Comparison of ancestral and traditional methods for elaborating sparkling wines.

This work compares the ancestral method for elaborating sparkling wines with the most widely used traditional method. Ancestral method is a single fermentation procedure in which the fermenting grape must is bottled before the end of alcoholic fermentation whereas traditional method involves a second fermentation of a base wine inside a bottle. Macabeo grapes were used to elaborate a traditional sparkling wine and two ancestral sparkling wines, one with a low yeast population and one with a high yeast population. The findings indicate that ancestral sparkling wines have lower ethanol content and can be elaborated using lower sulphur dioxide levels. In general, ancestral sparkling wines showed similar protein concentration, higher polysaccharide content, similar or better foamability (HM) than the traditional sparkling wine., No differences were found in the foam stability (HS). In addition, the sensory analysis indicated that ancestral sparkling wines have smaller bubble size, lower CO2 aggressivity, they seemed to have longer ageing time and were scored better than the traditional sparkling wine. These results therefore indicate that the ancestral method is of great interest for the elaboration of high-quality sparkling wines.

Use of Glutathione, Pure or as a Specific Inactivated Yeast, as an Alternative to Sulphur Dioxide for Protecting White Grape Must from Browning.

One of the problems that most seriously affects oenology today is enzymatic browning, especially when grapes are infected by grey rot. We studied the capacity of glutathione (GSH) and a specific inactivated dry yeast rich in glutathione (IDY-GSH) to protect white grape must from browning compared to that of sulphur dioxide (SO2). The results indicate that SO2 drastically reduces the oxygen consumption rate (by around 72%), protects hydroxycinnamic acids from oxidation and prevents grape must against browning even in the presence of laccase. Specifically, the presence of SO2 reduced the colour's blue-yellow component (b*) by around 91% in control conditions and around 76% in the presence of laccase. GSH, pure or in the form of IDY-GSH, also reduces the oxygen consumption rate (by 23% and 36%, respectively) but to a lesser extent than SO2. GSH also favours the formation of grape reaction product (GRP) from hydroxycinnamic acids and effectively protects grape must against browning in healthy grape conditions. Specifically, the presence of GSH reduced b* by around 81% in control conditions. Nevertheless, in the presence of laccase, it was not effective enough, reducing b* by around 39% in the case of pure GSH and 24% in the case of IDY-GSH. Therefore, both forms of GSH can be considered as interesting alternative tools to SO2 for preventing browning in white grape must, but only when the grapes are healthy.

Impact of rare yeasts in Saccharomyces cerevisiae wine fermentation performance: Population prevalence and growth phenotype of Cyberlindnera fabianii, Kazachstania unispora, and Naganishia globosa.

Saccharomyces cerevisiae is a highly fermentative species able to complete the wine fermentation. However, the interaction with other non-Saccharomyces yeasts can determine the fermentation performance of S. cerevisiae. We have characterised three rare non-Saccharomyces yeasts (Cyberlindnera fabianii, Kazachstania unispora and Naganishia globosa), studying their impact on S. cerevisiae fitness and wine fermentation performance. Using a wide meta-taxonomic dataset of wine samples, analysed through ITS amplicon sequencing, we show that about a 65.07% of wine samples contains Naganishia spp., a 27.21% contains Kazachstania spp., and only a 4.41% contains Cyberlindnera spp; in all cases with average relative abundances lower than 1% of total fungal populations. Although the studied N. globosa strain showed a limited growth capacity in wine, both K. unispora and C. fabianii showed a similar growth phenotype to that of S. cerevisiae in different fermentation conditions, highlighting the outstanding growth rate values of K. unispora. In mixed fermentations with S. cerevisiae, the three yeast species affected co-culture growth parameters and wine chemical profile (volatile compounds, polysaccharides and proteins). K. unispora DN201 strain presents an outstanding capacity to compete with S. cerevisiae strains during the first stage of wine fermentation, causing stuck fermentations in both synthetic and natural grape musts.

Oxygen consumption rate of lees during sparkling wine (Cava) aging; influence of the aging time.

