Yeast diversity during the spontaneous fermentation of wine with only the microbiota on grapes cultivated in Japan.

Making wine via spontaneous fermentation without sulfur dioxide and commercial yeast (spontaneous winemaking) is increasing in recent year, but there is scant research regarding microbial communities present in Japan during spontaneous winemaking using culture-independent molecular methods. We analyzed fungal communities and populations during laboratory-scale spontaneous winemaking using sterilized labware to avoid winery-resident microbes. In the spontaneous fermentation of four grape varieties (Pinot Noir, Riesling, Koshu, and Koshusanjaku) grown in the same Japanese vineyard, our analysis of yeast and other fungal species by next-generation sequencing based on the ITS1 region demonstrated that Saccharomyces cerevisiae was eventually dominant in seven of 12 fermentation batches (three replications for each grape variety), whereas non-Saccharomyces species (e.g., Schizosaccharomyces japonicus, Lachancea dasiensis, and Hanseniaspora valbyensis) became dominant in four batches at the end of fermentation. In another batch, lactic acid bacteria (LAB) became dominant and the fermentation remained incomplete. Diverse microbes were involved in the spontaneous fermentation (particularly in Koshusanjaku), indicating that residual sugar remained and lactic and acetic acid largely increased. Compared to the control wine made with SO2 and commercial yeast, the concentration of lactic acid was 47-fold higher in the must dominated by L. dasiensis, and the concentrations of acetic acid and lactic acid were 10-fold and 20-fold higher in the must dominated by LAB, respectively. Even when indigenous S. cerevisiae became dominant, the finished wines obtained high sensory-analysis scores for complexity but low scores for varietal typicality, indicating the risk of fermentation with unselected wild yeast on the grapes grown in Japan.

Chemometric approaches for determining the geographical origin of Japanese Chardonnay wines using oxygen stable isotope and multi-element analyses.

Determining the geographical origin of wines is a major challenge in wine authentication, but little information is available regarding non-parametric statistical approaches for wines. In this study, we collected 33 domestic Chardonnay wines vinified on a small scale from grapes cultivated in Japan, and 42 Chardonnay wines imported from 8 countries, for oxygen stable isotope and multi-element analyses. Non-metric multidimensional scaling (NMDS), kernel principal component analysis (KPCA) and principal component analysis (PCA) were applied to the oxygen stable isotopic compositions (δ18O) and the concentrations of 18 elements in the wines to compare the extractions by parametric and non-parametric methods. The non-parametric methods, NMDS and KPCA, separated domestic from imported Chardonnay wines better than the parametric method, PCA. Of 19 variables, 18 were important for geographical discrimination, with the δ18O value being the most significant in all statistic methods. Non-parametric multivariate analyses will help discriminate domestic from imported Chardonnay wines.

Prediction method for determining the carbon stable isotopic composition of berry sugars in the original must of Chardonnay wines.

The carbon stable isotopic composition, as indicated by the δ13C value, of wine ethanol is inherited from berry sugars, but little is known about the variation in sugar δ13C values of Japanese grapes relative to overseas grapes. This study found a large variation in sugar δ13C values of Chardonnay grapes grown in Japan (-27.2 ± 0.9‰, mean ± standard deviation, n = 33), with sugar δ13C values depending on the δ13C values and content of monosaccharides. After complete fermentation, the carbon isotope discrimination between berry sugars and wine ethanol was 1.5 ± 0.1‰. Ethanol δ13C values and carbon isotope discrimination enabled prediction of sugar δ13C values in the original must. Imported wines had higher sugar δ13C values than those of wines made from Japanese grapes, suggesting drier overseas viticulture conditions. The determination of sugar δ13C values in grape berries provides valuable information for viticulture and wine authentication.

Variation in the mineral composition of wine produced using different winemaking techniques.

It has been reported that the concentrations of minerals in wines can be used to discriminate their geographical origin. However, some winemaking techniques may also affect the mineral concentration of the final product. In this study, we examined the effects of various winemaking techniques, including (i) fining, (ii) aging with oak tips, (iii) maceration with grape skins, (iv) chaptalization and acidification, and (v) yeast nutrient addition for alcohol fermentation, on the concentration of 18 minerals (Li, B, Na, Mg, Si, P, S, K, Ca, Mn, Co, Ni, Ga, Rb, Sr, Mo, Ba and Pb) in a total of 154 wine samples using grapes from different production areas. Among the various winemaking techniques, maceration with grape skin had the largest effect on mineral content, significantly changing the concentrations of 17 or 18 elements (B, Na, Mg, Si, P, S, K, Ca, Mn, Co, Ni, Ga, Rb, Sr, Mo, Ba and Pb). Fining treatment using bentonites had the second largest effect, altering the concentrations of 14 elements (Li, B, Na, Si, P, S, Ca, Co, Ga, Rb, Sr, Mo, Ba and Pb). However, in principal component analysis using all data (n = 154), the wine samples were clearly clustered according to grapes used in the experiments rather than the winemaking technique. In conclusion, some winemaking techniques significantly altered the concentration of some minerals in wine; however, the principal factor influencing wine mineral composition seems to be its geographical origin.

