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Most commercially available red wines undergo alcoholic fermentation by Saccharomyces yeasts, followed by a second fermentation with the lactic acid bacteria Oenococcus oeni once the initial process is complete. However, this traditional approach can encounter complications in specific scenarios. These situations pose risks such as stalled alcoholic fermentation or the growth of undesirable bacteria while the process remains incomplete, leaving residual sugars in the wine. To address these challenges and the issue of low acidity prevalent in warmer viticultural regions, several novel alternatives are available. The alternatives involve the combined use of Lachancea thermotolerans to increase the acidity of the musts, lactic acid bacteria (Oenococcus oeni and Lactiplantibacillus plantarum) to ensure malic acid stability during early alcoholic fermentation stages, and Saccharomyces cerevisiae to properly complete alcoholic fermentation. The study showed variations in the final chemical parameters of wines based on the microorganisms used.

The study explores diverse strains of Lachancea thermotolerans in single-inoculum wine fermentation conditions using synthetic grape must. It aims to analyze the role of the species without external influences like other microorganisms or natural grape must variability. Commercial strains and selected vineyard isolates, untested together previously, are assessed. The research evaluates volatile and non-volatile chemical compounds in final wine, revealing significant strain-based variations. L. thermotolerans notably produces lactic acid and consumes malic acid, exhibiting moderate ethanol levels. The volatile profile displays strain-specific impacts, affecting higher alcohol and ester concentrations compared to S. cerevisiae. These effects vary based on the specific compounds. Using a uniform synthetic must enables direct strain comparisons, eliminating grape-related, environmental, or timing variables in the experiment, facilitating clearer insights into the behavior of L. thermotolerans in wine fermentation. The study compares for the first time all available commercial strains of L. thermotolerans.

Hydrogen can drastically degrade the mechanical properties of a variety of metallic materials. The so-called hydrogen environment embrittlement of austenitic CrNi-type steels is usually accompanied by the formation of secondary surface cracks, which can be investigated in order to assess the embrittlement process. The occurrence of hydrogen-induced cracks is often related to element segregation effects that locally impact the austenite stability. Since there is as yet a lack of investigation methods that can visualize both structures three-dimensionally, the present study investigates the imageability of hydrogen-induced cracks and element segregation structures in austenitic CrNi-steel via micro-computed tomography (CT). In order to improve the X-ray visibility of segregation structures, modified versions of the reference steel, X2CrNi18-9, that contain W and Si are designed and investigated. The investigations demonstrated that small differences in the X-ray attenuation, caused by the W or Si modifications, can be detected via CT, although segregation structures could not be imaged due to their small size scale and image noise. Hydrogen-induced cracks were characterized successfully; however, the detection of the smaller cracks is limited by the resolution capability.

Chitosan exerts a significant influence on various chemical parameters affecting the quality of wine produced using multiple strains of Lachancea thermotolerans. The impact of chitosan on these parameters varies depending on the specific strain studied. We observed that, under the influence of chitosan, the fermentation kinetics accelerated for all examined strains. The formation of lactic acid increased by 41% to 97% across the studied L. thermotolerans strains, depending on the specific strain. This effect also influenced acidity-related parameters such as total acidity, which increased by 28% to 60%, and pH, which experienced a decrease of over 0.5 units. The consumption of malic acid increased by 9% to 20% depending on the specific strain of L. thermotolerans. Nitrogen consumption also rose, as evidenced by all L. thermotolerans strains exhibiting a residual value of Primary Amino Nitrogen (PAN) of below the detection limit, and ammonia consumption increased by 90% to 100%, depending on the strain studied. However, certain parameters such as acetic acid, succinic acid, and glycerol showed contradictory results depending on the strain under investigation. In terms of volatile composition, chitosan supplementation led to increased production of i-butanol by 32% to 65%, 3-methylbutanol by 33% to 63%, and lactic acid ethyl ester by 58% to 91% across all studied strains of L. thermotolerans. Other analyzed aroma compounds exhibited varying changes depending on the specific strain of L. thermotolerans.

