Fine wine recognition and appreciation: It is time to change the paradigm of wine tasting.

Wine quality maybe understood under two perspectives: (a) commercial quality, intended to satisfy overall consumers, and (b) fine wine quality, aimed at achieving a product with aesthetic value. The current food sensory techniques (e.g. Descriptive Analysis) have been successfully applied to develop wines accepted worldwide and characterized by pleasant sweetish flavours and smooth mouthfeel. On the contrary, these techniques are not suited to characterize fine wines given their dependence on sensory properties with aesthetic value. The conventional tasting approaches follow the sequence of vision, smell (orthonasal), taste and mouthfeel, ending by an overall evaluation. The sensory descriptors tend to be analytic (e.g. different aromas and tastes) or synthetic (e.g. body, structure) and the quality judgement is left for the final step. Some synthetic attributes may have an aesthetic significance (e.g. complexity, harmony, depth) and are more valued when the analytic or synthetic descriptors are highly praised (e.g. oakiness, silkiness, body, minerality). Consequently, these highly praised attributes are regarded as surrogates of fine wine quality. However, commercial wines are frequently judged of higher quality than fine wines irrespective of the taster expertise. We argue that the conventional sensory analysis sequence makes the overall evaluation secondary in relation to the previous analytical steps blurring the assessment of wine's aesthetic properties. Probably due to top-down processing, the initial evaluation of colour or flavours governs the final overall quality evaluation that may be inconsistently rated. Then, to promote the recognition of fine wines, tasting should begin by first acknowledging the aesthetic properties and only proceed to the analytical steps if necessary. A tasting method is proposed to consumer educational programs where emotional responses are used to explain the differences between commercial and fine wine styles. Furthermore, cultural aspects should be included to appreciate the wholeness of wine. Hopefully, this holistic perspective would turn wine appreciation more approachable and facilitate the recognition of fine wines among consumers, increasing their appreciation and enjoyment.

The Socioeconomic Factors of Street Food Vending in Developing Countries and Its Implications for Public Health: A Systematic Review.

The sale of ready-to-eat (RTE) street food represents an important source of income in many developing countries. However, these foods are frequently implicated in outbreaks of gastrointestinal diseases. Street food vendors face several constraints that hamper improvement in the microbiological quality of their products. The aim of this review was to update knowledge about the main causes of foodborne illnesses in developing countries, including the growing concern with the microbial transmission of antibiotic resistance. Following PRISMA guidelines, this systematic review was conducted on original articles published from January 2010 to July 2023. The search was carried out using Scopus, PubMed, Web of Science, Food Science and Technology Abstracts (FSTA), the International Information System for Agricultural Sciences and Technology (AGRIS), as well as isolated searches of relevant articles from Google Scholar. The initial search identified 915 articles, 50 of which were included in this systematic review. The results indicate that, in the majority of the 15 countries examined, women constitute the predominant segment of street food vendors, representing more than 55% of the total number of these vendors. In 11 countries, street food vendors under the age of 18 were identified. Most vendors had a low level of education and, consequently, were unaware of good hygiene practices when handling food. The combination of factors such as poor hygiene practices on the part of food handlers and the lack of facilities, namely, the absence of available potable water, were frequently listed as the main causes of food contamination. Enterobacteriaceae such as Escherichia coli (61.9%), Salmonella (30.1%), and Shigella spp. (9.5%), as well as Staphylococcus aureus (30.1%) and Listeria monocytogenes (14.3%), were the most common pathogens found in RTE street foods. In 22 studies from 13 developing countries, 59% (13/22) reported high multidrug resistance in Enterobacteriaceae (40% to 86.4% in E. coli, 16.7 to 70% in Salmonella, and 31 to 76.4% in S. aureus). To address the challenges faced by street vendors and improve their economic activities, it is necessary for government entities, consumers, and vendors to work together collaboratively.

Multidrug Resistance in Enterococci Isolated from Cheese and Capable of Producing Benzalkonium Chloride-Resistant Biofilms.

