Abstract
In this study, the ethanolic extract, and the dichloromethane fraction from L. origanoides were assessed for their antibacterial activity alone or in combination with fluoroquinolones against a S. aureus strain resistant to hydrophilic fluoroquinolones due to overexpression of the norA gene encoding the NorA efflux pump, aiming to investigate the occurrence of efflux pump inhibitors in this plant. Minimal inhibitory concentrations (MICs) and modulatory activity on resistance to fluoroquinolones of the extracts were determined by a micro-dilution assay. The ethanolic extract showed weak anti-staphylococci activity, while the dichloromethane extract demonstrated moderate antimicrobial activity against the strains tested. The addition of ethanolic and dichloromethane extracts to the growth medium at sub-inhibitory concentrations enhanced the activity of norfloxacin and ciprofloxacin, as well as ethidium bromide, against S. aureus SA1199-B overproducing NorA. L. origanoides can be a source of secondary metabolites able to inhibit the S. aureus NorA efflux pump.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Availability of data and materials
All data generated or analyzed during this study are included in the article.
Abbreviations
- MRSA:
-
Methicillin resistant S. aureus
- LOEE:
-
Ethanol extract of Lippia origanoides
- LODCM:
-
Dichloromethane fraction of Lippia origanoides
- ATCC:
-
American Type Culture Collection
- BHI:
-
Brain hearth infusion
- MICs:
-
Minimal inhibitory concentrations
- ANOVA:
-
Analysis of variance
References
Aldred KJ, Kerns RJ, Osheroff N (2014) Mechanism of quinolone action and resistance. Biochemistry 53:1565–1574. https://doi.org/10.1021/bi5000564
Baldissera MD, Souza CF, Mourão RHV, Silva LVF, Monteiro SG (2017) Trypanocidal action of Lippia alba and Lippia origanoides essential oils against Trypanosoma evansi in vitro and in vivo used mice as experimental model. J Parasit Dis 41:345–351. https://doi.org/10.1007/s12639-016-0800-7
Barreto HM, Cerqueira FF, Oliveira AP, Arcanjo DDR, Santos BHC, Abreu APL, Coutinho HDM, Silva RAC, Sousa TO, Medeiros MGF, Citó AMGL, Lopes JAD (2014a) Phytochemical prospection and modulation of antibiotic activity in vitro by Lippia origanoides HBK in methicillin resistant Staphylococcus aureus. Biomed Res Int 2014:305610. https://doi.org/10.1155/2014/305610
Barreto HM, de Lima IS, Coelho KMRN, Osório LR, de Almeida MR, Santos BHC, Coutinho HDM, Abreu APL, Medeiros MGF, Citó AMGL, Lopes JAD (2014b) Effect of Lippia origanoides H.B.K. essential oil in the resistance to aminoglycosides in methicillin resistant Staphylococcus aureus. Eur J Integr Med 6:560–564. https://doi.org/10.1016/j.eujim.2014.03.011
Barreto HM, Coelho BRC, Menezes-Silva SMP, Siqueira-Júnior JP, Coutinho HDM, Lemos IZC, Sousa TO, Silva RAC, Medeiros MGF, Citó AMGL, Lopes JAD (2014c) Light-mediated antibacterial activity of Lippia origanoides H.B.K. in vitro. Photochem Photobiol Sci 13:1650–1654. https://doi.org/10.1039/c4pp00334a
Beelders T, De Beer D, Stander MA, Joubert E (2014) Comprehensive phenolic profiling of Cyclopia genistoides (L.) Vent. by LC-DAD-MS and -MS/MS reveals novel xanthone and benzophenone constituents. Molecules 19:11760–11790. https://doi.org/10.3390/molecules190811760
Beytur A, Yakupogullari Y, Oguz F, Otlu B, Kaysadu H (2014) Oral Amoxicillin-Clavulanic Acid treatment in urinary tract infections caused by extended-spectrum beta-lactamase–producing organisms. Jundishapur J Microbiol 8:e13792. https://doi.org/10.5812/jjm.13792