The development of new fermentative strategies exploiting the potential of different wine-related species is of great interest for new winemaking conditions and consumer preferences. One of the most promising non-conventional approaches to wine fermentation is the combined use of deacidifying and acidifying yeasts. Lachancea thermotolerans shows several other properties besides lactic acid production; among them, high malic acid consumption is of great interest in the production of red wines for avoiding undesirable refermentations once bottled. The combination of a L. thermotolerans strain that is able to consume malic acid with a Schizosaccharomyces pombe strain helps to ensure malic acid elimination during alcoholic fermentation while increasing the final acidity by lactic acid production. To properly assess the influence of this alternative strategy, we developed combined fermentations between specific strains of L. thermotolerans and S. pombe under sequential inoculation. Both species showed a great performance under the studied conditions, influencing not only the acidity but also the aromatic compound profiles of the resulting wines. The new proposed biotechnological strategy reduced the final concentrations of ethanol, malic acid and succinic acid, while it increased the concentrations of lactic acid and esters.

Gaining insight into the many agents that determine the underlying material microstructure is essential to engineer new and efficient solutions. Tool steels resistant to abrasive wear are particularly interesting because it is possible to tailor their macroscopic properties by adjusting some of the primary carbide phase features: micromechanical properties, volume fraction, size, and shape. For many popular ledeburitic cold-work and high-speed tool steel alloys, there is a generally good understanding of the effects of the HIP temperature, pressure, and holding time on these traits. Nevertheless, there is still no thorough investigation on the influence of powder size distribution on the primary carbide phase. To that end, we employ in this work novel microstructural characterization methods that shed light on the nature and extent of its influence. We show that powder size has an enduring effect on primary carbide geometric features associated with the solidification process during powder atomization. This work contributes significant process-structure links, which uncover new opportunities for microstructural design.

Abstract This study researched the winemaking performance of new biotechnology involving the cooperation of Lachancea and Schizosaccharomyces genera in the production of wine. In all fermentations where Lachancea thermotolerans was involved, higher lactic acid concentrations appeared, while all fermentations where Schizosaccharomyces pombe was involved, lower levels in malic acid concentration took place. The sensorial properties of the final wines varied accordingly. Differences in mouthfeel properties and acidity occurred in the different fermentation trials. Fermentations with the highest concentration of hydrolyzed mannose showed the highest mouthfeel properties, but the lack of acidity reduced their overall impression. Wines made from a combination of L. thermotolerans and S. pombe showed the highest overall impression and were preferred by the tasters due to the balance between mouthfeel properties and acidity.

The current study highlights the effects of intra- and interspecific hybrid yeasts of the genus Saccharomyces (S.) on the alcoholic fermentation and formation of aroma compounds in cool climate Riesling wines. Three different hybrid yeasts: S. cerevisiae × S. paradoxus (SC × SP), S. cerevisiae × S. kudriavzevii (SC × SK) and S. cerevisiae var. cerevisiae × S. cerevisiae var. bayanus (SC × SB) were investigated. The species S. cerevisiae var. bayanus (SB) was chosen as control variant. It has been demonstrated that the hybrid yeasts have the ability to preserve positive properties while, suppressing undesired properties from the parental yeast species. The hybrid SC × SK showed an increase of desired acetate esters and monoterpenes. The concentrations of higher alcohols were higher in wines fermented by SC × SP, compared to the other variants. SC × SP fermentations resulted in decreased concentrations of l-malate and sulphites. Keywords: Wine flavour, Aroma, Riesling, Hybrid yeasts, Saccharomyces cerevisiae var. bayanus, Saccharomyces kudriavzevii, Saccharomyces paradoxus, Monoterpenes

