title: Unexplored endemic fruit species from Brazil: Antibiofilm properties, insights into mode of action, and systemic toxicity of four Eugenia spp.
abstract: Brazilian endemic fruit species have aroused attention due to their highly valuable, yet unexplored, agro-industrial, food and therapeutic potential. Herein, we describe the antifungal activity of four Eugenia spp. against Candida albicans biofilms, and further demonstrate insights into their potential mode(s) of action and toxicity in vitro and in vivo. Extracts from different parts (seeds, pulps, leaves) of E. leitonii (EL), E. brasiliensis (EB), E. myrcianthes (EM) and E. involucrata (EI) were obtained (S23°23',W45°39') and chemically characterized by GC/MS. The active extracts were tested against C. albicans biofilm viability and architecture, as well as mode of action, and toxicology using RAW 264.7 macrophages and Galleria mellonella larvae. The MIC values ranged from 15.62 to >2000 μg/mL. The most active extracts were EL (seed, 15.62 μg/mL) and EB (leaf and seeds, 31.25 and 15.62 μg/mL, respectively). Treatment with these extracts at 10xMIC reduced biofilm viability by 54-55% (P < 0.0001) as compared to 42% by nystatin. At 10xMIC, all extracts caused damages to biofilm architecture and integrity, and fewer hyphae remained attached to treated biofilms. None of them was found to interfere with cell wall biosynthesis or complexation with ergosterol. The extracts had low toxicity against macrophages in vitro (P > 0.05) and G. mellonella larvae, with mean in vivo LD50of 1500 mg/kg (EL, seeds); 2500 mg/kg (EB, seeds); and 1250 mg/kg (EB, leaf). The phenolic compounds epicatechin and gallic acid were the major constituents in the extracts. Our findings may open avenues for the application of these yet unexplored native fruits in the food and pharmaceutical industry.
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas, Piracicaba, SP, Brazil.
authors.forename: Janaina de Cássia Orlandi
authors.initials: JC
authors.lastname: Sardi
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas, Piracicaba, SP, Brazil.
authors.forename: Irlan Almeida
authors.initials: IA
authors.lastname: Freires
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas, Piracicaba, SP, Brazil.
authors.forename: Josy Goldoni
authors.initials: JG
authors.lastname: Lazarini
authors.affiliation: Department of Agri-Food Industry, Food and Nutrition, "Luiz de Queiroz" College of Agriculture, University of São Paulo, Piracicaba, SP, Brazil.
authors.forename: Juliana
authors.initials: J
authors.lastname: Infante
authors.affiliation: Department of Agri-Food Industry, Food and Nutrition, "Luiz de Queiroz" College of Agriculture, University of São Paulo, Piracicaba, SP, Brazil.
authors.forename: Severino Matias
authors.initials: SM
authors.lastname: de Alencar
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas, Piracicaba, SP, Brazil. Electronic address: rosalen@fop.unicamp.br.
authors.forename: Pedro Luiz
authors.initials: PL
authors.lastname: Rosalen
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects*; D001938:Brazil; D002176:Candida albicans / Q000187:drug effects; D002473:Cell Wall / Q000378:metabolism; D004875:Ergosterol / Q000378:metabolism; D000068856:Eugenia / Q000737:chemistry*; D005638:Fruit / Q000737:chemistry*; D005707:Gallic Acid / Q000737:chemistry; D051379:Mice; D008826:Microbial Sensitivity Tests; D009761:Nystatin / Q000494:pharmacology; D010636:Phenols / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity*; D018515:Plant Leaves / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry; D010947:Plants, Toxic / Q000737:chemistry*; D000067996:RAW 264.7 Cells; D012639:Seeds / Q000737:chemistry
chemical_list: D000935:Antifungal Agents; D010636:Phenols; D010936:Plant Extracts; D009761:Nystatin; D005707:Gallic Acid; D004875:Ergosterol
keywords: Antimicrobial activity; Biofilms; Candida albicans; Eugenia spp.