Anti-biofilm property of essential oils from Cymbopogon sp. against pathogenic bacteria in single-culture and co-culture

Alessandra Farias Millezi, Vanessa Schuh, Janaina Schuh, Taciara Penno do Amaral


In this study, we evaluated whether the essential oils (EOs) from Cymbopogon flexuosus and Cymbopogon martinii can prevent production of biofilms either in single or combined culture of Staphylococcus aureus and Pseudomonas aeruginosa. Biofilm formation was assessed by microtiter-plate test with further quantification of viable cells and biofilm biomass. The evaluated EOs at 0.78 % significantly (P < 0.05) reduced only the viable cells of S. aureus that inhabited biofilm. However, in single-and co-culture assays, both oils significantly (P > 0.05) decreased the amount of biofilm biomass. Biofilm reductions between 52-83% and 60-93% were achieved for the treatments with EOs from C. flexuosus and C. martinii, respectively. Although the biomass reductions of simgle and co-cultivated biofilms were significant, the same was not true for viable cells, except for S. aureus. Considering that the remaining colony forming units can reconstitute the EPS matrix, studies with higher concentrations than those used in this research are suggested in order to obtain greater logarithmic reductions of viable bacterial cells.


Biofilm; Staphylococcus aureus; Pseudomonas aeruginosa.

Full Text:



ADUKWU EC, ALLEN SCH, PHILLIPS CA. The anti-biofilm activity of lemongrass (Cymbopogon flexuosus) and grapefruit (Citrus paradisi) essential oils against five strains of Staphylococcus aureus. J. Appl. Microbiol. 2012;1217–1227.

ADUKWU EC, BOWLES M, EDWARDS-JONES V, BONE H. Antimicrobial activity, cytotoxicity and chemical analysis of lemongrass essential oil (Cymbopogon flexuosus) and pure citral. Appl. Microbiol. Biotechnol. 2016;100(22):9619-9627.

ATSHAN SS, SHAMSUDIN MN, SEKAWI Z, THIAN LUNG LT, BARANTALAB F, LIEW YK et al. Comparative proteomic analysis of extracellular proteins expressed by various clonal types of Staphylococcus aureus and during planktonic growth and biofilm development. Front Microbiol. 2015;524.

BAKKALI F, AVERBECK S, AVERBECK D, IDAOMAR M. Biological effects of essential oils: a review. Food Chem Toxicol. 2008;46(2):446–475.

BRÕZEL VS, STRYDOM GM, CLOETE TEE. A method for the study of de novo protein synthesis in Pseudomonas aeruginosa after attachment. Biofouling. 1995;195–210.

BURT S. Essential oils: their antibacterial properties and potential applications in foods—a review. Int J Food Microbiol. 2004;94(3):223–253.

CAROVIC-STANKO K, ORLIC´ S, POLITEO O, STRIKIC´ F, KOLAK I, MILOS M et al. Composition and antibacterial activities of essential oils of seven Ocimum taxa. Food Chem. 2010;196-201.

CLSI (Clinical and Laboratory Standards Institute). Performance standards for antimicrobial disk susceptibility tests: approved standard. 8.ed. 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA, 2003. (CLSI document M2-A8 [ISBN 1-56238-485-6]).

COPE TA. Poaceae: Flora of Pakistan. Karachi: University of Karachi; 1982.

COSGROVE SE, KAYE KS, ELIOPOULOUS GM, CARMELI Y. Health and economic outcomes of the emergence of third-generation cephalosporin resistance in Enterobacter species. Arch Intern Med. 2002;162(2):185–190.

DONLAN RM, COSTERTON JW. Biofilm Formation: A Clinically Relevant Microbiological Process. Clin. Microbiol. Rev.2001;15(8), 1387-1392

DONLAN, R. Biofilms: microbial life on surfaces. Emerging infectious diseases. Emerg Infect Dis. 2002;8(9):881-90.

DUBERN JF, DIGGLE SP. For a recent review. Mol. BioSyst. 2008;4(9):882-8.

ESPINA L, PAGÁN R, LÓPEZ D, GARCÍA-GONZALO D. Individual constituents from essential oils inhibit biofilm mass production by multi-drug resistant staphylococcus aureus. Molecules. 2015;19-20(6):11357-72.

EL-SAYED AM. The Pherobase: Database of Pheromones and Semiochemicals. 2018. [accessed 2018 August 24].