The interest in Lachancea thermotolerans, a yeast species with unusual characteristics, has notably increased in all ecological, evolutionary, and industrial aspects. One of the key characteristics of L. thermotolerans is the production of high quantities of lactic acid compared to other yeast species. Its evolution has mainly been driven by the influence of the environment and domestication, allowing several metabolic traits to arise. The molecular regulation of the fermentative process in L. thermotolerans shows interesting routes that play a complementary or protective role against fermentative stresses. One route that is activated under this condition is involved in the production of lactic acid, presenting a complete system for its production, showing the involvement of several enzymes and transporters. In winemaking, the use of L. thermotolerans is nowadays mostly focused in early–medium-maturity grape varieties, in which over-ripening can produce wines lacking acidity and with high concentrations of ethanol. Recent studies have reported new positive influences on quality apart from lactic acid acidification, such as improvements in color, glutathione production, aroma, malic acid, polysaccharides, or specific enzymatic activities that constitute interesting new criteria for selecting better strains. This positive influence on winemaking has increased the availability of commercial strains during recent years, allowing comparisons among some of those products. Initially, the management of L. thermotolerans was thought to be combined with Saccaharomyces cerevisiae to properly end alcoholic fermentation, but new studies are innovating and reporting combinations with other key enological microorganisms such as Schizosaccharomyces pombe, Oenocous oeni, Lactiplantibacillus plantarum, or other non-Saccharomyces.

Most commercialized red wines are produced through alcoholic fermentation performed by yeasts of the Saccharomyces genus, and a second fermentation performed by lactic bacteria of the Oenococus oeni species once the first is completely finished. However, the classical process can suffer complications, of which the risks can increase in grape juices with high contents of sugar and pH. Due to climate change, these situations are becoming more common in the winemaking industry. The main risks in those scenarios are alcoholic-fermentation stops or sluggish and undesirable bacteria development while alcoholic fermentation is not finished yet and wine still contains residual sugars. The study propose a novel alternative that offers a solution or reduces the risk of those scenarios while increasing acidity, which is another serious problem of warm viticulture regions. The alternative consists of the combined use of Lachancea thermotolerans to reduce the pH of musts that suffer from a lack of acidity, Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) to achieve malic acid stability during the first stages of alcoholic fermentation, and Saccharomyces bayanus to complete the alcoholic fermentation in difficult wines of high potential alcohol degree of over 15% (v/v). The new proposed biotechnology produced wines with higher final concentrations in lactic acid, glycerol, color intensity, ethyl lactate and 2-phenyl ethyl acetate in 2.39 g/L, 0.52 g/L, 21%, 48% and 37% respectively than the classical methodology where Saccharomyces genus performs alcoholic fermentation and later Oenococus oeni performs malolactic fermentation. Additionally, the new alternative produced wines with lower concentration in ethanol, pH, acetic acid, ethyl acetate, diacetyl and 1-propanol in 0.37% (v/v), 0.26, 0.08 g/L, 22%, 69% and 28% respectively than the classic method.

This study evaluates the influence of Lachancea thermotolerans on low-acidity Airén grape must from the south of Spain. For this purpose, combined fermentations with Lachancea thermotolerans and Saccharomyces cerevisiae were compared to a single fermentation by S. cerevisiae. Results of all developed analyses showed significant differences in several parameters including acidity, population growth kinetics, concentration of amino acids, volatile and non-volatile compounds, and sensorial parameters. The Airén wine quality increased mainly due to the acidification by L. thermotolerans. The acidification process caused a lactic acid increment of 3.18 g/L and a reduction of 0.22 in pH compared to the control fermentation, performed by S. cerevisiae.

This study investigates the influence of the antimicrobial agent chitosan on a selected Schizosaccharomyces pombe strain during the alcoholic fermentation of ultra-pasteurized grape juice with a high concentration of malic acid. It also studies a selected Saccharomyces cerevisiae strain as a control. The study examines several parameters relating to wine quality, including volatile and non-volatile compounds. The principal aim of the study is to test the influence of chitosan on the final chemical composition of the wine during alcoholic fermentation, and to compare the two studied fermentative yeasts between them. The results show that chitosan influences the final concentration of acetic acid, ethanol, glycerol, acetaldehyde, pyruvic acid, α-ketoglutarate, higher alcohols, acetate esters, ethyl esters, and fatty acids, depending on the yeast species.