This study aimed to investigate enterococci recovered from eight Portuguese cheeses made with raw ewe's milk, regarding antibiotic resistance, virulence genes, minimum inhibitory concentration (MIC) of benzalkonium chloride (BAC), biofilm formation capacity, and biofilm eradication (MBEC) by BAC. Antimicrobial resistance against seven antibiotics of five groups was evaluated using the disk diffusion method. The presence of the genes that encode resistance to the antibiotics penicillin (blaZ), erythromycin (ermA, ermB, and ermC), vancomycin (vanA and vanB), aminoglycoside (aac(6')-Ie-aph(2″)-Ia), and ß-lactam (pbp5) and the genes that encode virulence factors, frsB, cylA, gelE, esp, and agg, were investigated via multiplex PCR. The susceptibility of planktonic cells to BAC was evaluated by the MIC and MBC values of the isolates, using the broth microdilution method. To assess the biofilm-forming ability and resistance of biofilms to BAC, biofilms were produced on stainless steel coupons, followed by exposure to BAC. The results showed a high resistance to the antibiotics vancomycin (87.5%), erythromycin (75%), tetracycline (50%), and penicillin (37.5%). Multidrug resistance was observed in 68.8% of the isolates. Genes encoding the virulence factors FrsB (frsB) and gelatinase E (gelE) were detected in all isolates. The esp and cylA genes were found in 56.3% and 37.5% of the isolates, respectively. All isolates exhibited a biofilm-forming ability, regardless of incubation time and temperature tested. However, after 72 h at 37 °C, E. faecium and E. faecalis biofilms showed significant differences (p ≤ 0.05). Although most isolates (62.5%) were susceptible to BAC (MIC ≤ 10 mg/L), biofilms of the same isolates were, generally, resistant to the higher concentration of BAC (80 mg/mL) tested. This study using Enterococcus isolates from a ready-to-eat food, such as cheese, reveals the high percentages of vancomycin resistance and multidrug resistance, associated with the presence of virulence genes, in isolates also capable of producing biofilms resistant to BAC, an important active ingredient of many disinfectants. These results emphasize the need for effective control measures to ensure the safety and quality of dairy products.

Proximate Composition, Physicochemical and Microbiological Characterization of Edible Seaweeds Available in the Portuguese Market.

BACKGROUND: The aim of this work was the study of the proximate composition and profile of fatty acids, minerals, and some microbiological aspects of four edible seaweed species (Chondrus crispus, Palmaria palmata, Porphyra sp., and Ulva sp.) available in the Portuguese market for food consumption, and produced in a national Integrated Multi-Trophic System (IMTA). METHODS: Moisture, ash, and total lipids were determined gravimetrically. Crude protein was analysed by Duma's combustion procedures. The total carbohydrate content was assayed by the phenol/sulphuric acid method. The assessment of the fatty acids methyl esters (FAMEs) was determined through GC-MS. Characterization of elemental analysis was performed by ICP-AES. Different standard microbiological methods were applied for microorganisms. Statistics were performed using the non-parametric Mann-Whitney U test to assess significant differences between samples. RESULTS: Lipid contents (n = 3) were very low (1.6-2.3%), particularly in Palmaria palmata, and Chondrus crispus (1.6-1.7%). The protein content (n = 4) varied from 14.4% in P. palmata to 23.7% in Porphyra sp. Carbohydrates (n = 3) were the major constituent of most seaweeds (31-34%), except in Porphyra sp., with higher content in proteins than carbohydrates. Regarding the fatty acid content (n = 4), in general, saturated fatty acids (SFAs) were the most abundant followed by polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs). Among macro and trace elements (n = 3), Chondrus crispus shows the highest average content in Zn (71.1 mg ⋅ kg - 1 D.W.), Palmaria palmata the highest average content in K (124.8 g ⋅ kg - 1 D.W.), Porphyra sp. the highest average content in P (2.1 g ⋅ kg - 1 D.W.), and Ulva sp. the highest average content of Ca (5.5 g ⋅ kg - 1 D.W.), Mg (55.8 g ⋅ kg - 1 D.W.), and Fe (336.3 mg ⋅ kg - 1 D.W.). In general, Na and K were the most abundant elements among analysed seaweed. Additionally, the microbiological results (n = 4) comply with the Portuguese guidelines (subgroup 2D) on the application of general principles of food hygiene in ready-to-eat foods. CONCLUSIONS: Overall, the results highlight the potential of using these seaweeds as an alternative and sustainable source of elements and bioactive compounds to produce enriched food products with a beneficial potential for human nutrition.

