Density gradient separation of active and non-active cells from natural environments

AS Whiteley, MR Barer, A.G. O'Donnell

    Research output: Contribution to journalArticlepeer-review

    11 Citations (Scopus)

    Abstract

    We present a method for the selective, physical separation of active and non-active bacterial cells from natural communities. The method exploits the reduction of tetrazolium salts to form insoluble formazan crystals intracellularly in response to the addition of different oxidisable substrates. The intracellular deposition of formazan alters the bouyant density of active cells enabling them to be separated by density gradient centrifugation. The method has been successfully applied to the fractionation and collection of large whole cell sub-populations of active and non-active cells from sea-water samples. Removal of the bands from the density gradient, followed by PCR amplification and DGGE analyses showed distinct differences in the PCR amplicon diversity associated with the active and non-active cell fractions; an indication of changes in bacterial community structure in response to the addition of oxidisable substrate. Thus, based on their in situ respiration potential, the approach enables the cytochemical enrichment and molecular characterisation of mixed bacterial populations in natural environments.
    Original languageEnglish
    Pages (from-to)173-177
    JournalANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY
    Volume77
    DOIs
    Publication statusPublished - 2000

    Fingerprint

    Dive into the research topics of 'Density gradient separation of active and non-active cells from natural environments'. Together they form a unique fingerprint.

    Cite this