Incomplete cell disruption of resistant microbes

Biomolecules for OMIC analysis of microbial communities are commonly extracted by bead-beating or ultra-sonication, but both showed varying yields. In addition to that, different disruption pressures are necessary to lyse bacteria and fungi. However, the disruption efficiency and yields comparing be...

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Bibliographic Details
Published in:Scientific reports 2019-04, Vol.9 (1), p.5618, Article 5618
Main Authors: Starke, Robert, Jehmlich, Nico, Alfaro, Trinidad, Dohnalkova, Alice, Capek, Petr, Bell, Sheryl L., Hofmockel, Kirsten S.
Format: Article
Language:English
Subjects:
45/22
631/326/2565/855
631/337
Bacteria - genetics
Bacterial Proteins - chemistry
Bacterial Proteins - isolation & purification
Cell disruption
Deoxyribonucleic acid
DNA
DNA - chemistry
DNA - isolation & purification
DNA, Bacterial - genetics
Drug Resistance, Bacterial - genetics
Drug Resistance, Fungal - genetics
Ecological studies
Fungal Proteins - chemistry
Fungal Proteins - isolation & purification
Fungi
Fungi - genetics
Genomes
Gram-negative bacteria
Humanities and Social Sciences
Microbial activity
Microorganisms
Microspheres
multidisciplinary
Science
Science (multidisciplinary)
Sonication
Sonication - methods
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Summary:Biomolecules for OMIC analysis of microbial communities are commonly extracted by bead-beating or ultra-sonication, but both showed varying yields. In addition to that, different disruption pressures are necessary to lyse bacteria and fungi. However, the disruption efficiency and yields comparing bead-beating and ultra-sonication of different biological material have not yet been demonstrated. Here, we show that ultra-sonication in a bath transfers three times more energy than bead-beating over 10 min. TEM imaging revealed intact gram-positive bacterial and fungal cells whereas the gram-negative bacterial cells were destroyed beyond recognition after 10 min of ultra-sonication. DNA extraction using 10 min of bead-beating revealed higher yields for fungi but the extraction efficiency was at least three-fold lower considering its larger genome. By our critical viewpoint, we encourage the review of the commonly used extraction techniques as we provide evidence for a potential underrepresentation of resistant microbes, particularly fungi, in ecological studies.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-019-42188-9