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The fermentation characteristics of five apple and five pear varieties from non-fertilized meadow orchards (“Streuobstwiesen”) was studied. While some varieties fermented without problems, some varieties showed sluggish and stuck fermentation with high residual sugar concentration at the end of the fermentation period. A potential reason for these fermentation problems could be a poor nitrogen supply. The effect of diammonium phosphate (DAP) supplementation on the fermentation dynamics was studied over two consecutive years, with 200.0 mg/L DAP added on day 0 or day 3 of fermentation. DAP effectively prevented stuck fermentations and accelerated sluggish fermentations. In particular, in the 2021 harvest, early DAP addition reduced the fermentation time by up to 6 days. The studied pear varieties indicated a greater demand for additional nitrogen compared to apple mashes. In general, nutrient addition proved beneficial for yeast cell growth of all ten fruit mashes. These findings suggest that the correct use of DAP could enhance the fermentation process in fruit mashes with low nutrient supply, which might be crucial for producing high-quality distillates from meadow orchard fruits.

Combined with real-time monitoring of density and temperature, the control of the redox potential provides a new approach to influencing cell metabolism during growth, cell viability and non-growing yeast activity in wine fermentations. Prior research indicates that the problem of sluggish and incomplete fermentation can be alleviated by maintaining a constant redox potential during the ethanol fermentation. A secondary trait of hydrogen sulfide formation from elemental sulfur also seems to be associated with the development of low redox potentials during fermentation and this might be prevented by the deliberate control of redox potentials in a certain range. While the control of the redox potential during wine fermentations has been demonstrated previously at the research scale (100 L), the ability to control it in larger volumes typically seen in commercial conditions remained unanswered. Wine fermentations from the same load of Cabernet Sauvignon grapes from the 2021 harvest were conducted at three volumes: 100 L and 1500 L in a research winery and 10,000 L in a commercial winery. Using only pulses of air delivery, the redox potential was successfully controlled to −40 mV referenced to a silver/silver chloride electrode throughout the fermentations, at all scales. This appears to be the first published result of a controlled fermentation trial that includes the commercial scale and demonstrates the scalability of control of redox potential in wine fermentations.

Societal Impact Statement Climate warming and demand for fuller‐bodied wines has resulted in increased grape sugar contents and, therefore, increased alcohol in wines. Concurrently, there are increased warnings about the health risks of alcohol, and consumers seek low‐alcohol options as part of a healthy lifestyle. High alcohol levels can also impact the flavor balance of wines, resulting in economic losses. However, exploration of native vineyard yeasts shows that fermentation with novel species can result in both alcohol reduction and improved wine flavor. Our findings show that native yeasts have the potential to bring the North American cool‐climate wine industry into a healthier future. Summary There is recent interest in developing wines fermented with non‐traditional yeasts that reflect the microbial terroir of the grape‐growing region. These native yeasts inhabit grape skins and can produce wines with distinctive flavor profiles, and when used in concert with traditional wine yeast, the risk of incomplete fermentation is minimized. Non‐traditional yeasts can also produce wines with relatively low alcohol content, an attractive characteristic given the health and societal issues associated with overconsumption. Yeasts isolated from L'Acadie and Pinot Noir grapes grown at Nova Scotia vineyards were identified by DNA sequencing and characterized regarding fermentative properties. Promising isolates of Saccharomyces uvarum, Hanseniaspora uvarum, Wickerhamomyces anomalus, and Zygotorulaspora florentina were selected for mixed and sequential fermentations with Saccharomyces cerevisiae based on their ethanol and SO2 tolerance, β‐glucosidase activity, and low H2S production. The resulting wines were evaluated for sensory properties by 15 experienced panelists. Alcohol levels of many of the fermentations that included non‐traditional yeasts were significantly lower than those made with traditional yeast alone, while fructose was higher, and total phenolic contents were similar. Experienced panelists also found that these fermentations resulted in wines with novel fruit and floral attributes. Our results demonstrate that native yeasts from cool‐climate wine regions can be used in concert with traditional wine yeast to produce novel, lower alcohol wines with a low risk of fermentation failure. This unlocks the potential of naturally occurring yeasts from these vineyards to create wines with flavor profiles that reflect the local wine‐making regions. Climate warming and demand for fuller‐bodied wines has resulted in increased grape sugar contents, and therefore increased alcohol in wines. Concurrently, there are increased warnings about the health risks of alcohol, and consumers seek low‐alcohol options as part of a healthy lifestyle. High alcohol levels can also impact the flavour balance of wines, resulting in economic losses. However, exploration of native vineyard yeasts shows that fermentation with novel species can result in both alcohol reduction and improved wine flavour. Our findings show that native yeasts have the potential to bring the North American cool‐climate wine industry into a healthier future.

