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A doença periodontal (DP) é consequência da atividade bacteriana e deposição de cálculo dentário, causando danos teciduais e processo inflamatório na cavidade oral. É uma enfermidade que atinge mais de 80% dos cães adultos e comumente está ligada ao aparecimento de doenças sistêmicas. A análise do estresse oxidativo sérico gerado na doença periodontal pode ser um biomarcador de lesão tecidual e agravamento da enfermidade. Este trabalho teve como objetivo a quantificação da peroxidação lipídica (TBARS) e da capacidade antioxidante total (TEAC) sérica de cães com periodontite. Os testes foram realizados em um grupo de cães saudáveis (n = 9) e um grupo de cães com DP severa (n = 8). Após a realização dos ensaios de metabolismo redox nos cães com DP, estes animais passaram por procedimento cirúrgico, remoção de cálculo dentário e medicação pós-cirúrgica de 10 dias, que incluía antibioticoterapia com amoxicilina e clavulanato de potássio e meloxicam como medicação anti-inflamatória. Após 30 dias da cirurgia, os animais foram novamente avaliados. Os resultados mostraram que houve um aumento significativo da peroxidação lipídica (43,81%) e uma diminuição da capacidade antioxidante total (8%) nos cães com DP. Após o tratamento, não houve redução dos níveis de estresse oxidativo, mas observou-se um aumento (8%) da capacidade antioxidante total. Estes dados colaboram para o melhor entendimento do metabolismo redox decorrente dos processos infecciosos e inflamatórios característicos da DP. O estudo demonstrou que o estresse oxidativo está presente na doença periodontal em cães, e apesar da não redução do dano oxidativo em 30 dias, a capacidade antioxidante total sérica, que estava diminuída, foi restaurada.

Problems can often arise at the beginning of the second fermentation (prise de mousse) of sparkling wines, such as no start, a long lag period or slow fermentation. These problems are generally associated with yeast stress when inoculated in a base wine with high ethanol content and low pH. However, few studies focus on sulphites, which are often added to base wines to prevent malolactic fermentation, microbiological instability, and wine oxidation. This study aimed to evaluate the joint effect of ethanol and sulfur dioxide on yeasts during the second fermentation. For this purpose, yeasts (Saccharomyces cerevisiae EC1118) were subjected to ethanol, sulfur dioxide and ethanol/sulfur dioxide at the beginning of fermentation, and their vitality and viability, as well as the accumulation of intracellular reactive oxygen species and intracellular pH, were evaluated by flow cytometry. Furthermore, the expression of genes involved in sulfur transport and metabolism was determined. The results showed high mortality, ROS accumulation and intracellular pH reduction in fermentations with both ethanol and sulfur dioxide. The negative effect of ethanol, sulfur dioxide and ethanol/sulfur dioxide on yeasts was found to be dose-dependent and high in those commonly found in some base wines. Cells treated with ethanol/sulfur dioxide showed over-expression of genes involved in sulphite transport (SUL1 and SUL2), efflux pump (SSU1 and FZF1) and metabolism of sulfur amino acids (MET14). Altogether, our data indicate that ethanol and sulfur dioxide have a synergistic effect on yeasts, which may be the root cause of the problems encountered at the beginning of the second fermentation of sparkling wines, and should thus be seriously taken into consideration by winemakers.

In this study, we followed the yeast and wine behaviour during the second fermentation and subsequent lees ageing of sparkling wines produced by Traditional and Charmat methods at an industrial scale. During this period, we conducted physicochemical, microbiological, gene expression, and marker analyses of characteristics related to wine ageing. Our results show that the yeast behaviour during the fermentation is similar in both methods. However, after fermentation, there is a faster decrease in yeast vitality and viability in the Charmat method, together with an increase in the expression of autophagy-related genes (AMS1, APE1, and ATG8). We relate these factors to ageing with the continuous homogenization of the liquid practised in the Charmat method, and static ageing with the lees concentrated at the bottom of the bottle performed in the Traditional method. Despite the variation in yeast viability during ageing, there are no differences in soluble proteins, free amino nitrogen, total phenols, antioxidant activity, and colour evolution between the wines produced by the two methods, assuming few differences over the time that monitoring was conducted.

