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Título

Evaluating the Flow-Cytometric Nucleic Acid Double-Staining Protocol in Realistic Situations of Planktonic Bacterial Death

AutorFalcioni, Tania ; Papa, Stefano; Gasol, Josep M.
Fecha de publicaciónmar-2008
EditorAmerican Society for Microbiology
CitaciónApplied and Environmental Microbiology 74(6): 1767-1779 (2008)
ResumenSince heterotrophic prokaryotes play an important biogeochemical role in aquatic ecosystems and have a high capacity to survive in extreme environments, easy-to-perform protocols that probe their physiological states and the effects of environmental variables on those states are highly desired. Some methodologies combine a general nucleic acid stain with a membrane integrity probe. We calibrated one of these, the nucleic acid double-staining (NADS) protocol (G. Gre ́gori, S. Citterio, A. Ghiani, M. Labra, S. Sgorbati, S. Brown, and M. Denis, Appl. Environ. Microbiol. 67:4662–4670, 2001), determining the optimal stain concentrations in seawater and the response to conditions that generate prokaryote death (such as heat) and to conditions that are known to produce death in plankton, such as nutrient limitation or flagellate grazing. The protocol was validated by comparison to two methods used to detect viability: active respiration by 5-cyano 2,3-ditolyl tetrazolium chloride (CTC) and incorporation of tritiated leucine. We show that concentrations in the range of 5 to 20
Descripción13 pages, 3 tables, 5 figures, http://icm.csic.es/bio/projects/icmicrobis/pdf
Versión del editorhttp://dx.doi.org/10.1128/AEM.01668-07
URIhttp://hdl.handle.net/10261/16714
DOI10.1128/AEM.01668-07
ISSN0099-2240
E-ISSN1098-5336
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