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Lippia grata Essential Oil Acts Synergistically with Ampicillin Against Staphylococcus aureus and its Biofilm

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Abstract

Antimicrobial resistance (AMR) presents a global challenge as microorganisms evolve to withstand the effects of antibiotics. In addition, the improper use of antibiotics significantly contributes to the AMR acceleration. Essential oils have garnered attention for their antimicrobial potential. Indeed, essential oils extracted from plants contain compounds that exhibit antibacterial activity, including against resistant microorganisms. Hence, this study aimed to evaluate the antimicrobial and antibiofilm activity of the essential oil (EO) extracted from Lippia grata and its combination with ampicillin against Staphylococcus aureus strains (ATCC 25923, ATCC 700698, and JKD6008). The plant material (leaves) was gathered in Mossoro, RN, and the EO was obtained using the hydrodistillation method with the Clevenger apparatus. The antimicrobial activity of the EO was assessed through minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Antibiofilm activity was evaluated by measuring biomass using crystal violet (CV) staining, viable cell counting, and analysis of preformed biofilms. In addition, the synergistic effects of the EO in combination with ampicillin were examined by scanning electron and confocal microscopy. The EO displayed a MIC value of 2.5 mg/mL against all tested S. aureus strains and an MBC only against S. aureus JKD6008 at 2.5 mg/mL. L. grata EO caused complete biofilm inhibition at concentrations ranging from 10 to 0.312 mg/mL against S. aureus ATCC 25923 and 10 to 1.25 mg/mL against S. aureus ATCC 700698 and S. aureus JKD6008. In the viable cell quantification assay, there was a reduction in CFU ranging from 1.0 to 8.0 logs. The combination of EO with ampicillin exhibited a synergistic effect against all strains. Moreover, the combination showed a significantly inhibiting biofilm formation and eradicating preformed biofilms. Furthermore, the EO and ampicillin (individually and in combination) altered the cellular morphology of S. aureus cells. Regarding the mechanism, the results revealed that L. grata EO increased membrane permeability and caused significant membrane damage. Concerning the synergy mechanism, the results revealed that the combination of EO and ampicillin increases membrane permeability and causes considerable membrane damage, further inhibiting bacteria synergistically. The findings obtained here suggest that L. grata EO in combination with ampicillin could be a viable treatment option against S. aureus infections, including MRSA strain.

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Acknowledgements

The authors are grateful to Central Analítica-UFC/CT-INFRA/MCTI-SISNANO/Pró-Equipamentos CAPES.

Funding

This research was funded by FAPEMIG (Grant#: APQ-00224-2 and APQ-02972-22 for Mayron Alves de Vasconcelos).

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Authors and Affiliations

  1. Faculdade de Ciências Exatas E Naturais, Universidade do Estado do Rio Grande do Norte, Mossoró, RN, Brazil

    Anderson Paulo Rodrigues Lopes, Francisco Fábio Mesquita Oliveira, Cynthia Cavalcanti de Albuquerque & Mayron Alves de Vasconcelos

  2. Laboratório Integrado de Biomoléculas, Departamento de Patologia E Medicina Legal, Universidade Federal do Ceará, Fortaleza, CE, Brazil

    Alexandre Lopes Andrade, Aryane de Azevedo Pinheiro, Leonardo Silva de Sousa, Ellen Araújo Malveira, Edson Holanda Teixeira & Mayron Alves de Vasconcelos

  3. Centro Universitário Inta – UNINTA, Itapipoca, CE, Brazil

    Aryane de Azevedo Pinheiro

  4. Faculdade de Educação de Itapipoca, Universidade Estadual do Ceará, Itapipoca, CE, Brazil

    Mayron Alves de Vasconcelos

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  1. Anderson Paulo Rodrigues Lopes
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Contributions

APRL, ALA, AAP, LSS, and EAM—Contributed to antibacterial and antibiofilm activities and writing of the manuscript. FFMO—Contributed to the plant material collection and essential oil extraction. EHT, CCA, and MAV—Contributed to the analysis of the data, critical reading, draft of the manuscript, and design of the study, and supervised the laboratory work.

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Correspondence to Mayron Alves de Vasconcelos.

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Lopes, A.P.R., Andrade, A.L., Pinheiro, A.d. et al. Lippia grata Essential Oil Acts Synergistically with Ampicillin Against Staphylococcus aureus and its Biofilm. Curr Microbiol 81, 176 (2024). https://doi.org/10.1007/s00284-024-03690-0

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