Fungal control strategies based on the use of Bacillus have emerged in agriculture as eco-friendly alternatives to replace/reduce the use of synthetic pesticides. Bacillus sp. P1 was reported as a ...new promising strain for control of Aspergillus carbonarius, a known producer of ochratoxin A, categorized as possible human carcinogen with high nephrotoxic potential. Grape quality can be influenced by vineyard management practices, including the use of fungal control agents. The aim of this study was to evaluate, for the first time, the quality parameters of Chardonnay grapes exposed to an antifungal Bacillus-based strategy for control of A. carbonarius, supporting findings by genomic investigations. Furthermore, genomic tools were used to confirm that the strain P1 belongs to the non-pathogenic species Bacillus velezensis and also to certify its biosafety. The genome of B. velezensis P1 harbors genes that are putatively involved in the production of volatiles and hydrolytic enzymes, which are responsible for releasing the free form of aroma compounds. In addition to promote biocontrol of phytopathogenic fungi and ochratoxins, the treatment with B. velezensis P1 did not change the texture (hardness and firmness), color and pH of the grapes. Heat map and hierarchical clustering analysis (HCA) of volatiles evaluated by GC/MS revealed that Bacillus-treated grapes showed higher levels of compounds with a pleasant odor descriptions such as 3-hydroxy-2-butanone, 2,3-butanediol, 3-methyl-1-butanol, 3,4-dihydro-β-ionone, β-ionone, dihydroactinidiolide, linalool oxide, and β-terpineol. The results of this study indicate that B. velezensis P1 presents desirable properties to be used as a biocontrol agent.
•Bacillus-treated grapes did not significantly change the texture, color and pH.•B. velezensis P1 genome revealed 11 genes involved in volatile compounds formation.•Bacillus-treated grapes showed higher levels of 3-hydroxy-2-butanone and 2,3-butanediol.•B. velezensis P1 expressed terpene synthesis enzymes in response to A. carbonarius.
•GC×GC/MS and chemometrics allowed distinguishing volatiles of base and sparkling wine.•Potential aroma contributions of volatile compounds of sparkling wine were discussed.•2nd dimension and ...spectral deconvolution improved separation of wine volatiles.•Chemometrics applied to GC×GC/MS data reduced the number of representative volatiles.
The main changes in the volatile profile of base wines and their corresponding sparkling wines produced by traditional method were evaluated and investigated for the first time using headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry detection (GC×GC/TOFMS) and chemometric tools. Fisher ratios helped to find the 119 analytes that were responsible for the main differences between base and sparkling wines and principal component analysis explained 93.1% of the total variance related to the selected 78 compounds. It was also possible to observe five subclusters in base wines and four subclusters in sparkling wines samples through hierarchical cluster analysis, which seemed to have an organised distribution according to the regions where the wines came from. Twenty of the most important volatile compounds co-eluted with other components and separation of some of them was possible due to GC×GC/TOFMS performance.
Compounds with toxic potential may occur in beer, such as carbonyl compounds (acetaldehyde, acrolein, ethyl carbamate EC and formaldehyde) and furan derivatives furfural and furfuryl alcohol (FA). ...The objective of this study was, for the first time, to validate a method based on headspace-solid phase microextraction using a PDMS-overcoated fibre and gas chromatography with mass spectrometric detection in selected ion monitoring mode (HS-SPME-GC/MS-SIM) to investigate target carbonyl compounds and furan derivatives in beers. Analytical curves showed proper linearity with r
2
ranging from 0.9731 to 0.9960 for acetaldehyde and EC, respectively. The lowest LOD was found for acetaldehyde (0.03 µg L
−1
), while the lowest LOQ value (1.0 µg L
−1
) was found for acetaldehyde and EC, formaldehyde and furfural. Recovery (90% to 105%), intermediate precision and repeatability (lower than 13%), limits of detection and quantification (values below 2.5 μg L
−1
) showed that the method is suitable to simultaneously quantify these compounds. EC was detected in only two samples (1 lager and 1 ale). Furfural was found in 37% and 82% of ale and lager beers, respectively. Acetaldehyde, acrolein, formaldehyde and FA were detected in all samples. However, acrolein was the only compound found in the commercial samples at a concentration capable of causing health risk. Besides furfural and FA, four other furan-containing compounds (5-methyl-2-furan methanethiol, acetylfuran, 5-methylfurfural and γ-nonalactone) were also found in beers, however, at levels low enough not to impose potential health risk.
