Incomplete cell disruption of resistant microbes

Sci Rep. 2019 Apr 4;9(1):5618. doi: 10.1038/s41598-019-42188-9.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteria / genetics
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / isolation & purification*
  • DNA / chemistry
  • DNA / isolation & purification*
  • DNA, Bacterial / genetics
  • Drug Resistance, Bacterial / genetics
  • Drug Resistance, Fungal / genetics
  • Fungal Proteins / chemistry
  • Fungal Proteins / isolation & purification*
  • Fungi / genetics
  • Microspheres
  • Sonication / methods

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Fungal Proteins
  • DNA