Mead or honey wine is an alcoholic beverage traditionally produced by the fermentation of a mixture of honey and water. Mead fermentation faces several problems that include sluggish and stuck ferm...

Many computational methods aim to improve the prediction and recognition of transcription elements in prokaryotes. Despite this, the natural features of those elements make their prediction and recognition remain as an open field of research. In this paper, we compared the open-access tools BacPP, BPROM, bTSSfinder, CNNPromoter_b, iPro70-PseZNC, NNPP2, PePPer, and PromPredict. First, we listed the overall functionalities of each tool and the resources available on their web pages. Later, we carried out a comparison of prediction results using 206 intergenic regions. When evaluating the prediction using intergenic regions containing a single promoter within each, NNPP2 and BacPP obtained >90% correct predictions, with NNPP2 obtaining the highest values of match between predicted promoter location and location indicated by RegulonDB. Overall, many discrepancies were observed among the results. They may be explained by the differences in the methodologies that each tool applies for promoter prediction, not excluding the natural features of promoters as a factor as well. In any case, the results highlight the necessity to continue the efforts to improve promoter prediction, perhaps combining multiple approaches. Through said efforts, some of the challenges of the postgenomic era may be tackled as well.

In the present postgenomic era, the capacity to generate big data has far exceeded the capacity to analyze, contextualize, and make sense of the data in clinical, biological, and ecological applica...

Yeasts of the genus

Pyment is a type of mead that is produced from the alcoholic fermentation of a honey solution with the addition of grape juice. Due to the demand for new beverages, pyment can be a profitable alternative for both grape and honey producers. Therefore, this work aimed to characterize the aromatic and physicochemical composition of pyments. The pyments were prepared with addition of 10, 20 and 30% of Moscato juice, and compared with Moscato wine and traditional mead. The results showed an increase in the fermentation rates of Moscato-pyments, indicating that the addition of Moscato juice reverses the low fermentative vigor often reported in mead fermentations. Physicochemical parameters showed an increase in total acidity and a decrease in residual sugar and alcohol, depending on Moscato juice concentration. Moscato-pyments showed an intermediate concentration of volatile compounds between the traditional mead and Moscato wine, with a better balance between fruity, floral and buttery, manifesting characteristic aromas of wines made with Moscato grapes and simultaneously, exposing characteristic aromas of honey. The sensory analysis reveals a significant difference between mead, pyments and Moscato wine. In general, pyments were considered, by the panelists, as the most equilibrated with intermediary aroma intensity, floral, fruity and honey aromas, and good persistence in the mouth.

OBJECTIVE: To describe the evolution of seropositivity in the State of Rio Grande do Sul, Brazil, through 10 consecutive surveys conducted between April 2020 and April 2021. METHODS: Nine cities covering all regions of the State were studied, 500 households in each city. One resident in each household was randomly selected for testing. In survey rounds 1–8 we used the rapid WONDFO SARS-CoV-2 Antibody Test (Wondfo Biotech Co., Guangzhou, China). In rounds 9–10, we used a direct ELISA test that identifies IgG to the viral S protein (S-UFRJ). In terms of social distancing, individuals were asked three questions, from which we generated an exposure score using principal components analysis. RESULTS: Antibody prevalence in early April 2020 was 0.07%, increasing to 10.0% in February 2021, and to 18.2% in April 2021. In round 10, self-reported whites showed the lowest seroprevalence (17.3%), while indigenous individuals presented the highest (44.4%). Seropositivity increased by 40% when comparing the most with the least exposed. CONCLUSIONS: The proportion of the population already infected by SARS-Cov-2 in the state is still far from any perspective of herd immunity and the infection affects population groups in very different levels.

Yeast pigments adsorption can modify wine color intensity and is considered an important trait in wine yeasts. The existing methods for the evaluation of yeast adsorption are laborious, time consuming, need confirmation experiments, or are difficult to apply for a large number of strains generated in breeding programs. In this study, a new test is proposed to measure yeast pigments adsorption and wine color in a single experiment. The optimized method included the microfermentation of grape musts obtained by thermal extraction, digital determination of yeast biomass color, and spectrophotometric wine intensity evaluation. Results showed significant negative correlation between yeast pigments adsorption and wine color intensity. Pigments adsorption occurs from the middle to the end of fermentation, indicating cell wall changes and/or anthocyanins modifications over the process. Significant differences were observed on anthocyanins adsorption and wine intensity among yeast strains independent of the grape variety, and yeasts could be grouped as low, medium and high adsorption strains. The proposed method showed high reproducibility and allows the concomitant screening of hundreds of yeast strains in a short period of time.

