Evaluation of the Antibacterial Effect of Propolis Extract and its Use in Mouthwash Production

  • Rahman Nazeri Faculty of Dentistry, AJA University of Medical Sciences, Tehran, Iran
  • Marzieh Ghaiour Department of Pediatric Dentistry and Research Committee, Faculty of Dentistry, Isfahan University of Medical Sciences, Isfahan, Iran
  • Shima Abbasi Mail Department of Oral and Maxillofacial Pathology, Faculty of Dentistry, AJA University of Medical Sciences, Tehran, Iran
Anti-bacterial Agents, Propolis, Mouthwash


Objectives: The purpose was to determine the antibacterial properties of propolis and to evaluate its use as an antibacterial mouthwash with minimal complications
Materials and Methods: In this experimental laboratorial study, the alcoholic extract of Propolis was first prepared. Then the Minimum Inhibitory Consentration(MIC) was calculated for four bacterial species of staphylococcus aureus, Streptococcus mutans, Lactobacillus acidophilus and Enterococcus faecalis using the agar dilution test. Using the MIC, the Propolis antibacterial mouthwas was produced and compared to water, chlorhexidine and Listerine in laboratory rats for clinical examination. Salivary specimens of rats were collected 12 hours, 1 week and 2 weeks after using the mouthwash and examined by the real time PCR. Data were analyzed by SPSS using one-way ANOVA and the independent t-test (α=0.05).
Results: The results of agar dilution test showed the lowest MIC for Staphylococcus aureus and the highest for Lactobacillus acidophilus. On the other hand, the results of this study showed that water alone had no effect on the level of oral bacteria. In the case of Streptococcus mutans, Enterococcus faecalis, and Lactobacillus Acidophilus mouthwash, the Propolis mouthwash showed a significant difference with chlorhexidine and listerine (P <0.05), while chlorhexidine and listerine had lower effect in comparison. As for Staphylococcus aureus, there was no significant difference between chlorhexidine and Propolis bacteria (P >0.05), but Listerine had the lower effect than either (P <0.05).
Conclusions: According to the results, propolis mouthwash was more efficient against the studied oral bacteria compared to CHX and Listerine.


Wilder RS, Bray KS. Improving periodontal outcomes: Merging clinical and behavioral science. Periodontol 2000. 2016;71 65–81.

Teles RP, Teles FRF. Antimicrobial agents used in the control of periodontal biofilms: Effective adjuncts to mechanical plaque control? Braz Oral Res. 2009;23(Special Issue 1):39–48.

Borgnakke WS. Does treatment of periodontal disease influence systemic disease? Dent Clin North Am. 2015;59(4):885–917.

Asadoorian J. Therapeutic oral rinsing with commercially available products: Position paper and statement from the Canadian Dental Hygienists Association. Can J Dent Hyg. 2016;50(3):126–39.

Shetty PR, Setty SB, Kamat SS, Aldarti AS, Shetty SN. Comparison of the antigingivitis and antiplaque efficacy of the herbaoral (Herbal extract) mouthwash with Chlorhex-idine and Listerine mouthwashes: A Clinical Study. Pak Oral Dental J 2013;33(1):76-82.

Osso D, Kanani N. Antiseptic Mouth Rinses: an update on comparative effectiveness, risk and recommendations. J Dent Hygiene. 2013; 87(1):10-8.

Sykes LM, Comley M, Kelly L. Availability, indications for use and main ingredients of mouthwashes in six major supermarkets in Gauteng. South African Dental Journal. 2016;71(7):308-13.

Gill S, Kapoor D, Singh J, Nanda T. Comparison of antiplaque efficacy of commercially available HiOra (herbal) mouthwash with Listerine mouthwash: a clinical study. J Periodontol Implant Dent. 2017;9(2):53-57.

Pereira EM, da Silva JL, Silva FF, De Luca MP, Ferreira EF, Lorentz TC, et al. Clinical Edvidence of the efficacy of a mouthwash containing propolis for the control of plaque and gingivitis: a phase II study. Evidence-Based Complimentary and Alternative Medicine. 2011; 1-7.

Hooper SJ, Lewis MA, Wilson MJ, Williams DW. Antimicrobial activity of Citrox bioflavonoid preparations against oral microorganisms. Br Dent J. 2011;210(1):1–5.

Walker E, Nowacki AS. Understanding equivalence and noninferiority testing. J Gen Intern Med. 2011;26(2):192–96.

World Health Organization. WHO Traditional Medicine Strategy 2014–2023. Geneva: WHO; 2013. Available from: apps.who.int/ iris/bitstream/10665/92455/1/9789241506090_eng.pdf.

