Evaluation of the alkaline methylene blue staining method for yeast activity determination

doi:10.1016/0922-338X(94)90292-5

Journal of Fermentation and Bioengineering

Volume 78, Issue 3, 1994, Pages 212-216

Journal of Fermentat...

https://doi.org/10.1016/0922-338X(94)90292-5 Get rights and content

Abstract

To ensure reliable manufacture of high quality beer, the brewing yeast must be maintained in a highly active state. The conventional methylene blue staining method, which is very simple but not highly sensitive, has been improved to allow sensitive measurement of yeast activity. Two kinds of yeasts with different activity levels were stained with methylene blue solution with pH levels between 4 and 12. The results showed that highly sensitive measurement of yeast activity was possible when the staining solution was alkaline. When cells were stained with methylene blue solution with a pH of 10.6 at 25°C for 15 min (the alkaline methylene blue staining method), the percentage of unstained cells was consistent with viability measurements made using the slide culture method. The alkaline methylene blue staining method makes possible sensitive, quick, and simple measurement of yeast activity, which is essential to the management of fermentation process.

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There are more references available in the full text version of this article.

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To examine whether the inclusion of Δmnn9 in the diet has immunostimulatory benefits on C. gigas juveniles (5.94 ± 0.67 mm shell length; 33.1 ± 5.9 mg ind−1 (mean ± SD; N = 50)), five pre-challenge diets were tested for 24 h: the bi-algal diet (100A) and its substituted version with Δmnn9 at four levels, 5%, 10%, 25% and 50% (95A5Y, 90A10Y, 75A25Y, 50A50Y, respectively). In order to verify whether oysters can digest Δmnn9 cells, faeces from treatments 50A50Y and 75A25Y were collected during the pre-challenge feeding period and stained with methylene blue (MB) according to Sami et al. [35]. The staining technique is based on the assumption that MB is able to enter cells of which the selective permeability of the plasma membrane has been destroyed or severely compromised, allowing the discrimination between living/viable (unstained) and dead/unviable (blue-stained) Δmnn9 cells [35,36].
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Removal of methanol vapors in a jet-loop bioscrubber equipped with a venturi injector
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Citation Excerpt :
During the last stages of the JLR operation, the occurrence of yeast cells was observed. The yeast concentration was determined with a serial dilution procedure and trough counting in a Neubauer chamber after alkaline methylene blue staining [26]. The JLR was operated with coalescing media (tap water) and non-coalescing media (mineral medium).
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The pelleted TMR was stored for one month before the start of the formal animal experiment. At the beginning of the formal experimental period, agglomerated LY and feed samples were collected in triplicate for LY counts using microscopy in the alkaline methylene blue staining method (Sami et al., 1994). The survival rate of LY is defined as the percentage of LY counts in feed after pelleting in the total counts of LY supplied to TMR before pelleting.
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In the presence of all three Ns i.e. RuCTAC, RuDDA, and RuHEXA, S. cerevisiae cells were observed to be both viable and non-viable indicating the effect of Ns on cells (Fig. 5b, c, & d), arrow specifies the viable cells. Viable cells also attain the dye but stain interacts with the active enzymes i.e. oxidoreductase presents within them leading to the reduction of the methylene dye, making live S. cerevisiae cells appear colorless (Sami et al., 1994). FESEM investigation was further done to observe the Ns effect.
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Citation Excerpt :
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Journal of Fermentation and Bioengineering

Abstract

References (15)

Ion transport in yeast

Biochim. Biophys. Acta

Proton pumps of the plasmalemma of the yeast Rhodotorula gracilis their coupling to fluxes of potassium and other ions

Biochim. Biophys. Acta

Analytica microbiologica. Method 2. 2. 2. 3

J. Inst. Brew.

Method of analysis

Report of the group

J. Inst. Brew.

Analysis committee report on measurement of yeast viability

J. Inst. Brew.

Method of analysis

Cited by (65)

Controlling the growth of yeast by culturing in high magnetic fields

β-1,3-Glucan/chitin unmasking in the Saccharomyces cerevisiae mutant, Δmnn9, promotes immune response and resistance of the Pacific oyster (Crassostrea gigas) to Vibrio coralliilyticus infection

Removal of methanol vapors in a jet-loop bioscrubber equipped with a venturi injector

Agglomerated live yeast (Saccharomyces cerevisiae) supplemented to pelleted total mixed rations improves the growth performance of fattening lambs

Toxicity profiling of metallosurfactant based ruthenium and ruthenium oxide nanoparticles towards the eukaryotic model organism Saccharomyces cerevisiae

Efficient drop reactor processing of methylene blue degradation with silver nanowire catalysts