Borges AR, Aires JRDA, Higino TMM, Medeiros MGF, Citó AMGL, Lopes JAD, Figueiredo RCBQ (2012) Trypanocidal and cytotoxic activities of essential oils from medicinal plants of Northeast of Brazil. Exp Parasitol 132:123–128. https://doi.org/10.1016/j.exppara.2012.06.003
Brito A, Ramirez JE, Areche C, Sepúlveda B, Simirgiotis MJ (2014) HPLC-UV-MS profiles of phenolic compounds and antioxidant activity of fruits from three citrus species consumed in Northern Chile. Molecules 19:17400–17421. https://doi.org/10.3390/molecules191117400
Cáceres M, Hidalgo W, Stashenko EE, Torres R, Ortiz C (2023) Metabolomic analysis of the effect of Lippia origanoides essential oil on the inhibition of quorum sensing in Chromobacterium violaceum. Molecules 12:814. https://doi.org/10.3390/antibiotics12050814
Chua KYL, Howden BP, Jiang J-H, Stinear T, Peleg AY (2014) Population genetics and the evolution of virulence in Staphylococcus aureus. Infect Genet Evol 21:554–562. https://doi.org/10.1016/j.meegid.2013.04.026
Coelho AG, Lima Neto JS, Moura AKS, de Sousa TO, Morais ICPS, Carvalho GD, Cunha FVM, Medeiros MGF, Vasconcelos EAF, Oliveira AP, Arcanjo DDR, Nunes LCC, Citó AMGL (2015) Optimization and standardization of extraction method from Lippia origanoides H.B.K.: focus on potential anti-hypertensive applications. Ind Crops Prod 78:124–130. https://doi.org/10.1016/j.indcrop.2015.10.033
Cuyckens F, Claeys M (2004) Mass spectrometry in the structural analysis of flavonoids. J Mass Spectrom 39:1–15. https://doi.org/10.1002/jms.585
Cuyckens F, Ma YL, Pocsfalvi G, Claeys M (2000) Tandem mass spectral strategies for the structural characterization of flavonoid glycosides. Analysis 28:888–895. https://doi.org/10.1051/analusis:2000280888
Dantas DM, Morais LP, Silva AA, Silva RER, Dias FJ, Amorim TS, Cruz-Martins N, Coutinho HDM, Barbosa R (2023) Pharmacological screening of species from the Lippia genus, content in terpenes and phenylpropanoids, and their vasorelaxing effects on human umbilical artery. Curr Pharm Des 29:535–542. https://doi.org/10.2174/1381612829666221124101321
De Lastours V, Chau F, Roy C, Larroque B, Fantin B (2014) Emergence of quinolone resistance in the microbiota of hospitalized patients treated or not with a fluoroquinolone. J Antimicrob Chemother 69:3393–3400. https://doi.org/10.1093/jac/dku283
Diniz-Silva HT, Magnani M, Siqueira S, Souza EL, Siqueira-Júnior JP (2016) Fruit flavonoids as modulators of norfloxacin resistance in Staphylococcus aureus that overexpresses norA. Food Sci Technol 85:324–326. https://doi.org/10.1016/j.lwt.2016.04.003
Du D, Wang-Kan X, Neuberger A, van Veen HW, Pos KM, Piddock LJV, Luisi BF (2018) Multidrug efflux pumps: structure, function and regulation. Nat Rev Microbiol 16:523–539. https://doi.org/10.1038/s41579-018-0048-6
Falcão-Silva VS, Silva DA, de Souza MFV, Siqueira-Junior JP (2009) Modulation of drug resistance in Staphylococcus aureus by a kaempferol glycoside from Herissantia tiubae (Malvaceae). Phytother Res 23:1367–1370. https://doi.org/10.1002/ptr.2695
Feketeová L, Barlow CK, Benton TM, Rochfort SJ, O’Hair RAJ (2011) The formation and fragmentation of flavonoid radical anions. Int J Mass Spectrom 301:174–183. https://doi.org/10.1016/j.ijms.2010.08.017
Funari CS, Eugster PJ, Martel S, Carrupt PA, Wolfender JL, Silva DHS (2012) High resolution ultra high pressure liquid chromatography-time-of-flight mass spectrometry dereplication strategy for the metabolite profiling of Brazilian Lippia species. J Chromatogr A 1259:167–178. https://doi.org/10.1016/j.chroma.2012.03.069
Gade N, Qazi M (2013) Fluoroquinolone therapy in Staphylococcus aureus infections: where do we stand? J Lab Phys 5:109–112. https://doi.org/10.4103/0974-2727.119862
