Evaluation of the Fluorescent Dye 1-Anilino-8-Naphthalene Sulfonic Acid for Yeast Viability Determination

McCaig, Robert

doi:10.1094/asbcj-48-0022

Cited by 17 publications

(15 citation statements)

References 7 publications

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“…While this method has been used for many years to determine viability in wine fermentations (Boulton et al, 1996), several studies with brewer's yeast have found that methylene blue staining may overestimate the live cell concentration, especially at low viabilities (Chilver et al, 1978;King et al, 1981;McCaig, 1990). However, we (Schreiber and Block, 2001) and others (Morrisey, 1998) have found the same extended viability in problem fermentations using direct colony counts on plates as well.…”

Section: Discussionmentioning

confidence: 69%

Kinetic model for nitrogen‐limited wine fermentations

Cramer

Vlassides

Block

2001

Biotech & Bioengineering

114

143

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A physical and mathematical model for wine fermentation kinetics has been developed to predict sugar utilization curves based on experimental data from wine fermentations with various initial nitrogen and sugar concentrations in the juice. The model is based on: (1) yeast cell growth limited by nitrogen; (2) sugar utilization rates and ethanol production rates proportional solely to the number of viable cells; and (3) a death rate for cells proportional to alcohol content. All but one parameter in the model can be estimated from existing data. However, experiments to find this final parameter, a constant describing cell death, indicate that cell death may not be the critical factor in determining fermentation kinetics as cell viability remains significant until sugar utilization has ceased. The model, nevertheless, predicts a transition from normal to sluggish to stuck fermentations as initial nitrogen levels decrease. It also predicts that fermentations with high initial Brix levels may go to completion when supplemented with nitrogen in the form of ammonia. Therefore, we hypothesize that the model is valid but that ethanol causes the yeast cells to become inactive while remaining viable. Experimental verification of the model has been performed using flask-scale experiments. The model has also been used to evaluate the possibility of using nitrogen or viable cell additions to avoid or correct problem (i.e., sluggish or stuck) fermentations.

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Section: Discussionmentioning

confidence: 69%

Kinetic model for nitrogen‐limited wine fermentations

Cramer

Vlassides

Block

2001

Biotech & Bioengineering

114

143

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“…The commonly used viability tests are based on the bright-field stains methylene blue (MB) (Chilver et al, 1978) and methylene violet (MV) (Smart et al, 1999) and the fluorescent dye 1-anilino-8-naphthalenesulphonic acid (MgANS) (McCaig, 1990). MgANS differentiates dead cells clearly by entering nonviable cells and binding to cytoplamic proteins to generate yellow/green fluorescence (McCaig, 1990). The colourless 'leuco' form is thought to be the result of the slow uptake of the dye into viable cells to be oxidized whereas the dead cells cannot exclude the dye or perform this reaction.…”

Section: Cellars Fermentation Systemsmentioning

confidence: 99%

The yeastSaccharomyces cerevisiaeâ the main character in beer brewing

Lodolo¹,

Kock

Axcell³

et al. 2008

FEMS Yeast Research

204

169

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Historically, mankind and yeast developed a relationship that led to the discovery of fermented beverages. Numerous inventions have led to improved technologies and capabilities to optimize fermentation technology on an industrial scale. The role of brewing yeast in the beer-making process is reviewed and its importance as the main character is highlighted. On considering the various outcomes of functions in a brewery, it has been found that these functions are focused on supporting the supply of yeast requirements for fermentation and ultimately to maintain the integrity of the product. The functions/processes include: nutrient supply to the yeast (raw material supply for brewhouse wort production); utilities (supply of water, heat and cooling); quality assurance practices (hygiene practices, microbiological integrity measures and other specifications); plant automation (vessels, pipes, pumps, valves, sensors, stirrers and centrifuges); filtration and packaging (product preservation until consumption); distribution (consumer supply); and marketing (consumer awareness). Considering this value chain of beer production and the 'bottle neck' during production, the spotlight falls on fermentation, the age-old process where yeast transforms wort into beer.

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“…Fluorescence microscopy detection of yeast has been studied extensively in the past [11,15,22,32], but the majority of reported work involved use of pure yeast samples. To understand how the Xuorescence of yeast behaves in a corn mash medium, initial experiments were performed by staining YCM provided by Lincolnway Energy with various commercially available dyes such as SYTO 9 (Invitrogen), carboxyXuoroscein diacetate (CFDA, Invitrogen), calcein AM (Invitrogen), calcoXuor (Invitrogen), acridine orange (Biotium), propidium iodide (Biotium), ethidium bromide (Biotium), and 4',6-diamidino-2-phenylindole (DAPI) (Biotium) (Fig.…”

Section: Introductionmentioning

confidence: 99%

Direct concentration and viability measurement of yeast in corn mash using a novel imaging cytometry method

Chan¹,

Lyettefi²,

Pirani³

et al. 2010

J Ind Microbiol Biotechnol

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Worldwide awareness of fossil-fuel depletion and global warming has been increasing over the last 30 years. Numerous countries, including the USA and Brazil, have introduced large-scale industrial fermentation facilities for bioethanol, biobutanol, or biodiesel production. Most of these biofuel facilities perform fermentation using standard baker's yeasts that ferment sugar present in corn mash, sugar cane, or other glucose media. In research and development in the biofuel industry, selection of yeast strains (for higher ethanol tolerance) and fermentation conditions (yeast concentration, temperature, pH, nutrients, etc.) can be studied to optimize fermentation performance. Yeast viability measurement is needed to identify higher ethanol-tolerant yeast strains, which may prolong the fermentation cycle and increase biofuel output. In addition, yeast concentration may be optimized to improve fermentation performance. Therefore, it is important to develop a simple method for concentration and viability measurement of fermenting yeast. In this work, we demonstrate an imaging cytometry method for concentration and viability measurements of yeast in corn mash directly from operating fermenters. It employs an automated cell counter, a dilution buffer, and staining solution from Nexcelom Bioscience to perform enumeration. The proposed method enables specific fluorescence detection of viable and nonviable yeasts, which can generate precise results for concentration and viability of yeast in corn mash. This method can provide an essential tool for research and development in the biofuel industry and may be incorporated into manufacturing to monitor yeast concentration and viability efficiently during the fermentation process.

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Evaluation of the Fluorescent Dye 1-Anilino-8-Naphthalene Sulfonic Acid for Yeast Viability Determination

Cited by 17 publications

References 7 publications

Kinetic model for nitrogen‐limited wine fermentations

Kinetic model for nitrogen‐limited wine fermentations

The yeastSaccharomyces cerevisiaeâ the main character in beer brewing

Direct concentration and viability measurement of yeast in corn mash using a novel imaging cytometry method

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Evaluation of the Fluorescent Dye 1-Anilino-8-Naphthalene Sulfonic Acid for Yeast Viability Determination

Cited by 17 publications

References 7 publications

Kinetic model for nitrogen‐limited wine fermentations

Kinetic model for nitrogen‐limited wine fermentations

The yeastSaccharomyces cerevisiaeâ the main character in beer brewing

Direct concentration and viability measurement of yeast in corn mash using a novel imaging cytometry method

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Product

Resources

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The yeastSaccharomyces cerevisiaeâ the main character in beer brewing