Uncovering the Counterfeit: A study of whiskey authenticity through volatile organic compound fingerprinting, aroma and color sensory analysis.

This study presents a method employing gas chromatography coupled with mass spectrometry and headspace solid-phase microextraction (HS-SPME-GC-MS), supplemented with chemometrics (Soft independent modelling of class analogies - SIMCA), to analyze volatile organic compound (VOCs) profiles in suspect whiskey samples. Furthermore, a sensory analysis of aroma and color was conducted with a panel of 52 non-trained volunteers to evaluate their ability to discriminate and preference for counterfeit whiskeys. The HS-SPME-GC-MS method successfully distinguished 41 seized samples from authentic beverages. Interestingly, sensory analysis revealed that panelists could differentiate between counterfeit and authentic samples with a reference standard but did not consistently show a preference for aroma. In some cases, there was even a preference for the color of counterfeit whiskeys. The findings suggest that sensorial tests alone may not effectively distinguish counterfeit from authentic whiskeys, especially for non-expert consumers, highlighting the need for analytical instrumentation methods in fraud detection.

Revealing the influence of exogenously inoculated Bacillus spp. on the microbiota and metabolic potential of medium-temperature Daqu: A meta-omics analysis.

To determine the unique contribution of the bioturbation to the properties of the medium-temperature Daqu, we investigated the differences in microbiota and metabolic composition using the meta-omics approach. Bioturbation increased the amounts of microbial specie and influenced the contribution of the core microbiota to the metabolome. Specifically, inoculated synthetic microbiota (MQB) enhanced the abundance of Bacillus amyloliquefaciens, while Bacillus licheniformis (MQH) increased the abundance of the two Aspergillus species and four species level of lactic acid bacteria. These changes of the microbial profiles significantly increased the potentials of carbohydrate metabolism, amino acid metabolism, and biosynthesis of ester compounds. Consequently, both patterns significantly increased the content of volatile compounds and free amino acids, which were 27.61% and 21.57% (MQB), as well as 15.14% and 17.83% (MQH), respectively. In addition, the contents of lactic acid in MQB and MQH decreased by 65.42% and 42.99%, respectively, closely related to the up- or down-regulation of the expression of their corresponding functional enzyme genes. These results suggested that bioturbation drove the assembly of the core microbiota, rather than becoming critical functional species. Overall, our study provides new insights into the functional role of exogenous isolates in the Daqu microecosystem.

Pediococcus spp.-mediated competition interaction within Daqu microbiota determines the temperature formation and metabolic profiles.

Fermented microbiota is critical to the formation of microenvironment and metabolic profiles in spontaneous fermentation. Microorganisms generate a diverse array of metabolites concurrent with the release of heat energy. In the case of Daqu fermentation, the peak temperature exceeded 60°C, forming a typical high-temperature fermentation system known as high-temperature Daqu. However, microorganisms that cause the quality variation in Daqu and how they affect the functional microbiota and microenvironment in the fermentation process are not yet clear. This study adopted high-throughput sequencing and monitored the dynamic fluctuations of metabolites and environmental factors to identify the pivotal microorganism responsible for the alterations in interaction patterns of functional keystone taxa and quality decline in the fermentation system of different operational areas during the in situ fermentation process that had been mainly attributed to operational taxonomic unit (OTU)_22 (Pediococcus acidilactici). Additionally, we used isothermal microcalorimetry, plate inhibition experiments, and in vitro simulation fermentation experiments to explore the impact of Pediococcus spp. on heat generation, microorganisms, and metabolite profiles. Results showed the heat peak generated by Pediococcus spp. was significantly lower than that of Bacillus spp., filamentous fungi, and yeast. In addition, the preferential growth of P. acidilactici strain AA3 would obviously affect other strains to colonize through competition, and its metabolites made a significant impact on filamentous fungi. The addition of P. acidilactici strain AA3 in simulated fermentation would cause the loss of pyrazines and acids in metabolites. These evidences showed that the overgrowth of Pediococcus spp. greatly influenced the formation of high temperatures and compounds in solid-state fermentation systems. Our work illustrated the vital impact of interaction variability mediated by Pediococcus spp. for microbial assembly and metabolites, as well as in forming temperature. These results emphasized the functional role of Daqu microbiota in metabolites and heat production and the importance of cooperation in improving the fermentation quality.IMPORTANCEThe stable and high-quality saccharifying and fermenting starter in traditional solid-state fermentation was the prerequisite for liquor brewing. An imbalance of microbial homeostasis in fermentation can adversely impact production quality. Identification of such critical microorganisms and verifying their associations with other fermentation parameters pose a challenge in a traditional fermentation environment. To enhance the quality of spontaneous fermented products, strategies such as bioaugmentation or the control of harmful microorganisms would be employed. This work started with the differences in high-temperature Daqu metabolites to explore a series of functional microorganisms that could potentially contribute to product disparities, and found that the differences in interactions facilitated directly or indirectly by Pediococcus spp. seriously affected the development of microbial communities and metabolites, as well as the formation of the microenvironment. This study not only identified functional microbiota in Daqu that affected fermentation quality, but also demonstrated how microorganisms interact to affect the fermentation system, which would provide guidance for microbial supervision in the actual production process. Besides, the application of isothermal microcalorimetry in this study was helpful for us to understand the heat production capacity of microorganisms and their adaptability to the environment. This study presented a commendable framework for improving and controlling the quality of traditional fermentation and inspired further investigations in similar systems.