Bacillus-based biocontrol agents have emerged as a strategy to eliminate or reduce the use of synthetic fungicides that are detrimental to health and the environment. In vineyards, a special concern ...arises from the control of Aspergillus carbonarius, a fungus known for its potential to produce ochratoxins. Ochratoxin A (OTA) is the most toxic form among ochratoxins and its maximum limit in wine has been established in Europe and Brazil as 2 μg/kg. Wine quality, especially the volatile profile, may be influenced by the antifungal strategies, since fungicide residues are transferred from grapes to must during winemaking. The objective of this study was to evaluate, for the first time, the impact of a biocontrol strategy containing Bacillus velezensis P1 on the volatile profile and occurrence of ochratoxins when grapes infected with A. carbonarius were used in winemaking. The evaluation of ochratoxins was carried out by liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QToF-MS), and volatile compounds were analyzed using comprehensive two-dimensional gas chromatography coupled to quadrupole mass spectrometry (GC × GC/qMS). Six ochratoxins were identified in must prepared with Chardonnay grapes inoculated with A. carbonarius (ochratoxin α, ochratoxin β, ochratoxin α methyl-ester, ochratoxin α amide, N-formyl-ochratoxin α amide and OTA). Although winemaking causes a decrease in the levels of all forms of ochratoxins, the co-occurrence of these mycotoxins was verified in wine made with grapes containing A. carbonarius. B. velezensis P1 prevented the occurrence of ochratoxins in must, ensuring the safety of wines. Regarding the volatile profile, a predominant presence of terpenic compounds was verified in samples treated with B. velezensis when compared with those not treated with the biocontrol strategy, whereas the presence of A. carbonarius resulted in a higher concentration of volatile compounds with an odor described as fatty/waxy, possibly compromising wine quality. Therefore, B. velezensis P1 is a new biofungicide possibility to produce ochratoxin-free grapes and high-quality wines.
•Ochratoxins were found in the vinification of grapes inoculated with A. carbonarius.•B. velezensis P1 prevented ochratoxin A and other 5 ochratoxins in wines.•B. velezensis P1 reduced fatty/waxy odor compounds produced by A. carbonarius.•Wine made using grapes treated with B. velezensis P1 were richer in terpenes.
A uva é uma das frutas convencionais mais produzidas e valorizadas no setor agrícola, por ser economicamente rentável e seu consumo estar associado a benefícios à saúde. São os compostos bioativos ...presentes na fruta, que apresentam efeito antioxidante, antimicrobiano, que previne câncer e retarda o envelhecimento precoce. No entanto, a concentração e composição desses compostos são dependentes de diversos fatores, como manejo do produtor, clima, solo. O município de Marialva-PR é o maior produtor de uva fina do estado do Paraná. A viticultura representa mais da metade da produção agrícola do município, sendo determinante para a economia, desenvolvimento e progresso da cidade. Reconhecendo a importância do cultivo da fruta para a região e os compostos bioativos para a alimentação humana, o presente trabalho tem como objetivo efetuar um planejamento para otimizar as extrações de compostos bioativos presentes na casca de uva da variedade Brasil, oriunda desse município. Para a avaliação da melhor condição de extração foi realizado um planejamento fatorial 23 com triplicata no ponto central. Os fatores proporção de solvente: água/álcool etílico, pH e concentração do substrato do meio extrator foram observados. O ponto ótimo para a extração ocorreu na proporção água/álcool etílico 60:40 (v/v) respectivamente, pH 2,0 e concentração 10g/L.
The human nose has been used as a detector in gas chromatography analysis to evaluate odoriferous compounds related to aroma and quality of wine. Several olfactometric techniques are available to ...access the description, intensity, and/or duration of the odor of each compound. Olfactometry can be associated with one‐dimensional gas chromatography or multidimensional gas chromatography, including heart‐cut gas chromatography and comprehensive two‐dimensional gas chromatography. Multidimensional gas chromatography may help to resolve coeluted compounds and detect important trace components for the aroma. The identification of odor‐active compounds may help to differentiate wines according to terroir, grapes cultivars used in winemaking or types of aging, understand the role of fungal infection of grapes for wine quality, find the best management practices in vineyard and vinification to obtain the greatest quality. In addition, when the instrumental techniques are combined with sensory analysis, even more accurate information may be obtained regarding the overall wine aroma. This review discloses the state of the art of olfactometric methods and the analytical techniques used to investigate odor‐active compounds such as one‐dimensional gas chromatography, multidimensional gas chromatography, and comprehensive two‐dimensional gas chromatography. The advances in knowledge of wine aroma achieved with the use of these techniques in the target and profiling approaches were also discussed.
Carbonyl compounds and furan derivatives may form adducts with DNA and cause oxidative stress to human cells, which establishes the carcinogenic potential of these compounds. The occurrence of these ...compounds may vary according to the processing characteristics of the beer. The objective of this study was, for the first time, to investigate the free forms of target carbonyl compounds acetaldehyde, acrolein, ethyl carbamate (EC) and formaldehyde and furan derivatives furfural and furfuryl alcohol (FA) during the brewing stages of ale and lager craft beers. Samples were evaluated using headspace-solid phase microextraction and gas chromatography with mass spectrometric detection in selected ion monitoring mode (HS-SPME-GC/MS-SIM). Acetaldehyde, acrolein, formaldehyde and furfuryl alcohol were found in all brewing stages of both beer types, while EC and furfural concentrations were below the LOD and LOQ of the method (0.1 and 0.01 µg L
−1
, respectively). Boiling and fermentation of ale brewing seem to be important steps for the formation of acrolein and acetaldehyde, respectively, while boiling resulted in an increase of FA in both types of beer. Conversely, pasteurisation and maturation reduced the levels of these compounds in both types of beer. An increase in concentration of acrolein has not been verified in lager brew probably due to the difference in boiling time between these two types of beer (60 and 90 min for ale and lager, respectively).