Ion exchange resins are polymers with high physical and chemical stability, which, upon activation, acquire positive charge (H+) ions which interchange with existing cations during musts or wines treatment. The chemical composition of the grape must, and wines are highly affected by said modifications. This study aimed to evaluate the impact of the cationic resin treatments on the physicochemical and organoleptic characteristics of base white wines for sparkling wine production. Thus for, Chardonnay must was exposed to a cation exchange resin specific for oenological usage, and fermented to obtain wines. Resulting wines included an untreated control sample with pH 3.15 and treated samples with pH 3.08, 3.02 and 2.97. Physicochemical and volatile compounds analyses as well as quantitative sensory evaluations were conducted. The results showed that, on the scenarios tested, treatments with cationic resins before fermentation resulted in wine with higher oxidative stability, with significative variation on the concentration of several volatile compounds that influenced organoleptic perceptions.

The antifungal activity of 20 monoterpenes, currently found as main compounds in many essential oils, were evaluated against the model yeast Saccharomyces cerevisiae. Minimal inhibitory concentration (MIC), Minimal fungicidal concentration (MFC), and the time/dosage effect of selected monoterpenes were determined. The results showed that oxygenated monoterpenes exhibited higher fungistatic and fungicidal activity than hydrocarbons. Among oxygenated monoterpenes, the more effectives were citral, geraniol, citronellol, and citronellal, with MIC and MFC values between 0.64 and 3.68 mM, and 1.56 and 6.25 mM, respectively. Time response experiments showed that the selected monoterpenes rapidly reduce yeast cell viability in a time and dose‐dependent manner. Moreover, the reduction of viability was associated with loss of cell membrane integrity. These results may aid in the selection of essential oils for the control of undesirable yeasts or fungi, and serve as a basis for the study of chemical structure influence on the mode of action of monoterpenes. NOVELTY IMPACT STATEMENT: Oxygenated monoterpenes have more fungicidal activity against S. cerevisiae. Citral, citronellol, citronellal, and geraniol were the most efficient monoterpenes. Monoterpenes' fungicidal activity is involved with cell membrane damages.

Essential oils and their main components, monoterpenes, have been proven to be important alternatives for the control of pathogenic and spoiling microorganisms, but the mode of action of these compounds is poorly understood. This work aimed to determine the mode of action of citral and geraniol on the model yeast Saccharomyces cerevisiae using a flow cytometry approach. Exponentially growing yeast cells were treated with different concentrations of citral and geraniol for 3 h, and evaluated for cell wall susceptibility to glucanase, membrane integrity, reactive oxygen species (ROS) accumulation, mitochondrial membrane potential, and metacaspase activity. Results provide strong evidence that citral and geraniol acute fungicidal activity against Saccharomyces cells involves the loss of membrane and cell wall integrity resulting in a dose-dependent apoptotic/necrotic cell death. However, yeast cells that escape this first cell membrane disruption, particularly evident on sub-lethal concentration, die by metacaspase-mediated apoptosis induced by the accumulation of intracellular ROS. The deleted mutant on the yca1 gene showed high tolerance to citral and geraniol.

The aim of this study is to evaluate Muscat sparkling wines from the traditional Asti region (Italy) and the new geographical indication “IG Farroupilha” (Brazil). We analyzed 23 commercial Muscat sparkling wines from Asti and Farroupilha. Sample characterization of the aromatic profile by headspace-solid phase microextraction and gas chromatography detector yielded a total of 32 targeted components in Muscat wines. The multivariate analysis of data differentiates the Muscat sparkling wines based on origin: Brazilian or Italian. The main quantitative differences between wines are residual sugar, sulfite content, isoamyl acetate, hexyl acetate, limonene, rose oxide, linalool, and citronellol. The sensory analysis confirmed that Brazilian wines were less sweet with freshener aroma, whereas the Italian wines were more intense in aroma, complex, and sweet. These peculiarities can be associated with terroir, including technological processes that may be tailored to fit different wine styles for particular markets. Practical applications: The chemical and sensorial characterizations of sparkling wines produced in regions with geographical indication are important to differentiate products made in different regions.

