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Title Phylogenetic analysis of atmospheric halotolerant bacterial communities athigh altitude in an Asian dust ( KOSA ) arrival region

The microbial communities transported by Asian desert dust (KOSA) events have attracted much attention as bioaerosols because the transported microorganisms are thought to influence the downwind ecosystems in Korea and Japan. However, the atmospheric microbial community has not been investigated at high attitude in the KOSA arrival area. In this study, to estimate the viability and diversity of atmospheric halotolerant bacteria, which are expected to resist to various environmental stresses as well as high salinities, bioaerosol samples were collected at 10 and 600 m above the ground within the KOSA arrival area, Suzu City, Japan, during KOSA events. During the sampling period, the particle numbers at 600 m were higher than those at less than 20 m, suggesting that large particles of aerosol fall from the high attitude of 600 m to the ground surface. The microorganisms in bioaerosol samples grew in media containing up to 15 % NaCl concentrations demonstrating the viability of the halotorerant bacteria in bioaerosol samples. The PCR-DGGE analysis using 16S rDNA revealed that the bacterial species in NaClamended cultures were similar to the bacteria detected from the genomic DNA directly extracted from the bioaerosol samples. The 16S rDNA sequences of bacterial communities in bioaerosol samples were classified into 4 phylotypes belonging to the B. cereus or B. subtilis group. The bioaerosol samples collected at 600 m included 2 phylotypes belonging to B. subtilis, and one phylotype among all 4 phylotypes was identical between the samples at 10 and 600 m. In the atmosphere at 600 m, the halotolerant bacterial community would remain viable, and the species composition would include a few species of the genus Bacillus. During this investigation period, these atmospheric bacteria may have been vertically transported to the ground surface, where the long-range KOSA particle transport from China is frequently observed. Suggested Reviewers: Huey-Jen Su Department of Environmental and Occupational Health, Medical College, Natinal Cheng Kung University hjsu@mail.nck.edu.tw His research group is investigating the increase of atmospheric microorganisms in Taiwan during the KOSA events. Ruprecht Jaenicke Institut für Physik der Atmosphäre, Universität Mainz jaenicke@uni-mainz.de His research group monitored the abundance of cellular material and proteins in the atmosphere, demonstrating that the bioaerosol would occupy the total amount of aerosol. Kimberly A. Prather Department of Chemistry and Biochemistry, University of California kprather@ucsd.edu His research group demonstrated that bioaerosols at high altitudes may act as ice nuclei and cloud condensation nuclei affecting ice-cloud processes, and that tehe bioaerosols were transported to high attitude and were important factor for establishing the aerosol models. Kenneth Sassen Geophysical Institute, University of Alaska Fairbanks ksassen@gi.alaska.edu His research group is investigating the bioaerosol attached on KOSA particles using the LIDAR (Light Detection and Ranging) . Dale W Griffin Center for Coastal and Watershed Studies, U.S. Geological Survey dgriffin@usgs.gov His research group is investigating the bioaerosol transported by African dust using the culture technique.