Inhibition of NorA efflux pump of Staphylococcus aureus by anacardic acids isolated from the cashew nutshell liquid of Anacardium occidentale L.

The rising of diseases caused by multidrug-resistant bacteria has encouraged researchers to explore more antimicrobial substances, as well as chemicals capable of potentiating the action of existing ones against multidrug-resistant bacteria. Anacardium occidentale produces a fruit known as cashew nut, filled with a dark, almost black, caustic, and flammable liquid called cashew nutshell liquid (CNSL). The goal of the study was to evaluate the intrinsic antimicrobial activity of the major compounds present in CNSL, called anacardic acids (AA), as well as their possible modulatory action as an adjuvant of Norfloxacin against a Staphylococcus aureus strain overproducing the NorA efflux pump (SA1199B). Microdilution assays were performed to determine the minimum inhibitory concentration (MIC) of AA against different microbial species. Norfloxacin and Ethidium Bromide (EtBr) resistance modulation assays were performed in the presence or absence of AA against SA1199-B. AA showed antimicrobial activity against Gram-positive bacterial strains tested but no activity against Gram-negative bacteria or yeast strains. At subinhibitory concentration, AA reduced the MIC values for Norfloxacin and EtBr against the SA1199-B strain. Furthermore, AA increased the intracellular accumulation of EtBr in this NorA overproducer strain, indicating that AA are NorA inhibitors. Docking analysis showed that AA probably modulates Norfloxacin efflux by spatial impediment at the same binding site of NorA.

Mechanism of the lethal effect of Riparin E against bacterial and yeast strains.

Riparins are alkamides naturally found in the fruits of Aniba riparia (Nees) Mez, but currently synthetic molecules as Riparin E (Rip-E) can be obtained. Potential biological of Rip-E as schistosomicidal agent against Schistosoma mansoni worms, as well as against Staphylococcus aureus strains has already been described. However, the mechanism of action related to antimicrobial activity of Rip-E against bacterial or fungi species has not yet been reported. This study had as objective to evaluate the Rip-E antimicrobial activity against Gram-positive and Gram-negative bacteria, as well as against yeast species of clinical importance. Minimal inhibitory concentrations of the compound against bacterial and yeast strains were determined by microdilution method. To verify if a possible lethal effect caused by Rip-E were related to plasma membrane damage, microbial cells treated with Rip-E were stained with 7-aminoactinomycin D (7-AAD) and analyzed by flow cytometry. Rip-E showed a bactericide effect against Gram-positive species S. aureus and S. epidermidis, as well as, against Gram-negative species Escherichia coli and Salmonella enterica Typhimurium, but was inactive against Pseudomonas aeruginosa. Moreover, Rip-E showed activity against fungi species Candida albicans and C. tropicalis. S. aureus, E. coli and C. albicans cells treated with Rip-E were marked with 7-aminoactinomycin D (7-AAD) indicating that Rip-E can cause plasma membrane damage, acting as a potential microbicide agent for prevention or treatment of infectious diseases.