Exploring the regulated effects of solid-state fortified Jiuqu and liquid-state fortified agent on Chinese Baijiu brewing.

Zaopei is the direct source of Chinese liquor (Baijiu). Adding functional strains to Zaopei is a potential strategy to regulate Baijiu brewing, mainly including the two ways of solid-state fortified Jiuqu (SFJ) and liquid-state fortified agent (LFA). Here, to explore their regulated details, the response patterns of Zaopei microecosystem and the changes in the product features were comprehensively investigated. The results showed that SFJ more positively changed the physicochemical properties of Zaopei and improved its ester content, from 978.57 mg/kg to 1078.63 mg/kg over the fermentation of 30 days, while LFA decreased the content of esters, alcohols, and acids. Microbial analysis revealed that SFJ significantly increased Saccharomycopsis and Aspergillus from the start of fermentation and induced a positive interaction cluster driven by the added functional Paenibacillus, while LFA exhibited a community structure near that of the original microecosystem and led to a simpler network with the reduced microbial nodes and correlations. Metabolism analysis found that both SFJ and LFA weakened the flavor-producing metabolism by suppressing some key enzyme pathways, such as EC 3.2.1.51, EC 4.2.1.47, EC 1.1.1.27, EC 1.1.1.22, EC 1.5.1.10, EC 1.14.11.12. As a result, SFJ improved the raw liquor yield by 28.5 % and endowed the final product with a more fragrant aroma, mainly through ethyl (E)-cinnamate, ethyl isovalerate, ethyl phenacetate with the higher odor activity values, while LFA promoted the yield by 13.2 % and resulted in a purer and less intense aroma through the aroma-active ß-damascenone, ethyl heptoate, ethyl phenacetate. These results facilitated the regulated mechanism of SFJ and LFA on Baijiu brewing and indicated that the used functional strains in this study could be applicated in SFJ way for the further industrial-scale application.

Rice grain structural characteristics of sake rice cultivar Hakutsurunishiki for daiginjo-shu brewing.

Hakutsurunishiki is a sake rice cultivar bred using Yamadabo (seed parent) and Wataribune 2 (pollen parent), equivalent to a Yamadanishiki sibling. This study evaluated the structural characteristics of the Hakutsurunishiki rice grain that contribute to the brewing characteristics of daiginjo-shu, via a comparison with Yamadanishiki. Hakutsurunishiki brown rice was a little heavy and had a large white core. Observing a cross-section of white rice after soaking revealed that the rice grain structure of Hakutsurunishiki was different from that of Yamadanishiki. Hakutsurunishiki white rice showed fewer voids than Yamadanishiki, promoting a slower water absorption rate. Glucose distribution in rice koji obtained by mass spectrometry imaging showed that Hakutsurunishiki rice koji, like Yamadanishiki, is tsuki-haze type, suggesting that its grain structure is suitable for making rice koji for daiginjo-shu. With these observations, we were able to clarify the structural characteristics of Hakutsurunishiki rice grain.

Analysis of differences in microorganisms and aroma profiles between normal and off-flavor pit mud in Chinese strong-flavor Baijiu.