Nature Chemical Biology. Editorial [Internet]. Springer Nature; 2007. Available from: ature.com/nchembio/journal/v3/n7/full/nchembio0707-351.html.

Balappanavar A, Sardana V, Singh M. Comparison of the effectiveness of 0.5% tea, 2% neem and 0.2% chlorhexidine mouthwashes on oral health: a randomized control trial. Indian J Dent Res. 2013;24(1):26–34.

Akca AE, Akca G, Topcu FT, Macit E, Pikdoken L, Ozgen IS. The Comparative Evaluation of the Antimicrobial Effect of Propolis with Chlorhexidine against Oral Pathogens: An In Vitro Study. Biomed Res Int. 2016;2016(10):3627463.

Clinical and Laboratory Standards Institute, “Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically,” Approved Standard M7-A7, CLSI,Wayne, Pa, USA,2007.

InstituteCaLS, “Methods for antimicrobial susceptibility testing of anaerobic bacteria,” in Approved Standard M11-A7, CLSI,Wayne, Pa, USA, 2007.

Yokoyama M, Fukui M, Masuda K, et al. Measurment of the total number of bacteria in saliva using quantitative real-time PCR (qPCR): evalution of the oral hygiene status. J Dent Health. 2009; 59: 183-189.

Chen L, Mao T, Du M, Yang Y, Xu Q, Fan M. Caries status and quantification of four bacteria in saliva of Chinese preschool children: a cross-sectional study. J Dent Sci. 2014;9(3):283-8.

Morel AS, Dubourg G, Prudent E, Edouard S, Gouriet F, Casalta JP, et al. Complementarity between targeted real-time specific PCR and conventional broad-range 16S rDNA PCR in the syndrome-driven diagnosis of infectious diseases. Eur J Clin Microbiol Infect Dis. 2015 Mar;34(3):561-70.

Kuribayashi M, Kitasako Y, Matin K, Sadr A, Shida K, Tagami J. Intraoral pH measurement of carious lesions with qPCR of cariogenic bacteria to differentiate caries activity. J Dent. 2012;40:222e8.

Song JJ, Twumasi-Ankrah P, Salcido R. Systematic review and meta-analysis on the use of honey to protect from the effects of radiation-induced oral mucositis. Adv Skin Wound Care 2012;25:23-8. doi: 0.1097/01.ASW.0000410687.14363.a3

Bardy J, Slevin NJ, Mais KL, Molassiotis A. A systematic review of honey uses and its potential value within oncology care. J Clin Nurs 2008;17:2604-23. doi: 10.1111/j.1365-2702.2008.02304.x

Steinberg D, Kaine G, Gedalia I. Antibacterial effect of propolis and honey on oral bacteria. Am J Dent 1996;9:236-9.

Diba K, Mousavi B, Mahmoudi M, Hashemi J. In-vitro anti fungal activity of Propolis alcoholic extract on Candida spp. and Aspergillus spp. Tehran University Medical Journal 2010;68:80-6[In persian].

Ozan F, Sümer Z, Polat ZA, Ozan U, Deger O. Effect of Mouthrinse Containing Propolis on Oral Microorganisms and Human Gingival Fibroblasts. Eur J Dent. 2007; 1(4): 195-201.

Daugsch A, Fort P, Park YK. Brazilian Red Propolis Chemical Composition and Botanical Origin. Evid Based Complement Alternat Med. 2008; 5(4): 435-441.

Lotfy M. Biological activity of bee propolis in health and disease. Asian Pac J Cancer Prev 2006;7:22-31.

Carneiro FC, Nadanovsk P. Dentística Ultraconservativa: fundamentos e técnicas de tratamento da cárie em dentina. 1ª edição. São Paulo: Santos; 2003. p. 23.

Kidd EAM, Nyvab B. Controle da cárie dentária para cada paciente. In: Fejerskov O, Kidd E. Cárie dentária - A doença e seu tratamento clínico. São Paulo: Santos; 2005. p. 303-311.

Soares MP, Valenca AMV. Avaliação clínica do potencial terapêutico do gel e verniz fluoretado na remineralização de lesões cariosas incipientes. Pesquisa Brasileira em Odontopediatria e Clínica Integrada. 2003; 3 (2): 35 – 41.

Rozier RG. Effectiveness of methods used by dental professionals for the primary prevention of dental caries. Journal of Dental Education. 2001; 65(10): 1063-1072.

