Carla Monteiro Leal¹, Stella Martins Castro¹, Gabrielle Lavor de Queiroz¹, Marilia dos Reis Nery Chen¹, Diego Rodrigues², Anderson da Rocha Gripp², Rodrigo Lemes², Fernanda Sampaio Cavalcante³, Henriette Uthe⁴, Jörg Ziegler⁴, Maria Isabel Sampaio Dos Santos¹, Ludger A. Wessjohann⁴, Ivana Correa Ramos Leal¹

Introduction: The National Park of Jurubatiba Sandbank is a Federal conservation unit, located along the north coast of the RJ State, showing xeromorphic aspect of vegetation and flooding soil. The Restinga has different phytophysiognomic formations, as follows: Clusia Open Shrub Formation (COSF), Ericacea Open Shrub Formation (EOSF), Post-beach Formation (PBF) and Periodically Floodedforest Formation (PFF). Among the plants found in Restinga, Andira legalis (Fabaceae) is common in COSF and PBF. The literature is scarce regarding the chemical profile of this species, impelling this study for on deep investigations. The aim of the study is to search for antibacterial and antibiofilm compounds of Andira legalis extracts from different phytophysiognomic formations and seasons. Methods: Leaves of Andira legalis (n= 20; COSF and PBF) was collected in 2024, in the summer (-S) and autumn (-A) seasons, and was stored at -80ºC. The dried and crushed material (20 mg) was extracted with 1 mL MeOH:H2O (8:2). The chemical analyses were performed in the Agilient 1290 infinity II HPLC system coupled to timsTOFBruker® with ESI source, in the positive ionization mode. Data processing was performed in MetaboScape® while the data analysis in the Sirius 5.8, Metlgel, GNPS2, and Metaboanalyst 6.0. First, the minimum inhibitory concentration (MIC) of the extracts was measured using the Steers replicator inoculation method (solid medium), at 128 to 512 μg/mL, in duplicate. There were tested 32 multidrug-resistant bacteria, such as S. aureus, S. haemolyticus, and S. epidermidis isolated from nasal colonization of neonates, and ATCC strains of S. simulans and S. epidermidis (CAAE: 25808819.4.0000.5699). The minimum bactericidal concentration (MBC) and MIC (liquid medium) were also evaluated at concentrations varying from 16 to 512 μg/mL. The active extracts were further evaluated for their ability to prevent biofilm formation at sub-MIC concentrations (<256 μg/mL) against the bacterial strains that demonstrated higher susceptibility. Results: Catechin, epicatechin, epigallocatechin, procyanidin A- and B-types, O- and C-glycosyde flavonoids were anottated in Andira legalis specimens extracts collected in COSF and PBF, during summer and autumn. The Metlgel chemical classification showed the flavonoids as the majority compounds. Andira legalis specimens from COSF and PBF, collected in summer (-S) and autumn (-A), inhibited S. haemolyticus at 128µg/mL. One specimen (COSF-S) inhibited S. epidermidis (ATCC) at 128 µg/mL and S. aureus at 256 µg/mL, while one specimen COSF-A inhibited S. aureus at 128 µg/mL. By the MIC assay, in liquid medium, these same specimens inhibited S. haemolyticus, S. epidermidis (ATCC) and S. aureus at 32 µg/mL,32 µg/mL and 128 µg/mL, respectively. All extracts showed to be bacteriostatic. In the antibiofilm activity evaluation, two Andira legalis specimens from COSF-S, showed 39% and 44% inhibition of S. haemolyticus at 256 μg/mL. Conclusion: Through the untargeted metabolomics it was possible to highlight phenolic acids, as well as flavonoids, as the major compounds present in Andira legalis in diferent phytophysiognomic, in summer and autumn seasons. In the future, LC-HRMS/MS data will be correlated with environmental conditions, such as soil mineral components, temperature, pH and humidity, by the use of local sensors.

Agradecimentos: Faperj, CNPq and CAPES for financial support. Faperj for scholarship. Leibniz Institute of Plant Biochemistry (Germany) for scientific support in the development of the research.