Over the last decade, several non-Saccharomyces species have been used as an alternative yeast for producing wines with sensorial properties that are distinctive in comparison to those produced using only Saccharomyces cerevisiae as the classical inoculum. Among the non-Saccharomyces wine yeasts, Metschnikowia is one of the most investigated genera due to its widespread occurrence and its impact in winemaking, and it has been found in grapevine phyllospheres, fruit flies, grapes, and wine fermentations as being part of the resident microbiota of wineries and wine-making equipment. The versatility that allows some Metschnikowia species to be used for winemaking relies on an ability to grow in combination with other yeast species, such as S. cerevisiae, during the first stages of wine fermentation, thereby modulating the synthesis of secondary metabolites during fermentation in order to improve the sensory profile of the wine. Metschnikowia exerts a moderate fermentation power, some interesting enzymatic activities involving aromatic and color precursors, and potential antimicrobial activity against spoilage yeasts and fungi, resulting in this yeast being considered an interesting tool for use in the improvement of wine quality. The abovementioned properties have mostly been determined from studies on Metschnikowia pulcherrima wine strains. However, M. fructicola and M. viticola have also recently been studied for winemaking purposes.

The combined use of Lachancea thermotolerans and Schizosaccharomyces pombe is a new winemaking biotechnology that aims to solve some modern industrial oenology problems related to warm viticulture regions. These areas are characterized for producing musts with high levels of sugar that can potentially be converted into wines with elevated ethanol contents, which are usually associated with high pH levels. This biotechnology was reported for the first time in 2015, and since then, several scientific articles have been published regarding this topic. These reported scientific studies follow an evolution similar to that performed in the past for Saccharomyces cerevisiae and Oenococcus oeni; they start by reporting results for basic winemaking parameters at the beginning, later continuing with more advanced parameters. This review compares the results of different researchers that have applied this new biotechnology and have studied wine quality parameters such as ethanol, glycerol, malic acid, lactic acid, amino acids, aroma compounds, or anthocyanins. It is shown that the new biotechnology is repeatedly reported to solve specific winemaking problems such as the lack of acidity, biogenic amines, ethyl carbamate, or undesirable color losses. Such results highlight this biotechnology as a promising option for warm viticulture areas.

This study was performed with the aim of characterizing the fermentative performance of three commercial strains of Torulaspora delbrueckii and their impact on the production of volatile and non-volatile compounds. Laboratory-scale single culture fermentations were performed using a commercial white grape juice. The addition of commercial nutrient products enabled us to test the yeasts under two different nutrient conditions. The addition of nutrients promoted fermentation intensity from 9% to 20 % with significant differences (p < 0.05) among the strains tested. The strain diversity together with the nutrient availability influenced the production of volatile compounds.

Since the beginning of enology and fermentation research, wine quality has been parametrized from a chemical and sensory point of view [...]

Most red wines commercialized in the market use the malolactic fermentationprocess in order to ensure stability from a microbiological point of view. In this secondfermentation, malic acid is converted into L-lactic acid under controlled setups. Howeverthis process is not free from possible collateral effects that on some occasions produceoff-flavors, wine quality loss and human health problems. In warm viticulture regions suchas the south of Spain, the risk of suffering a deviation during the malolactic fermentationprocess increases due to the high must pH. This contributes to produce wines with highvolatile acidity and biogenic amine values. This manuscript develops a new red winemakingmethodology that consists of combining the use of two non-Saccharomyces yeast strains asan alternative to the traditional malolactic fermentation. In this method, malic acid is totallyconsumed by Schizosaccharomyces pombe, thus achieving the microbiological stabilizationobjective, while Lachancea thermotolerans produces lactic acid in order not to reduce andeven increase the acidity of wines produced from low acidity musts. This technique reducesthe risks inherent to the malolactic fermentation process when performed in warm regions.The result is more fruity wines that contain less acetic acid and biogenic amines than thetraditional controls that have undergone the classical malolactic fermentation.

