Analysis of the antimicrobial activity of propolis: A narrative review of in-vitro studies

LAZO, GLENDA L.; LEMES, DANIELI CRISTINA; PAULETTI, PATRÍCIA M.; BASTOS, JAIRO K.; AMBRÓSIO, SÉRGIO RICARDO; VENEZIANI, RODRIGO CASSIO S.; PIRES, REGINA HELENA

doi:10.1590/0001-3765202520241302

person GLENDA L. LAZO

methodology · formal analysis · investigation · writing—original draft preparation · writing—review and editing

schoolUniversidade de Franca, Programa de Pós-Graduação em Promoção de Saúde, Av. Dr. Armando Salles de Oliveira, 201, 14404-600 Franca, SP, BrazilschoolAgrarian University of Ecuador, Faculty of Veterinary Medicine and Zootechnics, 25 de Julio y Pio Jaramillo Ave., 090104 Guayaquil, Ecuador

0000-0002-3328-6776

person DANIELI CRISTINA LEMES

methodology · formal analysis

schoolUniversidade de Franca, Programa de Pós-Graduação em Promoção de Saúde, Av. Dr. Armando Salles de Oliveira, 201, 14404-600 Franca, SP, Brazil

0000-0002-3650-2198

person PATRÍCIA M. PAULETTI

formal analysis

schoolUniversidade de Franca, Programa de Pós-Graduação em Promoção de Saúde, Av. Dr. Armando Salles de Oliveira, 201, 14404-600 Franca, SP, Brazil

0000-0001-1831-9686

person JAIRO K. BASTOS

data curation · writing—review and editing

schoolUniversidade de São Paulo, Faculdade de Ciências Farmacêuticas, Avenida Professor Doutor Zeferino Vaz, 14040-093 Ribeirão Preto, SP, Brazil

0000-0001-8641-9686

person SÉRGIO RICARDO AMBRÓSIO

data curation

schoolUniversidade de Franca, Programa de Pós-Graduação em Promoção de Saúde, Av. Dr. Armando Salles de Oliveira, 201, 14404-600 Franca, SP, Brazil

0000-0001-5032-3930

person RODRIGO CASSIO S. VENEZIANI

investigation · funding acquisition

schoolUniversidade de Franca, Programa de Pós-Graduação em Promoção de Saúde, Av. Dr. Armando Salles de Oliveira, 201, 14404-600 Franca, SP, Brazil

0000-0001-7057-9706

person REGINA HELENA PIRES

conceptualization · writing—review and editing · funding acquisition

schoolUniversidade de Franca, Programa de Pós-Graduação em Promoção de Saúde, Av. Dr. Armando Salles de Oliveira, 201, 14404-600 Franca, SP, Brazil

0000-0002-6004-5951

Correspondence to: Regina Helena Pires E-mail: regina.pires@unifran.edu.br

Figures (6)
Tables (1)

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Figure 1 Selection of studies included in the study.

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Figure 2 Main antimicrobial components of propolis and the frequency of their identification across different regions. The bar chart highlights the diversity of propolis compounds with antibacterial and antifungal properties, reflecting regional variation in chemical composition.

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Figure 3 Molecular structures of various compounds found in propolis with known antibacterial activity.

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Figure 4 Correlation between various extraction methods and the relative yield of phenolic compounds in propolis. Hydroalcoholic and ultrasound-assisted extractions demonstrate superior effectiveness, yielding the highest concentrations of phenolic compounds and highlighting their potential for optimizing bioactive compound recovery. In contrast, aqueous extraction results in significantly lower yields, underscoring the importance of selecting an appropriate extraction method to maximize the therapeutic potential of propolis.

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Figure 5 Antibacterial action of propolis based on the findings of Silva et al. (2012), Barbosa et al. (2014), Campos et al. (2015), Ristivojević et al. (2016), Moncayo Luján et al. (2018), Veloz et al. (2019), Petruzzi et al. (2020), Rodríguez-Pérez et al. (2020), Navarro-Pérez et al. (2021), Sa-Eed et al. (2023), and Campos et al. (2023). Propolis components, particularly phenolic compounds, interact with bacterial cell structures through multiple mechanisms. Key actions include damage to the plasma membrane, DNA gyrase inhibition, disruption of ATP synthesis, and interference with protein synthesis. These combined actions hinder bacterial growth and demonstrate the potential of propolis as an antibacterial agent.

