Abstract
Undue exposure to antimicrobials has led to the acquisition and development of sophisticated bacterial resistance mechanisms, such as efflux pumps, which are able to expel or reduce the intracellular concentration of various antibiotics, making them ineffective. Therefore, inhibiting this mechanism is a promising way to minimize the phenomenon of resistance in bacteria. In this sense, the present study sought to evaluate the activity of the Carvacrol (CAR) and Thymol (THY) terpenes as possible Efflux Pump Inhibitors (EPIs), by determining the Minimum Inhibitory Concentration (MIC) and the association of these compounds in subinhibitory concentrations with the antibiotic Norfloxacin and with Ethidium Bromide (EtBr) against strains SA-1199 (wild-type) and SA-1199B (overexpresses NorA) of Staphylococcus aureus. In order to verify the interaction of the terpenes with the NorA efflux protein, an in silico molecular modeling study was carried out. The assays used to obtain the MIC of CAR and THY were performed by broth microdilution, while the Efflux Pump inhibitory test was performed by the MIC modification method of the antibiotic Norfloxacin and EtBr. docking was performed using the Molegro Virtual Docker (MVD) program. The results of the study revealed that CAR and THY have moderate bacterial activity and are capable of reducing the MIC of Norfloxacin antibiotic and EtBr in strains of S. aureus carrying the NorA efflux pump. The docking results showed that these terpenes act as possible competitive NorA inhibitors and can be investigated as adjuvants in combined therapies aimed at reducing antibiotic resistance.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
Geopark Araripe; FUNCAP; CAPES.
Funding
This study was funded by the Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico—FUNCAP [BP4-0172-00168.01.00/20]; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior — CAPES.
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Francisco Assis Bezerra da Cunha, Saulo Relison Tintino, Henrique Douglas Melo Coutinho; Conceptualization, Supervision, Coordinated, Cristina Rodrigues dos Santos Barbosa; Investigation, Writing, Original Draft Preparation, Jackelyne Roberta Scherf, Raimundo Luiz Silva Pereira, Joycy Francely Sampaio dos Santos, Sarah Silva Patrício de Jesus, Thais Pereira Lopes, Zildene de Sousa Silveira; Performed the Antibacterial Assays, Irwin Rose Alencar de Menezes; Performed the in Silico Analysis, Thiago Sampaio de Freitas, Cícera Datiane de Morais Oliveira-Tintino; Performed the Statistical Analysis, José Pinto Siqueira Júnior; Contributed With the Resources for the Work.
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dos Santos Barbosa, C.R., Scherf, J.R., de Freitas, T.S. et al. Effect of Carvacrol and Thymol on NorA efflux pump inhibition in multidrug-resistant (MDR) Staphylococcus aureus strains. J Bioenerg Biomembr 53, 489–498 (2021). https://doi.org/10.1007/s10863-021-09906-3
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DOI: https://doi.org/10.1007/s10863-021-09906-3