García TH, Quintino da Rocha C, Dias MJ, Pino LL, del Barrio G, Roque A, Pérez CE, Campaner dos Santos L, Spengler I, Vilegas W (2017) Comparison of the qualitative chemical composition of extracts from Ageratina havanensis collected in two different phenological stages by FIA-ESI-IT-MS n and UPLC/ESI-MSn: antiviral activity. Nat Prod Commun 12:31–34. https://doi.org/10.1177/1934578X1701200
Garcia-Fernández A, Gallina S, Owczarek S, Dionisi AM, Benedetti I, Decastelli L, Luzzi I (2015) Emergence of ciprofloxacin-resistant Salmonella enterica Serovar Typhi in Italy. PLoS ONE 10:e0132065. https://doi.org/10.1371/journal.pone.0132065
Guillín Y, Cáceres M, Stashenko EE, Hidalgo W, Ortiz C (2023) Untargeted metabolomics for unraveling the metabolic changes in planktonic and sessile cells of Salmonella Enteritidis ATCC 13076 after treatment with Lippia origanoides essential oil. Antibiotics (basel) 12:899. https://doi.org/10.3390/antibiotics12050899
Holetz FB, Pessini GL, Sanches NR, Cortez DAG, Nakamura CV, Dias-Filho BP (2002) Screening of some plants used in the Brazilian folk medicine for the treatment of infectious diseases. Mem Inst Oswaldo Cruz 97:1027–1031. https://doi.org/10.1590/s0074-02762002000700017
Kaatz G, Seo SM (1993) Efflux-mediated fluoroquinolone resistance in Staphylococcus aureus. Antimicrob Agents Chemother 37:1086–1094. https://doi.org/10.1128/aac.37.5.1086
Kaatz GW, Seo SM (1995) Inducible NorA-mediated multidrug resistance in Staphylococcus aureus. Antimicrob Agents Chemother 39:2650–2655. https://doi.org/10.1128/aac.39.12.2650
Lambert RJW, Skandamis PN, Coote PJ, Nychas G-JE (2001) A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol. J Appl Microbiol 91:453–462. https://doi.org/10.1046/j.1365-2672.2001.01428.x
Leitão SG, Leitão GG, Vicco DKT, Pereira JPB, de Morais SG, Oliveira DR, Celano R, Campone L, Piccinelli AL, Rastrelli L (2017) Counter-current chromatography with off-line detection by ultra high performance liquid chromatography/high resolution mass spectrometry in the study of the phenolic profile of Lippia origanoides. J Chromatogr A 1520:83–90. https://doi.org/10.1016/j.chroma.2017.09.004
Levine DP (2006) Vancomycin: a history. Clin Infect Dis 42:5–12. https://doi.org/10.1086/491709
Macêdo CAF, Paiva GO, Menezes PMN, Ribeiro TF, Brito MC, Vilela DAD, Duarte Filho LAMS, Ribeiro FPRA, Lucchese AM, Lima JT, Silva FS (2022) Lippia origanoides essential oil induces tocolytic effect in virgin rat uterus and inhibits writhing in a dysmenorrhea mouse model. J Ethnopharmacol 90:115099. https://doi.org/10.1016/j.jep.2022.115099
Maia GLDA, Falcão-Silva VDS, Aquino PGV, De Araújo-Júnior JX, Tavares JF, Silva MS, Rodrigues LC, Siqueira-Júnior JP, Barbosa-Filho JM (2011) Flavonoids from Praxelis clematidea R.M. King and Robinson modulate bacterial drug resistance. Molecules 16:4828–4835. https://doi.org/10.3390/molecules16064828
Markham P, Westhaus E, Lyachko KK, Johnson ME, Neyfakh A (1999) Multiple novel inhibitors of the NorA multidrug transporter of Staphylococcus aureus. Antimicrob Agents Chemother 43:2404–2408. https://doi.org/10.1128/AAC.43.10.2404
Martínez A, Stashenko EE, Sáez RT, Zafra G, Ortiz C (2023) Effect of essential oil from Lippia origanoides on the transcriptional expression of genes related to quorum sensing, biofilm formation, and virulence of Escherichia coli and Staphylococcus aureus. Antibiotics (basel) 12:845. https://doi.org/10.3390/antibiotics12050845
Melo ARP, Higino TMM, Oliveira ADPR, Fontes A, Silva DCN, Castro MCAB, Lopes JAD, Figueiredo RCBQ (2020) Lippia sidoides and Lippia origanoides essential oils affect the viability, motility and ultrastructure of Trypanosoma cruzi. Micron 129:102781. https://doi.org/10.1016/j.micron.2019.102781