The unique cellar fermentation process of Chinese strong-flavor Baijiu is the reason for its characteristic cellar aroma flavor. The types, abundance, community structure and metabolic activity of microorganisms in the pit mud directly affect the microbial balance in the white spirit production environment, promoting the formation of typical aromas and influencing the quality of CFSB. During the production process, the production of off-flavor in the cellar may occur. The aim of this study is to elucidate the differences in microbiota and flavor between normal pit mud and abnormal pit mud (pit mud with off-flavor). A total of 46 major volatile compounds were identified, and 24 bacterial genera and 21 fungal genera were screened. The esters, acids, and alcohols in the abnormal pit mud were lower than those in the normal pit mud, while the aldehydes were higher. 3-Methyl indole, which has been proven to be responsible for the muddy and musty flavors, was detected in both types of pit mud, and for the first time, high levels of 4-methylanisole was detected in the pit mud. The microbial composition of the two types of pit mud showed significant differences in the bacterial genera of Sporosarcina, Lactobacillus, Garciella, Anaerosalibacter, Lentimicrobium, HN-HF0106, Petrimonas, Clostridium_sensu_stricto_12 and Bacillus, and the fungal genera of Millerozyma, Penicillium, Mortierella, Monascus, Saccharomyces, Issatchenkia, Pithoascus, Pseudallescheria, and Wickerhamomyces. Additionally, we speculate that Sporosarcina is the predominant bacterial genus responsible for the imbalance of microbiota in pit mud.

Effect of S-adenosyl-methionine accumulation on hineka odor in sake brewed with a non-Kyokai yeast.

Hineka is a type of off-flavor of sake and is attributed to the presence of several compounds, including a major one called dimethyl trisulfide (DMTS). The production of the main precursor of DMTS involves yeast methionine salvage pathway. The DMTS-producing potential (DMTS-pp) of sake brewed using the Km67 strain, a non-Kyokai sake yeast, is lower than that of sake brewed using Kyokai yeast; however, the detailed mechanism is unclear. We focused on S-adenosyl-methionine (SAM) and aimed to elucidate the mechanism that prevents DMTS production in sake brewed using the Km67 strain. We revealed that SAM is involved in DMTS production in sake, and that the conversion of SAM to the DMTS precursor occurs through an enzymatic reaction rather than a chemical reaction. Based on previous reports on ADO1 and MDE1 genes, sake brewing tests were performed using the Km67 Δmde1, Δado1, and Δmde1Δado1 strains. A comparison of the SAM content of pressed sake cakes and DMTS-pp of sake produced using the Km67 Δado1 strain showed an increase in both SAM content and DMTS-pp compared to those produced using the parent strain. However, the Km67 Δmde1Δado1 strain showed little increase in DMTS-pp compared to the Km67 Δmde1 strain, despite an increase in SAM content. These results suggest that SAM accumulation in yeast plays a role in the production of DMTS in sake through the methionine salvage pathway. Moreover, the low SAM-accumulation characteristic of the Km67 strain contributes to low DMTS production in sake.

The initial composition and structure of microbial community determined the yield and quality of Baijiu during the spontaneous fermentation / La composición inicial y la estructura de la comunidad microbiana determinaron el rendimiento y la calidad del Baijiu durante la fermentación espontánea

The microbiota during pit mud fermentation is a crucial factor in Baijiu brewing since it determines the yield and flavor. However, the impact of the microbial community during the initial fermentation stage on Baijiu quality remains uncertain. Herein, high-throughput sequencing was employed to investigate the microbial diversities and distribution during Baijiu fermentation in individual pit mud workshops at both initial and late stages. During the initial fermentation stage, the bacterial community exerted a more pronounced effect on Baijiu quality than the fungal community. And the high-yield pit mud workshop exhibited lower richness and evenness, as well as greater Bray-Curtis dissimilarity during Baijiu fermentation. Lactobacillus was the dominant genus and biomarker in high-yield pit mud, and it constituted the only genus within the bacterial association network during the late fermentation stage. Fungal communities tended to maintain a simple association network with selected core species. Based on the correlation network, Rhizopus and Trichosporon were identified as biomarkers in Baijiu fermentation process. Together, Lactobacillus and Rhizopus could serve as bio-indicators for Baijiu quality during the initial fermentation stage. Therefore, these findings provided novel insights into microbiota interactions during fermentation and the impact of initial microbiota on final Baijiu quality.(AU)

Japanese sake making using wild yeasts isolated from natural environments.

