| Summary: | The genus <i>Vaccinium</i> contains about 400 species distributed worldwide, but only a few species and varieties have had their phenolic composition and biological activity documented. In this study, phenols, flavonoids, anthocyanins, antioxidant and antimicrobial activity of methanolic extracts of <i>Vaccinium stenophyllum</i> Steud. fruits: 1-totally immature, 2-immature, 3-immature/close to maturity and 4-mature, were determined using UV-Visible spectrometry and HPTLC. The totally immature fruit extract showed the highest content of total phenols (19.153 ± 0.175 mg GAE/g DW), chlorogenic acid (20.867 ± 0.240 mg CAE/g DW), and the highest antioxidant activity by ABTS<sup>●+</sup> (196.761 ± 0.641 µM TE/g DW) and DPPH<sup>●</sup> (146.580 ± 6.466 µM TE/g DW). Immature, immature/close to maturity and mature fruits extracts, exhibited the lowest MIC (9.37 mg/mL) and MBC (18.75 mg/mL) against <i>Escherichia coli</i>, <i>Salmonella choleraesuis</i>, and <i>Shigella flexneri</i>. The mature fruits extract exhibited the highest content of total anthocyanins (0.141 ± 0.004 mg CE/g DW) and cyanidin-3-glucoside (19.230 ± 0.309 mg CGE/g DW). The content of phenols, flavonoids and anthocyanins was higher than that reported for other <i>Vaccinium</i> species. These results showed the relevance of <i>Vaccinium stenophyllum</i> Steud. for breeding purposes to enhance the bioactivity of cultivars, or as a source of natural additives for the food industry, among others.
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