Vaishampayan, Parag and Probst, Alexander J. and La Duc, Myron T. and Bargoma, Emilee and Benardini, James N. and Andersen, Gary L. and Venkateswaran, Kasthuri (2013) New perspectives on viable microbial communities in low-biomass cleanroom environments. ISME JOURNAL, 7 (2). pp. 312-324. ISSN 1751-7362, 1751-7370
Full text not available from this repository. (Request a copy)Abstract
The advent of phylogenetic DNA microarrays and high-throughput pyrosequencing technologies has dramatically increased the resolution and accuracy of detection of distinct microbial lineages in mixed microbial assemblages. Despite an expanding array of approaches for detecting microbes in a given sample, rapid and robust means of assessing the differential viability of these cells, as a function of phylogenetic lineage, remain elusive. In this study, pre-PCR propidium monoazide (PMA) treatment was coupled with downstream pyrosequencing and PhyloChip DNA microarray analyses to better understand the frequency, diversity and distribution of viable bacteria in spacecraft assembly cleanrooms. Sample fractions not treated with PMA, which were indicative of the presence of both live and dead cells, yielded a great abundance of highly diverse bacterial pyrosequences. In contrast, only 1% to 10% of all of the pyrosequencing reads, arising from a few robust bacterial lineages, originated from sample fractions that had been pre-treated with PMA. The results of PhyloChip analyses of PMA-treated and -untreated sample fractions were in agreement with those of pyrosequencing. The viable bacterial population detected in cleanrooms devoid of spacecraft hardware was far more diverse than that observed in cleanrooms that housed mission-critical spacecraft hardware. The latter was dominated by hardy, robust organisms previously reported to survive in oligotrophic cleanroom environments. Presented here are the findings of the first ever comprehensive effort to assess the viability of cells in low-biomass environmental samples, and correlate differential viability with phylogenetic affiliation. The ISME Journal (2013) 7, 312-324; doi: 10.1038/ismej.2012.114; published online 11 October 2012
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PROPIDIUM MONOAZIDE; QUANTITATIVE PCR; ETHIDIUM MONOAZIDE; CLONE LIBRARY; DEEP-SEA; DIVERSITY; BACTERIAL; SPACECRAFT; MICROARRAY; CELLS; viability; microarray; 454 pyrosequencing; PMA; PhyloChip; 16S rRNA gene |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 24 Apr 2020 08:39 |
| Last Modified: | 24 Apr 2020 08:39 |
| URI: | https://pred.uni-regensburg.de/id/eprint/17174 |
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