Machado B, Pulcino TN, Silva AL, Melo DT, Silva RG, Mendonca IG. Propolis as an alternative in prevention and control of dental cavity. J Apither. 2016;1(2):47-50.

Vargas AC, Loguercio AP, Witt NM, Costa MM, Silva MS, Viana LR. Atividade antimicrobiana “in vitro” de extrato alcóolico de própolis. Ciência Rural. 2004, 34 (1): 159-163.

Boryana T, Popova M, Bankova V, Simova S, Marcucci MC, Miorin PL, et.al. Bioactive Constituents of Brazilian Red Propolis. Oxford Jounals. 2006; 3(2): 249-254.

Lustosa SR, Galindo AB, Nunes LCC, Randau KP, Pedro J, Neto. Própolis: atualizações sobre a química e a farmacologia. Revista Brasileira de Farmacognosia. 2008; 18(3): 447-454.

Park YK, Alencar SM, Pippa AR, Aguiar SCL. Própolis produzida no sul do Brasil, Argentina e Uruguai: Evidências fitoquímicas de sua origem vegetal. Ciência Rural. 2002; 32(6): 997-1003.

Sawaya AC, Palma AM, Caetano FM, Marcucci MC, Cunha IBS, Araujo CE, et al. Comparative study of in vitro methods used to analyse the activity of propolis extracts with different compositions against species of Candida. Lett Appl Microbiol. 2002 Sep;35(3):203-7.

Agüero MB, Svetaz L, Baroni V, Lima B, Luna L, Zacchino S, et al. Urban propolis from San Juan province (Argentina): Ethnopharmacological uses and antifungal activity against Candida and dermatophytes. Ind Crops Prod. 2014 Jun;57:166-73. doi: 10.1016/j.indcrop.2014.03.009.

Moran J.M., “Home-use oral hygiene products: mouthrinses,”Periodontol 2000;48( 1):42–53.

Pereira EM, da Silva JL, Silva FF, De Luca MP, Ferreira EF, Lorentz TC, et al. Clinical Evidence of the Efficacy of a Mouthwash Containing Propolis for the Control of Plaque and Gingivitis: A Phase II Study. Evid Based Complement Alternat Med. 2011;2011(10):750249.

Bürgers R, Witecy C, Hahnel S, Gosau M.The effect of various topical periimplantitis antiseptics Staphylococcus epidermidis, Candida albicans and Streptococcus sanguinis. Arch Oral Biol. 2012;57:940-47.

Karpinski TM, Szkaradkiewicz AK. Microbiology of dental caries. J Biol Earth Sci. 2013;3:M21-M24.

Loesche WJ. Role of Streptococcus mutans in human dental decay. Microbiol Rev 1986;50:353e80.

Acevedo AM, Ray MV, Socorro M, Rojas-Sanchez F. Frequency and distribution of Mutans Streptococci in dental plaque from caries-free and caries-affected Venezuelan children. Acta Odontol Latinoam 2009;22:15e20.

Karygianni L, Al-Ahmad A, Argyropoulou A, Hellwig E, Anderson AC, Skaltsounis AL. Natural Antimicrobials and Oral Microorganisms: A Systematic Review on Herbal Interventions for the Eradication of Multispecies Oral Biofilms. Front Microbiol. 2016; 16(1529): 1-17.

Love RM. Enterococcus faecalis-a mechanism for its role in endodontic failure. Int Endod J. 2001;34:399–405.

Choi EJ, Lee SH, Kim YJ. Quantitative real-time polymerase chain reaction for Streptococcus mutans and Streptococcus sobrinus in dental plaque samples and its association with early childhood caries. Int J Paediatr Dent 2009;19:141e7.

Becker MR, Paster BJ, Leys EJ, et al. Molecular analysis of bacterial species associated with childhood caries. J Clin Microbiol 2002;40:1001e9.

Tanner AC, Kent Jr RL, Holgerson PL, et al. Microbiota of severe early childhood caries before and after therapy. J Dent Res 2011;90:1298e305.

Luo AH, Yang DQ, Xin BC, Paster BJ, Qin J. Microbial profiles in saliva from children with and without caries in mixed dentition.Oral Dis 2012;18:595e601.

Goto I, Furudoi S, Akashi M, Komori T. Measurement of the Total Number of Bacteria in Saliva Using Quantitative Real-Time PCR During Treatment for Head and Neck Malignancy: A Series of Cases. Oral Health Dent Manag. 2015;14(2);85-89.