On-site small-scale sanitation is common in rural areas and areas without infrastructure, but the treatment of the collected fecal matter can be inefficient and is seldom directed to resource recovery. The aim of this study was to compare low-technology solutions such as composting and lactic acid fermentation (LAF) followed by vermicomposting in terms of treatment efficiency, potential human and environmental risks, and stabilization of the material for reuse in agriculture. A specific and novel focus of the study was the fate of native pharmaceutical compounds in the fecal matter. Composting, with and without the addition of biochar, was monitored by temperature and CO2 production and compared with LAF. All treatments were run at three different ambient temperatures (7, 20, and 38°C) and followed by vermicomposting at room temperature. Materials resulting from composting and LAF were analyzed for fecal indicators, physicochemical characteristics, and residues of ten commonly used pharmaceuticals and compared to the initial substrate. Vermicomposting was used as secondary treatment and assessed by enumeration of Escherichia coli, worm density, and physicochemical characteristics. Composting at 38°C induced the highest microbial activity and resulted in better stability of the treated material, higher N content, lower numbers of fecal indicators, and less pharmaceutical compounds as compared to LAF. Even though analysis of pH after LAF suggested incomplete fermentation, E. coli cell numbers were significantly lower in all LAF treatments compared to composting at 7°C, and some of the anionic pharmaceutical compounds were detected in lower concentrations. The addition of approximately 5 vol % biochar to the composting did not yield significant differences in measured parameters. Vermicomposting further stabilized the material, and the treatments previously composted at 7°C and 20°C had the highest worm density. These results suggest that in small-scale decentralized sanitary facilities, the ambient temperatures can significantly influence the treatment and the options for safe reuse of the material.

Tea is the most popular and widely consumed beverage worldwide, especially black tea. Summer tea has a bitter and astringent taste and low aroma compared to spring tea due to the higher content of polyphenols and lower content of amino acids. Microbial fermentation is routinely used to improve the flavor of various foods. This study analyzed the relationship between the quality of black tea, metabolic characteristics, and microbial communities after microbial stuck fermentation in summer black tea. Stuck fermentation decreased the bitterness, astringency sourness, and freshness, and increased the sweetness, mellowness, and smoothness of summer black tea. The aroma also changed from sweet and floral to fungal, with a significant improvement in overall quality. Metabolomics analysis revealed significant changes in 551 non-volatile and 345 volatile metabolites after fermentation. The contents of compounds with bitter and astringent taste were decreased. Sweet flavor saccharides and aromatic lipids, and acetophenone and isophorone that impart fungal aroma showed a marked increase. These changes are the result of microbial activities, especially the secretion of extracellular enzymes. Aspergillus, Pullululanibacillus, and Bacillus contribute to the reduction of bitterness and astringency in summer black teas after stuck fermentation, and Paenibacillus and Basidiomycota_gen_Incertae_sedis contribute positively to sweetness. In addition, Aspergillus was associated with the formation of fungal aroma. In summary, our research will provide a suitable method for the improvement of tea quality and utilization of summer tea, as well as provide a reference for innovation and improvement in the food industry.

Abstract Nitrogen limitation is one of the most common causes for stuck or sluggish fermentation. A broad range of values have been reported as the minimum nitrogen concentration necessary for the completion of alcoholic fermentation. We have analyzed the minimum nitrogen concentration required to yield the maximum biomass (nitrogen reference value) using a microwell plate reader to monitor fermentation with different nitrogen sources and sugar concentrations. The biomass yield was dependent on the amount of available nitrogen, the nature of nitrogen source, and the sugar concentration in the medium. Nevertheless, achieving the maximum biomass was not sufficient to ensure the completion of the alcoholic fermentation, because the fermentation of 280 g sugar L−1 stuck, regardless of the nature and concentration of nitrogen source. However, a mixture of five amino acids (Leu, Ile, Val, Phe and Thr) as the nitrogen source allowed for maximum sugar consumption. Analysis of cell vitality by impedance showed a significant improvement in the vitality for cells fermenting using this amino acid combination.