Sparkling wines elaborated with a traditional method need to age in the bottle in contact with wine lees because yeast autolysis enriches the wines in colloids and improves their effervescence, foam and aromatic complexity. It is generally considered that lees protect the wine against oxidation because they consume small amounts of oxygen that can permeate the crown cap. However, to our knowledge there is no specific study on this subject using lees from real sparkling wine. Therefore, the oxygen consumption rate (OCR) of the lees of sparkling wines from the first to the ninth year of aging time was measured using a noninvasive fluorescence measurement method. The results indicate that lees really consume oxygen and that their OCR tended to decrease with the wine aging time. These data suggest that the lees' capacity to protect against oxidation decreases over time, which could affect the ability of sparkling wines to age properly.

Influence of grape seeds on wine composition and astringency of Tempranillo, Garnacha, Merlot and Cabernet Sauvignon wines.

BACKGROUND: The aim of this work was to study the influence of grape berry morphology, especially the seed weight percentage, on the tannin concentration and astringency of red wine. Clusters of Tempranillo, Garnacha, Merlot, and Cabernet Sauvignon were characterized and their seeds were extracted and macerated in a model wine solution. In parallel, we elaborated three types of wines of each cultivar. One wine was made with only grape juice, one wine was made adding the appropriate proportion of seeds to the grape juice, and the last wine was elaborated with the complete destemmed and crushed berries. RESULTS: Merlot and Cabernet Sauvignon grapes, which have higher percentage of seed weight with respect to the berry weight than Tempranillo and Garnacha grapes originated wines with higher tannin concentration and astringency than Tempranillo and Garnacha wines. CONCLUSION: The main conclusion of this study is that the seed weight percentage with respect to the berry weight is one of the main determinants of the final tannin concentration and astringency of red wines.

Oenological tannins to prevent Botrytis cinerea damage in grapes and musts: Kinetics and electrophoresis characterization of laccase.

Enzymatic parameters (KM and Vmax), residual activity, effect of bentonite and electrophoresis characterization of laccase in the presence of different oenological tannins (OT) were investigated in relation to B. cinerea negative effects in grapes and musts. Five OT were tested (gallotannin, ellagitannin, quebracho, grape-skin and grape-seed) in comparison with ascorbic acid (AA), sulfur dioxide (SO2) and bentonite. We added OT, AA, SO2 and bentonite to botrytized must obtained by inoculation of grapes with B. cinerea strain 213. Laccase activity was measured by the syringaldazine method at different concentrations of substrate. Enzymatic parameters were determined using Michaelis-Menten and Lineweaver-Burk plots. The B. cinerea strain was also grown in a liquid medium for laccase production. Molecular weight of laccases and effect of OT upon these laccases were studied by SDS-PAGE. Results confirm that bentonite, contrary to OT, did not permit to reduce laccase activity. Regardless the tannin considered, Vmax, KM and laccase activity were reduced and gallotannin, grape-skin and grape-seed tannin presented the greatest ability. Efficiency of grape-seed tannin addition in order to reduce the laccase activity, was comparable to that of AA or SO2 at the typical doses employed in oenology for each one. Oenological tannins appear to be excellent processing aids to prevent laccase effects and contribute to reduce the use of SO2 in grapes and musts.

Occurrence and enological properties of two new non-conventional yeasts (Nakazawaea ishiwadae and Lodderomyces elongisporus) in wine fermentations.

