Terroir 2004 banner
IVES 9 IVES Conference Series 9 The use of epifluorescence versus plating to monitor the effect of different parameters on microorganisms in wine

The use of epifluorescence versus plating to monitor the effect of different parameters on microorganisms in wine

Abstract

The monitoring of the number of micro-orgranisms in wine is crucial for the wine producer. Traditional counting methods include microscopic enumeration and plating on selective media, which measures the culturability of the cells. The use of epifluorescence microscopy is, however, a method, which can measure both culturability and viability in wine. This method distinguishes between live and dead cells. Research showed that little difference existed between plating and epifluorescence numbers to enumerate lactic acid bacteria in wine. However, a difference exists between these two methods to distinguish between acetic acid bacteria numbers in wine. Plating counting numbers were lower than plate numbers for Acetobacter pasteurianus in wine under anaerobic conditions. This difference was, however, negated by the addition of oxygen to the wine. SO2 additions lowered the culturability of A. pasteurianus at dosages higher than 0.35mg/L molecular SO2, but higher dosages were required to lower epifluorescence intensity, which is an indication of viability. Brettanomyces bruxellensis culturability was inhibited at lower dosages, but total cell numbers according to epifluorescence microscopy were affected at higher molecular SO2 dosages. Epifluorescence microscopy and plating also showed that B. bruxellensis was drastically affected after 120 min after molecular SO2 addition and it’s culturability after only 30 min. An exposure time of 5 min to molecular SO2 reduced the cell’s viability drastically and 45 min completely inhibiting the viability after two days. The bonded form of sulphur dioxide did not affect both micro-organisms. Epifluorescence microscopy can thus be used as a quick alternative to assess micro-organisms numbers and culturability in wine. This technique has both advantages and disadvantages over traditional enumeration methods, which will also be discussed.

DOI:

Publication date: January 12, 2022

Issue: Terroir 2004

Type: Article

Authors

W.J. du Toit (1), I.S. Pretorius (1,2) and A. Lonvaud-Funel (3)

(1) Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, Victoria Street, ZA 7600 Stellenbosch, South Africa
(2)The Australian Wine Research Institute, Waite Road, Urrbrae, SA 5064 Adealide, Australia
(3) Faculté d’Oenologie, 351, Cours de la Libération, 33405 Talence, France

Contact the author

Tags

IVES Conference Series | Terroir 2004

Citation

Related articles…

Grapevine xylem embolism resistance spectrum reveals which varieties have a lower mortality risk in a future dry climate

Wine growing regions have recently faced intense and frequent droughts that have led to substantial economical losses, and the maintenance of grapevine productivity under warmer and drier climate will rely notably on planting drought-resistant cultivars. Given that plant growth and yield depend on water transport efficiency and maintenance of photosynthesis, thus on the preservation of the vascular system integrity during drought, a better understanding of drought-related hydraulic traits that have a significant impact on physiological processes is urgently needed. We have worked towards this end by assessing vulnerability to xylem embolism in 30 grapevine commercial varieties encompassing red and white Vitis vinifera varieties, hybrid varieties characterized by a polygenic resistance for powdery and downy mildew, and commonly used rootstocks. These analyses further allowed a global assessment of wine regions with respect to their varietal diversity and resulting vulnerability to stem embolism. Hybrid cultivars displayed the highest vulnerability to embolism, while rootstocks showed the greatest resistance. Significant variability also arose among Vitis vinifera varieties, with Ψ12 and Ψ50 values ranging from -0.4 to -2.7 MPa and from -1.8 to -3.4 MPa, respectively. Cabernet franc, Chardonnay and Ugni blanc featured among the most vulnerable varieties while Pinot noir, Merlot and Cabernet Sauvignon ranked among the most resistant. In consequence, wine regions bearing a significant proportion of vulnerable varieties, such as Poitou-Charentes, France and Marlborough, New Zealand, turned out to be at greater risk under drought. These results highlight that grapevine varieties may not respond equally to warmer and drier conditions, outlining the importance to consider hydraulic traits associated with plant drought tolerance into breeding programmes and modeling simulations of grapevine yield maintenance under severe drought. They finally represent a step forward to advise the wine industry about which varieties and regions would have the lowest risk of drought-induced mortality under climate change.

Organic recycled mulches in sustainable viticulture: assessment of spontaneous plants communities and weed coverage

In recent years, developing more efficient and sustainable viticulture management has been essential due to the impact of climate change in semiarid regions. For this reason, the use of recycled organic mulching (ROM) in the vineyard has become an interesting strategy to cope with water stress, isolated soil from extreme temperatures and improving soil humidity, control the presence of weeds and therefore reduce the inputs of herbicides and improve soil fertility. This work aimed to analyse the effect of three different organic mulches [straw (S), grape pruning debris (GPD) and spent mushroom compost (SMC)] and two traditional soil management techniques [herbicide (H) and interrow (IN)] on weed coverage and the spontaneous plant communities’ presence. Data sampling was collected throughout the vine vegetative cycle of 2021 in La Rioja, Spain. The different soil management techniques had a clear effect on weed coverage and his development during the vine vegetative cycle. SMC and H were the treatments with the highest and the lowest coverage percentage, respectively. IN had a delayed weed emergence at the beginning of the vine vegetative cycle, but finally it reached maximum values nearby SMC. GPD and S had similar effects on weed emergence, reaching 25-30% of the maximum coverage values. A total of 29 herbaceous species were identified during the vegetative cycle, some of them very isolated and occasional. Principal component analysis (PCAs) showed a good association between spontaneous species and treatments, furthermore, specific species-treatment associations were found. Moreover, three clear groups of herbaceous communities were identified by cluster analysis. This study provides interesting information about the effect of different alternative soil management on herbaceous plant coverage and weed species communities which could contribute to making more sustainable viticulture.