Saccharomyces cerevisiae is one of the most important microorganisms for the food industry, including Japanese sake, beer, wine, bread, and other products. For sake making, Kyokai sake yeast strains are considered one of the best sake yeast strains because these strains possess fermentation properties that are suitable for the quality of sake required. In recent years, the momentum for the development of unique sake, which is distinct from conventional sake, has grown, and there is now a demand to develop unique sake yeasts that have different sake making properties than Kyokai sake yeast strains. In this minireview, we focus on "wild yeasts," which inhabit natural environments, and introduce basic research on the wild yeasts for sake making, such as their genetic and sake fermentation aspects. Finally, we also discuss the molecular breeding of wild yeast strains for sake fermentation and the possibility for sake making using wild yeasts.

Charentaise distillation of cognac. Part II: Process simulation and impact of recycling practices on the aroma composition of freshly distilled spirit.

A growing number of studies over the years has successfully employed computer simulation tools to understand, optimize and design spirit distillations. Amongst distilled spirits, cognac is a reputed wine spirit resulting from a double batch distillation process known as Charentaise distillation. This complex operation comprises the wine distillation (WD) and the brouillis distillation (BD), which are carried out in copper alembics. The distillate produced in each batch is fractionated and some of those fractions are recycled in subsequent batches. To improve the current understanding of the behavior of aroma compounds during the process, computer simulation modules were built in this work for a WD and a BD and the results were compared with experimental data. Of the 62 aroma compounds detected in the samples over time, 52 could be represented in the simulations, including 37 using the NRTL thermodynamic model to calculate vapor-liquid equilibria and another 15 with the UNIFAC model. Half of those had their concentration profiles and their partitioning accurately described by the simulation, most of which were modeled with NRTL. This highlights the need for reliable vapor-liquid equilibrium data for aroma compounds that were poorly represented or absent from the simulation as well as kinetic data for chemical reactions occurring during distillation. Furthermore, the impact of the recycling operation on the composition in aroma compounds of freshly distilled cognac was investigated. To represent a steady state, a mathematical model was employed to implement the recycling of distillate fractions during 8 successive Charentaise distillation cycles. The operation was shown to improve the extraction of ethanol and of all volatile compounds in the heart, reaching a pseudo steady state after 3 to 5 cycles. The recycling of the second fraction had a higher influence on the extraction of alcohols and terpenes, while for most esters and norisoprenoids the recycled head fractions played a bigger role.

Case study on the influence of serving temperature on the aroma release and perception of Huangjiu, a fermented alcoholic beverage.

Serving temperature plays a crucial role in influencing the sensory experience of consumers. In this context, this study investigated the influence of serving temperature on the aroma release and perception of a typical fermented alcoholic beverage named Huangjiu. A quantitative sensory description analysis was conducted, determining serving temperature significantly influenced the 17 sensory attributes in both semi-dry and semi-sweet Huangjiu. The variation in the contents of 41 volatiles in the Huangjiu with temperature was investigated using gas chromatography-ion mobility spectrometry, resulting in volatile content significantly increasing above 30 ℃. The partial least squares discriminant analysis was conducted to predict the variable importance for the projection (VIP) of volatiles, and 22 volatiles (VIP > 1) were screened. These 22 volatiles were confirmed as key odorants influenced by serving temperature though aroma addition experiments. The findings would provide a reference for the effects of serving temperature on the flavor perception of fermented alcoholic beverages.

Charentaise distillation of cognac. Part I: Behavior of aroma compounds.

The Charentaise distillation plays an essential role in designing cognac aroma by extracting and selectively concentrating aroma compounds from the wine along with ethanol, in addition to promoting compound formation or degradation through different chemical reactions. This traditional mode of distillation still relies heavily on empirical knowledge and the impact of its different parameters on the composition of cognac is not fully elucidated. In this context, this study aimed to broaden the current knowledge on the behavior of aroma compounds throughout the two steps of the Charentaise distillation and to investigate the formation of aroma compounds during the operation, an aspect which is seldom considered. The concentration profiles of 62 aroma compounds were represented over time for a wine and a brouillis distillation in usual scale (25 hL) with recycling. A classification system was then proposed to group compounds based on their volatilities at different ethanol concentrations in the boiling liquid, their concentration profiles and their chemical properties. This could help identify how chemical characteristics of aroma compounds affect their volatilities in hydroalcoholic media during distillation. In addition, several compounds appear to be formed during distillation, most of which are terpenes, norisoprenoids and aldehydes. Finally, to highlight the importance of different compounds to the aroma of freshly distilled cognac, their odor activity values (OAV) in the heart fraction were estimated, revealing isobutanol and (E)-ß-damascenone to be the most odorant compounds. These results provided additional elements of understanding for different aspects of the Charentaise distillation for the production of cognac, several of which can be transposed, at least in part, to different modes of distillation pertaining to other distilled beverages.