The microbial diversity of wine alcoholic fermentation is not restricted to the presence and activity of Saccharomyces yeast strains. Some non-Saccharomyces species have been described as part of the fermentative microbiota, specially found in the initial steps of wine fermentations. These species may play roles from wine spoilage to flavor quality enhancement. From a large number of wine fermentations (429 wine samples), analyzed by ITS-amplicon sequencing to define their mycobiome, 2 non-conventional yeast species (Nakazawaea ishiwadae and Lodderomyces elongisporus) were detected, in a very limited number of samples but in significant levels of relative abundance. One strain of each species was isolated and their technological and enological potential have been characterized in this work. Compared with the Saccharomyces cerevisiae Viniferm Revelacion wine strain, the studied N. ishiwadae BMK17.1 and L. elongisporus BMK12.5 strains showed, as expected, a lower tolerance and growth fitness in high ethanol concentrations. However, N. ishiwadae BMK17.1 was able to grow also at 15% ethanol and L. elongisporus BMK12.5 at 10% reaching, in the latter case, slightly higher efficiency rates than S. cerevisiae at this level. Contrary to most non-Saccharomyces yeasts, these species were able to growth in presence of high doses of potassium-metabisulfite, reaching in both cases higher efficiency rates than S. cerevisiae. A notable affinity of L. elongisporus BMK12.5 for high pH values was clearly observed. Their fermentation kinetics and the final chemical-analytical characterization were studied in micro-fermentation assays, using synthetic grape must. L. elongisporus BMK12.5 was able to complete, in single inoculation, the sugar fermentation after 19 days, but, N. ishiwadae BMK17.1 left about 80 g/L sugars at this time. Co-inoculation assays (in a 1:100 proportion of S. cerevisiae:non-Saccharomyces strains) finished sugar consumption with similar kinetics than the S. cerevisiae single inoculation, in the case of L. elongisporus BMK12.5 co-inoculation, and with lower kinetics when using N. ishiwadae BMK17.1. A remarkable malic acid consumption and a low acetic acid production was associated with L. elongisporus BMK12.5 fermentations, together with a high production of 3-methyl-1-butanol and 2-phenylethanol, and the release of high amounts of proteins into the wines. N. ishiwadae BMK17.1, although unable to finish the fermentation itself, showed a high production of oligosaccharides and volatile compounds such as isobutanol or isobutyric acid. This work reports, for the first time, the occurrence and enological potential of two strains pertaining to the non-conventional yeast genera Lodderomyces and Nakazawaea.

Copigmentation of Malvidin-3-O-Monoglucoside by Oenological Tannins: Incidence on Wine Model Color in Function of Botanical Origin, pH and Ethanol Content.

The effect of the botanical origin, pH level and ethanol content of different oenological tannins on the color of malvidin-3-O-monoglucoside solution, including their effectiveness as copigments, was studied. Briefly, a model wine solution (4 g/L of tartaric acid, pH 3.5 and 12% ethanol) containing 50 mg/L of malvidin-3-O-monoglucoside was prepared and supplemented with 0.1, 0.2 and 0.4 g/L of commercial tannins using (-)-epicatechin as reference control copigment. Six additional model wine solutions (12% ethanol at pH 3.1, 3.5 or 3.9, and 10%, 12% or 14% ethanol at pH 3.5) were prepared as previously described. Samples were stored under airtight conditions. After a week the full absorbance spectrum in the visible range (400-800 nm) was measured and CIELAB color space was determined. These measurements, including an increase in a* (redness), a decrease in b* (yellowness) and a decrease in L* (lightness), indicated that all oenological tannins had a clear positive effect on color copigmentation. Moreover, hydrolysable tannins appeared to be better copigments than condensed tannins as the copigmentation effectiveness (Cp) was found to be between two to four times higher. The effects of these tannins were dose-dependent because a higher addition resulted in a greater impact on copigmentation. In general, an increase in pH and ethanol content resulted in a decrease of the effect of tannins on color. Independent of intrinsic wine conditions, hydrolysable tannins, more specifically gallotannin, remain the most effective in increasing red wine color. These results prove that supplementation with oenological tannins, especially hydrolysable tannins, could be an interesting tool for the improvement of the red wine color.

Chemical characterization, antioxidant properties and oxygen consumption rate of 36 commercial oenological tannins in a model wine solution.

The chemical composition (CC), antioxidant capacity (AC) and oxygen consumption rate (OCR) of 36 different commercial tannins were measured. The CC was analyzed by total polyphenol index, Bate-Smith, methyl-cellulose, Folin-Ciocalteu, OIV official method and phloroglucinolisis. The AC was measured by different methods (ABTS, CUPRAC, DPPH, FRAP, ORAC) using Trolox as standard. The OCR was measured using a non-invasive method based on luminescence. The results indicate that it is possible to obtain differentiation between procyanidins/prodelphinidins, profisetinidins/prorobinetidins, gallotannins and ellagitannins by PCA based on their CC data. It is also possible to separate condensed from hydrolysable tannins by PCA based on their AC data. The results show that ellagitannins are the fastest oxygen consumers of the various oenological tannins, followed in descending order by condensed tannins and finally gallotannins. The combination of CC, AC and OCR analyses enable to classify tannins according to their effectiveness in protecting wines against oxidation.

