## 3 Results and Discussion
### Microbiological Analysis  
Identification of isolated yeast colonies reveals the presence of three non-_Saccharomyces_ species, _T. delbrueckii_, _L. thermotolerans_, and _Hanseniapora oputtiae_, before the start of alcoholic fermentation, detecting 4 \(\times\) 10\({}^{3}\), 1.1 \(\times\) 10\({}^{3}\), and 2.8 \(\times\) 10\({}^{2}\) colony-forming units (CFU)/mL of each species, respectively. Figure 3 shows the relative abundance of the detected yeast during the fermentation process. In SF condition, non-_Saccharomyces_ yeast appeared from the beginning of the alcoholic fermentation until the eighth day, _T. delbrueckii_ the most common yeast species, with 75% of isolates on the first day and _L. thermotolerans_ and _H. oputtiae_ counting the 20% and 5%, respectively; however, its presences decrease as the fermentation process progresses with no detection of _H. oputtiae_ after the second day and with no isolation of _L. thermotolerans_ after the sixth day. Last, _T. delbrueckii_ was isolated until the eighth day, after which only _S. cerevisiae_ was found in the SF condition. In the VC10 and VC12 conditions, where the _W. anomalous_ strains were inoculated at the beginning of the alcoholic fermentation, only this non-_Saccharomyces_ species was isolated, with the exception of _T. delbrueckii_, on the eighth day (1.8% of relative abundance), Figure 3. _W. anomalous_ was only isolated in the VC10 and VC12 conditions; this yeast species was not found either in the grape must before alcoholic fermentation or during the sampling in the SF condition, indicating correct implantation of the inoculated strains in the grape must. _S. cerevisiae_ yeast dominates the alcoholic fermentation at the final stage of the three conditions, being the most abundant species from the sixth day in SF. In the VC10 and VC12 conditions, _S. cerevisiae_ was isolated from the eighth day, displacing _W. anomalous_ as the most  
Figure 3: Relative abundance of _S. cerevisiae_ (dark brown dashed line), non-_Saccharomyces_ yeasts, excluding _W. anomalous_ (blue dots) and _W. anomalous_ (green line) during the alcoholic fermentation of grape must in each condition (**A**): Spontaneous fermentation (SF), (**B**): _W. anomalous_ strain VC10, (**C**): _W. anomalous_ strain VC12. abundant yeast on days 10 and 11, respectively. The emergence of _S. cerevisiae_ matches with the highest fermentation rates (gCO\({}_{2}\)/day) in all conditions, which highlights the higher fermentation power of this species. This occurrence is common when non-_Saccharomyces_ yeasts, such as _M. pulcherrina_ or _Pichi kluyveri_, are used in alcoholic fermentation to increase wine complexity because of their lower fermentation power, with expectations such as _L. thermotolerans_, which is capable of surviving when _S. cerevisiae_ dominates the fermentation process [5; 7; 18]. Microbiological analysis and fermentation kinetics data indicate that both VC10 and VC12 _W. anomalus_ strains influence the indigenous microbiota of the grape must, displacing non-_Saccharomyces_ yeasts and slowing down the emergence and dominance of _S. cerevisiae_. These results match with the studied in Cabernet Sauvignon grape must, where the alcoholic fermentations with _W. anomalus_ strains were slower than those carried out with _S. cerevisiae_[2]. In fact, _W. anomalus_ has less fermenting power than the other yeast identified in this work; however, his relative abundance does not decrease on the seventh day, being the most common yeast in the VC10 and VC12 conditions until days 9 and 10, respectively. This dominance may be related not only to the initial inoculum of 5 \(\times\) 10\({}^{6}\) cells/mL but also to the killer toxin and ethyl acetate production [25; 26; 27]. _W. anomalus_ killer toxins production has been mainly tested against spoil yeast such as the genus Brettanomyces; however, the antimicrobial activity of this species has also been reported against other non-_Saccharomyces_ species during the early stages of grape fermentation and _S. cerevisiae_[1]. _W.