Genetic and Technological Characterisation of Vineyard-and Winery-Associated Lactic Acid Bacteria

Vineyard-and winery-associated lactic acid bacteria (LAB) from two major PDO regions in Greece, Peza and Nemea, were surveyed. LAB were isolated from grapes, fermenting musts, and winery tanks performing spontaneous malolactic fermentations (MLF). Higher population density and species richness were detected in Nemea than in Peza vineyards and on grapes than in fermenting musts. Pediococcus pentosaceus and Lactobacillus graminis were the most abundant LAB on grapes, while Lactobacillus plantarum dominated in fermenting musts from both regions. No particular structure of Lactobacillus plantarum populations according to the region of origin was observed, and strain distribution seems random. LAB species diversity in winery tanks differed significantly from that in vineyard samples, consisting principally of Oenococcus oeni. Different strains were analysed as per their enological characteristics and the ability to produce biogenic amines (BAs). Winery-associated species showed higher resistance to low pH, ethanol, SO 2 , and CuSO 4 than vineyard-associated isolates. The frequency of BA-producing strains was relatively low but not negligible, considering that certain winery-associated Lactobacillus hilgardii strains were able to produce BAs. Present results show the necessity of controlling the MLF by selected starters in order to avoid BA accumulation in wine

Next-Generation Sequencing Reveals Significant Bacterial Diversity of Botrytized Wine

While wine fermentation has long been known to involve complex microbial communities, the composition and role of bacteria other than a select set of lactic acid bacteria (LAB) has often been assumed either negligible or detrimental. This study served as a pilot study for using barcoded amplicon next-generation sequencing to profile bacterial community structure in wines and grape musts, comparing the taxonomic depth achieved by sequencing two different domains of prokaryotic 16S rDNA (V4 and V5). This study was designed to serve two goals: 1) to empirically determine the most taxonomically informative 16S rDNA target region for barcoded amplicon sequencing of wine, comparing V4 and V5 domains of bacterial 16S rDNA to terminal restriction fragment length polymorphism (TRFLP) of LAB communities; and 2) to explore the bacterial communities of wine fermentation to better understand the biodiversity of wine at a depth previously unattainable using other techniques. Analysis of amplicons from the V4 and V5 provided similar views of the bacterial communities of botrytized wine fermentations, revealing a broad diversity of low-abundance taxa not traditionally associated with wine, as well as atypical LAB communities initially detected by TRFLP. The V4 domain was determined as the more suitable read for wine ecology studies, as it provided greater taxonomic depth for profiling LAB communities. In addition, targeted enrichment was used to isolate two species of Alphaproteobacteria from a finished fermentation. Significant differences in diversity between inoculated and uninoculated samples suggest that Saccharomyces inoculation exerts selective pressure on bacterial diversity in these fermentations, most notably suppressing abundance of acetic acid bacteria. These results determine the bacterial diversity of botrytized wines to be far higher than previously realized, providing further insight into the fermentation dynamics of these wines, and demonstrate the utility of next-generation sequencing for wine ecology studies

Yeast biodiversity in vineyard environments is increased by human intervention

One hundred and five grape samples were collected during two consecutive years from 33 locations on seven oceanic islands of the Azores Archipelago. Grape samples were obtained from vineyards that were either abandoned or under regular cultivation involving common viticultural interventions, to evaluate the impact of regular human intervention on grape yeast biota diversity in vineyards. A total of 3150 yeast isolates were obtained and 23 yeast species were identified. The predominant species were Hanseniaspora uvarum, Pichia terricola, Starmerella bacillaris and Issatchenkia hanoiensis. The species Barnettozyma californica, Candida azymoides and Pichia cecembensis were reported in grapes or wine-associated environments for the first time. A higher biodiversity was found in active vineyards where regular human intervention takes place (Shannon index: 1.89 and 1.53 in the first and second years, respectively) when compared to the abandoned ones (Shannon index: 0.76 and 0.31). This finding goes against the assumptions that human intervention can destroy biodiversity and lead to homogeneity in the environment. Biodiversity indices were considerably lower in the year with the heaviest rainfall. This study is the first to report on the grape yeast communities from several abandoned vineyards that have undergone no human intervention.Joao Drumonde Neves is the recipient of a fellowship of the Azorean Government (M321/006/F/2008) and PROEMPREGO. This work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalizacao (POCI), and by national funds through FCT by the projects FCOMP-01-0124-008775, PTDC/AGR-ALI/103392/2008 and PTDC/AGR-ALI/121062/2010.info:eu-repo/semantics/publishedVersio

Microbiological and chemical monitoring of Marsala base wine obtained by spontaneous fermentation during large-scale production