Residual behavior of dinotefuran and its metabolites during Huangjiu fermentation and their effects on flavor.

Yellow rice wine (Huangjiu) is a traditional Chinese alcoholic beverage. However, there is a risk of pesticide residues in Huangjiu due to pesticide indiscriminate use. In this study, the residues of dinotefuran and its metabolites during Huangjiu fermentation and their effects on flavor substances were studied. The initial concentrations of dinotefuran ranged from 856.3 to 1874.9 µg/L, and its half-life was no more than 3.65 d. At 24 d of Huangjiu fermentation, the terminal residues of dinotefuran, 1-methyl-3-(tetrahydro-3-furylmethyl)urea (UF) and 1-methyl-3-(tetrahydro-3-furylmethyl)guanidine (DN) were 195.1-535.3 µg/L, 38.33-48.70 µg/L and 37.8-74.1 µg/L, respectively. Twenty potential degradation compounds were identified by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS), and their toxicity was evaluated. Finally, the effect of dinotefuran on physicochemical properties and total phenol content of Huangjiu were analyzed. The risk of rancidity was significantly increased and bitter amino acids were formed. These findings provide a guidance and the safe production of Huangjiu.

Chemical characterization and sensory properties of apple brandies aged with different toasted oak chips and ultra-high-pressure treatments.

Aging is an important processing step of producing high quality apple brandy. In this study, apple brandies aged by traditional method and using three different toasted oak chips combined with or without ultra-high-pressure (UHP) treatment were prepared to compare their differences in chemical characterization and sensory properties. The results indicated that the brandies aged with toasted oak chip increased the levels of total acidity, volatile acidity and phenolic compounds. It also had the desirable color and taste. The brandy aged with toasted oak chip combined with UHP reached the highest levels of total acidity (1.06 g/L), total phenolic content (284.92 mg/L) and aromatic esters (49.37 %). Therefore, the aging with high toasted oak chip combined with UHP treatment could cut the traditional aging time to meet the same quality as traditional aging method. The results are very useful to develop a fast and efficient aging technique for brandy production.

Characterization of Key Odorants in Chinese Texiang Aroma and Flavor Type Baijiu (Chinese Liquor) by Means of a Molecular Sensory Science Approach.

The popularity of texiang aroma type baijiu (liquor), known for its unique production technology and multilayered flavor, has been steadily increasing among consumers. So far, no research has determined its key aroma compounds for characterizing the unique flavor, which is the purpose of this paper. Using gas chromatography-olfactometry (GC-O) coupled with the GC-mass spectrometry (GC-MS) method, 87 aroma-active compounds were identified and screened out with intensity values ≥2.0. 1,1,6-Trimethyl-1,2-dihydronaphthalene (TDN) was first confirmed by GC-O in Chinese liquor. According to a quantitative study and odor activity values (OAVs), 42 odorants were determined as important aroma compounds. These odorants were recombined based on quantitative concentrations, successfully simulating the overall aroma profile. Omission experiments verified ethyl hexanoate, ß-damascenone, and 2-furfuryl ethyl ether as the key aroma compounds and revealed that ethyl acetate, furfural, and ethyl 2-phenylacetate were important aroma compounds to the overall flavor of texiang aroma and flavor type liquor.

Distribution and Sensory Impact of (2E,4E,6Z)-nonatrienal and Trans-4,5-epoxy-(E)-2-decenal in Wines and Spirits.

This study reports the distribution of (2E,4E,6Z)-nonatrienal (1) and trans-4,5-epoxy-(E)-2-decenal (2) in wines and spirits. We validated a quantification method using solid-phase extraction (SPE) and negative chemical ionization (NCI, NH3) gas chromatography-mass spectrometry (GC-MS) analysis. Both were identified for the first time in wines and spirits from different grape varieties and raw materials. Their olfactory detection thresholds (ODTs) were 16 and 60 ng/L, respectively. Analysis of 66 wines showed that the highest levels of (1) (441.3 ng/L) and (2) (386.5 ng/L) were found in red and white wines, respectively. At these levels, they modify the balance of the fruity expression of red (fresh to cooked fruits) and white (vegetal/green hazelnut nuance) wines. Similar quantitative and sensory analyses were conducted in spirits. With ODT estimated at 500 and 400 ng/L and concentrations ranging from trace amounts to 1.1 and 2.4 µg/L respectively, (1) and (2) can contribute directly to the aroma of spirits.