A high level of antibiotic resistance in Klebsiella and Aeromonas isolates from street water sold in Mozambique, associated with the prevalence of extended-spectrum and AmpC ß-lactamases.

This study aims to evaluate the resistance profile and the prevalence of antibiotic resistance genes in 30 isolates of Klebsiella spp. and Aeromonas spp. recovered from water sold in the streets of Maputo. Susceptibility profiles to 15 antibiotics were performed according to Clinical Laboratory Standard Institute guidelines with antibiotic disks on Mueller-Hinton agar plates. Multiplex PCRs were performed targeting 10 ß-lactamase genes, five ESBL (blaTEM-variants, blaOXA-variants, BlaSHV-variants, MCTX-M Group 1 and Group 9 variants) and five AmpC (ACC variants, FOX variants, MOX variants, CIT variants and DHA variants). The results showed a high prevalence of Klebsiella resistance to ß-lactam antibiotics, such as amoxicillin/clavulanic acid (62.5%), amoxicillin (56.3%), ampicillin (50%), cefoxitin (43.8%), and cefotaxime (43.8%). Aeromonas showed resistance to cefoxitin and ampicillin (71.4%), amoxicillin/clavulanic acid (57.1%) and imipenem (42.9%). ESBL blaOXA-variants, blaSVH-variants, MCTX-M Group 1 variants, and MCTX-M Group 9 variants were the most prevalent b-lactam genes, followed by the b-lactams AmpC, ACC variants and FOX variants. It is extremely important to improve waterborne disease control strategies, especially in terms of public awareness of the potential health implications of multidrug-resistant strains of Klebsiella and Aeromonas, which are often neglected.

Wine Spoilage Control: Impact of Saccharomycin on Brettanomyces bruxellensis and Its Conjugated Effect with Sulfur Dioxide.

The yeast Brettanomyces bruxellensis is one of the most dangerous wine contaminants due to the production of phenolic off-flavors such as 4-ethylphenol. This microbial hazard is regularly tackled by addition of sulfur dioxide (SO2). Nevertheless, B. bruxellensis is frequently found at low levels (ca 103 cells/mL) in finished wines. Besides, consumers health concerns regarding the use of sulfur dioxide encouraged the search for alternative biocontrol measures. Recently, we found that Saccharomyces cerevisiae secretes a natural biocide (saccharomycin) that inhibits the growth of different B. bruxellensis strains during alcoholic fermentation. Here we investigated the ability of S. cerevisiae CCMI 885 to prevent B. bruxellensis ISA 2211 growth and 4-ethylphenol production in synthetic and true grape must fermentations. Results showed that B. bruxellensis growth and 4-ethylphenol production was significantly inhibited in both media, although the effect was more pronounced in synthetic grape must. The natural biocide was added to a simulated wine inoculated with 5 × 102 cells/mL of B. bruxellensis, which led to loss of culturability and viability (100% dead cells at day-12). The conjugated effect of saccharomycin with SO2 was evaluated in simulated wines at 10, 12, 13 and 14% (v/v) ethanol. Results showed that B. bruxellensis proliferation in wines at 13 and 14% (v/v) ethanol was completely prevented by addition of 1.0 mg/mL of saccharomycin with 25 mg/L of SO2, thus allowing to significantly reduce the SO2 levels commonly used in wines (150-200 mg/L).