The present work was undertaken to evaluate the effect of the natural winemaking on the microbial and chemical composition of Marsala base wine. To this purpose, a large-scale vinification process of Grillo grape cultivar was monitored from harvesting to the final product. Total yeasts (TY) showed a rapid increase after must pressing and reached values almost superimposable to those registered during the conventional winemakings. Lactic acid bacteria (LAB) were registered at the highest levels simultaneously to yeast growth at the beginning of the process. Saccharomyces cerevisiae was the species found at the highest concentrations in all samples analysed. Several strains (n= 16) was registered at high levels during the alcoholic fermentation and/or aging of wine; only two of them were detected on the grape surface. Lactobacillus plantarum was the LAB species most frequently isolated during the entire vinification process. Ethanol content was approximately 14% (v/v) at the end of vinification. The value of pH did not greatly vary during the process and the volatile acidity (VA) was detected at low concentrations during the entire transformation. The concentration of malic acid rapidly decreased during the AF; on the other hand, lactic acid showed an irregular trend during the entire process. trans-caffeil tartaric acid was the most abundant hydroxycinnamoyl tartaric acid and volatile organic compounds (VOC) were mainly represented by isoamylic alcohol and isobutanol

Eating disorders in children with cerebral palsy

Malnutrition is among the main causes of growth disorder in children with cerebral palsy (CP), and nutritional status is an important parameter in the assessment of their overall functionality and quality of life. We reviewed the international literature on the etiology and impact of malnutrition in children with CP and the nutritional support guidelines for this group. Severity of the motor impairment, secondary problems (such as dysphagia and gastroesophage-al reflux disease) antiepileptic medication are among the factors that determine the energy needs of these children. Dietary intervention depends on the nutritional status, the ability of the child to consume adequate amounts of solid food and fluids, and the risk of aspiration. Monitoring of the diet and growth should be included in the holistic care of children with CP, as poor nutrition is a risk factor for morbidity and mortality. Intervention should aim not only at promoting linear growth, but also at enhancing the functional capacity. Obesity among CP children and adolescents is showing an increasing tendency and this is another risk factor that requires intervention, and relevant information should be provided for the caregivers. There are no general guidelines regarding malnutrition in children with CP; personalization of advice is necessary and every case needs to be considered individually. © Athens Medical Society

Understanding Wine through Yeast Interactions

Wine is a product of microbial activities and microbe–microbe interactions. Yeasts are the principal microorganisms responsible for the evolution and fulfillment of alcoholic fermentation. Several species and strains coexist and interact with their environment and with each other during the fermentation course. Yeast–yeast interactions occur even from the early stages of fermentation, determining yeast community structure and dynamics during the process. Different types of microbial interactions (e.g., mutualism and commensalism or competition and amensalism) may exert positive or negative effects, respectively, on yeast populations. Interactions are intimately linked to yeast metabolic activities that influence the wine analytical profile and shape the wine character. In this context, much attention has been given during the last years to the interactions between Saccharomyces cerevisiae (SC) and non-Saccharomyces (NS) yeast species with respect to their metabolic contribution to wine quality. Yet, there is still a significant lack of knowledge on the interaction mechanisms modulating yeast behavior during mixed culture fermentation, while much less is known about the interactions between the various NS species or between SC and Saccharomyces non-cerevisiae (SNC) yeasts. There is still much to learn about their metabolic footprints and the genetic mechanisms that alter yeast community equilibrium in favor of one species or another. Gaining deeper insights on yeast interactions in the grape–wine ecosystem sets the grounds for understanding the rules underlying the function of the wine microbial system and provides means to better control and improve oenological practices

Use of Vitamin D Bolus in Fortified Juice for Improving Vitamin D Status in Children with Cerebral Palsy

Children with cerebral palsy (CP) are at risk of poor nutrition due to a number of factors. Feeding, eating, drinking, and swallowing (FEDS) problems are common in these children and may result in protein-calorie malnutrition usually accompanied by micronutrient deficiencies. Vitamin D is among the elements whose uptake is obstructed. Insufficient exposure to solar radiation in children and adolescents with CP adds to further decreasing serum vitamin D levels thus potentially affecting growth, bone density, and muscle function. Since maintaining long-term adherence to daily oral administration of vitamin D in this population is often difficult, bolus therapy by using vitamin D-fortified products could be an alternative way of effective and safe vitamin D intake. Purpose: Assessing the efficacy of administration of bolus vitamin D in fortified juice for increasing 25(OH)D levels in a group of 15 children with CP. Results: The juice was well tolerated, and a significant increase in 25(OH)D levels was observed from 54.1 to 110.3 nmol/L (p < 0.0001) 4 weeks after the administration without any case of hypercalcemia. Conclusion: Bolus therapy with vitamin D3-fortified juice is well tolerated and effectively increases 25(OH)D levels in children with CP. © 2021, The Author(s), under exclusive license to Springer Nature Switzerland AG

Yeast Populations Residing on Healthy or Botrytis-Infected Grapes from a Vineyard in Attica, Greece

The yeast flora associated with healthy and Botrytis-infected grapes was assessed. Molecular identification methods assigned isolates to six genera and nine species. For the first time Hanseniaspora opuntiae was encountered as an inhabitant of the grape ecosystem. By using DraI, an informative restriction fragment length polymorphism pattern was generated to distinguish H. opuntiae from the closely related organism Hanseniaspora guilliermondii. Botrytis infection resulted in a larger population and greater diversity of yeasts enriched with fermentative or spoilage species