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Figure 6 Antifungal action of propolis based on the studies of Campos et al. (2015), Rufatto et al. (2018), Correa et al. (2020), Petruzzi et al. (2020), Rodríguez-Pérez et al. (2020), Sokolonski et al. (2021), Fernández-Calderón et al. (2021), Mateo Aldama et al. (2020), Campos et al. (2023), and Rivera-Yañez et al. (2022). Propolis components, particularly phenolic compounds, exert antifungal effects by interacting with the cell wall, increasing its permeability; disrupting the plasma membrane, causing ion leakage (Na+, K+, H+); and altering mitochondrial function, which interferes with the electron transport chain and energy production. Prolonged exposure can also induce apoptosis, followed by secondary necrosis. These combined actions underscore propolis’s potential as a powerful antifungal agent.

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Table I Characteristics of the selected studies: study location, microorganism, type of propolis extract used, method employed, and principal results.

COUNTRY	REFERENCE	MICROORGANISM	PROPOLIS EXTRACT TYPE/Color	METHOD	RESULTS
Brazil	Bastos et al. (2011)	Escherichia coli ATCC 25922	Ethanolic extract/ Brown	Disk diffusion and broth microdilution	The zones of inhibition ranged from 10 to 11.3 mm, and the MIC values for 42.9% of the samples were 250 mg/mL.
Colombia	Alves Ferreira Bastos et al. (2011)	Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Candida albicans ATCC 36802	Colombian propolis extract/ Green	Disk diffusion	The inhibition zones for E. coli ranged from 8 to 12 mm, while for S. aureus, they ranged from 8.3 to 23.5 mm. No activity was observed against C. albicans.
Brazil	Probst et al. (2011)	Clinical isolates of Staphylococcus aureus and Escherichia coli	Ethanolic extract with essential oils from Caryophyllus aromaticus, Zingiber officinale, Cinnamomum zeylanicum and Mentha piperita/ Brown	Agar diffusion	The combinations of EEP with Z. officinale and M. piperita essential oils, as well as with C. zeylanicum, Z. officinale and C. aromaticus essential oils, exhibited bacteriostatic and bactericidal effects, respectively, on the growth of S. aureus.
Brazil	Dantas de Almeida et al. (2012)	Candida albicans ATCC 76618, Candida krusei ATCC 6538, Candida tropicalis ATCC 13803	Propolis tincture/ Brown	Agar diffusion	The pure propolis tincture demonstrates antifungal activity against C. krusei and C. tropicalis, with no observed effect against C. albicans.
Portugal	Silva et al. (2012)	ATCC strains of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans and clinical isolates of the same organisms resistant to methicillin, imipenem cephalosporin and fluconazole, respectively	Aqueous, methanolic, and hydro-alcoholic extracts/Brown	Broth microdilution	The propolis showed greater activity against Gram-positive bacteria than against Gram-negative. In contrast, no activity was observed against C. albicans.
Brazil	Barbosa et al. (2014)	Propionibacterium acnes ATCC 1969	Propolis tincture/ Brown	Broth macrodilution - CLSI	The propolis tincture exhibited activity at concentrations from 10% to 0.625% against P. acnes.
Algeria	Benhanifia et al. (2014)	S. aureus ATCC 25923, S. aureus ATCC 43300 and 1 clinical isolate, Bacillus cereus ATCC 11778. Bacillus subtilis ATCC 6633, E. coli ATCC 25922, P. aeruginosa ATCC 27853 and 01 clinical isolate	Ethanolic extract/ Dark brown	Disk diffusion	Activity only against Gram-positive bacteria. Propolis sample with the lowest phenolic content showed better effect.