This work examines the physiology of Schizosaccharomyces pombe (represented by strain 938) in the production of red wine, as the sole fermentative yeast, and in mixed and sequential fermentations with Saccharomyces cerevisiae 796. For further comparison, fermentations in which Saccharomyces cerevisiae was the sole fermentative yeast were also performed; in these fermentations a commercial lactic acid bacterium was used to perform malolactic fermentation once alcoholic fermentation was complete (unlike S. cerevisiae, the Sc. pombe performs maloalcoholic fermentation and therefore removes malic acid without such help). Relative density, acetic, malic and pyruvic acid concentrations, primary amino nitrogen and urea concentrations, and pH of the musts were measured over the entire fermentation period. In all fermentations in which Sc. pombe 938 was involved, nearly all the malic acid was consumed from an initial concentration of 5.5 g/L, and moderate acetic acid concentrations below 0.4 g/L were formed. The urea content of these wines was notably lower, showing a tenfold reduction when compared with those that were made with S. cerevisiae 796 alone. The sensorial properties of the different final wines varied widely. The wines fermented with Sc. pombe 938 had maximum aroma intensity and quality, and they were preferred by the tasters.

In recent years, studies have reported the positive influence of non-Saccharomyces yeast on wine quality. Many grape varieties under mixed or sequential inoculation show an overall positive effect on aroma enhancement. A potential impact by non-Saccharomyces yeast on volatile and non-volatile compounds should benefit the flavor of Riesling wines. Following this trend, four separate sequential fermentations (using the non-Saccharomyces yeasts Torulaspora delbrueckii, Metschnikowia pulcherrima, Pichia kluyveri, and Lachancea thermotolerans with Saccharomyces cerevisiae) were carried out on Riesling must and compared to a pure culture of S. cerevisiae. Sequential fermentations influenced the final wine aroma. Significant differences were found in esters, acetates, higher alcohols, fatty acids, and low volatile sulfur compounds between the different trials. Other parameters, including the production of non-volatile compounds, showed significant differences. This fermentation process not only allows the modulation of wine aroma but also chemical parameters such as glycerol, ethanol, alcohol, acidity, or fermentation by-products. These potential benefits of wine diversity should be beneficial to the wine industry.

In the past, some microbiological studies have considered most non-Saccharomyces species to be undesirable spoilage microorganisms. For several decades, that belief made the Saccharomyces genus the only option considered by winemakers for achieving the best possible wine quality. Nevertheless, in recent decades, some strains of non-Saccharomyces species have been proven to improve the quality of wine. Non-Saccharomyces species can positively influence quality parameters such as aroma, acidity, color, and food safety. These quality improvements allow winemakers to produce innovative and differentiated wines. For that reason, the yeast strains Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, Schizosaccharomyces pombe, and Pichia kluyveri are now available on the market. Other interesting species, such as Starmerella bacillaris, Meyerozyma guilliermondii, Hanseniospora spp., and others, will probably be available in the near future.

The undesirable effects of some hazardous compounds involved in the different steps of the winemaking process may pose health risks to consumers; hence, the importance of compliance with recent international food safety standards, including the Hazard Analysis and Critical Control Point (HACCP) standards. In recent years, there has been a rise in the development of new technologies in response to the hazardous effects of chemical compounds detected during the winemaking process, whether naturally produced or added during different winemaking processes. The main purpose was to reduce the levels of some compounds, such as biogenic amines, ethyl carbamate, ochratoxin A, and sulfur dioxide. These technological advances are currently considered a necessity, because they produce wines free of health-hazardous compounds and, most importantly, help in the management and prevention of health risks. This review shows how to prevent and control the most common potential health risks of wine using a HACCP methodology.