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•Cabernet Sauvignon grape was more susceptible to A. carbonarius than Moscato Italico.•Must of Cabernet Sauvignon contained more ochratoxin A than Moscato Italico must.•Higher OTA ...level was verified in Moscato Italico wine than Cabernet Sauvignon.•Higher level of acids and terpenes was found in grape that contained A. carbonarius.•A. carbonarius has not interfered in the antioxidant activity of wines.
Aspergillus carbonarius can produce a possibly carcinogenic mycotoxin named ochratoxin A (OTA). The metabolism of this fungus can also impact grape and wine quality as it influences the volatile and phenolic profiles, which are related to aroma and antioxidant activity, respectively. The objective of this study was to evaluate the effect of A. carbonarius on OTA levels and for the first time on volatile profile and antioxidant activity of grapes and their respective wines. Cabernet Sauvignon (CS, red) grapes presented higher susceptibility to A. carbonarius than Moscato Italico (MI, white) grapes and OTA levels in their respective musts were in accordance with this same trend. However, vinification of red grapes resulted in 67% reduction of OTA, while the reduction observed with white wines was 45%. The presence of acids (hexanoic, octanoic, nonanoic and decanoic, fatty odor) was found to be an indicative of the fungus incidence in grapes. These acids were precursors of esters that might impart negative aroma (methyl nonanoate and isoamyl octanoate, fatty odor) or provide desirable fruity characteristics (ethyl hexanoate, ethyl octanoate and methyl octanoate) for wine. In addition, terpenes were detected only in wines produced with grapes (CS and MI) inoculated with A. carbonarius. The presence of A. carbonarius increased the antioxidant activity of CS grapes. For MI grapes and both wines (CS and MI) no differences were verified in the antioxidant activity of the samples affected or not affected by this fungus. Although A. carbonarius occurrence has shown no influence on the antioxidant activity of wines, it produced OTA and has negatively influenced the wine odor profile, due to the production of some volatiles that impart a deleterious effect on wine aroma.
Femoral fractures and severe bleeding frequently occur in old patients showing a delayed healing. As there are no studies investigating the combined effect of high age and severe blood loss on ...fracture healing, this was examined radiographically and biomechanically in this study. Therefore, young and old male mice were randomly assigned to three operation groups. In the fracture group (Fx), external fixator and osteotomy were applied to the femur. The combined trauma group (THFx) additionally received a pressure-controlled hemorrhage. Sham animals were only implanted with arterial catheter and external fixator. Sacrifice was performed after three weeks and bone healing was evaluated radiologically via µCT, as well as biomechanically using a three-point bending test. A decreased share of callus/total bone volume was observed in old mice with blood loss compared to old Fx. Hemorrhagic shock also reduced the trabecular number in old mice compared to Fx and young THFx. Moreover, a lower elastic limit in old Sham mice without fracture was revealed. Fracture combined with a high loss of blood further reduced the elastic limit in old mice compared to isolated Fx in old animals. In conclusion, this study showed that severe blood loss has a higher negative effect in old mice compared to young ones.
•Terpenes decreased their contribution to aroma during Moscatel sparkling winemaking.•Esters increased their contribution to aroma during Moscatel sparkling winemaking.•GC×GC/MS resolved 1D ...co-elutions resulting in 66 compounds out of 69 separated in 2D.•173 volatiles were tentatively identified by GC×GC and 43 by GC in Moscatel sparkling wines.
This study reports, for the first time, the main changes that occur with some important aroma compounds of Moscatel sparkling wines during winemaking, measured using headspace solid-phase microextraction, one-dimensional and comprehensive two-dimensional gas chromatography (GC×GC) with mass spectrometry detection (MS). The best conditions of volatile extraction included the use of PDMS/DVB fibre, 2mL of wine, 30% of NaCl, 40°C for 30min. The chromatographic profile of sparkling wines showed decreasing amounts of monoterpenes (limonene, 4-terpineol, terpinolene, citronellol, α-terpineol, linalool, hotrienol, and nerol oxide), increasing amounts of esters (terpenyl esters, ethyl octanoate, ethyl decanoate and hexyl acetate) and alcohols (1-nonanol and 2-phenylethanol). Sixty-nine compounds co-eluted in the first dimension; only six co-eluted in the second dimension. GC×GC/TOFMS allows more detailed study of the volatile profile of sparkling wines.
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