Immobilization of Phosphatidylserine by Ethanol and Lysozyme on the Cell Surface for Evaluation of Apoptosis-Like Decay in Activated-Sludge Bacteria.
activated sludge
annexin V-FITC staining
apoptosis-like decay
bacterial viability
ethanol-lysozyme treatment
Journal
Applied and environmental microbiology
ISSN: 1098-5336
Titre abrégé: Appl Environ Microbiol
Pays: United States
ID NLM: 7605801
Informations de publication
Date de publication:
02 07 2020
02 07 2020
Historique:
received:
14
02
2020
accepted:
30
04
2020
pubmed:
18
5
2020
medline:
11
11
2020
entrez:
17
5
2020
Statut:
epublish
Résumé
Accurate determination of microbial viability can be crucial in microbe-dominated biosystems. However, the identification of metabolic decay in bacterial cells can be elaborate and difficult. We sought to identify apoptosis-like bacterial processes by using annexin V-fluorescein isothiocyanate (FITC) (AVF), a probe typically used to stain phosphatidylserine (PS) on exposed cell membranes. The bacterial cell wall provides a barrier that is responsible for low efficiency of direct PS staining of decayed bacterial cells. This can be overcome by pretreatment of the bacteria with 70% ethanol, which fixates the bacteria and preserves the PS status, combined with lysozyme treatment to hydrolyze the cell wall. That treatment improved the efficiency of AVF staining considerably, as shown for pure strains of an
Identifiants
pubmed: 32414801
pii: AEM.00345-20
doi: 10.1128/AEM.00345-20
pmc: PMC7357489
pii:
doi:
Substances chimiques
Phosphatidylserines
0
Sewage
0
Ethanol
3K9958V90M
Muramidase
EC 3.2.1.17
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2020 American Society for Microbiology.
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