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Yeast Viability and Vitality

  • Graham G. Stewart
Chapter
Part of the The Yeast Handbook book series (YEASTHDB)

Abstract

Despite the frequent use of the terms yeast viability and yeast vitality in both the brewing and distilling literature, the application of these terms is often confused. There are a number of analyses to show that yeast is alive; the relationships between a cell’s environment and the methods employed for its cultivation are complex! These methods include the ability of cells to grow on solid or in liquid media and strain-based systems; some are based on changes in the charges and functionality of the cell membrane, and others are dyes that penetrate into live and/or dead cells. The methods for assessing cell viability only provide information on live and dead cells in a whole population. Methods for yeast vitality determination are based on various physiological and metabolic aspects of cells. These methods include:
  • Intracellular ATP content based on the luciferin reaction

  • Determination of mitochondrial membrane potential

  • Acidification power test following the addition of glucose

  • Magnesium release test based on the fact that low molecular weight ions such as magnesium and others are released by yeast following addition of glucose to the medium

  • Determination of the activity of enzymes such as esterase, oxidoreductases or several different redox enzymes

Recent studies have aimed to develop a clear classification to develop methods for the analysis of both the viability and vitality of yeast.

References

  1. Aeschbacher M, Reinhardt CA, Zbinden G (1986) A rapid cell membrane permeability test using fluorescent dyes and flow cytometry. Cell Biol Toxicol 2:247–255CrossRefPubMedGoogle Scholar
  2. Ansehn S, Nilsson L (1984) Direct membrane-damaging effect of ketoconazole and tioconazole on Candida albicans demonstrated by bioluminescent assay of ATP. Antimicrob Agents Chemother 26:22–25CrossRefPubMedPubMedCentralGoogle Scholar
  3. Austriaco NR (1996) Review: To bud until death: the genetics of ageing in the yeast Saccharomyces. Yeast 12:623 630CrossRefGoogle Scholar
  4. Bamforth C, Lentini A (2009) The flavor instability of beer. In: Bamforth CW (ed) Beer: a quality perspective. Elsevier, Boston, MA, pp 85–109CrossRefGoogle Scholar
  5. Bapat P, Nandy SK, Wangikar P, Venkatesh KV (2006) Quantification of metabolically active biomass using Methylene Blue dye Reduction Test (MBRT): measurement of CFU in about 200 s. J Microbiol Methods 65:107–116CrossRefPubMedGoogle Scholar
  6. Barker MG, Smart KA (1996) Morphological changes associated with the cellular ageing of a brewing yeast strain. J Am Soc Brew Chem 54:121–126Google Scholar
  7. Bochner BS, McKelvey AA, Schleimer RP, Hildreth JEK, DW MG Jr (1989) Flow cytometric methods for the analysis of human basophil surface antigens and viability. J Immunol Methods 125:265–271CrossRefPubMedGoogle Scholar
  8. Bolat I (2008) The importance of trehalose in brewing yeast survival. Innov Roman Food Biotech 2:1–10Google Scholar
  9. Botstein D, Fink GR (2011) Yeast: an experimental organism for 21st century biology. Genetics 189:685–704CrossRefGoogle Scholar
  10. Boulton C, Quain D (2001) Brewing yeast and fermentation. Blackwell Science, OxfordGoogle Scholar
  11. Breeuwer P, Jean-Louis D, Bunschoten N (1995) Characterization of uptake and hydrolysis of fluorescein diacetate and carboxyfluorescein diacetate by intracellular esterases in Saccharomyces cerevisiae, which result in accumulation of fluorescent product. Appl Environ Microbiol 61:1614–1619PubMedPubMedCentralGoogle Scholar
  12. Carvell JP, Turner K (2003) New applications and methods utilizing radio-frequency impedance measurements for improving yeast management. MBAA Tech Quart 40:30–38Google Scholar
