, for risk assessment in work environments. With direct counting of microorganisms by EtBr concentrations of total cells were obtained in a shorter time than plate culture, the traditional method for detection of microorganisms. Total cell concentrations (cells/m3) were about 100 times greater than colonized cell concentrations (CFU/m3) in all samples. In some microscopic fields, macro conidia produced from some kinds of fungi were observed, but they were not detected by plate culture. Airborne bacterial 16S rDNA amplified by PCR were determined for their base sequences by DNA sequence analysis and classified by sequence-based homologies. Base sequences from 2 samples each contained 6 common groups of phylum. The combination of filter sampling and direct counting by EtBr staining was shown to be a better method for detecting and evaluating total cell concentrations in the risk assessment of sick building syndrome and allergy.