| Summary: | There is a lack of studies evaluating the metabolic contribution of non-<i>Saccharomyces</i> yeasts in early fermentation phases. This study aimed to investigate the volatile aroma profiles produced by various non-<i>Saccharomyces</i> yeasts just before sequential inoculation with <i>Saccharomyces cerevisiae</i> to provide an insight into the particular effects they induce at this stage. The grape must of Malvazija istarska was inoculated with monocultures of <i>Torulaspora delbrueckii</i>, <i>Metschnikowia pulcherrima</i>, <i>Pichia kluyveri</i>, <i>Lachancea thermotolerans</i>, and <i>Schizosaccharomyces pombe</i>, alongside a <i>S. cerevisiae</i> control. Eighty volatile compounds were quantified via headspace solid-phase microextraction and gas chromatography–mass spectrometry, and the data were statistically elaborated. Volatile profiles of non-<i>Saccharomyces</i> yeasts differed significantly from the <i>S. cerevisiae</i> control. Most treatments caused increases in linalool and β-damascenone, decreases in higher alcohols and fatty acids, and improved synthesis of odoriferous esters. <i>Torulaspora delbrueckii</i> and <i>M. pulcherrima</i> produced compounds not commonly found in <i>S. cerevisiae</i> fermented wines. Multivariate statistical analysis linked the investigated yeasts to specific, particularly abundant compounds. Future studies should explore to what degree these contributions persist after sequential inoculation with <i>S. cerevisiae</i> in diverse grape must matrices.
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