High Fecal Contamination and High Levels of Antibiotic-Resistant Enterobacteriaceae in Water Consumed in the City of Maputo, Mozambique.

In the city of Maputo, Mozambique, food and water are often sold on the streets. Street water is packaged, distributed, and sold not paying attention to good hygienic practices, and its consumption is often associated with the occurrence of diarrheal diseases. Coincidentally, the increase of diarrheal diseases promotes the inappropriate use of antibiotics that might cause the emergence of antibiotic-resistant bacterial strains. In this context, the present study aimed to assess the microbiological quality of water sold on the streets of Maputo, as well as the antibiotic resistance profile of selected Enterobacteriaceae isolates. The 118 water samples analyzed were from street home-bottled water (n = 81), municipal water distribution systems (tap water) (n = 25), and selected supply wells in several neighborhoods (n = 12). The samples were analyzed for total mesophilic microorganisms, fecal enterococci, fecal coliforms, Escherichia coli, and Vibrio spp. The results showed a high level of fecal contamination in all types of water samples. In home-bottled water, fecal coliforms were found in 88% of the samples, and E. coli in 66% of the samples. In tap water, fecal coliforms were found in 64%, and E. coli in 28% of the samples. In water from supply wells, fecal coliforms and E. coli were found in 83% of the samples. From 33 presumptive Vibrio spp. colonies, only three were identified as V. fluvialis. The remaining isolates belonged to Aeromonas spp. (n = 14) and Klebsiella spp. (n = 16). Of 44 selected Enterobacteriaceae isolates from water samples (28 isolates of E. coli and 16 isolates of Klebsiella spp.), 45.5% were not susceptible to the beta-lactams ampicillin and imipenem, 43.2% to amoxicillin, and 31.8% to amoxicillin/clavulanic acid. Regarding non-beta-lactam antibiotics, there was a high percentage of isolates with tolerance to tetracycline (52.3%) and azithromycin (31.8%). In conclusion, water in Maputo represents a risk for human health due to its high fecal contamination. This situation is made more serious by the fact that a relatively high percentage of isolates with multidrug resistance (40%) were found among Enterobacteriaceae. The dissemination of these results can raise awareness of the urgent need to reduce water contamination in Maputo and other cities in Mozambique.

Historical Wines of Portugal: The Classification, Consumer Associations and Marketing Implications.

Geographical origin, use of traditional varieties and ancestral viticulture/oenology practices characterize wines classified as Historical Wines of Portugal (HWP). This study identifies the authenticity attributes consumers associate with this classification and assesses the relative strength of associations. A review of brand authenticity research and interviews with Portuguese wine producers (n = 3) and consumers (n = 12) were conducted to identify HWP classification attributes. Strength of attribute association was subsequently evaluated in an online questionnaire with a convenience sample of Portuguese wine consumers (n = 641), which included a measure of general wine knowledge and questions about the adequacy of different contexts for HWP purchase and consumption. Wine knowledge markedly affected the nature and strength of consumer associations. Compared to Aspirational Explorers, wine connoisseurs emerged as Heritage Gatekeepers, associating origin, cultural heritage, quality, production and at-home consumption more strongly with HWP, and tradition, wine age and out-of-home consumption less strongly. Market recognition of HWP as a novel and distinctive table wine classification, with well-defined and unique attributes, is thus likely to depend on consumers' general wine knowledge. Related promotional activities targeting wine novices should first focus on educating them on HWP classification, whereas those directed at savvier consumers should emphasize wine authenticity cues instead.

Consumers Associate High-Quality (Fine) Wines with Complexity, Persistence, and Unpleasant Emotional Responses.