Study of the Interaction of Anthocyanins with Phenolic Aldehydes in a Model Wine Solution.

Aldehydes may be present in wines as a result of metabolic processes during wine fermentation or through oxidation and extraction from wood during wine aging in oak barrels. Apart from acetaldehyde, the most abundant aldehyde in wine, other aldehydes such as furfural and more recently vanillin have shown to contribute to the formation of more stable pigments. The copigmentation effect of phenolic molecules, including flavanols and anthocyanins themselves, has been previously evaluated in wine and model solutions, and even the effect of aldehydes related to wine aging has been documented at different pHs and molar ratios. The copigmentation phenomenon is observed by hyperchromic effects and bathochromic shifts of λmax, and, in the same time, the presence of larger molecular weight pigments, potentially less susceptible to degradation, was followed up. This experimental work intended to evaluate the potential of five different aldehydes, all of which are safe for human consumption and are used in the food industry, to the formation of pyranoanthocyanin-like and polymeric pigments in the model solution.

Oxygen consumption rates by different oenological tannins in a model wine solution.

The kinetics of oxygen consumption by different oenological tannins were measured in a model wine solution using the non-invasive method based on luminiscence. The results indicate that the oxygen consumption rate follows second-order kinetics depending on tannin and oxygen concentrations. They also confirm that the oxygen consumption rate is influenced by temperature in accordance with Arrhenius law. The indications are that ellagitannins are the fastest oxygen consumers of the different oenological tannins, followed in decreasing order by quebracho tannins, skin tannins, seed tannins and finally gallotannins. This methodology can therefore be proposed as an index for determining the effectiveness of different commercial tannins in protecting wines against oxidation.

Improved method for the extraction and chromatographic analysis on a fused-core column of ellagitannins found in oak-aged wine.

A new method for the analysis of ellagitannins observed in oak-aged wine is proposed, exhibiting interesting advantages with regard to previously reported analytical methods. The necessary extraction of ellagitannins from wine was simplified to a single step of solid phase extraction (SPE) using size exclusion chromatography with Sephadex LH-20 without the need for any previous SPE of phenolic compounds using reversed-phase materials. The quantitative recovery of wine ellagitannins requires a combined elution with methanol and ethyl acetate, especially for increasing the recovery of the less polar acutissimins. The chromatographic method was performed using a fused-core C18 column, thereby avoiding the coelution of main ellagitannins, such as vescalagin and roburin E. However, the very polar ellagitannins, namely, the roburins A, B and C, still partially coeluted, and their quantification was assisted by the MS detector. This methodology also enabled the analysis of free gallic and ellagic acids in the same chromatographic run.

The effect of supplementation with three commercial inactive dry yeasts on the colour, phenolic compounds, polysaccharides and astringency of a model wine solution and red wine.

BACKGROUND: Nowadays supplementing red wines with commercial inactive dry yeasts is a widespread practice in winemaking because it leads to better balanced wines through increased mouthfeel and smooth astringency. The aim of this article is to study, in a red wine and in a model wine solution, how supplementation with three commercial inactive dry yeasts affects chemical composition and astringency. This will give us a better understanding of the action mechanism involved. RESULTS: The results suggest that this action mechanism is related to two different phenomena. The first is that inactive yeasts release polysaccharides and oligosaccharides which can increase mouthfeel and inhibit interactions between salivary protein and tannins. The second is that they have a direct effect on the precipitation or absorption of proanthocyanidins, especially the larger polymers, which have been described as the most astringent. CONCLUSION: It can be concluded that supplementation with inactive yeasts is indeed a useful tool for smoothing the astringency of red wines. © 2016 Society of Chemical Industry.

Neisseria meningitidis and Streptococcus pneumoniae as leading causes of pediatric bacterial meningitis in nine Mexican hospitals following 3 years of active surveillance.