Pruned vine biomass exclusion from a clay loam vineyard soil – examining the impact on physical/chemical properties

The wine industry worldwide faces increasing challenges to achieve sustainable levels of carbon emission mitigation. This project seeks to establish the feasibility of harvesting winter pruned vineyard biomass (PVB) for potential use in carbon footprint reduction, through its use as a renewable biofuel for energy production. In order to make this recommendation, technical issues such as the potential environmental impact, chemical composition and fuel suitability, and logistical challenges of harvesting biomass needs to be understood to compare with the results from similar studies. Of particular interest is the role PVB plays as a carbon source in vineyard soils and what effect annual removal might have on soil carbon sequestration. A preliminary trial was established in the Waite Campus vineyard (University of Adelaide) to test current management strategies. Vines are grown in a Eutrophic, Red Dermosol clay loam soil with well managed midrow swards. A comparison was undertaken of mid-row treatments in two 0.25 Ha blocks (Shiraz and Semillon), including annual cultivation for seed bed preparation, the deliberate exclusion of PVB (25 years) and incorporation of PVB (13 years) at an average of 3.4 and 5.5 Mg/Ha-1 for Shiraz and Semillon respectively. In both 0-10cm and 10-30cm soil core sample depths, combined soil carbon % measures in the desired range of 1.80 to 3.50, were not significantly different between treatments or cultivars and yielded an estimated 42 Mg/ha-1 of sequestered soil carbon. Other key physical and chemical measures were likewise not significantly different between treatments. Preliminary results suggest that in a temperate zone vineyard, managed such as the one used in this study, there is no long term negative impact on soil carbon sequestration through removing PVB. This implies that growers could confidently harvest PVB for use in several end fates including as a bio fuel.

Comparison of imputation methods in long and varied phenological series. Application to the Conegliano dataset, including observations from 1964 over 400 grape varieties

A large varietal collection including over 1700 varieties was maintained in Conegliano, ITA, since the 1950s. Phenological data on a subset of 400 grape varieties including wine grapes, table grapes, and raisins were acquired at bud break, flowering, veraison, and ripening since 1964. Despite the efforts in maintaining and acquiring data over such an extensive collection, the data set has varying degrees of missing cases depending on the variety and the year. This is ubiquitous in phenology datasets with significant size and length. In this work, we evaluated four state-of-the-art methods to estimate missing values in this phenological series: k-Nearest Neighbour (kNN), Multivariate Imputation by Chained Equations (mice), MissForest, and Bidirectional Recurrent Imputation for Time Series (BRITS). For each phenological stage, we evaluated the performance of the methods in two ways. 1) On the full dataset, we randomly hold-out 10% of the true values for use as a test set and repeated the process 1000 times (Monte Carlo cross-validation). 2) On a reduced and almost complete subset of varieties, we varied the percentage of missing values from 10% to 70% by random deletion. In all cases, we evaluated the performance on the original values using normalized root mean squared error. For the full dataset we also obtained performance statistics by variety and by year. MissForest provided average errors of 17% (3 days) at budbreak, 14% (4 days) at flowering, 14.5% (7 days) at veraison, and 17% (3 days) at maturity. We completed the imputations of the Conegliano dataset, one of the world’s most extensive and varied phenological time series and a steppingstone for future climate change studies in grapes. The dataset is now ready for further analysis, and a rigorous evaluation of imputation errors is included.

Assessment of climate change impacts on water needs and growing cycle on grapevine in three DOs of NE Spain

This study assessed the suitability of grapevine growing in three DOs (Empordà, Pla de Bages and Penedès) of Catalonia (NE Spain) over the 21st century. For this purpose, an estimation of water needs and agroclimatic and phenological indicators was made. Climate change impacts were estimated at 1 km pixel resolution using temperature and precipitation projections from several general circulation models (GCM) and two climate change scenarios: RCP 4.5 (stabilization scenario) and RCP 8.5 (worst-case scenario). Potential crop evapotranspiration (following FAO procedure) and a daily water balance considering soil water holding capacity were used to estimate actual evapotranspiration of vines and, finally, water needs. Dynamics would be similar in the three DOs studied although the magnitude of impact differs. Water needs would be 2 and 3 times greater (ranging from 0 to more than 1500 m3/ha) than current water needs at both climate change scenarios. Moreover, blooming date would advance from 3 to 6 weeks, harvest date from 1 to 2.5 months, resulting in growing cycles from 10 to 80 days shorter. It should also be noted that frost risk would decrease from 6 to 76%, the number of days with temperatures above 30ºC during ripening would rise from 48 to 500% and tropical nights (minimum temperature >20ºC) at ripening would increase from 28 to 150%, depending on the scenario and the DOs. The impacts of climate change in the three DOs could result in significant limitations for grapevine cultivation and wine production if adaptive strategies are not applied. This result could serve as a basis for the design of specific and particular adaptation strategies to improve and maintain vineyards in the DOs studied and could be extrapolated to similar DOs and regions.