The conventional method for the sensory evaluation of wine is based on visual, olfactory and gustatory perceptions described by a domain-specific language. This is a complex task, requiring extensive training, which is not feasible from a consumer perspective. The objective of this study was to apply a wine tasting sheet, including sensory and emotional responses, to simplify the recognition of fine white wines by consumers. First, a panel of 15 semi-trained judges evaluated eight sensory attributes through Optimized Descriptive Profile (ODP) methodology. Then, a group of 104 consumers evaluated five white wines with different sensory characteristics using an improved emotional wine tasting sheet. The emotions and sensations most frequently associated with white wines were obtained through the Check-All-That-Apply (CATA) approach. The eight sensory attributes were significant (p-value < 0.05) in the distinction of wines by the ODP. Likewise, the distinction of the wines also provided significant differences in all the emotional and sensory attributes (p-value < 0.05). The different wine styles could be distinguished by Principal Component Analysis (PCA) using the semi-trained judges or the consumer responses. The highest score in the "global evaluation" was given to two young, fruity wines characterized by high aromatic "initial impression". The two fine wines, including a 2004 Burgundy Pouilly-Fuissé, were the lowest rated in "initial impression" and "global evaluation", although they were considered by the consumers among the most complex and persistent. These wines were also most frequently associated with unpleasant emotions by the CATA test. The recognition of these fine wine attributes and their incongruity with emotional responses can be used in a rapid way by professionals to explain the different wine styles to consumers.

Off-Flavours and Unpleasantness Are Cues for the Recognition and Valorization of Organic Wines by Experienced Tasters.

The drivers of consumer acceptance concerning organic wines are not well understood. In particular, among wine professionals, there are anecdotal evidences claiming that consumers accept off-flavours that would not be tolerated if the wines were conventionally produced. Therefore, the aim of this study was to shed further light on this issue by tasting blind wines of both types of production using a tasting panel comprised by experienced individuals of several nationalities. The tasted wines were both conventional and organic and were with and without off-flavours. The same wines were evaluated in three tasting sessions where the given information was: (1) all wines were conventional, (2) all wines were organic, and (3) tasters were asked to guess the mode of production. A group of untrained tasters also rated the same organic wines in an informed session. The results showed that wines were significantly better scored and were given a higher willingness to pay value in the "organic" session. In addition, the experienced tasting panel produced a list of the most frequent sensory descriptors. When tasters were asked to guess the mode of production, wines that were supposed to be organic received a higher citation of off-flavours, such as "oxidized", "reductive", and "animal/undergrowth". Moreover, an overall emotional response of unpleasantness was associated with the recognition of organic wines in the "guess" session. Untrained tasters rated the same organic wines with lower liking scores and were willing to pay less. In conclusion, off-flavours and their unpleasantness worked as a cue to identify wines supposed to be organic by experienced tasters. Their corresponding higher valorization could be explained by the psychological halo effect induced by the organic label. Contrarily, consumers did not show this halo effect, depreciating wines with unpleasant flavours irrespective of their mode of production.

Wine tasting based on emotional responses: An expedite approach to distinguish between warm and cool climate dry red wine styles.

In this study, we improved an empirical tasting sheet including emotional responses and common sensory attributes. An Optimized Descriptive Profile (ODP) was run to characterize different red wines according to sensory descriptors used in the improved sheet. A total of 5 wines were evaluated by a Consumer Panel (CP) of 103 subjects (36 females, 67 males) using the improved sheet and a Check-All-That-Apply (CATA) list of 25 emotions. In the ODP, the panel identified the main discriminating sensory attributes as "Complexity", "Astringency" and "Duration of the wine fragrance". However, this analysis did not allow for differentiating very distinct dry red wine styles originating from warmer or cooler regions. On the contrary, Principal Component Analysis of emotional attributes demonstrated that these two wine styles could be easily distinguished. In particular, wine with a red-brick color, complex smell and aggressive mouthfeel consistent with those from cooler regions was less liked by the CP than a warm climate gold-awarded wine. Although receiving lower scores considering its color and smell, the former wine was regarded as the most "surprising" in the CATA.

Patterns of sweetness preference in red wine according to consumer characterisation.

