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Title: Experimental parameters defining ultra-low biomass bioaerosol analysis
Authors: Luhung, Irvan
Uchida, Akira
LiLim, Serene Boon Yuean
Gaultier, Nicolas Eugene
Kee, Carmon
Lau, Kenny Jia Xu
Gusareva, Elena S.
Heinle, Cassie Elizabeth
Wong, Anthony
Premkrishnan, Balakrishnan N. V.
Purbojati, Rikky Wenang
Acerbi, Enzo
Kim, Hie Lim
Junqueira, Ana Carolina M.
Longford, Sharon Rae
Lohar, Sachin R.
Yap, Zhei Hwee
Panicker, Deepa
Koh, Yanqing
Kushwaha, Kavita K.
Ang, Poh Nee
Putra, Alexander
Drautz-Moses, Daniela Isabel
Schuster, Stephan Christoph
Keywords: Science::Biological sciences
Issue Date: 2021
Source: Luhung, I., Uchida, A., LiLim, S. B. Y., Gaultier, N. E., Kee, C., Lau, K. J. X., Gusareva, E. S., Heinle, C. E., Wong, A., Premkrishnan, B. N. V., Purbojati, R. W., Acerbi, E., Kim, H. L., Junqueira, A. C. M., Longford, S. R., Lohar, S. R., Yap, Z. H., Panicker, D., Koh, Y., ...Schuster, S. C. (2021). Experimental parameters defining ultra-low biomass bioaerosol analysis. Npj Biofilms and Microbiomes, 7(1), 37-.
Project: MOE2013-T3-1-013 
Journal: npj Biofilms and Microbiomes 
Abstract: Investigation of the microbial ecology of terrestrial, aquatic and atmospheric ecosystems requires specific sampling and analytical technologies, owing to vastly different biomass densities typically encountered. In particular, the ultra-low biomass nature of air presents an inherent analytical challenge that is confounded by temporal fluctuations in community structure. Our ultra-low biomass pipeline advances the field of bioaerosol research by significantly reducing sampling times from days/weeks/months to minutes/hours, while maintaining the ability to perform species-level identification through direct metagenomic sequencing. The study further addresses all experimental factors contributing to analysis outcome, such as amassment, storage and extraction, as well as factors that impact on nucleic acid analysis. Quantity and quality of nucleic acid extracts from each optimisation step are evaluated using fluorometry, qPCR and sequencing. Both metagenomics and marker gene amplification-based (16S and ITS) sequencing are assessed with regard to their taxonomic resolution and inter-comparability. The pipeline is robust across a wide range of climatic settings, ranging from arctic to desert to tropical environments. Ultimately, the pipeline can be adapted to environmental settings, such as dust and surfaces, which also require ultra-low biomass analytics.
ISSN: 2055-5008
DOI: 10.1038/s41522-021-00209-4
Rights: © 2021 The Author(s). Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit
Fulltext Permission: open
Fulltext Availability: With Fulltext
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