Brazil	Campos et al. (2015)	Clinical isolates and Klebsiella pneumoniae ATCC 4352, Pseudomonas aeruginosa ATCC 15442, Staphyococcus aureus ATCC 43300, and Staphylococcus epidermidis ATCC 1228, Enterococcus faecalis ATCC 43300, Proteus mirabilis ATCC 43300, C. glabrata ATCC 90030, C. albicans ATCC 90028	Ethanolic extract/ Brown	Broth microdiluiton	The propolis from T. fiebrigi presented greater activity against the Gram-positive bacteria than against Gram-negative bacteria. S. aureus was the most sensitive bacteria. It was also observed fungicidal activity.
Serbia	Ristivojević et al. (2016)	Aeromonas hydrophila ATCC 49140, Shigella flexneri ATCC 9199, Listeria monocytogenes ATCC 19111, Bacillus subtilis ATCC 6633, Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, Enterobacter cloacae ATCC 49141, Escherichia coli ATCC 25922, Proteus mirabilis ATCC 25933, Pseudomonas aeruginosa ATCC 15422 and Salmonella enteritidis ATCC 13076, Micrococcus luteus ATCC 7468, Streptococcus equisimilis ATCC 12394	. Methanolic extracts/ blue and orange	Disk-diffusion and broth microdilution	The orange type of propolis shows higher antimicrobial activity compared to the blue type. The most sensitive strain of Gram positive bacteria was L. monocytogenes, with MIC values from 0.1 to 1.9 mg/mL for orange and 0.4 to 10.6 mg/mL mL for blue propolis. Presence of phenolic compounds influencing the antimicrobial activity.
Mexico	Bucio-Villalobos & Martínez-Jaime (2017)	Escherichia coli ATCC 10536, Salmonella typhimurium ATCC 13311, Listeria monocytogenes ATCC 19115, and Staphylococcus aureus ATCC 11632	Aqueous and Ethanolic Extracts / Brown	Agar diffusion	The aqueous extract of propolis did not affect any of the four bacteria evaluated. The ethanolic extract used showed antimicrobial activity against S. aureus and L. monocytogenes.
Venezuela	Joya et al. (2017)	Clinical isolates and ATCC strains of Candida albicans, C. guillermondi, C. krusei and C. tropicalis	Ethanolic extracts /Brown	Broth macrodilution	C. tropicalis is the most resistant to propolis, and C. guilliermondii the most sensitive species. Propolis from Germany and Italy are the most effective against species of Candida isolated from Venezuelan patients.
Chile	Maureira et al. (2017)	Candida spp.	Ethanolic extract/ Brown	Agar diffusion	The extract at 0.4 µg/mL inhibited the growth of 90.32% of Candida spp.
Brazil	Rufatto et al. (2018)	Bacillus subtilis CIP 52.62, Escherichia coli CIP 54127, Pseudomonas aeruginosa CIP 82118, and Staphylococcus aureus CIP 4.83	Ethanolic extract/Red propolis	Agar diffusion	Tthe Gram-positive bacteria (S. aureus and B. subtilis) were more sensitive than Gram-negative bacteria (E. coli and P. aeruginosa).
México	Moncayo Luján et al. (2018)	Staphylococcus aureus ATCC 35556, Escherichia coli ATCC 25922, Salmonella typhi ATCC 14028, Proteus mirabilis ATCC 9150	Ethanolic extracts / Brown	Disk diffusion	The least sensitive microorganism was S. typhi, and the most sensitive was E. coli.
Germany, Ireland, and Czech Republic	Al-Ani et al. (2018)	32 ATCC strains (Gram-positive bacteria, Gram-negative bacteria, and fungi) and clinical strains of Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant enterococci (VRE), Gram-negative bacteria and fungi (Candida species)	Ethanolic extracts / Brown	Broth macrodilution (CLSI) and Checkerboard methodology with antibiotics	Propolis samples showed moderate antibacterial effect against Gram-positive microorganisms with MIC ranging from 0.08 mg/mL to 2.5 mg/mL. Additionally, they displayed moderate antifungal activity (MIC values between 0.6–2.5 mg/mL).
Brazil	Silva et al. (2019)	Escherichia coli ATCC 25972, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Candida albicans ATCC 10231, Candida krusei ATCC 6258	Ethanolic extracts / Red	Disk diffusion	Propolis demonstrated no activity against Gram-negative bacteria, but it exhibited inhibitory activity against S. aureus. Additionally, it displayed antifungal activity against C. krusei.