The preference for sweet taste in red wine was examined according to consumer categories of age, gender, drinking experience and personality type (Big-5 personality-test). A total of 114 subjects revealed their preferences for sweetness after tasting dry red wine spiked with equal concentrations of glucose and fructose at 2g/L, 4g/L, 8g/L, 16g/L and 32g/L, following an ascending forced choice paired comparison method (2-AFC). The overall preference for sweetness was shown within the range of 4.8 to 21.9g/L, with maximal liking at 8g/L. Three patterns of response to sweetness were observed (sweet dislikers, sweet likers and indifferent to sweet) according to the different categories of consumers. Differences (p>0.05) were not found in sweetness preference among the categories up to 16g/L sugar except for the trait extraversion at 8g/L, where low extraverts showed a higher proportion of responses preferring the sweeter sample. Most significant differences were found only under the highest tasted concentration (32g/L). Females and novices preferred sweeter samples (p<0.05) when compared with the response of males and experienced consumers, respectively.

Influence of Brettanomyces ethylphenols on red wine aroma evaluated by consumers in the United States and Portugal.

Brettanomyces may add complexity to wine at low concentrations but at high concentrations, can result in objectionable wines. The objective of this study was to determine the concentrations at which consumers from two different locations were able to detect Brettanomyces volatile compounds present in a red wine. A red wine blend, used in both countries, was spiked to create five treatments containing different concentrations of 4-ethylphenol (4-EP), 4-ethylguiacol (4-EG), and 4-ethylcatechol (4-EC) in a 5:1:1 ratio, respectively. These treatments were evaluated by consumers in the United States and Portugal (n=121) using a difference from control test. Consumers were also classified as having low, medium, or high wine knowledge. Among the spiked samples, the greatest degree of difference was found between the second and third treatments, corresponding to reported detection and recognition threshold ranges of 4-EP and 4-EG. For some treatments, consumers from Portugal classified in the medium or high knowledge level reported significantly higher mean differences from the control than those in the low knowledge group (p<0.05). Results demonstrated consumers' ability to detect differences in red wines due to Brettanomyces volatile compounds. Results provide useful context on how wine knowledge and cultural variants may affect the detection of Brettanomyces.

Effect of Ethanol, Sulfur Dioxide and Glucose on the Growth of Wine Spoilage Yeasts Using Response Surface Methodology.

Response surface methodology (RSM) was used to study the effect of three factors, sulfur dioxide, ethanol and glucose, on the growth of wine spoilage yeast species, Zygosaccharomyces bailii, Schizosaccharomyces pombe, Saccharomycodes ludwigii and Saccharomyces cerevisiae. Seventeen central composite rotatable design (CCRD) trials were designed for each test yeast using realistic concentrations of the factors (variables) in premium red wine. Polynomial regression equations were fitted to experimental data points, and the growth inhibitory conditions of these three variables were determined. The overall results showed Sa. ludwigii as the most resistant species growing under high ethanol/free sulfur dioxide concentrations, i.e., 15% (v/v)/20 mg L-1, 14% (v/v)/32 mg L-1 and 12.5% (v/v)/40 mg L-1, whereas other yeasts did not survive under the same levels of ethanol/free sulfur dioxide concentrations. The inhibitory effect of ethanol was primarily observed during longer incubation periods, compared with sulfur dioxide, which showed an immediate effect. In some CCRD trials, Sa. ludwigii and S. cerevisiae showed growth recovery after a short death period under the exposure of 20-32 mg L-1 sulfur dioxide in the presence of 11% (v/v) or more ethanol. However, Sc. pombe and Z. bailii did not show such growth recovery under similar conditions. Up to 10 g L-1 of glucose did not prevent cell death under the sulfur dioxide or ethanol stress. This observation demonstrates that the sugar levels commonly used in wine to sweeten the mouthfeel do not increase wine susceptibility to spoilage yeasts, contrary to the anecdotal evidence.

New insights into the ecological interaction between grape berry microorganisms and Drosophila flies during the development of sour rot.