Background: Rio Grande do Sul, the southernmost state in Brazil, had a slow start in the pandemic compared to other regions of the country, but the number of new cases rose sharply in December 2020, with a huge spike in March 2021, when the number of new cases was close to 800 per million people. In this article, we describe the pandemic in the State through the lens of 10 consecutive surveys conducted between April 2020 and April 2021. Methods: We used a sentinel city approach, covering all regions of the State. In each city we selected 50 urban census tracts with probability proportional to size. In each tract, 10 households were selected using a systematic approach. Within each selected household, all residents were listed, and one was selected randomly. In the first rounds of the study, we used the rapid point-of-care lateral- flow WONDFO SARS-CoV-2 Antibody Test (Wondfo Biotech Co., Guangzhou, China). In rounds nine and 10, we used an in-house direct ELISA test that identifies the presence of IgG to the viral spike (S) protein from dried blood spot samples (S-UFRJ). In terms of social distancing, individuals were asked three questions, from which we generated an exposure score through principal components analysis. Findings: Antibody prevalence in early April 2020 was 0.07 (95% CI 0.01-0.32), increasing to 10.0% (95% CI 9.1-11.0) in February 2021, and to 18.2% (95% CI 16.9-19.5) in April 2021. Self-reported whites showed the lowest seroprevalences (9.3%, 95% CI 8.3–10.4 in round 9 and 17.3%, 95% CI 15.9–18.8 in round 10), while indigenous individuals presented the highest levels (20.0%, 95% CI 8.5– 40.3 in round 9 and 44.4%, 95% CI 22.5–68.7 in round 10), followed by blacks (14.4%, 95% CI 11.3– 18.1 in round 9 and 21.6%, 95% CI 17.4–26.4 in round 10). We found an increase in seropositivity in relation to our exposure score; the most exposed 10% of the population presented a 40% higher seropositivity. Interpretation: The proportion of the population already infected by SARS-Cov-2 in the State is still far from any perspective of herd immunity, suggesting that herd immunity will only be achieved through vaccination. Funding: the Brazilian Ministry of Health, Banco do Estado do Rio Grande do Sul (BANRISUL), Instituto Cultural Floresta, UNIMED Porto Alegre, “Todos pela Saude” Group, Instituto Serrapilheira, Brazilian Collective Health Association (ABRASCO) and the JBS S.A. initiative “Fazer o Bem Faz Bem”. Declaration of Interest: None to declare. Ethical Approval: Ethical approval was obtained from the Brazilian’s National Ethics Committee (30415520.2.0000.5313), and we obtained written informed consent from all participants.

Chlorothalonil is a broad-spectrum fungicide largely used for the control of several diseases of grapevines. With a moderate persistence in plants, soil and, water, it can be carried to grape musts, particularly when applied to control grape rot diseases. This work aimed to determine the effect of chlorothalonil on Saccharomyces cerevisiae under fermentative conditions using a flow cytometry approach. Yeasts were cultivated in synthetic must with different concentrations of chlorothalonil (0 to 60 μM) and evaluated for culture-ability, membrane integrity, reactive oxygen species (ROS) accumulation, mitochondrial membrane potential, metacaspase activity, ATP, nonprotein SH and, SH-proteins. The results confirmed the oxidation of nonprotein SH, including glutathione, and the binding of the fungicide with sulfhydryl proteins, which led to changes in the cell and mitochondrial membranes that result in the necrotic death of part of the yeast population, and a reduction in metabolic activity. Moreover, the reduction in glutathione-SH concentration was responsible for the increase in ROS which in turn triggers metacaspase-dependent apoptotic cell death. Cells that escape death adapt and began to grow and ferment after a dose-dependent lag-phase period, exhibiting an almost normal fermentative behavior thereafter. Moreover, was observed unexpected protection of chlorothalonil sub-dosages on yeast cell membrane integrity during alcoholic fermentation. This study contributed insights into how chlorothalonil leads the non-target organism S. cerevisiae to cell death and explores the effect of the fungicide during alcoholic fermentation.