A State-of-the-Science Review of Alcoholic Beverages and Polycyclic Aromatic Hydrocarbons.

BACKGROUND: The association between alcohol and certain cancers is well established, yet beyond ethanol and its metabolite acetaldehyde, little is known about the presence of other carcinogenic compounds in alcoholic beverages, including polycyclic aromatic hydrocarbons (PAHs), such as benzo[a]pyrene (a Group I carcinogen). OBJECTIVES: We summarized the published literature on PAH levels in alcoholic beverages to identify potential gaps in knowledge to inform future research. METHODS: Medline and Scopus were searched for primary research published from January 1966 to November 2023 that quantified PAH levels among various types of alcoholic beverages, including whisky, rum, brandy, gin, vodka, wine, and beer. Studies that were not primary literature were excluded; only studies that quantified PAH content in the specified alcoholic beverages were included. RESULTS: Ten studies published from 1966 to 2019 met the criteria for review. Other than beverage type, no publication reported selection criteria for their samples of tested alcohol products. Studies used a variety of analytical methods to detect PAHs. Of the 10 studies, 7 were published after 2000, and 6 assessed <20 products. Of the studies, 7 examined spirits; 3, beer; and 4, wines. Benzo[a]pyrene was most prevalent among spirit products, particularly whisky, with values generally exceeding acceptable levels for drinking water. Some beer and wine products also contained PAHs, albeit at lower levels and less frequently than spirit products. DISCUSSION: PAHs are found in some alcohol products and appear to vary by beverage type. However, there is an incomplete understanding of their presence and levels among large, representative samples from the range of currently available alcohol products. Addressing this gap could improve understanding of alcohol-cancer relationships and may have important implications for public health and the regulation of alcohol products. In addition, novel methods, such as direct mass spectroscopy, may facilitate more thorough testing of samples to further investigate this relationship. https://doi.org/10.1289/EHP13506.

Impact on wine sales of removing the largest serving size by the glass: An A-B-A reversal trial in 21 pubs, bars, and restaurants in England.

BACKGROUND: Interventions that alter aspects of the physical environments in which unhealthy behaviours occur have the potential to change behaviour at scale, i.e., across populations, and thereby decrease the risk of several diseases. One set of such interventions involves reducing serving sizes, which could reduce alcohol consumption. The effect of modifying the available range of serving sizes of wine in a real-world setting is unknown. We aimed to assess the impact on the volume of wine sold of removing the largest serving size by the glass from the options available in licensed premises. METHODS AND FINDINGS: The study was conducted between September 2021 and May 2022 in 21 licensed premises in England that sold wine by the glass in serving sizes greater than 125 ml (i.e., 175 ml or 250 ml) and used an electronic point of sale till system. It used an A-B-A reversal design, set over 3 four-weekly periods. "A" represented the nonintervention periods during which standard serving sizes were served and "B" the intervention period when the largest serving size for a glass of wine was removed from the existing range in each establishment: 250 ml (18 premises) or 175 ml (3 premises). The primary outcome was the daily volume of wine sold, extracted from sales data. Twenty-one premises completed the study, 20 of which did so per protocol and were included in the primary analysis. After adjusting for prespecified covariates, the intervention resulted in -420·8 millilitres (ml) (95% confidence intervals (CIs) -681·4 to -160·2 p = 0·002) or -7·6% (95% CI -12·3%, -2·9%) less wine being sold per day. There was no evidence that sales of beer and cider or total daily revenues changed but the study was not powered to detect differences in these outcomes. The main study limitation is that we were unable to assess the sales of other alcoholic drinks apart from wine, beer, and cider, estimated to comprise approximately 30% of alcoholic drinks sold in participating premises. CONCLUSIONS: Removing the largest serving size of wine by the glass from those available reduced the volume of wine sold. This promising intervention for decreasing alcohol consumption across populations merits consideration as part of alcohol licensing regulations. TRIAL REGISTRATION: ISRCTN https://doi.org/10.1186/ISRCTN33169631; OSF https://osf.io/xkgdb.