In this work, we studied the ecological interactions between grape berry microorganisms and Drosophila sp. flies involved in sour rot disease during grape ripening. After veráison the total microbial counts of grape berries affected by sour rot increased from about 2 log CFU/g of berries to more than 7 log CFU/g. Berry damage provoked a clear shift in yeast diversity from basidiomycetes to ascomycetous fermentative species. The latter were mostly Pichia terricola, Hanseniaspora uvarum, Candida zemplinina, and Zygoascus hellenicus. However, these species were not able to produce the metabolites characteristic of sour rot (gluconic and acetic acids) in inoculated berries. On the contrary, the acetic acid bacteria Gluconacetobacter saccharivorans produced high levels of these acids, mainly when berries were incubated in the presence of the insect Drosophila sp. Sour rot was not observed when grape bunches were physically separated from insects, even when berries were artificially injured. The wounds made in berry skin healed in the absence of insects, thus preventing the development of sour rot. Therefore, in the vineyard, the induction of sour rot depends on the contamination of wounded berries by a microbial consortium--yeasts and acetic acid bacteria--transported by drosophilid insects which disseminate sour rot among damaged berries. In the absence of these insects, plant defense mechanisms are effective and lead to skin healing, preventing disease spread. Thus, we showed that Drosophila sp. act as a vector for microorganisms associated with grape sour rot disease.

Changes in sour rotten grape berry microbiota during ripening and wine fermentation.

This study investigated the microbiota of sour rotten wine grapes and its impact on wine fermentations. Yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB) were enumerated and identified on sound and sour rot grapes during the ripening stage. The alteration of the ecological balance induced by sour rot was particularly evidenced by the unequivocal increase of yeast and AAB counts on rotten grapes, since the beginning of ripening. Yeast and AAB species diversity in rotten grape samples were much higher than those found in sound grapes. LAB populations were low detected from both healthy and sour rotten grapes. The yeast species Issatchenkia occidentalis, Zygoascus hellenicus and Zygosaccharomyces bailii and the AAB species Gluconacetobacter hansenii, Gluconacetobacter intermedius and Acetobacter malorum, were recovered from damaged grapes and resulting grape juices in the winery. Acetobacter orleaniensis and Acetobacter syzygii were only recovered from sour rotten grapes. Dekkera bruxellensis and Oenococcus oeni were only recovered after wine fermentation induced by starter inoculation, irrespective of grape health, probably originating from cellar environment. After malolactic fermentation, racking and sulphur dioxide addition the only remaining species were the yeast Trigonopsis cantarellii and Saccharomyces cerevisiae, independently of the grape health status.

Analytical and sensorial characterization of the aroma of wines produced with sour rotten grapes using GC-O and GC-MS: identification of key aroma compounds.

In the present work, the aroma profiles of wines elaborated from sound and sour rot-infected grapes as raw material have been studied by sensory analysis, gas chromatography-olfactometry (GC-O), and gas chromatography-mass spectrometry (GC-MS), with the aim of determining the odor volatiles most likely associated with this disease. The effect of sour rot was tested in monovarietal wines produced with the Portuguese red grape variety Trincadeira and in blends of Cabernet Sauvignon and sour rotten Trincadeira grapes. Wines produced from damaged berries exhibited clear honey-like notes not evoked by healthy samples. Ethyl phenylacetate (EPhA) and phenylacetic acid (PAA), both exhibiting sweet honey-like aromas, emerged as key aroma compounds of sour rotten wines. Their levels were 1 order of magnitude above those found in controls and reached 304 and 1668 µg L(-1) of EPhA and PAA, respectively, well above the corresponding odor thresholds. Levels of γ-nonalactone also increased by a factor 3 in sour rot samples. Results also suggest that sour rot exerts a great effect on the secondary metabolism of yeast, decreasing the levels of volatiles related to fatty acids and amino acid synthesis. The highest levels of γ-decalactone of up to 405 µg L(-1) were also found in all of the samples, suggesting that this could be a relevant aroma compound in Trincadeira wine aroma.

The effect of sugar concentration and temperature on growth and volatile phenol production by Dekkera bruxellensis in wine.

