Molecular methods for the assessment of bacterial viability

Keer, Jacquie T.; Birch, L.

doi:10.1016/s0167-7012(03)00025-3

Cited by 345 publications

(241 citation statements)

References 67 publications

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“…Cell viability could also be reflected by changes in morphology, motility or the membrane or physiological state, including production of enzymes for healthy metabolism, such as esterase and oxidoreductase, the ability to pump out or exclude certain dyes or levels of macromolecules important for life, such as ATP, proteins and nucleic acids (reviewed by Breeuwer & Abee 2000; Keer & Birch 2003).…”

Section: Introductionmentioning

confidence: 99%

Applicability of the MTT assay for measuring viability of cyanobacteria and algae, specifically for Microcystis aeruginosa (Chroococcales, Cyanobacteria)

Li¹,

Song²

2007

Phycologia

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Several assay methods were screened for viability assessment in cyanobacteria using Microcystis aeruginosa FACHB 905. Compared with fluorescent diacetate (FDA), Evan's Blue and autofluorescence, the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, which was based on the ability of viable cells to reduce MTT to formazan, was found to be reliable and was selected for further study. MTT concentration, incubation time and temperature were optimized for M. aeruginosa. Improvements to the sensitivity and reproducibility of the MTT assay included performing it in the dark to reduce the effects of formazan light sensitivity when extracted in DMSO. Another improvement involved collecting viability data by cell by counting rather than colourimetrically, which was concluded from the fact that oxidoreductase activity, responsible for MTT reduction, would elevate or decrease under stress conditions. Half-life of oxidoreductase in dead cell was calculated to be 3 h. The MTT assay was also found to be applicable to other cyanobacteria and diatoms, including field samples, but not for algae belonging to Chlorophyta, Euglenophyta, Pyrrophyta or Chrysophyta. Based on the above results, we proposed an optimized procedure for the MTT method on Microcystis strains. The use of this assay may be of importance to better understand the dynamics of bloom and the fate of Microcystis under natural or disturbed conditions.

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Section: Introductionmentioning

confidence: 99%

Applicability of the MTT assay for measuring viability of cyanobacteria and algae, specifically for Microcystis aeruginosa (Chroococcales, Cyanobacteria)

Li¹,

Song²

2007

Phycologia

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“…In addition to these limitations, several days of incubation are often needed for bacteria to form countable colonies. Recently, new viability assays were developed for bacteria (Keer and Birch 2003). These tests were essentially based on respiratory activity (Bhupathiraju et al 1999;Rodriguez et al 1992), external membrane integrity (Nebe-von-Caron et al 2000), intracellular pH (Chitara et al 2000), and membrane potential (Deere et al 1995;Manson et al 1993Manson et al , 1994Manson et al , 1995.…”

Section: Introductionmentioning

confidence: 99%

Flow cytometry analysis of germinating Bacillus spores, using membrane potential dye

Laflamme

Veillette

et al. 2004

Arch Microbiol

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“…In recent years, various molecular techniques have been used for the lysis of L. casei by chemical and mechanical protocols (12). In chemical methods, the peptidoglycan can be lysed by cell wall hydrolase enzymes (e.g.…”

Section: Introductionmentioning

confidence: 99%

An Efficient DNA Extraction Method for Lactobacillus casei, a Difficult-to-Lyse Bacterium

Alimolaei¹,

Golchin²

2016

Int J Enteric Pathog

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Background: There are several protocols to extract DNA from Lactobacillus spp. In the case of L. casei it is harder because of its especial and thick cell wall. Objectives: In this study, nine DNA extraction protocols (by lysozyme treatment) were evaluated and compared in two categories (traditional and kit-based protocols) and an improved method was presented. Materials and Methods: DNA quantity and quality was determined by spectrophotometry, agarose gel electrophoresis and polymerase chain reaction (PCR). Results:The results revealed that the yield of extracted DNA differed by each protocol (5.8 -17.1 μg/100 μL), but provided appropriate DNA for PCR amplification. The modified protocol offered the best total DNA extraction method when both quality (DNA purity; 1.54 μg) and quantity (DNA yield; 17.1 μg) were considered. Conclusions: We suggest this protocol for effective and inexpensive DNA isolation from L. casei for downstream biological processes such as PCR.

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Molecular methods for the assessment of bacterial viability

Cited by 345 publications

References 67 publications

Applicability of the MTT assay for measuring viability of cyanobacteria and algae, specifically for Microcystis aeruginosa (Chroococcales, Cyanobacteria)

Applicability of the MTT assay for measuring viability of cyanobacteria and algae, specifically for Microcystis aeruginosa (Chroococcales, Cyanobacteria)

Flow cytometry analysis of germinating Bacillus spores, using membrane potential dye

An Efficient DNA Extraction Method for Lactobacillus casei, a Difficult-to-Lyse Bacterium

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