  13. Carvell JP, Austin G, Matthee A, Van de Spiegle K, Cunningham S, Harding C (2000) Developments in using off-line radio frequency impedance methods for measuring viable cell concentration in the brewery. J Am Soc Brew Chem 58:57–62Google Scholar
  14. Casey G, Chen E, Ingledew W (1985) High gravity brewing: production of high levels of ethanol without excessive concentrations of esters and fusel alcohols. J Am Soc Brew Chem 43:179–182Google Scholar
  15. Castro FAV, Mariani D, Panek AD, Eleutherio ECA, Pereira MD (2008) Cytotoxicity mechanism of two naphthoquinones (menadione and plumbagin) in Saccharomyces cerevisiae. PLoS One 3(12):e3999CrossRefPubMedPubMedCentralGoogle Scholar
  16. Chan L, Driscoll D, Kuksin D, Saldi S (2016) Measuring lager and ale yeast viability and vitality using fluorescence-based image cytometry. MBAA Tech Quart 53:49–54Google Scholar
  17. Chrzanowski TH, Crotty RD, Hubbard JG, Welch RP (1984) Applicability of the fluorescein diacetate method of detecting active bacteria in freshwater. Microb Ecol 10:179–185CrossRefPubMedGoogle Scholar
  18. Cooper DJ, Stewart GG, Bryce JH (1998) Some reasons why high gravity brewing has a negative effect on head retention. J Inst Brew 104:221–228CrossRefGoogle Scholar
  19. Cooper DJ, Stewart GG, Bryce JH (2000) Yeast proteolytic activity during high and low gravity wort fermentations and its effect on head retention. J Inst Brew 106:197–202CrossRefGoogle Scholar
  20. Cunningham (2000) The reaction of brewer’s yeast from different fermentation conditions to acid washing. PhD thesis, Heriot Watt University, EdinburghGoogle Scholar
  21. Cunningham S, Stewart GG (2000) Acid washing and serial repitching a brewing ale strain of Saccharomyces cerevisiae in high gravity wort and the role of wort oxygenation conditions. J Inst Brew 106:389–402CrossRefGoogle Scholar
  22. Czekanska EM (2011) Assessment of cell proliferation with resazurin-based fluorescent dye. Methods Mol Biol 740:27–32CrossRefPubMedGoogle Scholar
  23. D’Amore T, Russell I, Stewart GG (1989) Sugar utilization by yeast during fermentation. J Ind Microbiol 4:315–324CrossRefGoogle Scholar
  24. D’Amore T, Crumplen R, Stewart GG (1991) The involvement of trehalose in yeast stress tolerance. J Ind Microbiol 7:191–196CrossRefGoogle Scholar
  25. Davey HM (2011) Life, death, and in-between: meanings and methods in microbiology. Appl Environ Microbiol 77:5571–5576CrossRefPubMedPubMedCentralGoogle Scholar
  26. Dinsdale MG, Loyd D, Jarvis B (1995) Yeast vitality during cider fermentation: two approaches to the measurement of membrane potential. J Inst Brew 101:453–458CrossRefGoogle Scholar
  27. Ernandes JR, D’Amore T, Russell I, Stewart GG (1992) Regulation of glucose and maltose transport in strains of Saccharomyces. J Ind Microbiol 9:127–130CrossRefGoogle Scholar
  28. Ernandes JR, Williams JW, Russell I, Stewart GG (1993) Respiratory deficiency in brewing yeast strains – effects on fermentation, flocculation and beer flavour components. J Am Soc Brew Chem 51:16–20Google Scholar
  29. Fannjiang Y, Cheng WC, Lee SJ, Qi B, Pevsner J, McCaffery JM, Hill RB, Basañez G, Hardwick JM (2004) Mitochondrial fission proteins regulate programmed cell death in yeast. Genes Dev 18:2785–2797CrossRefPubMedPubMedCentralGoogle Scholar
  30. Gadd GM, Chalmers K, Reed RH (1997) The role of trehalose in dehydration resistance in Saccharomyces cerevisiae. FEMS Microbiol Lett 48:249–254CrossRefGoogle Scholar
  31. Galton F (1869) Heredity genius: an enquiry into its laws and consequences. Macmillan, LondonGoogle Scholar
  32. Gänzle MG (2014) Enzymatic and bacterial conversions during sourdough fermentation. Food Microbiol 37:2–10CrossRefPubMedGoogle Scholar
  33. Gilliland RB (1959) Determination of yeast viability. J Inst Brew 65:424–429CrossRefGoogle Scholar
  34. Grant HL (1999) Hops. In: McCabe JT (ed) The practical brewer. Master Brewers Association of the Americas, Wauwatosa, WI, pp 201–219Google Scholar
  35. Hayflick L (1965) The limited in vitro lifespan of human diploid cell strains. Exp Cell Res 37:614–636CrossRefPubMedGoogle Scholar