OBJECTIVES: Meningococcal meningitis is reported as a rare condition in Mexico. There are no internationally published studies on bacterial causes of meningitis in the country based on active surveillance. This study focuses on finding the etiology of bacterial meningitis in children from nine Mexican Hospitals. METHODS: From January 2010 to February 2013, we conducted a three years of active surveillance for meningitis in nine hospitals throughout Mexico. Active surveillance started at the emergency department for every suspected case, and microbiological studies confirmed/ruled out all potentially bacterial pathogens. We diagnosed based on routine cultures from blood and cerebrospinal fluid (not polymerase chain reaction or other molecular diagnostic tests), and both pneumococcal serotyping and meningococcal serogrouping by using standard methods. RESULTS: Neisseria meningitidis was the leading cause, although 75% of cases occurred in the northwest of the country in Tijuana on the US border. Serogroup C was predominant. Streptococcus pneumoniae followed Neisseria meningitides, but was uniformly distributed throughout the country. Serotype 19A was the most incident but before universal implementation of the 13-valent pneumococcal conjugate vaccine. Other bacteria were much less common, including Enterobacteriaceae and Streptococcus agalactiae (these two affecting mostly young infants). CONCLUSIONS: Meningococcal meningitis is endemic in Tijuana, Mexico, and vaccination should be seriously considered in that region. Continuous universal vaccination with the 13-valent pneumococcal conjugate vaccine should be nationally performed, and polymerase chain reaction should be included for bacterial detection in all cultures - negative but presumably bacterial meningitis cases.

Influence of Grape Seeds and Stems on Wine Composition and Astringency.

The aim of this paper is to study the real influence of seeds and stems on wine composition, astringency, and bitterness. A decolored grape juice and a grape juice macerated for 4 days from the same Cabernet Sauvignon grapes were fermented with or without supplementation with 100% seeds, 300% seeds, or 100% stems. Once alcoholic fermentation had finished, the wines were analyzed and tasted. The presence of seeds and stems increased the concentration of flavan-3-ol monomers with respect to the controls. However, the seeds mainly released (+)-catechin and (-)-epicatechin, whereas the stems mainly released (+)-catechin and (+)-gallocatechin. The seeds and stems also released proanthocyanidins; those from seeds have a lower mDP and a high percentage of galloylation, whereas those from stems have a higher mDP and a relatively high percentage of prodelphinidins. The presence of seeds and stems brought about a slight but significant increase in pH and lowered titratable acidity and ethanol content. The presence of seeds boosted color intensity, whereas stems had the opposite effect. Finally, both seeds and stems increased wine astringency and bitterness.

Contribution of yeast and base wine supplementation to sparkling wine composition.

BACKGROUND: The differential characteristic of sparkling wine is the formation of foam, which is dependent, among other factors, on yeast autolysis, aging and oenological practices. In this study, we analyzed the effects of yeast strain, nutrient supplementation to the base wine and aging process on the sparkling wine composition and its foamability. RESULTS: We determined that the addition of inorganic nitrogen promoted nitrogen liberation to the extracellular medium, while the addition of inactive dry yeast to the base wine caused an increase in the polysaccharide concentration and foaming properties of the sparkling wine. The use of synthetic and natural base wines allowed us to discriminate that the differences in high-molecular-weight polysaccharides and oligosaccharides could be attributed to the yeast cells and that the higher nitrogen content in the natural wine could be due to external proteolysis. CONCLUSION: The practices of nitrogen addition and supplementation of inactive dry yeast could modulate the main characteristics of the sparkling wine and be a critical element for the design of this kind of wine. © 2016 Society of Chemical Industry.

Oxygen consumption by oak chips in a model wine solution; Influence of the botanical origin, toast level and ellagitannin content.

The botanical origin, toast level and ellagitannin content of oak chips in a model wine solution have been studied in terms of their influence on oxygen consumption. French oak chips released significantly higher amounts of ellagitannins than American oak chips at any toast level. The release of ellagitannins by oak chips decreased as the toast level increased in the French oak but this trend was not so clear in American oak. Oxygen consumption rate was clearly related to the level of released ellagitannins. Therefore, oak chips should be chosen for their potential to release ellagitannins release should be considered, not only because they can have a direct impact on the flavor and body of the wine, but also because they can protect against oxidation.