The increased demand for pesticide-free foods has also increased the search for healthier and environmentally friendly alternatives in agriculture. Essential oils are known to possess natural antifungal properties, becoming a reliable alternative for commercial fungicides, especially for postharvest decay control. However, essential oils are volatile and photodegradable, which reduces their long-term activities. This work presents the development of a lemongrass essential oil-containing poly(lactic acid) nanocapsules. They have shown in vitro antifungal activity against Colletotrichum acutatum and Colletotrichum gloeosporioides with a MIC dosage of 0.1% (v/v) for both phytopathogens. In the in vivo assay with postharvest apples, the ones treated with encapsulated essential oil showed bitter rot lesions three times smaller than the ones treated with non-encapsulated essential oil, or in comparison to the apples in positive control. The methodology led to stable nanocapsules with spherical morphology, a mean diameter of 96.4 nm, and with an encapsulation efficiency of 99%.

This paper presents a prediction of Bacillus subtilis promoters using a Support Vector Machine system. In the literature, there is a lack of information on Gram-positive bacterial promoter sequences compared to Gram-negative bacteria. Promoter sequence identification is essential for studying gene expression. Initially, we collected the B. subtilis genome sequence from the NCBI database, and promoters were identified by their sigma factors in the DBTBS database. We then grouped the promoters according to 15 factors in 2 domains, corresponding to sigma 54 and sigma 70 of Gram-negative bacteria. Based on these data we developed a script in Python to search for promoters in the B. subtilis genome. After processing the data, we obtained 767 promoter sequences for B. subtilis, most of which were recognized by sigma SigA. To validate the data we found, we developed a software package called BacSVM+, which receives promoters as input and returns the best combination of parameters in a LibSVM library to predict promoter regions in the bacteria used in the simulation. All data gathered as well as the BacSVM+ software is available for download at http://bacpp.bioinfoucs.com/rafael/Sigmas.zip. Keywords: Promoter sequences, Bacillus subtilis, SVM

Dithianon is a broad-spectrum anthraquinone fungicide used to control several diseases of grapes, apples, and other fruits and vegetables. Its mode of action is described as multi-site and associated to thiol-reactivity. As other fungicides can affect non-phytopathogenic organisms as yeasts and bacteria, with impact on microbial population, diversity, and fermentation processes. In this context, we study the effect of dithianon on the model organism and fermentative yeast Saccharomyces cerevisiae in order to elucidate the mechanisms involved in yeast cell death., and explain its interference on wine fermentation kinetics. Thus for, we analyzed cellular protein and non-protein thiols, membrane and cell wall integrity, reactive oxygen species accumulation, mitochondrial membrane potential, and phosphatidylserine externalization. The results showed that when exponentially aerobic growing cells of S. cerevisiae are submitted to acute dithianon treatment they loss cell wall and membrane integrity, dying by necrosis, and this behavior is associated to a depletion of reduced proteic and non-proteic thiol groups. We also detected an important increase of cellular reactive oxygen species (ROS) associated to mitochondrial membrane potential modifications on dithianon treated cells. ROS accumulation was not associated to apoptotic cell death, but can be responsible for intracellular damages. Moreover, necrotic cell death induced by dithianon explains its effect on the kinetics of wine fermentations.

Background and Aims Moscato Branco is the most important cultivar used in the production of Brazilian muscat sparkling wines and is responsible for their sensory characteristics. The aim of this work was to evaluate the effect of monoculture and co‐fermentations by Torulaspora delbrueckii (TPI‐4) and Saccharomyces cerevisiae (EC1118) on the physicochemical, aromatic composition and sensory characteristics of Moscato Branco wines. Methods and Results Wines inoculated with axenic and co‐inoculated with S. cerevisiae and T. delbrueckii were elaborated and subjected to physicochemical, chromatographic and sensory analysis. The monoculture fermentation with T. delbrueckii, and co‐fermentation with 25%EC1118/75%TPI‐4 showed the lowest concentration of volatile fatty acids, considered as undesirable compounds and the highest concentration of 2‐phenylethanol, acetates and ethyl esters, with fruity and flowery descriptors. Based on odour activity values, the main positive contribution of T. delbrueckii was on the concentration of isoamyl acetate, 2‐phenylethanol, 2‐phenylethyl acetate, ethyl hexanoate and linalyl acetate. Conclusions Wines produced by co‐fermentation with 25%EC1118/75%TPI‐4 obtained the highest general quality, aroma quality and flavour intensity scores in the sensory evaluation, reflecting the positive contribution of T. delbrueckii on the sensory attributes of Moscato Branco wines. Significance of the Study This work showed that co‐inoculation with T. delbrueckii and S. cerevisiae improves the quality of muscat wines.