  36. Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains. Exp Cell Res 25:585–621CrossRefPubMedGoogle Scholar
  37. Heggart H, Margaritis A, Pilkington H, Stewart RJ, Dowhanick TM, Russell I (1999) Factors affecting yeast viability and vitality characteristics: a review. MBAA Tech Quart 36:383–406Google Scholar
  38. Heggart H, Margaritis A, Pilkington H, Stewart RJ, Sobczak H, Russell I (2000) Measurement of brewing yeast viability and vitality: a review of methods. MBAA Tech Quart 37:409–430Google Scholar
  39. Imai T, Ohno T (1994) Measurement of yeast intracellular pH by image processing and the change it undergoes during growth phase. J Biotechnol 38:165–172CrossRefGoogle Scholar
  40. Imai T, Ohno T (1995) The relationship between viability and intracellular pH in the yeast Saccharomyces cerevisiae. Appl Environ Microbiol 61:3604–3608PubMedPubMedCentralGoogle Scholar
  41. Imai T, Nakajima I, Ohno T (1994) Development of a new method of evaluation of yeast vitality by measuring intracellular pH. J Am Soc Brew Chem 52:5–8Google Scholar
  42. Jamieson DJ (1998) Oxidative stress responses of the yeast Saccharomyces cerevisiae. Yeast 14:1511–1527CrossRefPubMedGoogle Scholar
  43. Jenkins CL, Kennedy AI, Hodgson JA, Thurston P, Smart KA (2003) Impact of serial repitching on lager brewing yeast quality. J Am Soc Brew Chem 61:1–9Google Scholar
  44. Jett BD, Hatter KL, Huycke MM, Gilmore MS (1997) Simplified agar plate method for quantifying viable bacteria. BioTechniques 23:648–650PubMedGoogle Scholar
  45. Jiménez J, Bru S, Ribeiro M, Clotet J (2015) Live fast, die soon: cell cycle progression and lifespan in yeast cells. Microb Cell 2:62–67CrossRefPubMedPubMedCentralGoogle Scholar
  46. Jones M, Pierce JS (1964) Absorption of amino acids from wort by yeasts. J Inst Brew 70:307–315CrossRefGoogle Scholar
  47. Kaneda H, Tokashio M, Tomaki T, Osawa T (1997) Influence of pH on flavour staling during beer storage. J Inst Brew 103:21–23CrossRefGoogle Scholar
  48. Kara BV, Simpson WJ, Hammond JRM (1988) Prediction of the fermentation performance of brewing yeast with the acidification power test. J Inst Brew 94:153–158CrossRefGoogle Scholar
  49. Kennedy AI, Taidi B, Dola JL, Hodsgon JA (1997) Optimisation of a fully defined medium for yeast fermentation studies. Food Technol Biotechnol 35:261–265Google Scholar
  50. Kirsop BH (1974) Oxygen in brewery fermentation. J Inst Brew 80:252–259CrossRefGoogle Scholar
  51. Knudsen FB (1999) Fermentation, principles and practices. In: McCabe JT (ed) The practical brewer. Master Brewers Association of the Americas, Wauwatosa, WI, pp 235–261Google Scholar
  52. Kotyk A (1963) Folia. Microbiol (Prague) 8:27–31CrossRefGoogle Scholar
  53. Kwolek-Mirek M, Zadrag-Tecza R (2014) Comparison of methods used for assessing the viability and vitality of yeast cells. FEMS Yeast Res 14:1068–1079PubMedGoogle Scholar
  54. Kwolek-Mirek M, Bednarska S, Bartosz G, Bilinski T (2009) Acrolein toxicity involves oxidative stress caused by glutathione depletion in the yeast Saccharomyces cerevisiae. Cell Biol Toxicol 25:363–378CrossRefPubMedGoogle Scholar
  55. Kwolek-Mirek M, Bednarska S, Zadrąg-Tęcza R, Bartosz G (2011) The hydrolytic activity of esterases in the yeast Saccharomyces cerevisiae is strain dependent. Cell Biol Int 35:1111–1119CrossRefPubMedGoogle Scholar
  56. Layfield JB, Sheppard JD (2015) What brewers should know about viability, vitality, and overall brewing fitness: a mini-review. MBAA Tech Quart 52:132–140Google Scholar
  57. Lekkas C, Stewart GG, Hill AE, Taidi B, Hodgson J (2007) Elucidation of the role of nitrogenous wort components in yeast fermentation. J Inst Brew 113:3–8CrossRefGoogle Scholar
  58. Lentini A (1993) A review of the various methods available for monitoring the physiological status of yeast: yeast viability and vitality. Fermentation 6:321–327Google Scholar
