Effectiveness of SYTOX Green Stain for Bacterial Viability Assessment

Lebaron, Philippe; Catala, Philippe; Parthuisot, Nathalie

doi:10.1128/aem.64.7.2697-2700.1998

Cited by 167 publications

(114 citation statements)

References 19 publications

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“…Heating of samples is used for verification of viability methods (Deere et al 1995;Roth et al 1997;Lebaron et al 1998), and the present results confirmed that membranes, nucleic acids and certain enzymes are cellular sites of heat injury (Teixeira et al 1997).…”

Section: Light-irradiatedsupporting

confidence: 71%

“…It was recently reported (Lebaron et al 1998) that the SYTOX Green assay under-estimates the fraction of dead cells within starved populations of E. coli and Salmonella typhimurium. Several permeabilized (dead) cells had damaged nucleic acids and could therefore not accumulate more SYTOX Green nucleic acid stain than the less permeable, living cells.…”

Section: Effect Of Pre-uv Light Irradiation and Starvationmentioning

confidence: 99%

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Effect of u.v. light irradiation, starvation and heat on Escherichia colibetabeta-D-galactosidase activity and other potential viability parameters

Fiksdal

Tryland

1999

J Appl Microbiol

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The effect of u.v. light irradiation and two other types of stress (heat and starvation) on cellular functions of Escherichia coli have been studied. The severe reduction of the culturable cell number (cfu) and the direct viable count (DVC) after exposure to moderate u.v. light doses (48 mWs cm–2), was not reflected by the dehydrogenase activity (5‐cyano‐2,3‐ditolyl tetrazolium chloride (CTC)‐positive cells), the membrane integrity (SYTOX Green‐negative cells), the membrane potential (bis‐(1,3‐dibutylbarbituric acid) trimethine oxonol (DiBAC4[3]) (OXONOL)‐negative cells), and the β‐ d‐galactosidase activity. All parameters were affected by high u.v. light doses. Cellular activities (CTC, SYTOX, OXONOL, β‐ d‐galactosidase activity) were intact in non‐culturable cells with presumably severe damage to DNA, and the activities seemed not to be appropriate for detection of viable E. coli after u.v. light irradiation. Heating for 20–30 min at 63 °C was required to cause a severe loss of the β‐ d‐galactosidase activity and the numbers of CTC‐ positive, SYTOX Green‐negative or OXONOL‐negative cells. A large portion (≥38%) of pre‐ irradiated (190 mWs cm–2) cells maintained their ability to reduce CTC and exclude SYTOX Green and OXONOL after 51 d of starvation (dark, 7 °C) in phosphate‐buffered saline.

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Section: Light-irradiatedsupporting

confidence: 71%

Section: Effect Of Pre-uv Light Irradiation and Starvationmentioning

confidence: 99%

Effect of u.v. light irradiation, starvation and heat on Escherichia colibetabeta-D-galactosidase activity and other potential viability parameters

Fiksdal

Tryland

1999

J Appl Microbiol

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“…Sytox green constitutes a high affinity nucleic acid stain that does not cross the membranes of live cells, but it enters into the membranes of cells with compromised plasma membrane. This dead cell marker has shown its effectiveness as an indicator of assessment after the exposure of cells to antimicrobial agents (Roth et al 1997;Lebaron et al 1998). Thus, concentrated supernatants from Lact.…”

Section: Discussionmentioning

confidence: 99%

Effects of concentrated supernatants recovered fromLactobacillus plantarumonEscherichia coligrowth and on the viability of a human promyelocytic cell line

Puertollano

Cruz-Chamorro

et al. 2009

Journal of Applied Microbiology

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Aims: The ability of concentrated supernatants from Lactobacillus plantarum to produce a disruption of plasma membrane in eukaryotic and prokaryotic cells has been examined. Methods and Results: A strain of Lact. plantarum (tolerant to acid and bile salts and resistant to several antibiotics) was used. It inhibited the growth of pathogenic Escherichia coli and L. monocytogenes. Supernatants from Lact. plantarum were concentrated by centrifugation. Either E. coli or HL‐60 cells (a human promyelocytic cell line) were treated in the presence of the concentrated supernatants. The effect of concentrated supernatants from Lact. plantarum on E. coli growth demonstrated a bacteriostatic activity and a loss of cell viability measured by sytox green staining. Concentrated supernatants were capable of disturbing plasma membrane in E. coli and of promoting a cytotoxic and lyctic action on HL‐60 cells and on human erythrocytes, respectively. Conclusions: These results suggest that Lact. plantarum release an effective compound responsible for an important effect in the disruption of E. coli plasma membrane and for a cytototoxic activity on promyelocytic leukaemia cells. Significance and Impact of the Study: This is the first in vitro study about the antimicrobial and biological activities of concentrated supernatants from Lact. plantarum.

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“…Although SYTOX Green (S-7020, Invitrogen-Molecular Probes, Eugene, OR) has been used extensively with eukaryotes and heterotrophic bacterial cells (Lebaron et al, 1998;Brussaard et al, 2001;Veldhuis et al, 2001;Klauth et al, 2004), there are few examples of its use with cyanobacteria . Therefore, trial testing was neces-sary with many different strains to determine the effectiveness of SYTOX Green and the appropriate concentration.…”

Section: Sytox Green Analysismentioning

confidence: 99%

Effect of exogenous extracellular polysaccharides on the desiccation and freezing tolerance of rock-inhabiting phototrophic microorganisms

Knowles¹,

Castenholz²

2008

FEMS Microbiology Ecology

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Two major stresses that threaten rock-inhabiting microbial communities are desiccation and freezing; both result in a loss of liquid water in the cells. The mechanisms necessary to tolerate these extremes may be similar, but are not well understood. In both cases extracellular polysaccharides (EPS) seem to play an important role. This study examines whether the EPS released by a rock-inhabiting phototroph can have a protective effect on other members of similar and neighboring microbial communities. This interaction was modeled by adding EPS isolated from the cryptoendolithic cyanobacterium Nostoc sp. to cells of the cryptoendolithic green alga Chlorella sp. and to cells of the epilithic cyanobacterium Chroococcidiopsis sp. The cells were then subjected to desiccation and freezing and the survival rates were determined by vital staining, using membrane integrity as a measure of viability. The results clearly demonstrate the importance of exogenous EPS in the desiccation tolerance of both species, while mixed results were found for the freezing trials.

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Effectiveness of SYTOX Green Stain for Bacterial Viability Assessment

Cited by 167 publications

References 19 publications

Effect of u.v. light irradiation, starvation and heat on Escherichia colibetabeta-D-galactosidase activity and other potential viability parameters

Effect of u.v. light irradiation, starvation and heat on Escherichia colibetabeta-D-galactosidase activity and other potential viability parameters

Effects of concentrated supernatants recovered fromLactobacillus plantarumonEscherichia coligrowth and on the viability of a human promyelocytic cell line

Effect of exogenous extracellular polysaccharides on the desiccation and freezing tolerance of rock-inhabiting phototrophic microorganisms

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