Exploring the anti-virulence potential of plants used in traditional Mayan medicine to treat bacterial infections

Ethnopharmacological relevance: While the antimicrobial activity of a number of plants used in traditional Mayan medicine against infectious diseases has been documented, their potential to inhibit quorum sensing (QS) as means of discovering novel anti-virulence agents remains unexplored. Aim of the study: To evaluate the anti-virulence potential of plants used in traditional Mayan medicine by determining their inhibition of QS- regulated virulence factors in Pseudomonas aeruginosa. Methods: A group of plants used in traditional Mayan medicine against infectious diseases was selected, and their methanolic extracts were evaluated at 10 mg/mL for their antibacterial and anti-virulence activity using the reference strain P. aeruginosa PA14WT. The broth microdilution method was used to determine antibacterial activity (MIC), while anti-virulence activity was evaluated by measuring the anti-biofilm effect and the inhibition of pyocyanin and protease activities. The most bioactive extract was fractionated using a liquid-liquid partition procedure and the semipurified fractions were evaluated at 5 mg/mL for antibacterial and anti-virulence activity. Results: Seventeen Mayan medicinal plants traditionally used to treat infection-associated diseases were selected. None of the extracts exhibited antibacterial activity, whereas anti-virulence activity was detected in extracts of Bonellia flammea, Bursera simaruba, Capraria biflora, Ceiba aesculifolia, Cissampelos pareira and Colubrina yucatanensis. The most active extracts (74% and 69% inhibition) against biofilm formation were from C. aesculifolia (bark) and C. yucatanensis (root), respectively. Alternatively, the extracts of B. flammea (root), B. simaruba (bark), C. pareira (root), and C. biflora (root), reduced pyocyanin and protease production (50-84% and 30-58%, respectively). Fractionation of the bioactive root extract of C. yucatanensis allowed the identification of two semipurified fractions with anti-virulence activity. Conclusions: The anti-virulence activity detected in the crude extracts of B. flammea, B. simaruba, C. biflora, C. aesculifolia, C. pareira, and C. yucatanensis, confirms the efficacy and traditional use of these medicinal plants against infectious diseases. The activity of the extract and semipurified fractions of C. yucatanensis indicates the presence of hydrophilic metabolites capable of interfering with QS in P. aeruginosa. This study represents the first report of Mayan medicinal plants with anti-QS properties and suggests they represent an important source of novel anti-virulence agents.