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Plants in the Australian genus Eremophila (Scrophulariaceae) have attracted considerable recent attention for their antimicrobial compounds, which possess a wide range of chemical structures. As they are typically associated with the oily-waxy resin layer covering leaves and green branchlets, and Eremophila lucida is prominent among the species containing a pronounced sticky resin layer, this species was considered of interest for assessing its antibacterial constituents. The n -hexane fraction of the crude acetone extract of the leaves exhibited antibacterial activity against Staphylococcus aureus . Isolation led to the known compounds cembratriene, (3 Z , 7 E , 11 Z )-15-hydroxycembra-3,7,11-trien-19-oic acid (1), the sesquiterpenoid, farnesal (2) and the viscidane diterpenoid, 5α-hydroxyviscida-3,14-dien-20-oic acid (3). The purified compounds were tested for antibacterial activity with 2 and 3 showing moderate antibacterial activity against Gram-positive bacteria.

Plant metabolites that have shown activity against bacteria and/or environmental fungi represent valuable leads for the identification and development of novel drugs against clinically important human pathogenic fungi. Plants from the genus Eremophila were highly valued in traditional Australian Aboriginal medicinal practices, and E. alternifolia was the most prized among them. As antibacterial activity of extracts from E. alternifolia has been documented, this study addresses the question whether there is also activity against infectious fungal human pathogens. Compounds from leaf-extracts were purified and identified by 1- and 2-D NMR. These were then tested by disk diffusion and broth microdilution assays against ten clinically and environmentally relevant yeast and mould species. The most potent activity was observed with the diterpene compound, 8,19-dihydroxyserrulat-14-ene against Cryptococcus gattii and Cryptococcus neoformans, with minimum inhibition concentrations (MIC) comparable to those of Amphotericin B. This compound also exhibited activity against six Candida species. Combined with previous studies showing an antibacterial effect, this finding could explain a broad antimicrobial effect from Eremophila extracts in their traditional medicinal usage. The discovery of potent antifungal compounds from Eremophila extracts is a promising development in the search for desperately needed antifungal compounds particularly for Cryptococcus infections., Competing Interests: The authors declare no conflict of interest.

Ethnopharmacological Relevance: For traditional medicinal purposes Aboriginal Australians have utilised numerous plant species, Eremophila alternifolia is among the most prominent. Traditionally, fresh leaves, leaf-infusions and handmade leaf-pastes have been used as both external and internal preparations to provide relief from a variety of conditions. Preparations of the species have been used to treat various infections of skin, eyes and throat including the treatment of septic wounds. These usages suggest that the plant contains antibacterial compounds; however, to date they have not been isolated and identified., Aim of the Study: The present study aimed to identify antibacterial compounds from this important traditionally recorded medicinal species., Materials and Methods: Bioassay-guided fractionation was used to isolate compounds from the crude leaf-extract. Antibacterial activity of pure compounds was assessed through broth microdilution method by determining both minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs). Structure elucidation was performed using spectroscopic techniques such as 1D and 2D nuclear magnetic resonance spectroscopy and high resolution mass spectrometry., Results: Four compounds have been isolated from the leaf-extract; they include previously known flavanones [pinobanksin (1), pinobanksin-3-acetate (2) and pinobanksin-3-cinnamate (3)] and a serrulatane diterpene, 8-hydroxyserrulat-14-en-19-oic acid (4). While compound 4 had been found in other Eremophilas, flavanones 2 and 3 are identified for the first time from the genus Eremophila. The flavanone 3 is the most promising antibacterial compound with significant activity (10-20µM) against strains of the Gram-positive bacterium Staphylococcus aureus including methicillin resistant and biofilm forming strains. No activity was observed for any isolated compounds against the Gram-negative bacterium Escherichia coli., Conclusion: The antibacterial activity of the crude extract of E. alternifolia and of the isolated compounds against Gram-positive bacteria provides a Western scientific explanation of the therapeutic modality of this plant species in traditional Aboriginal medicinal practice., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)