FAGERLUND A, LANGSRUD S, HEIR E, MIKKELSEN MI, AND MORETRO T. Biofilm matrix composition affects the susceptibility of food associated staphylococci to cleaning and disinfection agents. Front Microbiol. 2016;856.

GACCHE RN, SHAIKH RU, CHAPOLE SM, JADHAV AD, JADHAV SG. Kinetics of inhibition of monoamine oxidase using Cymbopogon martinii (Roxb.) Wats.: a potential antidepressant herbal ingredient with antioxidant activity. Indian J Clin Biochem. 2011;26(3):303–308.

GHADYALE V, TAKALIKAR S, HALDAVNEKAR V, ARVINDEKAR A. Effective control of postprandial glucose level through inhibition of intestinal alpha glucosidase by Cymbopogon martinii (Roxb.). Evid Based Complement Altern Med 2012;1-6.

TYAGI BK, SHAHI AK, KAUL BL. Evaluation of repellent activities of Cymbopogon essential oils against mosquito vectors of malaria, filariasis and dengue fever in India. Phytomedicine 1998;5(4):324-9.

GÖTZ, F. Staphylococcus and biofilms. Mol Biotechnol.2002; 43(6):1367-78.

GUTIÉRREZ D, DELGADO S, VÁZQUEZ-SÁNCHEZ D, MARTÍNEZ B, CABO ML, RODRÍGUEZ A. et al. Incidence of Staphylococcus aureus and analysis of associated bacterial communities on food industry surfaces. Appl Environ Microbiol. 2012;78(24):8547–8554.

DA SILVA GS, DE SOUZA ME, QUATRIN PM, KLEIN B, WAGNER R, GÜNDEL A et al. Nanoemulsions containing Cymbopogon flexuosus essential oil: Development, characterization, stability study and evaluation of antimicrobial and antibiofilm activities. Microbial Pathogenesis. 2018;268–276.

HERIGSTAD B, HAMILTON M, HEERSINK J. How to optimize the drop plate method for enumerating bacteria. J Microbiol Methods. 2001;44(2):121-9.

JARAMILLO-COLORADO B, OLIVERO-VERBEL J, STASHENKO EE, WAGNER-DÖBLER I, KUNZE B. Anti-quorum sensing activity of essential oils from Colombian plants. Nat Prod Res. 2012;26(12):1075-86.

KARKALA S, GANJEWALA D. Antimicrobial activity of essential oils of four lemongrass (Cymbopogon flexuosus Steud) varieties. Medicinal and Aromatic Plant Science and Biotechnology. 2009;3(1) 107-109.

KUMARAN AM, D’SOUZA P, AGARWAL A, BOKKOLLA RM, BALASUBRAMANIAM M. Geraniol, the putative anthelmintic principle of Cymbopogonmartinii. Phytother Res. 2003;17(8):957.

LEWIS, K. Riddle of biofilm resistance. Antimicrob Agents Chemother. 2001;45(4):999-1007.

LINDSAY D, BRÖZEL VS, MOSTERT JF, VON HOLY A. Differential efficacy of a chlorinedioxide containing sanitizer against singles pecies and binary biofilms of adairy-associated Bacillus cereus and a Pseudomonas fluorescens isolate. J. Appl Microbiol. 2002;92(2):352-61.

MARQUES SC, REZENDE JGOS, ALVES LAF, SILVA BC, ALVES E, DE ABREU, L. R. et al. Formation of biofilms by Staphylococcus aureus on stainless steel and glass surfaces and its resistance to some selected chemical sanitizers. Braz J Microb. 2007;38(3):538–543.

MILLEZI AF, ROSSONI DF, CANO IA, PICCOLI RH. Sensibilidade de bactérias patogênicas em alimentos a óleos essenciais de plantas medicinais e condimentares. Hig Ali. 2016;30(254-255):117-122.

MILLEZI AF, DALLA COSTA KA, OLIVEIRA JM, LOPES SP, PEREIRA MO, PICCOLI RH. Antibacterial and anti-biofilm activity of cinnamon essential oil and eugenol. Ciên. Rural. 2019;49(01):1-7.

MASÁK J, ČEJKOVÁ A, SCHREIBEROVÁ O, ŘEZANKA T. Pseudomonas biofilms: Possibilities of their control. FEMS Microbiol Ecol. 2014;89(1):1-14.