The wine spoilage yeast Dekkera bruxellensis was evaluated for the production of 4-ethylphenol under low concentrations (0.02-20 g L(-1)) of glucose and fructose in synthetic media. Measurable amounts of 4-ethylphenol were produced over 0.2 g L(-1) of each sugar. The yeast growth rate and amount of biomass formed increased from 0.2 to 20 g L(-1) of glucose or fructose, being accompanied by increasing production of 4-ethylphenol. In red wines, the production of 4-ethylphenol was only observed in the presence of growing populations of indigenous or inoculated strains of D. bruxellensis. The production rate of 4-ethylphenol varied between 22 and 93 mug day(-1) either with inoculated strains or wild populations in bottled wines. The production rate of 4-ethylphenol as a function of the increase in the number of cells varied from 349 to 1882 mug L(-1) per one log CFU mL(-1). The effect of temperature on cellular viability and 4-ethylphenol production was tested in red wines with indigenous or inoculated strains of D. bruxellensis. Incubation temperatures of 15, 20 and 25 degrees C allowed cellular growth and volatile phenol production. Increasing incubation temperatures to 36 degrees C induced full viability loss of 10 strains of D. bruxellensis within <12 h.

Sour rot-damaged grapes are sources of wine spoilage yeasts.

Yeast species of sound and sour rot-damaged grapes were analysed during fermentation and grape ripening in the vineyard, using general and selective culture media. During 2003 and 2004 vintages, microvinifications were carried out with sound grapes to which different amounts of grapes with sour rot were added. The wine spoilage species Zygosaccharomyces bailii was only recovered during fermentations with sour rot, reaching 5.00 log CFU mL(-1) (2003) and 2.48 log CFU mL(-1) (2004) at the end of fermentation. The study of yeast populations during the sour rot ripening process (2005 vintage) showed that the veraison-damaged grapes always exhibited higher total yeast counts and a much greater diversity of species. From a total of 22 ascomycetous species, 17 were present only in damaged grapes. The most frequent species were Issatchenkia occidentalis and Zygoascus hellenicus. The spoilage species Z. bailii and Zygosaccharomyces bisporus were consistently isolated exclusively from damaged grapes. This work demonstrates that one of the most dangerous wine spoilage species, Z. bailii, is strongly associated with sour rot grapes and survives during fermentation with Saccharomyces cerevisiae. The use of selective media provides a more accurate characterization of grape contamination species.

Molecular typing of the yeast species Dekkera bruxellensis and Pichia guilliermondii recovered from wine related sources.

A total of 63 strains of Dekkera bruxellensis and 32 strains of Pichia guilliermondii isolated from wine related environments were identified by restriction analysis of the 5.8S-ITS region of the rDNA. These strains were subjected to intraspecific discrimination using mtDNA restriction and RAPD-PCR analysis. The isolates identified as D. bruxellensis yielded 3 different molecular patterns of mtDNA restriction using the endonuclease HinfI. The pattern A was the most frequent (58 strains) among strains from different sources, regions and countries. Pattern B (4 strains) and C (one strain) were determined in isolates from Portuguese wines. The discrimination among the pattern A strains was achieved by a RAPD-PCR assay with 3 primers (OPA-2, OPA-3 and OPA-9). A total of 12 haplotypes were obtained with the combination of the patterns provided by the 3 OPAs. The pattern 2 was the most frequent and extensively distributed being found in strains from different countries and from different sources like wine, barrique wood and insects. The strains of P. guilliermondii were characterized with restriction of mtDNA using the endonuclease HinfI yielding 7 different restriction patterns. These patterns were associated with different efficiencies of 4-ethylphenol production. Patterns A to D corresponded to 19 strains producing low levels of 4-ethylphenol (<1 mg/l) while patterns F and G grouped 13 strains producing high levels of 4-ethylphenol (>50 mg/l), when grown in synthetic media supplemented with 100 mg/l of p-coumaric acid. The high degree of polymorphism observed shows that intraspecific typing is essential for accurate yeast dissemination studies in wine related environments.