Progress 10/01/07 to 09/30/12
Outputs
OUTPUTS: We have subjected 6 nutrient formulations to elemental, vitamin, fatty acid and amino acid analyses. In addition, glutathion, ammonium and physical characteristics were determined. Growth studies were carried out with the selected nutrients in a model wine. Significant differences are found between the nutrients with regards to vitamins, amino acids and the elemental composition. These differences reflect variations in medium constituents used for the propagation of yeast. Interestingly, none of the nutrients was able to support growth of any of the 3 bacterial strains tested in the model wine suggesting that they did not supply all essential nutrients. Based on our evaluations, deficiencies in divalent metals and vitamins are most likely to account for this observation. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Targeted supplementation of yeast extract based enological nutrients intended for malolactic fermentation is likely to improve growth of wine lactic acid bacteria and bacterial deacidifcation. Further studies should evaluate the practical consequences of such supplementations and consider the legal framework in various winemaking countries.
Publications
- Various presentations and abstracts were offered at national conferences, specifically the 2012 ASEV and 2012 ASEV-ES conferences.
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Progress 10/01/10 to 09/30/11
Outputs
OUTPUTS: In this year, we have obtained 6 nutritional formulations (commercially available) and started their physiochemical evaluation as well as their effect on the growth of wine lactic acid bacteria. A significant effect of nutrients on the growth and malolactic activity of bacteria was found. Rather than by amino acid content, the data suggests a strong correlation of certain cations, including manganese and magnesium on the performance of wine lactic acid bacteria. Further characterization of the nutrients with regards to the vitamin and fatty acid content will allow to establish stronger associations. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The current project focus will allow to identify those compositional factors that are mostly responsible for supporting growth and metabolic activity of wine lactic acid bacteria with the aim of supporting malolactic fermentation in difficult to ferment cool climate wines.
Publications
- Christen, S., Li, E., Dube Morneau, A., Jackowetz, N. and Mira de Orduna, R., (2011) Bacterial Metabolism of Acetaldehyde and other SO2 binding compounds. Symposium of the International Association for Enology, Management and Wine Marketing. Eurac Convention Center, Bozen, Sudtirol, Italy.
- Mira de Orduna, R., Terrade, N., Jackowetz, N., Dube Morneau, A., Li, E. (2011) Nutrition and metabolism of wine lactic acid bacteria: Recent advances. Presentation at Danisco France. Dange St. Romain, France
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Progress 10/01/09 to 09/30/10
Outputs
OUTPUTS: We have further developed a chemically defined medium for wine lactic acid bacteria and determined the essential nutrient needs of several wine related lactic acid bacteria by the single omission technique. Together with a better understanding of the amino acid composition of New York wines, the data can be used in order to assess malolactic fermentability of New York State wines. We have also determined the effect of the amino acids arginine and citrulline on growth of various wine lactic acid bacteria. Results from these studies have been presented at various conferences including local (NY), national and international symposia, and several manuscripts have been published in international peer reviewed journals. The results allow winemakers to appreciate the effect of nutrient additions to malolactic fermentability and microbiological stability of wines. They also allow the producers of bacterial starters and nutrients to improve their formulations in order to ensure product quality. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: The direct target audience of this project is the New York State wine industry. However, project results are of interest to the global wine industry. The final objective of this project is a reduction in failed malolactic fermentations and improved wine quality, especially with regards to reduced biogenic amine concentrations, which can cause adverse effects in sensitive consumers. Accordingly, wine consumers can be regarded as the eventual target audience, as well. Finally, companies that produce microbial starters for malolactic fermentation and nutritional formulations to support the latter, will benefit from increased knowledge about the specific needs of the microorganisms. It is expected that know-how generated in this study be implemented in new formulations. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
We have presented a new chemically defined medium for the investigation of wine lactic acid bacteria. The medium compares well to traditional complex laboratory media and was successfully used for the growth of various wine lactic acid bacteria of the genera Oenococcus, Lactobacillus and Pediococcus. This medium was the basis for the comprehensive nutritional studies of wine lactic acid bacteria. The aim of the latter work was to investigate the essential nutrient requirements of Oenococcus and Lactobacillus wine lactic acid bacteria in order to assess suitability of musts and wines to support growth of lactic acid bacteria and thus, malolactic fermentation, but also to cause spoilage. It was found that Oenococcus and Lactobacillus strains could be grouped based on their nutritional requirements. Specifically, oenococci required more amino acids than the Lactobacillus strains studied. The research suggests that nutritional deficiencies are more likely to affect oenococci than lactobacilli and hence, proper bacterial nutrition may require specific attention to conduct malolactic fermentation with pure Oenococcus oeni strains. With regards to the metabolism of amino acids (Objective 2) it has been shown previously that arginine degradation by wine lactic acid bacteria during malolactic fermentation can contribute to the formation of the carcinogenic ethyl carbamate precursor citrulline. If arginine also increased growth of wine lactic acid bacteria, this could lead to arginine degrading bacteria having an ecological advantage in wine causing further arginine degradation and citrulline formation. Our project results have shown that both Oenococcus oeni strains as well as non-Oenococcus bacteria degraded arginine, but only non-Oenococcus bacteria (i.e. Lactobacillus spp.) were stimulated by arginine underlining the potential of arginine to support non-oenococcal populations in wines. It was also shown that arginine degradation by oenococci can be avoided if the bacteria are inactivated after malic acid depletion. In order to reduce ethyl carbamate precursor citrulline formation, the results suggest to carry our malolactic fermentation with pure strains of Oenococcus oeni and to inactivate the biomass rapidly after depletion of malic acid. In the last reporting period, we have further improved the chemically defined medium in order to provide for buffering and, hence, increased growth.We have also investigated an improved method for the determination of essential nutrient requirements applying a miniaturized assay that uses so-called microtiter plates. This approach allows to measure the growth of 96 cultures at the same time and was compared to the traditional method applying test tubes. These modifications have been applied to the determination of the nutritional requirements of two new strains of Lactobacillus plantarum, which are promising additions to the pool of malolactic straters and provide new opportunities in the wine industry for carrying our malolactic fermentation in very acidic wines.