Increase in the oxygen stable isotopic composition of water in wine with low ethanol yield.

The stable isotopic composition of oxygen (δ18O) in wine is often analysed to determine the geographic origin of the wine and the amount of water dilution. However, little is known regarding the effects of two major winemaking techniques (the addition of acid (acidification) and sugar (chaptalization)) on the δ18O value of water in wine. Here we show that acidification and chaptalization have minor direct effects on the δ18O value but indirect effects based on the ethanol yield, which causes isotopic variation of up to 0.6‰. During fermentation, δ18O values increase at low ethanol yields, suggesting that yeast release water with a high δ18O value into wine when consuming sugars. Additionally, the ethanol yield is negatively correlated with the consumption of amino acids by the yeast, indicating that yeast growth decreases the ethanol yield. We therefore identify ethanol yield, which is decreased by the consumption of sugars by yeast for non-alcohol-fermentation processes as a potential factor leading to variations in the δ18O value of water during the winemaking process.

Discrimination of wine from grape cultivated in Japan, imported wine, and others by multi-elemental analysis.

Differences in mineral concentrations were examined among three types of wine in the Japanese market place: Japan wine, imported wine, and domestically produced wine mainly from foreign ingredients (DWF), where Japan wine has been recently defined by the National Tax Agency as domestically produced wine from grapes cultivated in Japan. The main objective of this study was to examine the possibility of controlling the authenticity of Japan wine. The concentrations of 18 minerals (Li, B, Na, Mg, Si, P, S, K, Ca, Mn, Co, Ni, Ga, Rb, Sr, Mo, Ba, and Pb) in 214 wine samples were determined by inductively coupled-plasma mass spectrometry (ICP-MS) and ICP-atomic emission spectrometry (ICP-AES). In general, Japan wine had a higher concentration of potassium and lower concentrations of eight elements (Li, B, Na, Si, S, Co, Sr, and Pb) as compared with the other two groups of wine. Linear discriminant analysis (LDA) models based on concentrations of the 18 minerals facilitated the identification of three wine groups: Japan wine, imported wine, and DWF with a 91.1% classification score and 87.9% prediction score. In addition, an LDA model for discrimination of wine from four domestic geographic origins (Yamanashi, Nagano, Hokkaido, and Yamagata Prefectures) using 18 elements gave a classification score of 93.1% and a prediction score of 76.4%. In summary, we have shown that an LDA model based on mineral concentrations is useful for distinguishing Japan wine from other wine groups, and can contribute to classification of the four main domestic wine-producing regions of Japan.

[A case of group G Streptococcus sepsis, chest wall abscess, and vertebral osteomyelitis mimicking a primary lung cancer with bone metastasis].

A 73-year-old woman who had been followed in our department of gynecology because of ovarian cancer since 2002, was admitted with liver dysfunction and complaining of back pain and light precordial chest pain. The chest radiograph on admission revealed a tumor in her left upper lung field, and chest CT revealed a tumor adjacent to the chest wall and mediastinum. FDG-positron emission tomography (PET) showed abnormal uptake in the tumor and Th6/7, and the subaortic lymph nodes. On the basis of these findings, primary lung cancer with bone metastasis was suspected. She had a high grade fever on admission, and blood cultures were positive for group G streptococcus. The treatment with intravenous penicillin was started. Percutaneous biopsy of the tumor in her left chest showed an abscess wall in the chest wall, but no evidence of malignancy. Transbronchial lung biopsy and CT-guided biopsy also showed no malignant cells. Since the tumor decreased in size and back pain improved gradually by only antibiotic treatment, a diagnosis of sepsis of group G streptococcus, chest wall abscess, and vertebral osteomyelitis was made. She was treated with intravenous penicillin for 4 weeks and oral amoxicillin for another 4 weeks. After 60 days of antibiotic treatment, the tumor vanished.