At present, wine is generally produced using Saccharomyces yeast followed by Oenococus bacteria to complete malolactic fermentation. This method has some unsolved problems, such as the management of highly acidic musts and the production of potentially toxic products including biogenic amines and ethyl carbamate. Here we explore the potential of the fission yeast Schizosaccharomyces pombe to solve these problems. We characterise an extensive worldwide collection of S. pombe strains according to classic biochemical parameters of oenological interest. We identify three genetically different S. pombe strains that appear suitable for winemaking. These strains compare favourably to standard Saccharomyces cerevisiae winemaking strains, in that they perform effective malic acid deacidification and significantly reduce levels of biogenic amines and ethyl carbamate precursors without the need for any secondary bacterial malolactic fermentation. These findings indicate that the use of certain S. pombe strains could be advantageous for winemaking in regions where malic acid is problematic, and these strains also show superior performance with respect to food safety.

The most popular methodology to make red wine is through the combined use of Saccharomyces cerevisiae yeast and lactic acid bacteria, for alcoholic fermentation and malolactic fermentation respectively. This classic winemaking practice produces stable red wines from a microbiological point of view. This study aims to investigate a recent red winemaking biotechnology, which through the combined use of Lachancea thermotolerans and Schizosaccharomyces pombe is used as an alternative to the classic malolactic fermentation. In this new methodology, Schizosaccharomyces pombe totally consumes malic acid, while Lachancea thermotolerans produces lactic acid, avoiding excessive deacidification of musts with low acidity in warm viticulture areas such as Spain. This new methodology has been reported to be a positive alternative to malolactic fermentation in low acidity wines, since it has the advantage to produce wines with a more fruity flavor, less acetic acid, less ethyl carbamate originators and less biogenic amines than the traditional wines produced via conventional fermentation techniques. The study focuses on unexplored facts related to this novel biotechnology such as color and anthocyanin profile.

The most common way to produce red wine is through the use of Saccharomyces cerevisiae strains for alcoholic fermentation and lactic acid bacteria for malolactic fermentation. This traditional winemaking methodology produces microbiologically stable red wines. However, under specific conditions off-flavours can occur, wine quality can suffer and human health problems are possible, especially after the second fermentation by the lactic acid bacteria. In warm countries, problems during the malolactic fermentation arise because of the high pH of the must, which makes it very difficult to properly control the process. Under such conditions, wines with high acetic acid and histamine concentrations are commonly produced. This study investigates a recent red wine-making technology that uses a combination of Lachancea thermotolerans and Schizosaccharomyces pombe as an alternative to the conventional malolactic fermentation. This work studies new parameters such as aroma compounds, amino acids, ethanol index and sensory evaluation. Schizosaccharomyces pombe totally consumes malic acid while Lachancea thermotolerans produces lactic acid, avoiding excessive deacidification of musts with low acidity in warm viticulture areas. This methodology also reduces the malolactic fermentation hazards in wines with low acidity. The main products are wines that contain less acetic acid, less biogenic amines and precursors and less ethyl carbamate precursors than the traditional wines produced via conventional fermentation techniques.

Today in the wine industry, oenological tannins are widely used to improve wine quality and prevent oxidation in wine aging. With the development of tannin products, new oenological tannins are developed with many specific functions, such as modifying antioxidant effect, colour stabilization and aroma modifications. The aim of this work is to investigate effects of pre-fermentative addition of oenological tannins on wine colour, anthocyanins, volatile compounds and sensorial properties. In this case, Syrah juice was extracted with classic flash thermovinification from fresh must in order to release more colour and tannins. Three types of oenological tannins, which are, respectively, derived from grape skin, seed (Vitis vinifera) and French oak (Quercus robur and Querrus petraea), were selected to carry out the experiments with seven treatments. Results indicated that tannin treatments significantly improved wine aroma complexity and sensorial properties. However, the concentration of some stable pigments such as Vitisin A, Vitisin A-Ac and Vitisin B was negatively affected by tannin additions. Nevertheless, by means of cluster analysis and principal component analysis, it was observed that higher alcohols were significantly promoted by grape seed tannin while most anthocyanins can be improved by addition of grape tannins. In conclusion, low amount of oenological tannin derived from grape seed is a promising method to be applied especially for young red wine making.