Publications
- 1. Mira de Orduna, R. (2010) Management of malolactic fermentations. Proceedings from the Viticulture 2010 and 39th New York State Wine Industry Workshop, Riverside Convention Center, Rochester, USA.
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Progress 10/01/08 to 09/30/09
Outputs
OUTPUTS: We have developed a chemically defined medium for wine lactic acid bacteria and determined the essential nutrient needs of these bacteria by the single omission technique. Together with a better understanding of the amino acid composition of New York wines, the data can be used in order to assess malolactic fermentability of New York State wines. We have also determined the effect of the amino acids arginine and citrulline on growth of various wine lactic acid bacteria. Results from these studies have been presented at various conferences and 3 manuscripts have been published in international peer reviewed journals. Current results allow winemakers to appreciate the effect of nutrient additions to malolactic fermentability and microbiological stability of wines. PARTICIPANTS: The collection of must samples was carried out in collaboration with Stephanie Mira de Orduna and the New Yorks State wine industry. Laboratory work regarding the nutritional requirements of wine lactic acid bacteria was carried out by PhD candidate Nicolas Terrade as well as the summer interns Ronan Noel and Romain Couillaud. TARGET AUDIENCES: The direct target audience of this project is the New York State wine industry. However, project results are of interest to the global wine industry. The final objective of this project is a reduction in failed malolactic fermentations and improved wine quality, especially with regards to reduced biogenic amine concentrations, which can cause adverse effects in sensitive consumers. Accordingly, wine consumers can be regarded as the eventual target audience, as well. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
We have presented a new chemically defined medium for the investigation of wine lactic acid bacteria. The medium compares well to traditional complex laboratory media and was successfully used for the growth of various wine lactic acid bacteria of the genera Oenococcus, Lactobacillus and Pediococcus. This medium was the basis for the comprehensive nutritional studies of wine lactic acid bacteria. The aim of the latter work was to investigate the essential nutrient requirements of Oenococcus and Lactobacillus wine lactic acid bacteria in order to assess suitability of musts and wines to support growth of lactic acid bacteria and thus, malolactic fermentation, but also to cause spoilage. It was found that Oenococcus and Lactobacillus strains could be grouped based on their nutritional requirements. Specifically, oenococci required more amino acids than the Lactobacillus strains studied. The research suggests that nutritional deficiencies are more likely to affect oenococci than lactobacilli and hence, proper bacterial nutrition may require specific attention to conduct malolactic fermentation with pure Oenococcus oeni strains. With regards to the metabolism of amino acids (Objective 2) it has been shown previously that arginine degradation by wine lactic acid bacteria during malolactic fermentation can contribute to the formation of the carcinogenic ethyl carbamate precursor citrulline. If arginine also increased growth of wine lactic acid bacteria, this could lead to arginine degrading bacteria having an ecological advantage in wine causing further arginine degradation and citrulline formation. Our project results have shown that both Oenococcus oeni strains as well as non-Oenococcus bacteria degraded arginine, but only non-Oenococcus bacteria (i.e. Lactobacillus spp.) were stimulated by arginine underlining the potential of arginine to support non-oenococcal populations in wines. It was also shown that arginine degradation by oenococci can be avoided if the bacteria are inactivated after malic acid depletion. In order to reduce ethyl carbamate precursor citrulline formation, the results suggest to carry our malolactic fermentation with pure strains of Oenococcus oeni and to inactivate the biomass rapidly after depletion of malic acid. In the last reporting period, we have elaborated a new ultra high pressure HPLC method for the determination of amino acids using sub 2 micron particle size reverse phase columns, that would allow rapid and efficient profiling of New York musts and wines, and is that is currently being validated. We have also commenced to determine the nutritional requirements of several strains of Lactobacillus plantarum, which will become more significant as starter organisms to induce malolactic fermentation in the future.