Background Traditional bioethanol fermentation industries are not operated under strict sterile conditions and are prone to microbial contamination. Lactic acid bacteria (LAB) are often pervasive in fermentation tanks, competing for nutrients and producing inhibitory acids that have a negative impact on ethanol-producing yeast, resulting in decreased yields and stuck fermentations. Antibiotics are frequently used to combat contamination, but antibiotic stewardship has resulted in a shift to alternative antimicrobials. Results We demonstrate that endolysin LysMP, a bacteriophage-encoded peptidoglycan hydrolase, is an effective method for controlling growth of LAB. The LysMP gene was synthesized based on the prophage sequence in the genome of Limosilactobacillus fermentum KGL7. Analysis of the recombinant enzyme expressed in E. coli and purified by immobilized metal chelate affinity chromatography (IMAC) showed an optimal lysis activity against various LAB species at pH 6, with stability from pH 4 to 8 and from 20 to 40 °C up to 48 h. Moreover, it retains more than 80% of its activity at 10% ethanol (v/v) for up to 48 h. When LysMP was added at 250 µg/mL to yeast corn mash fermentations containing L. fermentum , it reduced bacterial load by at least 4-log fold compared to the untreated controls and prevented stuck fermentation. In comparison, untreated controls with contamination increased from an initial bacterial load of 1.50 × 10 7  CFU/mL to 2.25 × 10 9  CFU/mL and 1.89 × 10 9  CFU/mL after 24 h and 48 h, respectively. Glucose in the treated samples was fully utilized, while untreated controls with contamination had more than 4% (w/v) remaining at 48 h. Furthermore, there was at least a fivefold reduction in lactic acid (0.085 M untreated contamination controls compared to 0.016 M treated), and a fourfold reduction in acetic acid (0.027 M untreated contamination controls vs. 0.007 M treated), when LysMP was used to treat contaminated corn mash fermentations. Most importantly, final ethanol yields increased from 6.3% (w/v) in untreated contamination samples to 9.3% (w/v) in treated contamination samples, an approximate 50% increase to levels comparable to uncontaminated controls 9.3% (w/v). Conclusion LysMP could be a good alternative to replace antibiotics for mitigation of LAB contamination in biofuel refineries.

For three consecutive years, an Italian winery in Apulia has dealt with sudden alcoholic stuck fermentation in the early stages of vinification process, i.e., typical defects addressable to bacterial spoilage. After a prescreening trial, we assessed, for the first time, the influence of the commercial fungicide preparation Ridomil Gold® (Combi Pepite), containing Metalaxyl-M (4.85%) and Folpet (40%) as active principles, on the growth of several yeasts (Saccharomyces cerevisiae and non-Saccharomyces spp.) and lactic acid bacteria of oenological interest. We also tested, separately and in combination, the effects of Metalaxyl-M and Folpet molecules on microbial growth both in culture media and in grape must. We recalled the attention on Folpet negative effect on yeasts, extending its inhibitory spectrum on non-Saccharomyces (e.g., Candida spp.). Moreover, we highlighted a synergic effect of Metalaxyl-M and Folpet used together and a possible inhibitory role of the fungicide excipients. Interestingly, we identified the autochthonous S. cerevisiae strain E4 as moderately resistant to the Folpet toxicity. Our findings clearly indicate the urgent need for integrating the screening procedures for admission of pesticides for use on wine grape with trials testing their effects on the physiology of protechnological microbes.

Chardonnay, being the predominant white wine-grape cultivar in the Australian wine sector, is subject to widely varying winemaking processes with the aim of producing a variety of wine styles. Therefore, juice composition might not always be ideal for optimal fermentation outcomes. Our aim was to better understand the composition of Chardonnay juice and how compositional parameters impact on fermentation outcomes. This was achieved through a survey of 96 commercially prepared Chardonnay juices during the 2009 vintage. Common juice variables were estimated using near infrared spectroscopy, and elemental composition was determined using radial view inductively coupled plasma optical emission spectrometry. The influence of elemental composition on fermentation outcomes was assessed by fermentation of a defined medium formulated to reflect the composition and range of concentrations as determined by the juice survey. Yeast (Saccharomyces cerevisiae) strain effects were also assessed. Key parameters influencing fermentation outcomes were verified by laboratory scale fermentation of Chardonnay juice. This exploration of Chardonnay juice identified interactions between juice pH and potassium concentration as key factors impacting on fermentation performance and wine quality. Outcomes differed depending on yeast strain.

During red wine fermentation, high temperatures may cause stuck fermentation by affecting the physiology of fermenting yeast. This deleterious effect is the result of the complex interaction of temperature with other physicochemical parameters of grape juice, such as sugar and lipid content. The genetic background of fermenting yeast also interacts with this complex matrix and some strains are more resistant to high temperatures than others. Here, the temperature tolerance of nine commercial starters was evaluated, demonstrating that, at high sugar concentrations, half of them are sensitive to temperature. Using a classical backcross approach, one thermo-sensitive commercial starter was genetically improved by introducing quantitative trait loci conferring resistance to temperature. With this breeding program it is possible to obtain a thermo-resistant strain sharing most of its genome with the initial commercial starter. The parental and improved strains were compared for population growth and fermentation ability in various conditions. Despite their common genetic background, these two strains showed slight physiological differences in response to environmental changes that enable identification of the key physiological parameters influencing stuck fermentation.