The physicochemical characteristics and yeasts diversity in honey samples from 17 species of stingless bees of the genera Nannotrigona, Melipona, Plebeia, Scraptotrigona, and Tetragonisca cultivated in Southern Brazil were determined. The sugar content, moisture, water activity, pH, reducing sugars/total sugar ratio, and total yeast population varied significantly among the honey from the different bee species. The highest yeast population was found in the Plebeia's honey samples and correlated with their high water-activity. Sixteen yeast species were identified based on the nuclear large subunit (26S) ribosomal RNA partial sequences. The genera Starmerella and Zygosaccharomyces were found predominant, with a high prevalence of Starmerella sp., S. etchellsii, and S. apicola. Some yeast species were only identified in honey samples from specific bee species indicating a close relationship between the yeasts and the insects. For the first time, Wickerhamomyces sydowiorum in honey is being reported. In general, the yeast species isolated from stingless bee honey samples demonstrated high osmotolerance and low sugar assimilation.

In the present postgenomic era, the capacity to generate big data has far exceeded the capacity to analyze, contextualize, and make sense of the data in clinical, biological, and ecological applications. There is a great unmet need for automation and algorithms to aid in analyses of big data, in biology in particular. In this context, it is noteworthy that computational methods used to analyze the regulation of bacterial gene expression have in the past focused mainly on

Yeasts contribute to anthocyanin extraction during red wine fermentation, but they also reduce wine color by adsorption of pigments on their cell walls. Saccharomyces cerevisiae strains vary on their pigment adsorptive properties, but the mechanisms involved in this phenomenon remain unclear. In this work, we evaluated high, medium and low pigment-adsorbing S. cerevisiae winemaking strains during red wine fermentation. Flow cytometry protocols were devised to measure different populations of yeast and their pigment adsorption in association with other variables such as fermentation rate, cell viability, and cell wall/membrane integrity. The results showed that the high pigment adsorbing strain accumulated anthocyanin towards the end of alcoholic fermentation observed as an increase in the population of fluorescent cells. By contrast, low adsorbing strain displayed a single low adsorbing cell population. Pigment adsorption was negatively correlated with cell viability and cell wall/membrane integrity. Independent of their adsorptive potential during wine fermentation, viable cells displayed a low capacity to adsorb anthocyanins, while permeabilized yeast cells exhibited high pigment adsorption capacity. Our findings suggest that yeast pigment adsorption requires a breach to the inner part of the cell wall, for pigments to access constitutively expressed pigment-binding factors, and that the phenotypic differences among strains are determined by their fermentative live-span and cell wall/membrane integrity. These findings will help to rationalize and optimize the match of winemaking strains and vines.

Background Essential oils (EOs) are volatile natural compounds produced by plant secondary metabolism, and some of them exhibit antimicrobial activity. The objective of the present study was to determine the chemical composition the EOs of Eucalyptus staigeriana and Eucalyptus globulus, and their effect in vitro and in vivo against Botrytis cinerea and Colletotrichum acutatum, the most important fungal rot diseases of grapes. Moreover, grapes collected from field experiments were used to evaluate the impact of the alternative control on the alcoholic fermentation and wine composition. Results The major compound of E. staigeriana EO were citral 30.91% (19.74% geranial, 11.17% neral), 1.8-cineole (24.59%) and limonene (19.47%), while 1.8-cineole represented 68.26% of E. globulus EO. The two EOs showed in vitro antifungal activity against both pathogens. Eucalyptus staigeriana EO exhibited the highest activity inhibiting mycelial growth (MG) and conidial germination at 0.5 μL mL-1 . Moreover, this EO was able to reduce the incidence and severity of grey rot caused by B. cinerea and the severity of ripe rot caused by C. acutatum The alternative control did not significantly influence alcoholic fermentation, the physicochemical characteristics, and the volatile composition of wines. Conclusion These results are promising and indicate that E. staigeriana EO might be further investigated as a natural alternative for the control of fungal rots on wine grapes. © 2019 Society of Chemical Industry.