NEIDIG A, YEUNG AT, ROSAY T, TETTMANN B, STREMPEL N, RUEGER M et al. TypA is involved in virulence, antimicrobial resistance and biofilm formation in Pseudomonas aeruginosa. BMC Microbiol. 2013;9(17):1-10.

NOSTRO A, SUDANO ROCCARO A, BISIGNANO G, MARINO A, CANNATELLI MA, PIZZIMENTI FC, et al. Effects of oregano, carvacrol and thymol on Staphylococcus aureus and Staphylococcus epidermidis biofilms. J Med Microb. 2007;56(4):519-23.

NOVICK, R. P. Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol. Microb. 2003;48(6):1429–1449.

O’TOOLE GA, KOLTER R. The initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceedsvia multiple, convergent signaling pathways: a genetic ana-lysis. Mol Microbiol. 1998; 28(3):449-61.

PRASHAR A, HILI P, VENESS RG, EVANS CS. Antimicrobial action of palmarosa oil (Cymbopogon martinii) on Saccharomyces cerevisiae. Phytochemistry. 2003;63(5):569-75.

Prasad CS, Shukla R, Kumar A, Dubey NK. in vitro and in vivo antifungal activity of essential oils of Cymbopogon martinii and Chenopodium ambrosioides and their synergism against dermatophytes. Mycoses 2010; 53: 123-129.

PRATT LA, KOLTER R. Genetic analysis of Escherichia coli biofilm formation: defining the roles of flagella, motility, chemotaxis and type I pili. Mol Microbiol. 1998; 30(2):285-93.

RAINA VK, SRIVASTAVA SK, AGGARWAL KK, SYAMASUNDAR KV, KHANUJA SPS. Essential oil composition of Cymbopogon martinii from different places in India. Flavour and Fragrance Journal. 2003;18(4):312–315.

SANCHEZ-VIZUETE P, ORGAZ B, AYMERICH S, LE COQD, BRIANDET R. Pathogens protection against the action of disinfectants in multispecies biofilms. Front Microbiol. 2015:705.

SCHERER R, WAGNER R, DUARTE MCT, GODOY HT. Composição e atividades antioxidante e antimicrobiana dos óleos essenciais de cravo-da-índia, citronela e palmarosa. Rev Bras Plantas Med. 2009;11(4):442–449.

SHARMA PR, MONDHE DM, MUTHIAH S, PAL HC, SHAHI AK, SAXENA AK et al. Anticancer activity of an essential oil from Cymbopogon flexuosus. Chem Biol Interact. 2009;179(2-3):160-8.

SIKKEMA J, DE BONT JAM, POOLMAN B. Mechanisms of membrane toxicity of hydrocarbons. Microbiol Rev. 1995;59(2):201–222

SIMÕES M, SIMÕES LC, VIEIRA MJ. Species association increases biofilm resistance to chemical and mechanical treatments. Water Res. 2009;43(1):229-37.

STEPANOVIĆ S, VUKOVIĆ D, DAKIĆ I, SAVIĆ B, ŠVABIĆ-VLAHOVIĆ M. A modified microtiter plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods. 2000;40(2):175-9.

TAMULI P, SAIKIA M, BORUAH P. Post-infectional biochemical changes in Cymbopogon martinii (Roxb.) Wats and Cymbopogon citratus (DC) Stapf. due to leaf rust disease. Am J Plant Sci. 2013:1666-1668.

TSAI M, LIN CC, LIN WC, YANG CS. Antimicrobial, Antioxidant, and Anti-Inflammatory Activities of Essential Oils from Five Selected Herbs. Biosci Biotechnol Biochem. 2011;75(10):1977-83.

WANG R, KALCHAYANAND N, SCHMIDT JW, HARHAY DM. Mixed Biofilm Formation by Shiga Toxin-Producing Escherichia coli and Salmonella enterica Serovar Typhimurium Enhanced Bacterial Resistance to Sanitization due to Extracellular Polymeric Substances. J Food Prot. 2013 Sep;76(9):1513-22.

WATNICK P, KOLTER R. Biofilm, city of microbes. J Bacteriol. 2000;182(10):2675-9.

WIMPENNY JWT, COLASANTI R. A unifyinghypothesis for the structure of microbial biofilms basedon cellular utomaton models. FEMS Microbiol Ecol. 1997;22(1)1–16.


Copyright (c) 2020 Ciência e Natura

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.