This study was performed to evaluate and compare the in vitro antioxidant, antimicrobial, and antimutagenic activities, and the polyphenolic content of the Nord-West Romanian wild bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.) leaves from three different natural habitats (Smida, Turda, Borsa). In the case of both species, the flavanols level was higher in Smida habitat (altitude 1100 m), whereas quercetin derivates were more abundant in Borsa habitat (altitude 850 m). The bilberry leaf extracts contained in the highest amounts the feruloylquinic acid (59.65 ± 0.44 mg/g for Borsa habitat) and rutin (49.83 ± 0.63 mg/g for Borsa habitat), and showed relevant 2,2-diphenyl-1-picrylhydrazyl (DPPH) antioxidant activity (271.65 mM Trolox/100 g plant material for Borsa habitat, 262.77 mM Trolox/100 g plant material for Smida habitat, and 320.83 mM Trolox/100 g plant material for Turda habitat), for all the three extracts. Gallocatechin was the dominant flavanol in lingonberry species, with the highest amount being registered for Smida habitat (46.81 ± 0.3 mg/g), revealing a DPPH antioxidant activity of 251.49 mM Trolox/100 g plant material. The results obtained in the antimicrobial tests showed that the best inhibitory effect among bilberry species was attributed to the Turda (altitude 436 m) and Smida locations, against both Gram-positive and Gram-negative bacterial strains. For lingonberry, the differences in habitat did not influence the antibacterial effect, but the antifungal effect, only in the case of Candida zeylanoides. A strong antimutagenic effect was registered by the bilberry leaves toward Salmonella typhimurium TA100. Our study may be able to provide a better understanding of the correlation between natural habitat conditions and the accumulation of secondary metabolites and their related bioactivities in studied leaves.

Blueberries have seen an ascending production line boosted by World Health Organization (WHO) approvals for their contributions to a healthy diet and the evidence that they act against different diseases. This increase resulted in significant amounts of discarded leaves, which could be a valuable source of bioactive compounds. In the present study, ultrasound-assisted extraction technology was used to determine and compare the chemical and biological profiles of leaves from six commercial blueberry (Vaccinium corymbosum L.) varieties. Feruloylquinic acid was the major compound identified, ranging from 19.23 ± 0.18 mg/g (at the lowest level, registered in the Spartan variety) to 49.62 ± 0.41 mg/g (at the highest level, registered in the Nelson variety). Rutin was the second major compound identified, for which Toro, Nelson, and Elliot leaves registered the highest values, with 35.77 ± 0.19 mg/g, 32.50 ± 0.20 mg/g, and 31.53 ± 0.1 mg/g, respectively. Even though analogous polyphenols were detected in the six cultivars, their concentrations and amounts were different. The leaf extracts of the cultivars Toro, Elliot, and Nelson appear to be good sources of antioxidants, registering high percentage inhibitions of DPPH radicals, of 70.41%, 68.42%, and 58.69%, respectively. The blueberry leaf extracts had a strong antibacterial activity and a low antifungal capacity, and a low-to-moderate antimutagenic capacity towards Salmonella typhimurium TA98 and TA100 strains, with Toro leaf being the best candidate. All of these biological activities indicate health-related benefits, recommending them as suitable candidates for medical and pharmaceutical applications. The present paper adds significant knowledge to the field of blueberry leaves via chemical and biological profiles, supporting the ultrasound-assisted extraction technique as a useful and green method to provide alternative sources of bioactive compounds.

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