Meneses R, Ocazionez RE, Martínez JR, Stashenko EE (2009) Inhibitory effect of essential oils obtained from plants grown in Colombia on yellow fever virus replication in vitro. Ann Clin Microbiol Antimicrob 8:8. https://doi.org/10.1186/1476-0711-8-8
Neyfakh AA, Borsch CM, Kaatz GW (1993) Fluoroquinolone resistance protein NorA of Staphylococcus aureus is a multidrug efflux transporter. Antimicrob Agents Chemother 37:128–129. https://doi.org/10.1128/AAC.37.1.128
Oliva ML, Uchima SMM, Suárez JP, Arango ACM, Pabón MCM (2023) Lippia origanoides derivatives in vitro evaluation on polymicrobial biofilms: Streptococcus mutans, Lactobacillus rhamnosus and Candida albicans. Arch Oral Biol 148:105656. https://doi.org/10.1016/j.archoralbio.2023.105656
Oliveira DR, Leitão GG, Bizzo HR, Lopes D, Alviano DS, Alviano CS, Leitão SG (2007) Chemical and antimicrobial analyses of essential oil of Lippia origanoides H.B.K. Food Chem 101:236–240. https://doi.org/10.1016/j.foodchem.2006.01.022
Oliveira AF, Costa Junior LM, Lima AS, Silva CR, Ribeiro MNS, Mesquista JWC, Rocha CQ, Tangerina MMP, Vilegas W (2017) Anthelmintic activity of plant extracts from Brazilian savanna. Vet Parasitol 236:121–127. https://doi.org/10.1016/j.vetpar.2017.02.005
Pascual ME, Slowing K, Carretero E, Sánchez Mata D, Villar A (2001) Lippia: traditional uses, chemistry, and pharmacology: a review. J Ethnopharmacol 76:201–214. https://doi.org/10.1016/s0378-8741(01)00234-3
Pereira WH, Scherbakov AM, Buravchenko GI, Mikhaevich EI, Leitão SG, Cos P, Shchekotikhin AE, Monzote L, Setzer WN (2022) In vitro pharmacological screening of essential oils from Baccharis parvidentata and Lippia origanoides growing in Brazil. Molecules 27:1926. https://doi.org/10.3390/molecules27061926
Pinto CP, Rodrigues VD, Pinto FP, Pinto RP, Uetanabaro APT, Pinheiro CSR, Gadea SFM, Silva TRS, Lucchese AM (2013) Antimicrobial activity of Lippia species from the Brazilian semiarid region traditionally used as antiseptic and anti-infective agents. Evid-Based Complement Altern Med 2013:614501. https://doi.org/10.1155/2013/614501
Redgrave LS, Sutton SB, Webber MA, Piddock LJV (2014) Fluoroquinolone resistance: mechanisms, impact on bacteria, and role in evolutionary success. Trends Microbiol 22:438–445. https://doi.org/10.1016/j.tim.2014.04.007
Ribeiro FP, Oliveira MS, Feitosa AO, Marinho PSB, Marinho AMR, Andrade EHA, Ribeiro AF (2021) Chemical composition and antibacterial activity of the Lippia origanoides Kunth essential oil from the Carajás National Forest Brazil. Evid-Based Complement Altern Med 2021:9930336. https://doi.org/10.1155/2021/9930336
Ruiz-Duran J, Torres R, Stashenko EE, Ortiz C (2023) Antifungal and antibiofilm activity of Colombian essential oils against different Candida strains. Antibiotics (basel) 12:668. https://doi.org/10.3390/antibiotics12040668
Santos BNG, Furtado MM, Freitas EE, Lima LR, Leão PVS, Oliveira FAA, Medeiros MGF, Andrade EM, Feitosa RCA, Tavares SJS, Coutinho HDM, Ferreira JHL, Barreto HM (2023) Chemical profile of the essential oil of Lippia origanoides Kunth and antibiotic resistance-modifying activity by gaseous contact method. J Herb Med 41:100703. https://doi.org/10.1016/j.hermed.2023.100703
Santos Filho LGA, Reis RB, Souza ASQ, Canuto KM, Brito ES, Castro KNC, Pereira AML, Diniz FM (2023) Chemical composition and biological activities of the essential oils from Lippia alba and Lippia origanoides. Ann Braz Acad Sci 95:e20220359. https://doi.org/10.1590/0001-3765202320220359