Predicting the final metabolic profile based on the succession-related microbiota during spontaneous fermentation of the starter for Chinese liquor making.

Microbial inoculation is an effective way to improve the quality of fermented foods via affecting the microbiota structure. However, it is unclear how the inoculation regulates the microbiota structure, and it is still difficult to directionally control the microbiota function via the inoculation. In this work, using the spontaneous fermentation of the starter (Daqu) for Chinese liquor fermentation as a case, we inoculated different microbiota groups at different time points in Daqu fermentation, and analyzed the effect of the inoculation on the final metabolic profile of Daqu. The inoculated microbiota and inoculated time points both significantly affected the final metabolites via regulating the microbial succession (P < 0.001), and multiple inoculations can promote deterministic assembly. Twenty-seven genera were identified to be related to microbial succession, and drove the variation of 121 metabolites. We then constructed an elastic network model to predict the profile of these 121 metabolites based on the abundances of 27 succession-related genera in Daqu fermentation. Procrustes analysis showed that the model could accurately predict the metabolic abundances (average Spearman correlation coefficients >0.3). This work revealed the effect of inoculation on the microbiota succession and the metabolic profile. The established predicted model of metabolic profile would be beneficial for directionally improving the food quality.IMPORTANCEThis work revealed the importance of microbial succession to microbiota structure and metabolites. Multi-inoculations would promote deterministic assembly. It would facilitate the regulation of microbiota structure and metabolic profile. In addition, we established a model to predict final metabolites based on microbial genera related to microbial succession. This model was beneficial for optimizing the inoculation of the microbiota. This work would be helpful for controlling the spontaneous food fermentation and directionally improving the food quality.

Nonconventional yeasts and hybrids for low temperature handcrafted sparkling ciders elaboration in Patagonia.

Yeasts play a crucial role in transforming apple must into cider. While Saccharomyces cerevisiae (Sc) has been traditionally associated to cider fermentations worldwide, cryotolerant species such as Saccharomyces uvarum (Su) as well as natural S. cerevisiae × S. uvarum (Sc×Su) hybrids have also been detected in ciders fermented at low temperatures. This study aimed to evaluate the ability of two Patagonian cryotolerant yeast strains (Su and Se) and their interspecific hybrids with a Sc to conduct handcrafted apple must fermentations and a second fermentation process (champenoise method). The main chemical parameters and sensory quality of the resulting sparkling beverages was also analysed. Firstly, Sc×Se and Sc×Su hybrids were evaluated in their fermentative features at laboratory scale. Hybrids were compared with their respective parental species evidencing significant differences in the physicochemical and aromatic composition of the obtained base ciders. Both Su parental strain and the hybrid Sc×Se were selected for performing pilot scale fermentations (250 L) using natural (non-sterilized) apple juice at two different temperatures: 20 °C and 13 °C. Sc parental strain was also evaluated for comparative purposes. All base ciders obtained were then subjected to a second fermentation. A high implantation capacity of both Su and the hybrid was evidenced at the lowest evaluated temperature, while commercial Sc strain was not detected at the final fermentation stage, independently from the temperature. All sparkling ciders exhibited distinct physicochemical profiles. Ciders inoculated with commercial Sc (but effectively fermented with local Sc strains) allowed the development of malolactic fermentation (MLF) in processes carried out at both temperatures. Contrarily, no MLF was observed in ciders inoculated with either Su or the hybrid. Sparkling ciders fermented with Su displayed the highest concentrations of 2-phenylethanol and 2-phenylethyl acetate, regardless of the fermentation temperature. Conversely, ciders fermented with the hybrid at 20 °C exhibited the highest concentrations of ethyl octanoate and ethyl decanoate, contributing to floral and fruity notes in the beverage. Sensory analysis conducted with untrained individuals revealed a preference for sparkling ciders produced with the hybrid strain at both 20 °C and 13 °C. The cider fermented at 20 °C exhibited floral notes, sweetness, and a full body, while ciders fermented at 13 °C displayed moderate acidity and a well-balanced profile. Conversely, a trained panel described the cider fermented at 20 °C with Su as a fruity and acidic beverage, whereas the ciders fermented at 13 °C exhibited intense bitterness and acidity. This study highlights the potential of cryotolerant Saccharomyces species and hybrids in the development of new starter cultures for producing artisanal sparkling ciders with distinctive properties.