Ripe rot disease caused by Colletotrichum (Glomerella) has become a serious problem for viticulture in Southern Brazil. Global warming contributes to the increase of this devastating and difficult to control disease. Several species of Colletotrichum, with different phytopathological characteristics, have been associated with ripe rot disease in different viticultural regions. In this article, a total of 63 fungi were isolated from grapes showing symptoms of ripe rot disease, and classified by sequencing of ITS region, and compared with the sequences deposited in GenBank. The isolates were included in three clades of Colletotrichum: 84.1% belonged to the “gloeosporioides” clade, 3.2% to the “boninense” clade, and 12.7% to the “acutatum” clade. Of the 53 isolates included in the “gloeosporioides” clade, 44.4% were classified as C. viniferum/C. ampelinum, 37.1% as C. fruticola, 13.0% as C. kahawae and 5.5% as a species related to C. fruticola. In turn, the two isolates of the “boninense” clade were classified as C. kartii/C. phyllanthi, and the six “acutatum” isolates were similar to C. acutatum and C. nymphaeae reference materials. The identified species were previously linked to ripe rot disease in other viticulture regions of the world, but the frequency of some species in southern Brazil is particularly different.

Moscato grapes give very aromatic musts and wines, characterized by the presence of monoterpenoids. The Farroupilha region situated in the highlands of South Brazil, is responsible for 50% of Brazilian Moscato wines production, and obtained its Geographical Indication (GI) in 2015. Brazilian Moscato wines are produced with several varieties, mainly Moscato Branco, Moscato Bianco R2 and Moscato Giallo. The objective of this study was to characterize the aromatic profile of wines produced with the three varieties. Microvinifications were conducted with grapes collected in three vineyards located in Farroupilha. Volatile compounds were evaluated using gas chromatography, and sensory characteristics were determined by a panel of enologists using a specific descriptive chard. Moscato Giallo wines exhibited the highest concentrations of ethyl acetate, 2-phenylethanol, isoamyl acetate, linalool, and α-terpineol; Moscato R2 wines showed the highest concentrations of ethyl decanoate and nerol, while Moscato Branco wines, the most representative variety of Brazilian sparkling Moscato wines, exhibited the highest concentrations of ethanal and ethyl hexanoate, and intermediary concentrations of the other compounds. In sensory analysis, the three varieties exhibited pear, pitanga, rosemary, and citric fruits aromas, but in general, Moscato Giallo and Moscato R2 were more intense, while Moscato Branco showed the highest acidity.

Essential oils and their main compounds, monoterpenoids, are considered as alternative control systems for phytopathogenic fungi, particularly those related to late diseases of fruits and vegetables, like anthracnose caused by Colletotrichum species. In this context, we studied the effect of twenty monoterpenoids on Colletotrichum fructicola and C. acutatum to elucidate their effectiveness and mechanisms of action. Thus, we analyzed mycelial growth and conidial inhibitory concentration, as well as the effect of selected monoterpenoids on membrane integrity and cell vitality, reactive oxygen species (ROS) accumulation, and mitochondrial membrane potential by flow cytometry. The results showed that oxygenated monoterpenoids (alcohols and aldehydes) exhibited higher antifungal activity than their corresponding hydrocarbons, esters, and cyclic counterparts. Indicating that OH- and O- radicals react with cellular components affecting fungal homeostasis. In this sense, selected monoterpenoids (citral, citronellol, geraniol, carvacrol, and thymol) inhibited conidial germination of C. acutatum in a dose-dependent manner. The inhibition of conidial germination is associated with a loss of membrane integrity, a decrease of cell metabolism, and a dose-dependent accumulation of ROS, which was non-directly associated with modifications on mitochondrial membrane potential. Membrane dysfunction and ROS accumulation may be responsible for the necrotic behavior induced by high monoterpenoids concentrations, and possible apoptotic response in sub dosages of these compounds.