  59. Levitz SM, Diamond RD (1985) A rapid colorimetric assay of fungal viability with the tetrazolium salt MTT. J Infect Dis 152:938–945CrossRefPubMedGoogle Scholar
  60. Lloyd D, Hayes AJ (1995) Vigour, vitality and viability of microorganisms. FEMS Microbiol Lett 133:1–7CrossRefGoogle Scholar
  61. Lodolo EJ, Kock JLF, Axcell BC, Brooks M (2008) The yeast Saccharomyces cerevisiae – the main character in beer brewing. FEMS Yeast Res 8:1018–1036. Special Issue: Thematic issue: Alcoholic fermentation: beverages to biofuelGoogle Scholar
  62. Longo VD, Shadel GS, Kaeberlein M, Kennedy B (2012) Replicative and chronological aging in Saccharomyces cerevisiae. Cell Metab 16:18–31CrossRefPubMedPubMedCentralGoogle Scholar
  63. Ludovico P, Sansonetty F, Corte-Real M (2001) Assessment of mitochondrial membrane potential in yeast cell populations by flow cytometry. Microbiology 147:3335–3343CrossRefPubMedGoogle Scholar
  64. Maneval WE (1929) Some staining methods for bacteria and yeasts. Stain Technol 4:21–25CrossRefGoogle Scholar
  65. Marchi E, Cavalieri D (2008) Yeast as a model to investigate the mitochondrial role in adaptation to dietary fat and calorie surplus. Genes Nutr 3:159–166CrossRefPubMedPubMedCentralGoogle Scholar
  66. McGahon AJ, Martin SJ, Bissonnette RP, Mahboubi A, Shi Y, Mogil RJ, Nishioka WK, Green DR (1995) The end of the (cell) line: methods for the study of apoptosis in vitro. In: Schwartz LM, Osborne BA (eds) Methods in cell biology: vol 46. Cell death. Academic Press, New York, pp 153–187Google Scholar
  67. Millard PJ, Roth BL, Thi HPT, Yue ST, Haugland RP (1997) Development of the FUN-1 family of fluorescent probes for vacuole labeling and viability testing of yeasts. Appl Environ Microbiol 63:2897–2905PubMedPubMedCentralGoogle Scholar
  68. Minois N, Frajnt M, Wilson C, Vaupel JW (2005) Advances in measuring lifespan in the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci USA 102:402–406CrossRefPubMedGoogle Scholar
  69. Mochaba FM, O’Connor-Cox ESC, Axcell BC (1997) A novel and practical yeast vitality method based on magnesium ion release. J Inst Brew 103:99–102CrossRefGoogle Scholar
  70. Mochaba FM, O’Connor-Cox ESC, Axcell BC (1998) Practical procedures to measure yeast viability and vitality prior to pitching. J Am Soc Brew Chem 56:1–6Google Scholar
  71. Murray CR, Barich T, Taylor D (1984) The effect of yeast storage conditions on subsequent fermentations. MBAA Tech Quart 21:189–194Google Scholar
  72. Nielsen O (2005) Control of the yeast propagation process – how to optimize oxygen supply and minimize stress. MBAA Tech Quart 42:128–132Google Scholar
  73. Nielsen O (2010) Status of the yeast propagation process and some aspects of propagation for re-fermentation. Cerevisia 35:71–74CrossRefGoogle Scholar
  74. Nikolova M, Savova I, Marinov M (2002) An optimised method for investigation of the yeast viability by means of fluorescent microscopy. J Cult Collect 3:66–71Google Scholar
  75. Novak S, D’Amore T, Stewart GG (1990) 2-Deoxy-d-glucose resistant yeast with altered sugar transport activity. FEBS Lett 269:202–204CrossRefPubMedGoogle Scholar
  76. O’Brien J, Wilson I, Orton T, Pognan F (2000) Investigation of the alamar blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem 267:5421–5426CrossRefPubMedGoogle Scholar
  77. Odumeru JA, D’Amore T, Russell I, Stewart GG (1992) Change in protein composition of Saccharomyces brewing in response to heat shock and ethanol stress. J Ind Microbiol 9:229–234CrossRefGoogle Scholar
  78. Painting K, Kirsop B (1990) A quick method for estimating the percentage of viable cells in a yeast population, using methylene blue staining. World J Microbiol Biotechnol 6:346–347CrossRefPubMedGoogle Scholar
  79. Piskur J, Rozpedowska E, Polakova S, Merico A, Compagno C (2006) How did Saccharomyces evolve to become a good brewer? Trends Genet 22:183–186CrossRefPubMedGoogle Scholar
  80. Powell CD, Van Zandycke SM, Quain DE, Smart KA (2000) Replicative ageing and senescence in Saccharomyces cerevisiae and the impact on brewing fermentations. Microbiology 146:1023–1034CrossRefPubMedGoogle Scholar