Influence of the botanical origin and toasting level on the ellagitannin content of wines aged in new and used oak barrels.

The influence of the botanical origin (French oak: Quercus petraea and American oak: Quercus alba) and different toasting levels (light, medium and heavy) on the ellagitannin content of wines aged in oak barrels has been studied. This took place in two consecutive vintages in order to study what happens after the barrels have been already used for one year with another wine. This study was carried out with two red wines (Cabernet Sauvignon) and with two decolored white wines (Macabeo) from vintages 2012 and 2013 in order to work with a simpler matrix which facilitates chemical analysis. The results show that the botanical origin, toasting level and the number of times that the barrels have been used exert a major influence on the final ellagitannin concentration. In general, the behavior of all the individual ellagitannins was very similar to that described for the total ellagitannins. Briefly, the levels of total ellagitannins concentration in the decolored white wine aged for 12months in new French barrels ranged between 31.2mg/L in the lightly toasted and 4.7mg/L in the heavy toasted. In contrast, these levels were quite lower in American new barrels ranging between 3.6mg/L and 0.9mg/L. Finally, the total ellagitannin concentration decreased an average of 63% in the wines aged in the one year used barrels.

[Gastro-bronchial fistula major complication of sleeve gastrectomy]. / Fístula gastrobronquial: complicación mayor de manga gástrica.

BACKGROUND: Gastrobronchial fistula is a rare complication in gastroesophageal surgical procedures. It is difficult to diagnose and handling is complex. Therefore, there have been developments for non-surgical alternatives to obliterate minor fistula mortality. Endoscopic treatment is an option for patients with small fistulas or in serious condition. CLINICAL CASE: A 38 year old woman with evidence of gastrobronchial fistula postoperated of gastric sleeve, diagnosed during the postoperative period due to clinical variegated was initially handled as infectious respiratory symptoms; once the fistulous path was established, the intention was to close the path using endoclips. However, adding hemopneumothorax, drainage tube placement and thoracotomy were required. It was decided to chose a prosthetic esophageal endoscopic management of polytetraflouroethylene and fibrin as last therapy, because the patient had systemic inflammatory response syndrome, with favorable response to endoscopic management. DISCUSSION: Bariatric surgery has shown satisfactory results, however, the complexity of the procedure favors severe complications such as the present case. Gastrobronchial fistulas represent a diagnostic and therapeutic challenge, this is considering from a conservative management to endoscopic procedures, as in our patient. CONCLUSION: Although gastrobronchial fistulas are a rare complication, the use of endoscopy in resolution should be a first class weapon in its management, since it offers a lower morbidity in a patient with habitual respiratory symptoms that are difficult to control, with satisfactory results in the medium and long term.

[Spontaneous cholecystocutaneous fistula]. / Fístula colecistocutánea espontánea.

BACKGROUND: Spontaneous cholecystocutaneous fistula is defined as a gallbladder communication with the external environment through the abdominal wall rupture; the first reports were written in the seventeenth century by Thilesus. During the past 50 years 25 cases have been reported. CLINICAL CASE: We report a case of a 30-year-old woman presented with a five-year history of biliary colic, six months prior to medical assessment presents outlet of biliary material and gallstones spontaneously in the right upper quadrant. Fistulogram was performed without evidence of obstruction, subsequently cholecystectomy and resection of the fistula was performed. DISCUSSION: The biliary fistulas are an abnormal communication from the gallbladder into another surface, is a rare condition in our day as it only occurs in 10% of patients with gallbladder lithiasis; while spontaneous cholecystocutaneous fistula clinic is more than evident. It is imperative perform studies like ultrasound, tomography, and fistulogram. The mainstay of treatment is cholecystectomy, resection of the fistula and repair of abdominal wall defect. CONCLUSION: The incidence of cholecystocutaneous fistula today is minimal and it seems that the current trend is to become an entity anecdotal, the approach subcostal abdominal examination remains as the first choice. The laparoscopic approach is an option reserved for the experienced surgeon.