In vivo Antibacterial Activity of Star Anise

Introduction: Star anise fruits (Illicium verum Hook.) have been used as an important treatment in traditional Chinese medicine. The previous studies reported the activity of the non-polar fractions as potential sources of antibacterial metabolites, and little was done concerning the polar fractions of star anise. Methods: The antibacterial activity of the star anise aqueous methanolic (50%) extract against multidrug-resistant Acinetobacter baumannii AB5057 and methicillin-resistant Staphylococcus aureus (MRSA USA300) was investigated in vitro (disc diffusion assay, minimum bactericidal concentration determination, anti-biofilm activity and biofilm detachment activity). The antibacterial activity was further tested in vivo using a murine model of MRSA skin infection. Ultra-performance liquid chromatography coupled to high-resolution mass spectrometry (UPLC/HRMS) approach was applied for the identification of the metabolites responsible for the antibacterial activity. The antioxidant potential was evaluated using five in vitro assays: TAC (total antioxidant capacity), DPPH, ABTS, FRAP (ferric reducing antioxidant power) and iron-reducing power. Results: In vitro, star anise aqueous methanolic extract showed significant inhibition and detachment activity against biofilm formation by the multidrug-resistant and highly virulent Acinetobacter baumannii AB5057 and MRSA USA300. The topical application of the extract in vivo significantly reduced the bacterial load in MRSA-infected skin lesions. The extract showed strong antioxidant activity using five different complementary methods. More than seventy metabolites from different classes were identified: phenolic acids, phenylpropanoids, sesquiterpenes, tannins, lignans and flavonoids. Conclusion: This study proposes the potential use of star anise polar fraction in antivirulence strategies against persistent infections and for the treatment of staphylococcal skin infections as a topical antimicrobial agent. To our knowledge, our research is the first to provide the complete polar metabolome list of star anise in an approach to understand the relationship between the chemistry of these metabolites and the proposed antibacterial activity. Keywords: Acinetobacter baumannii, antimicrobial resistance, antioxidant, biofilm, metabolome, polar methanolic extract