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In the search for new natural resources with promising biological activity, chemical composition as well as antibacterial, antifungal and antioxidant activities of the essential oil from Croatian indigenous cultivar of lavandin (Lavandula × intermedia‘Budrovka’) were studied for the first time and compared with those of lavender (L. angustifolia) cultivated under the same pedo-climatic conditions of central Croatia. Forty compounds were identified in the studied Lavandula essential oils by GC–MS analysis. Linalool (57.1%), linalyl acetate (9.8%) and 1,8-cineole (8.4%) were the most abundant compounds of lavandin ‘Budrovka’ oil while lavender oil contained mainly linalool (54.0%), linalyl acetate (11.6%) and lavandulol (6.6%). Both essential oils were characterized by a very low amount of camphor (≤0.3%). The antimicrobial activities of two Lavandula essential oils and their dominant constituents were evaluated against 30 medically important pathogens and microbial contaminants of food, cosmetic and pharmaceutical products using agar well diffusion and broth dilution methods. L. × intermedia ‘Budrovka’ essential oil showed a broad-spectrum of antibacterial (MICs 0.25–2.5 mg/mL) and antifungal (MICs 0.1–2 mg/mL) activities, being generally more than or at least as effective as those from L. angustifolia. Among the two dominant compounds which were also tested comparatively and found to exhibit antimicrobial effect, linalool has been pointed out as much more potent agent then linalyl acetate. Antioxidant studies based on DPPH assay indicated that the lavandin ‘Budrovka’ essential oil has significantly stronger antioxidant properties than lavender essential oil (IC50 21.6 mg/mL and 27.7 mg/mL, respectively). This study highlighted Croatian indigenous cultivar lavandin ‘Budrovka’ as a rich source of essential oil with pronounced antimicrobial properties due to abundance of linalool and indicated its potential use as biopreservative in food, pharmaceutical and cosmetic products or as natural remedy with promising health benefit.