Presence of Saccharomyces eubayanus in fermentative environments reveals a new adaptive scenario in Patagonia.

Patagonia (Argentina and Chile) harbors the highest Saccharomyces eubayanus genomic diversity and its widest predominance in natural environments. In this work, S. eubayanus was isolated for the first time from a fermentative environment. This species was found dominating both a traditional apple chicha fermentation as well as feral apple trees in the Andean region of Aluminé (Argentina). S. eubayanus was the only Saccharomyces species found in the isolation substrates, although it coexisted with other non-Saccharomyces species. The absence of strong fermentative competitors of the Saccharomyces genus (like Saccharomyces uvarum or Saccharomyces cerevisiae) in the feral apples could promote the development and implantation of S. eubayanus in a spontaneous apple must fermentation. Phylogeographic analyses revealed a high intraspecific diversity in S. eubayanus, enabling the characterization of strains belonging to the genomic subpopulations PA1, PA2, and PB1 according to the sequences obtained for the intFR gene region. This result evidence that the studied sampling area represents a natural habitat for the species. Being a novel finding, studying the causes that allowed this species to prosper in a fermentative environment becomes essential. Hence, the physiological profile of the new isolates, including their ability to grow at different temperature, nitrogen, and ethanol concentrations was evaluated in comparison with a set of S. eubayanus strains previously isolated from natural environment and representing different genomic subpopulations. Greater physiological diversity was evidenced when strains isolated from both natural and fermentative environments were analyzed overall. Furthermore, no direct relationship between genomic population and physiological behavior was observed; on the opposite, strains appeared to exhibit similar behavior, primarily grouped by isolation origin.

Contaminated Perry in Patagonia Argentina: A Case Study.

Perry is a beverage obtained by fermentation of pear juice, popular in the North Hemisphere. In Argentina it is an emerging market, particularly in the Patagonian region. The aim of this work is to describe and to evaluate the spoilage yeasts associated to six perry samples showing signs of microbiological contamination from a local craft perry company in North Patagonian region. Eighteen yeasts were isolated from four of the six perry samples where Brettanomyces custersianus, Brettanomyces bruxellensis and Zygosaccharomyces parabailii were identified. The growth capacity of these isolates in the presence of antimicrobial agents (sulfite and potassium sorbate) was analyzed in solid media. Growth parameters in sterile perry must was evaluated and the production of undesirable compounds were evaluated, products were characterized in terms of their aromatic and physicochemical features. The yeasts Z. parabailii NPCC1791 was able to grow on plates containing sulfite concentrations of up to 4 mM and produced high methanol concentrations in perry. Additionally, B. bruxellensis NPPC1792 was able to produce methanol as well as high concentrations of volatile phenols including 4-ethylphenol and 4-ethylguaiacol. These results demonstrate the potential of these species as perry contaminants. Given the lack of studies describing the contaminating yeasts in perry production, this work represents the first report about perry spoilage yeasts in Argentina, with this knowledge, control strategies can be developed to prevent microbiological contamination and minimize product loss.

Evaluation of cryotolerant yeasts for the elaboration of a fermented pear beverage in Patagonia: Physicochemical and sensory attributes.

The production of pome fruits as pears and apples, as well as their derived industries, is of great economic importance in North Patagonia. The elaboration of fermented beverages as cider or perry has evidenced a substantial diversification during the last years, with the evaluation of different fruit varieties, yeast starters and technological changes. In this work, two cryotolerant yeasts belonging to the species Saccharomyces uvarum were evaluated at laboratory and pilot scale in sterile and no-sterile pear must. One of the strains was originally isolated from apple chicha (strain NPCC1314) and the other from apple cider (strain NPCC1420) in Patagonia. Both physicochemical and sensory features of the fermented products were evaluated. Both strains were able to successfully complete the fermentations, although strain NPCC1420 showed the better kinetic properties including a faster sugar consumption than the strain NPCC1314. Both strains showed excellent implantation capacity, but the fermented products showed different chemical profiles. The perry fermented with the strain NPCC1314 was characterized by better sensory attributes as assessed by trained panelists and a greater acceptance for untrained public than the same fermented with the strain NPCC1420. The two strains were able to consume sorbitol, both in pear must and in agar-plates supplemented with sorbitol as the sole carbon source. This ability is described for the first time in S. uvarum, at least for the two strains evaluated in this work.

Saccharomyces uvarum isolated from patagonian ciders shows excellent fermentative performance for low temperature cidermaking.

Saccharomyces uvarum has been recovered from natural habitats and traditionally fermented beverages (apple chicha) in Patagonia. However, this species has never been obtained from industrially relevant beverages like wine or cider in the same region. In this work, different strains belonging to the cryotolerant species S. uvarum were recovered from spontaneous cider fermentations carried out at low temperature in Red Delicious apple must. The strain S. uvarum NPCC1420 obtained from this cider and selected for its physiological and technological features, evidenced a better adaptation to the cidermaking process than a previously selected strain obtained from a less industrialized product called apple chicha. Some differences, like a higher ethanol and sulphite tolerance, seemed to be associated with differential domestication pressures suffered by each different strain. Moreover, the most important fermentative features of the strain NPCC1420 were a higher competition capacity than the strain NPCC1314 in non-sterile apple must, as well as significantly higher amounts of glycerol, 2-phenylethanol and 2-phenylethyl acetate than the strain isolated from apple chicha.

Physiological characterization of Saccharomyces uvarum and Saccharomyces eubayanus from Patagonia and their potential for cidermaking.

A diversity of yeast strains belonging to the cryotolerant fermentative species S. uvarum and S. eubayanus have been recovered from natural habitats and traditional fermentations in North Patagonia. The aim of this work was to evaluate the most relevant physiological features in a set of Patagonian strains belonging to S. uvarum and S. eubayanus, in order to analyze their potentiality to be used as starter cultures for cidermaking elaborated at low temperature. We evidenced that S. uvarum strains isolated from natural habitats (Araucaria araucana bark) showed similar physiological features to S. eubayanus strains obtained from the same habitat, and different from S. uvarum strains from fermentative environments (apple chichas). We also confirm the capacity of S. uvarum to produce high glycerol levels, low acetic acid and elevated production of the higher alcohol 2-phenylethanol and 2-phenylethyl acetate and demonstrated similar properties in S. eubayanus. Finally, we evidenced for the first time the antagonistic activity of S. eubayanus and selected three strains (two S. uvarum and one S. eubayanus) bearing the best combination of features to be used as a starter culture in cidermaking.