Price RR, Viscount HB, Stanley MC, Leung KP. Targeted profiling of oral bacteria in human saliva and in vitro biofilms with quantitative real-time PCR. Biofouling 2007;23:203e13.

Childers NK, Osgood RC, Hsu KL, et al. Real-time quantitative polymerase chain reaction for enumeration of Streptococcus mutans from oral samples. Eur J Oral Sci 2011;119:447e54.

Haffajee AD, Yaskell T, Socransky SS. Antimicrobial effectiveness of an herbal mouthrinse compared with an essential oil and a chlorhexidinemouthrinse. J Am Dent Assoc. 2008;139(5):606-11.

Ercan N, Erdemir EO, Ozkan SY, Hendek MK. The comparative effect of propolis in two different vehicles; mouthwash and chewing-gum on plaque accumulation and gingival inflammation. Eur J Dent. 2015 Apr-Jun;9(2):272-6.

Pedrazzi V, Leite MF, Tavares RC, Sato S, do Nascimento GC, Issa JP. Herbal mouthwash containing extracts of Baccharis dracunculifolia as agent for the control of biofilm: clinical evaluation in humans. ScientificWorldJournal. 2015;2015(10):712683.

Demirel G, Eryılmaz M, Altanlar N, Gür G. In vitro Antimicrobial Activity of Various Mouth Rinses against Streptococcus mutans, Lactobacillus casei, L. acidophilus and Candida albicans. Br J Med Med Res. 2015;9(11):1-5.

Drumond MRS, et al. Evaluation of effect of propolis on dental plaque, gum disease and s.mutans levels in saliva of caries free. Odontologia Clín. Científ. 2006; (4): 313-319.

Anauate-Netto C, Anido-Anido A, Leegoy HR, Matsumoto R, Alonso RC, Marcucci MC, et al. Randomized, double-blind, placebo-controlled clinical trial on the effects of propolis and chlorhexidine mouthrinses on gingivitis. Braz Dent Sci. 2014;17(1):11-5.

Suleman T, van Vuuren S, Sandasi M, Viljoen AM. Antimicrobial activity and chemometric modelling of South African propolis. J ApplMicrobiol. 2015; 119 (4): 981-990.

Vasconcelos WA , Braga NMA , Chitarra VR , Santos VR, Andrade AL, Domingues RZ . Bioactive Glass-Green and Red Propolis Association: Antimicrobial Activity Against Oral Pathogen Bacteria. Nat Prod Chem Res. 2014; 2 (6): 1-5.

Santiago KB, Piana GM, Conti BJ, Cardoso EO, Murbach Teles Andrade BF, Zanutto MR, et al. Microbiological control and antibacterial action of a propolis-containing mouthwash and control of dental plaque in humans. Nat Prod Res. 2017 Jun 23;23:1-5.

Bazvand L, Aminozarbian MG, Farhad A, Noormohammadi H, Hasheminia SM, Mobasherizadeh S. Antibacterial effect of triantibiotic mixture, chlorhexidine gel, and two natural materials Propolis and Aloe vera against Enterococcus faecalis: An ex vivo study. Dent Res J (Isfahan). 2014 Jul;11(4):469-74.

Mohan PV, Uloopi KS, Vinay C, Rao RC. In vivo comparison of cavity disinfection efficacy with APF gel, Propolis, Diode Laser, and 2% chlorhexidine in primary teeth. Contemp Clin Dent. 2016 Jan-Mar;7(1):45-50.

Carbajal Mejia JB. Antimicrobial effects of calcium hydroxide, chlorhexidine, and propolis on Enterococcus faecalis and Candida albicans. J Investig Clin Dent. 2014 Aug;5(3):194-200.

Nagappan N, John J. Antimicrobial efficacy of herbal and chlorhexidine mouthrinse-a systematic review. J Dent Med Sci. 2012;2:5-10.

Malhotra N, Rao SP, Acharya S, Vasudev B. Comparative in vitro evaluation of efficacy of mouthrinses against Streptococcus mutans, Lactobacilli and Candida albicans. Oral Health Prev Dent.2011;9:261-68.

Bhandari S, T SA, Patil CR. An in Vitro Evaluation of Antimicrobial Efficacy of 2% Chlorhexidine Gel, Propolis and Calcium Hydroxide Against Enterococcus faecalis in Human Root Dentin. J Clin Diagn Res. 2014 Nov;8(11):ZC60-3.

How to Cite
Nazeri R, Ghaiour M, Abbasi S. Evaluation of the Antibacterial Effect of Propolis Extract and its Use in Mouthwash Production. Front Dent. 16(1):1-12.
Original Article