Evaluation of a high-volume portable bioaerosol sampler in laboratory and field environments.

UNLABELLED: This study investigated the physical and biological performances of a portable centrifugal sampler for viable bioaerosols, RCS High Flow. The performance of the test sampler in the laboratory and field environments was compared with that of a reference sampler, BioSampler. The laboratory experiments with non-biological particles of KCl, oleic acid, and polystyrene latex showed that the test sampler's collection efficiency is about 22% for 0.5-microm particles, 48% for 1.0-microm particles, and approximately 100% for particles of 2.5 microm and larger. These tests indicated that the sampler's cut-off size (d50) was 1.1 microm. The test sampler's physical performance when collecting the spores and vegetative cells of Bacillus subtilis var. niger (BG) was similar to that when collecting non-biological particles of the same size. In the laboratory tests, the RCS High Flow sampler was found to enumerate approximately 40% of BG spores and cells relative to the reference sampler, BioSampler. A similar ratio was found during testing in an indoor environment. This ratio decreased to below 10% when testing was performed in an outdoor environment. We hypothesize that the test sampler's underperformance compared with the BioSampler could be caused by the damage to sensitive microorganisms during the collection process, test sampler's sensitivity to wind direction and speed as well as break-up of particle aggregates during the impingement process in BioSampler, which resulted in more colony-forming units (CFUs) being counted by the reference sampler than by the test sampler. Overall, when the RCS High Plus is used to sample culturable airborne microorganisms, the results obtained may have to be adjusted to avoid potential underestimation of microorganism concentration in the air. PRACTICAL IMPLICATIONS: The laboratory testing of the RCS High Flow bioaerosol sampler showed that the sampler collects 1 microm particles and larger with an efficiency of 50% and higher; the efficiency reaches approximately 100% for particles of 2.5 microm and larger. When considering this result, most of the airborne fungal spores would be collected with an efficiency between 50 and 100%. The field testing, however, indicated that the RCS High Flow sampler recovered from 41 to 71% of microorganisms collected relative to the reference sampler, Biosampler, and this ratio dropped to below 5% during outdoor testing. Thus, while the RCS High Flow sampler offers certain advantages over other samplers for viable bioaerosols--it is lightweight, battery operated, and collects viable microorganisms at a high flow rate directly on agar media, the results obtained may have to be adjusted to avoid potential underestimation of microorganism concentration in the air, especially if sampling is performed outdoors.

Cloning and expression of thermophilic catechol 1,2-dioxygenase gene (catA) from Streptomyces setonii.

Streptomyces setonii (ATCC 39116) degrades various single aromatic compounds such as phenol or benzoate via an ortho-cleavage pathway using catechol 1,2-dioxygenase (C12O). A PCR using degenerate primers based on the conserved regions of known C12O-encoding genes amplified a 0.45-kbp DNA fragment from S. setonii total DNA. A Southern hybridization analysis and size-selected DNA library screening using the 0.45-kbp PCR product as a probe led to the isolation of a 6.4-kbp S. setonii DNA fragment, from which the C12O-encoding genetic locus was found to be located within a 1.4-kbp DNA fragment. A complete nucleotide sequencing analysis of the 1.4-kbp DNA fragment revealed a 0.84-kbp open reading frame, which showed a strong overall amino acid similarity to the known high-G+C Gram-positive (but significantly less to the Gram-negative) bacterial mesophilic C12Os. The heterologous expression of the cloned 1.4-kbp DNA fragment in Escherichia coli demonstrated that this C12O possessed a thermophilic activity within a broad temperature range (up to 65 degrees C) and showed a higher activity against 3-methylcatechol than catechol or 4-methylcatechol, but no activity against protocatechuate.