Please use this identifier to cite or link to this item: https://doi.org/10.1002/bit.20304
DC FieldValue
dc.titleMonitoring of active but non-culturable bacterial cells by flow cytometry
dc.contributor.authorSachidanandham, R.
dc.contributor.authorGin, K.Y.-H.
dc.contributor.authorPoh, C.L.
dc.date.accessioned2016-11-28T10:13:41Z
dc.date.available2016-11-28T10:13:41Z
dc.date.issued2005-01-05
dc.identifier.citationSachidanandham, R., Gin, K.Y.-H., Poh, C.L. (2005-01-05). Monitoring of active but non-culturable bacterial cells by flow cytometry. Biotechnology and Bioengineering 89 (1) : 24-31. ScholarBank@NUS Repository. https://doi.org/10.1002/bit.20304
dc.identifier.issn00063592
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/130875
dc.description.abstractFlow cytometric signatures (i.e., light scatter, red and green fluorescence) were obtained for the active but non-culturable (ABNC) cells of E. coli and a coliform isolate H03N1, in seawater microcosms using BacLight, a live-dead assay kit from Molecular Probes (Eugene/Portland, OR). Previous studies have reported that there are two major adaptations, which cells undergo during the formation of ABNC states: cell wall toughening and DNA condensation. Therefore, we hypothesized that the matured ABNC forms should be more resistant to extreme temperature treatments (i.e., by freezing in liquid nitrogen and thawing at room temperature) than the normal and transition populations. It was shown that the membrane-compromised cells (comprising of normal wild-type and dead cells which are less resistant to rapid freeze thaw) could be differentiated from the matured ABNC using BacLight staining and fluorescence detection by flow cytometry. The population of ABNC cells, which could not be cultured using m-FC media (for the enumeration of fecal coliforms), was resuscitated in phosphate buffer saline followed by growth in Luria broth. Flow cytometry was thus able to detect and differentiate the ABNC cells against a mixed population comprising of culturable cells, transition populations, and dead cells. The results also showed that the formation of ABNC is as early as 2 days in seawater microcosms. By directly comparing the coliform levels enumerated by the BacLight based flow cytometry assays and m-FC technique, it was shown that the presence of coliforms can be undetected by the membrane filtration method. © 2004 Wiley Periodicals, Inc.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1002/bit.20304
dc.sourceScopus
dc.subjectABNC
dc.subjectBacteria
dc.subjectDNA condensation
dc.subjectEscherichia coli
dc.subjectFlow cytometry
dc.subjectWater quality
dc.typeArticle
dc.contributor.departmentCIVIL ENGINEERING
dc.contributor.departmentMICROBIOLOGY
dc.description.doi10.1002/bit.20304
dc.description.sourcetitleBiotechnology and Bioengineering
dc.description.volume89
dc.description.issue1
dc.description.page24-31
dc.description.codenBIBIA
dc.identifier.isiut000226120900004
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