Schindler BD, Jacinto P, Kaatz GW (2013) Inhibition of drug efflux pumps in Staphylococcus aureus: current status of potentiating existing antibiotics. Future Microbiol 8:491–507. https://doi.org/10.2217/fmb.13.16
Silva HC, Leal ALAB, Oliveira MM, Barreto HM, Coutinho HDMC, Santos HS, Santiago GMP, Freitas TS, Lima IKC, Teixeira AMR, Nogueira CES (2020) Structural characterization, antibacterial activity and NorA efflux pump inhibition of flavonoid fisetinidol. S Afr J Bot 132:140–145. https://doi.org/10.1016/j.sajb.2020.03.023
Souza EL, Oliveira CEV, Stamford TLM, Conceição ML, Neto NJG (2013) Influence of carvacrol and thymol on the physiological attributes, enterotoxin production and surface characteristics of Staphylococcus aureus strains isolated from foods. Braz J Microbiol 44:29–35. https://doi.org/10.1590/S1517-83822013005000001
Spínola V, Pinto J, Castilho PC (2015) Identification and quantification of phenolic compounds of selected fruits from Madeira Island by HPLC-DAD-ESI-MSn and screening for their antioxidant activity. Food Chem 173:14–30. https://doi.org/10.1016/j.foodchem.2014.09.163
Stashenko EE, Martínez JR, Cala MP, Durán DC, Caballero D (2013) Chromatographic and mass spectrometric characterization of essential oils and extracts from Lippia (Verbenaceae) aromatic plants. J Sep Sci 36:192–202. https://doi.org/10.1002/jssc.201200877
Timóteo P, Karioti A, Leitão SG, Vincieri FF, Bilia AR (2015) A validated HPLC method for the analysis of herbal teas from three chemotypes of Brazilian Lippia alba. Food Chem 175:366–373. https://doi.org/10.1016/j.foodchem.2014.11.129
Tong SYC, Davis JS, Eichenberger E, Holland TL, Fowler VG (2015) Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev 28:603–661. https://doi.org/10.1128/CMR.00134-14
Vali L, Dashti AA, Jadaon MM, El-Shazly S (2015) The emergence of plasmid mediated quinolone resistance qnrA2 in extended spectrum β-lactamase producing Klebsiella pneumoniae in the Middle East. DARU J Pharm Sci 23:34. https://doi.org/10.1186/s40199-015-0116-7
Vicuña GC, Stashenko EE, Fuentes JL (2010) Chemical composition of the Lippia origanoides essential oils and their antigenotoxicity against bleomycin-induced DNA damage. Fitoterapia 81:343–349. https://doi.org/10.1016/j.fitote.2009.10.008
Zapata-Zapata C, Rojas-López M, Garcia LT, Quintero W, Trrón MC, Luque D, Mesa-Arango AC (2023) Lippia origanoides essential oil or thymol in combination with fluconazole produces damage to cells and reverses the azole-resistant phenotype of a Candida tropicalis strain. J Fungi 9:888. https://doi.org/10.3390/jof9090888
Acknowledgements
We are grateful to Dr. Glenn W. Kaatz (Wayne State University, USA) who kindly provided us with the SA1199-B strain.
Funding
This research was funded by the Fundação de Amparo à Pesquisa do Estado do Piauí (Grant Number: 050/2019), CNPq and Universidade Federal do Piauí.
Author information
Authors and Affiliations
Contributions
The authors confirm contribution to the paper as follows: study conception and design: HMB, HDMC, and JPSJ. Data collection: FAAO, GMDL, JNS, BCGVL, and JSLN. Analysis and interpretation of results: HMB, GSF, DRJF, JHLF, and JSLN. Draft manuscript preparation: HMB, AMGLC. All authors reviewed the results and approved the final version of the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest to report regarding the present study.
Ethical approval
Not applicable.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
de Alcantara Oliveira, F.A., Lemos, G.M.D., de Sousa, J.N. et al. Modulation of fluoroquinolone resistance by Lippia origanoides Kunth in Staphylococcus aureus. Vegetos (2024). https://doi.org/10.1007/s42535-023-00783-2
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s42535-023-00783-2