Cunila spicata is an endangered aromatic and medicinal plant of South Brazil. In the present paper, the ISSR technique was employed to study the intra- and inter-population genetic diversity of this species. Nine primers generated a total of 109 amplification products, most of which were polymorphics. Low genetic diversity at population level (HE= 0.053) and species level (HT=0.196), and high differentiation among populations (GST= 0.727) were detected in C. spicata. The genetic diversity, low estimated genetic flow and absence of correlation between genetic distances, geographic distances and chemical composition, indicates that genetic drift and inbreeding may be the main factors involved in the genetic structure of C. spicata. Based on these findings, strategies are proposed for the genetic conservation and management of this species.Â

Captan is one of the most widely used broad-spectrum fungicide applied to control several early and late diseases of grapes, apples, and other fruits and vegetables, and as other phthalimide fungicides is defined as a multisite compound with thiol-reactivity. Captan can affect non-target organisms as yeasts, modifying microbial populations and fermentation processes. In this study, we asked whether Captan thiol-reactivity and other mechanisms are involved in acute Captan-induced cell death on aerobic growing Saccharomyces cerevisiae. Thus for, we analyze cellular protein and non-protein thiols, cell membrane integrity, reactive oxygen species accumulation, phosphatidylserine externalization, and apoptotic mutants behavior. The results showed that when submitted to acute Captan treatment most cells lost their membrane integrity and died by necrosis due to Captan reaction with thiols. However, part of the cells, even maintaining their membrane integrity, lost their culture ability. These cells showed an apoptotic behavior that may be the result of non-protein thiol depletion and consequent increase of reactive oxygen species (ROS). ROS accumulation triggers a metacaspase-dependent apoptotic cascade, as shown by the higher viability of the yca1-deleted mutant. Together, necrosis and apoptosis are responsible for the high mortality detected after acute Captan treatment of aerobically growing cells of S. cerevisiae.

Traditional methods for synthesizing metallic nanoparticles include the use of toxic chemicals, which can have negative impacts on the environment and human health. A safer alternative to produce these nanomaterials is based on the concept of Green Chemistry, which uses less harmful chemicals from renewable sources. In this work, we describe a specific approach for the green synthesis of silver nanoparticles (AgNPs) using the extract of Ives cultivar (Vitis labrusca) pomace, their characterization, and their immobilization in a polymer matrix of chitosan aiming the disinfection of a raw wastewater sample. The influence of some experimental parameters on synthesis such as pH, stabilization time as well as the temperature has been investigated. The grape pomace extract was initially characterized regarding its content of phenolic compounds and then used as stabilizing-reducing agent. The obtained AgNPs were analyzed by ultraviolet-visible spectroscopy, transmission electron microscopy, zeta potential, scanning electron microscopy coupled with energy dispersive spectroscopy, and raman spectroscopy. In general, the AgNPs were spherical in shape and very stable. Aiming to explain the reaction, we also proposed a mechanism of formation of AgNPs based on gallic acid molecule. Concerning the antimicrobial properties, the results demonstrated the synergistic activity of chitosan-AgNPs pellets against Gram-positive and Gram-negative bacteria. Additionally, these materials led to a reduction of 47% of the bacterial count of Escherichia coli on the disinfection of the wastewater. Finally, it was concluded that the extract of Ives cultivar pomace is capable of producing stable AgNPs that can be used as auxiliaries in wastewater disinfection.

The industry of fine wines and also locally consumed table wines is emerging in Brazil with an increasing volume and economic impact. Enologists in this region currently lack information about the prevalence and characteristics of spoilage yeasts, which may contaminate and potentially undervalue Brazilian wines. Herein, we analyzed 50 local red wines including 27 fine wines (V. vinifera) and 23 table wines (V. labrusca). Presumptive spoilage yeasts were isolated on differential medium, and classified by RFLP-PCR and sequencing of the ITS1-5.8S-ITS2 and D1/D2 26S rDNA loci. The prevalence of spoilage yeasts in fine wines (11 %) was comparable to that reported in European and US wines, and significantly lower than that observed for local table wines (70 %). The majority of isolates belonged to Brettanomyces bruxelliensis, followed by Pichia guillermondii, and more rarely Candida wickerhamii and Trigonopsis cantarelli. The Brettanomyces isolates varied greatly in off-flavor production, displayed ethanol tolerance (>10 % by volume), tolerated sulfite (≥0.68 mg/l mSO2), and 39 % of them grew on ethanol as sole carbon source. We discuss the causes and consequences of spoilage yeasts in relation to the Brazilian wine industry.