The kinetics and success of an industrial fermentation are dependent upon the health of the microorganism(s) responsible. Saccharomyces sp. are the most commonly used organisms in food and beverage production; consequently, many metrics of yeast health and stress have been previously correlated with morphological changes to fermentations kinetics. Many researchers and industries use machine vision to count yeast and assess health through dyes and image analysis. This study assessed known physical differences through automated image analysis taken throughout ongoing high stress fermentations at various temperatures (30 °C and 35 °C). Measured parameters included sugar consumption rate, number of yeast cells in suspension, yeast cross-sectional area, and vacuole cross-sectional area. The cell morphological properties were analyzed automatically using ImageJ software and validated using manual assessment. It was found that there were significant changes in cell area and ratio of vacuole to cell area over the fermentation. These changes were temperature dependent. The changes in morphology have implications for rates of cellular reactions and efficiency within industrial fermentation processes. The use of automated image analysis to quantify these parameters is possible using currently available systems and will provide additional tools to enhance our understanding of the fermentation process.

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This research aimed to assess how the partial removal of carbon dioxide affects fermentations to provide a better understanding of how the manipulation of carbon dioxide concentration can be used to optimize industrial fermentations. To achieve this, fermentation kinetics, fermentation metabolic products, and yeast stress indicators were analyzed throughout ongoing brewing fermentations conducted under partial vacuum with atmospheric pressure controls. The partial vacuum reduced the solubility of carbon dioxide in the media and decreased the time necessary to reach carbon dioxide saturation. The effect was an increased rate of fermentation, and significantly more viable cells produced under vacuum pressure compared to controls. Ethanol, glycerol, and volatile organic compound concentrations were all significantly increased under partial vacuum while indicators of yeast stress (trehalose) were reduced Additionally, as the number of yeast cells were higher under partial vacuum, less sugar was consumed per volume of yeast cell. This study measured fermentation kinetics, metabolic products, and yeast health to holistically assess the effect of partial vacuum during a batch fermentation and found significant differences in each that can be individually exploited by researchers and industry. Summary An exploration of batch yeast fermentation in a low-pressure environment, with a focus on the health and productivity of the yeast cells.

In this study the combinatory effect of several extrinsic factors on reduced (vacuum) pressure fermentations was explored. Specifically, the pressure, temperature, and FAN levels of high gravity Saccharomyces cerevisiae fermentations were manipulated, while yeast morphology was assessed using automated multivariate image analysis. Fermentation attributes including yeast growth, viability, and ethanol production were monitored using standard methods. Across all FAN and temperature levels, reduced pressure (vacuum pressure) fermentations resulted in a greater than or equal number of cells in suspension, higher average viability, and greater ethanol production in comparison to atmospheric pressure fermentations; however, the magnitude of the effect varied with extrinsic factors. The image analysis revealed that while yeast size was extremely variable across all fermentations, the ratio of vacuole to cell area consistently decreased over each fermentation and could be used to predict the point where the yeast experienced a sharp decline in viability ending the fermentation. This study showed that a combination of traditional measurements and novel automated analyses can be used by brewers to anticipate performance and endpoints of their fermentations, and that reduced pressure can have significant effects upon the rate and final ethanol concentration of variable industrial fermentations.