Publications
- Terrade, N. and Mira de Orduna, R. (2009) Arginine and citrulline do not stimulate growth of two Oenococcus oeni strains in wine. FEMS Microbiol. Lett. 290, 98-104.
- Terrade, N. and Mira de Orduna, R. (2009) Determination of the essential nutrient requirements of wine-related bacteria from the genera Oenococcus and Lactobacillus. Int. J. Food Microbiol. 133, 8-13.
- Terrade, N., Noel, R., Couillaud, R. and Mira de Orduna, R. (2009) A new chemically defined medium for wine lactic acid bacteria. Food Res. Int. 42, 363-367.
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Progress 10/01/07 to 09/30/08
Outputs
OUTPUTS: We have collected 100 musts from over 30 wineries in order to analyze amino acid levels of New York State wineries. Must donations have been received from most regions of New York State including the Finger Lakes, Lake Erie, Hudson River and Long Island. Musts were sterile filtered and analysed for amino acids by HPLC. The data obtained requires further analysis and representation. Concurrently, we have developed a chemically defined medium for wine lactic acid bacteria and determined the essential nutrient needs of these bacteria by the single omission technique. Once the amino acid profiles are available, the data can be compared with the nutritional needs of bacteria in order to assess malolactic fermentability of New York State musts and wines. Finally, we have determined the effect of the amino acids arginine and citrulline on growth of various wine lactic acid bacteria. Results from these studies have been presented at various conferences and 3 manuscripts have been submitted recently, two of which have been accepted for publication (both in press). Results from the objectives of the project will allow wineries to assess the composition of their wines with regards to their fermentability and microbiological stability. PARTICIPANTS: The collection of must samples was carried out in collaboration with Stephanie Mira de Orduna. Laboratory work regarding the nutritional requirements of wine lactic acid bacteria was carried out by PhD candidate Nicolas Terrade as well as the summer interns Ronan Noel and Romain Couillaud. TARGET AUDIENCES: The direct target audience of this project is the New York State wine industry. However, project results are of interest to the global wine industry. The final objective of this project is a reduction in failed malolactic fermentations and improved wine quality, especially with regards to reduced biogenic amine concentrations, which can cause adverse effects in sensitive consumers. Accordingly, wine consumers can be regarded as the eventual target audience, as well. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
We have presented a new chemically defined medium for the investigation of wine lactic acid bacteria. The medium compares well to traditional complex laboratory media and was successfully used for the growth of various wine lactic acid bacteria of the genera Oenococcus, Lactobacillus and Pediococcus. This medium was the basis for the comprehensive nutritional studies of wine lactic acid bacteria. The aim of the latter work was to investigate the essential nutrient requirements of Oenococcus and Lactobacillus wine lactic acid bacteria in order to assess suitability of musts and wines to support growth of lactic acid bacteria and thus, malolactic fermentation, but also to cause spoilage. It was found that Oenococcus and Lactobacillus strains could be grouped based on their nutritional requirements. Specifically, oenococci required more amino acids than the Lactobacillus strains studied. The research suggests that nutritional deficiencies are more likely to affect oenococci than lactobacilli and hence, proper bacterial nutrition may require specific attention to conduct malolactic fermentation with pure Oenococcus oeni strains. With regards to the metabolism of amino acids (Objective 2) it has been shown previously that arginine degradation by wine lactic acid bacteria during malolactic fermentation can contribute to the formation of the carcinogenic ethyl carbamate precursor citrulline. If arginine also increased growth of wine lactic acid bacteria, this could lead to arginine degrading bacteria having an ecological advantage in wine causing further arginine degradation and citrulline formation. Our project results have shown that both Oenococcus oeni strains as well as non-Oenococcus bacteria degraded arginine, but only non-Oenococcus bacteria (i.e. Lactobacillus spp.) were stimulated by arginine underlining the potential of arginine to support non-oenococcal populations in wines. It was also shown that arginine degradation by oenococci can be avoided if the bacteria are inactivated after malic acid depletion. In order to reduce ethyl carbamate precursor citrulline formation, the results suggest to carry our malolactic fermentation with pure strains of Oenococcus oeni and to inactivate the biomass rapidly after depletion of malic acid. It is expected that the project results will lead to a lesser incidence of sluggish and stuck malolactic fermentations in New York State by matching amino acid profiles with nutritional requirements of malolactic bacteria. Further work on amino acid metabolism will allow to reduce the formation of biogenic amines, which can lead to negative health effects in sensitive consumers.
Publications
- Terrade, N. and Mira de Orduna, R. (2009) Arginine and citrulline do not stimulate growth of two Oenococcus oeni strains in wine. IN PRESS. FEMS Microbiol. Letters 290, 98-104
- Terrade, N., Noel, R., Couillaud, R. and Mira de Orduna, R. (2009) A new chemically defined medium for wine lactic acid bacteria. Food Res. Int. IN PRESS
- Terrade, N. and Mira de Orduna, R. (2009) Determination of the essential nutrient requirements of two Oenococcus oeni and two Lactobacillus spp. wine lactic acid bacteria. Submitted to J. Int. Food Microbiol. PENDING
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