  81. Powell CD, Quain DE, Smart KA (2003) The impact of brewing yeast cell age on fermentation performance, attenuation and flocculation. FEMS Yeast Res 3:149–157CrossRefPubMedGoogle Scholar
  82. Pratt-Marshall PL (2002) High gravity brewing—an inducer of yeast stress. Its effect on cellular morphology and physiology. Ph.D. thesis, Heriot-Watt University, Edinburgh, ScotlandGoogle Scholar
  83. Pratt-Marshall PL, Bryce JH, Stewart GG (2003) The effects of osmotic pressure and ethanol on yeast viability and morphology. J Inst Brew 109:218–228CrossRefGoogle Scholar
  84. Quain DE, Tubb RS (1982) The importance of glycogen in brewing yeasts. MBAA Tech Quart 19:29–33Google Scholar
  85. Russell I, Stewart GG (eds) (2014) Whisky: Technology, Production and Marketing, 2nd edn. Academic Press (Elsevier), Boston, MAGoogle Scholar
  86. Sheppard JD, Dawson PSS (1999) Cell synchrony and periodic behavior in yeast populations. Can J Chem Eng 77:893–902CrossRefGoogle Scholar
  87. Simpson WJ, Hammond JRM (1989) Cold ATP extractants compatible with constant light signal firefly luciferase reagents. In: Stanley PE, McCarthy BJ, Smither R (eds) ATP luminescence: rapid methods in microbiology. Society for Applied Bacteriology technical series, vol 26. Blackwell Scientific Publications, Oxford, pp 45–52Google Scholar
  88. Slavik J (1982) Intracellular pH of yeast cells measured with fluorescent probes. FEBS Lett 140:22–26CrossRefPubMedGoogle Scholar
  89. Smart A, Whisker S (1996) Effect of serial repitching on the fermentation properties and condition of brewing yeast. J Am Soc Brew Chem 54:41–44Google Scholar
  90. Smart KA, Chambers KM, Lambert I, Jenkins C (1999) Use of methylene violet staining procedures to determine yeast viability and vitality. J Am Soc Brew Chem 57:18–23Google Scholar
  91. Stewart GG (2006) Studies on the uptake and metabolism of wort sugars during brewing fermentations. MBAA Tech Quart 43:265–269Google Scholar
  92. Stewart GG (2010) High gravity brewing and distilling – past experiences and future prospects. J Am Soc Brew Chem 68:1–9Google Scholar
  93. Stewart GG (2014a) Brewing intensification. American Society of Brewing Chemists, St Paul, MNGoogle Scholar
  94. Stewart GG (2014b) The concept of nature-nurture applied to brewer’s yeast and wort fermentation. MBAA Tech Quart 51:69–80Google Scholar
  95. Stewart GG (2015) Seduced by yeast. J Am Soc Brew Chem 73:1–21Google Scholar
  96. Stewart GG, Murray J (2010) A selective history of high gravity and high alcohol beers. MBAA Tech Quart 47: TQ-47-2-0416-01Google Scholar
  97. Stewart GG, Russell I, Goring TE (1975) Nature-nurture anomalies – further studies in yeast flocculation. Am Soc Brew Chem Proc 33:137–147Google Scholar
  98. Valli M, Sauer M, Branduardi V, Borth N, Porro D, Mattanovich D (2005) Intracellular pH distribution in Saccharomyces cerevisiae cell populations, analyzed by flow cytometry. Appl Environ Microbiol 71:1515–1521CrossRefPubMedPubMedCentralGoogle Scholar
  99. Walker GM, Chandrasena G, Birch RM, Maynard A (1995) Proceedings of the 4th Aviemore malting, brewing and distilling conference, 185–192Google Scholar
  100. Zheng X, D’Amore T, Russell I, Stewart GG (1994a) Transport kinetics of maltose and maltotriose in strains of Saccharomyces. J Ind Microbiol Biotechnol 13:159–166Google Scholar
  101. Zheng X, D’Amore T, Russell I, Stewart GG (1994b) Factors influencing maltotriose utilization during brewery wort fermentations. J Am Soc Brew Chem 52:41–47Google Scholar
  102. Zitomer RS, Lowry CV (1992) Regulation of gene expression by oxygen in Saccharomyces cerevisiae. Microbiol Rev 56:1–11PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  • Graham G. Stewart
    • 1
    • 2
  1. 1.International Centre for Brewing and DistillingHeriot Watt UniversityEdinburghUK
  2. 2.GGStewart AssociatesCardiff, WalesUK

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