Background Strawberry is a pseudofruit mainly cultivated in temperate climate regions. Considering its high levels of ascorbic acid and phenolic compounds, the consumption of strawberry fruit can be beneficial to health. The Brazilian strawberry production revolves around 3000 tons per year, significantly influencing the food market and generating income to farmers. However, this production can be partially impaired by two-spotted spider mite (TSSM) Tetranychus urticae Koch infestations, due to decreases in the quality and quantity of fruit. Since there are no data in the literature about alterations caused by TSSM infestation in strawberry plants, our work aimed towards evaluating nutritional and physicochemical parameters of TSSM-infested strawberry plants, along with the related chemical treatment (CT) (acaricide) or biological treatment (predatory mite Phytoseiulus macropilis Banks). Results Strawberry fruit from TSSM-infested plants present the highest levels of acidity and exhibit low levels of anthocyanin and phenolic compounds, while fruit from TSSM-infested plants + biological control using predatory mite shows high levels of soluble solids, phenolic compounds and ascorbic acid, along with a high soluble solid content/titratable (SSC/TA) acidity ratio, which indicates high quality fruit. Conclusions Our results suggest that TSSM infestation decreases fruit quality and that the biological control of TSSM using a predatory mite is a suitable alternative to organic production, since the presence of predatory mite does not affect fruit quality and development.