Argentina	Cibanal et al. (2019)	Penicillium sp	Hydroalcoholic extract/ Brown	Agar diffusion	All treatments based on propolis extracts had an inhibitory effect greater than 99% on the development of fungal colony-forming units.
Chile	Veloz et al. (2019)	Clinical isolates of Streptococcus mutans	Ethanolic extract and its main components (apigenin, pinocembrin, quercetin and caffeic acid phenethyl ester -CAPE)/ Brown	Broth microdilution (CLSI) and biofilm formed on FluoroDish microplates	Quercetin and CAPE presented high MIC values when compared to natural propolis, apigenin and pinocembrin. Both quercetin and CAPE at a concentration of 25 µg/mL were shown to reduce the thickness of S. mutans biofilms by approximately 10 µm.
Italy	Petruzzi et al. (2020)	Isolates of Pseudomonas putida, P. fluorescens, Hafnia alvei, Enterobacter spp.Lactobacillus plantarum, Saccharomyces cerevisiae Debaryomyces hansenii and Fusarium oxysporum from food	Hydroalcoholic extract/ Brown	Total plate count and radial growth on agar plates	Higher concentrations of propolis did not lead to complete inhibition of bacterial growth. D. hansenii and Enterobacter spp. were identified as the most sensitive microorganisms, while Pseudomonas species exhibited the highest resistance to propolis. The radial growth of F. oxysporum was retarded by propolis at high concentrations.
Brazil	Correa et al. (2020)	Clinical isolates of Candida albicans, C. albicans ATCC 90028	Ethanolic extract/ Green	Broth microdilution (CLSI); total plate count; cinetic assay	The inhibitory activity was obtained with concentrations between 837 and 1675 μg/mL, and fungicidal activity at concentrations between 3350 and 6700 μg/mL.
Brazil	Barreiras et al. (2020)	Clinical isolates of Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebisiela pneumoniae and Pseudomonas aeruginosa	Hydroalcoholic extract/ Brown	Broth microdilution (CLSI)	Gram positive bacteria were sensitive to propolis at a concentration of 31.25 µg/mL. For Gram-negative bacteria, effectiveness was achieved with higher concentrations, except for K. pneumoniae, which was equivalent to Gram-positive bacteria.
México	Rodriguez- Pérez et al. (2020)	Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 8739, Candida albicans ATCC 10231	Ethanolic extracts/ Brown	Broth microdilution	For S. aureus, the MIC ranged from 0.19 to 30 mg/dL; for E. coli, it ranged between 3.75 and 15 mg/dL, and for C. albicans, from 1.5 to 30 mg/dL, depending on the collection site and, consequently, the percentage of chemical compounds.
México	Mateo-Aldama et al. (2020)	Clinical isolate of Cryptococcus neoformans	Ethanolic extract / Brown	Broth macrodilution	The Minimum Inhibitory Concentration (MIC) was determined to be 1.25 mg/mL, and the Minimum Fungicide Concentration (MFC) was found to be 2.5 mg/mL.
Perú	Checalla-Collatupa & Sanchez-Tito (2021)	Streptococcus mutans ATCC 25175	Ethanolic extract/ Brown	Disk diffusion	At a concentration of 25%, presenting an inhibition halo of 17.58 mm (±2.578), surpassing the effectiveness of concentrations at 50%, 75%, and 100%. Nevertheless, none of the concentrations achieved halos equal to or greater than the drug control, chlorhexidine.
Spain	Navarro-Pérez et al. (2021)	S. mutans ATCC 25175, S. sanguinis ATCC 10556	Ethanolic extract / Brown	Broth microdilution (CLSI)	For S. mutans, the values obtained for MIC and MBC were 240 and 480 µg/mL, respectively, while for S. sanguinis, 60 and 120 µg/mL were obtained.
Spain	Fernández-Calderón et al. (2021)	Clinical and ambiental isolates of Candida glabrata	Ethanolic extract / Brown	Agar diffusion for planktonic cells, and broth microdilution for biofilms	For planktonic cells, MIC50 values ranged from 60 to 240 μg/mL, while MIC90 ranged from 120 μg/mL. Subinhibitory concentrations of propolis extract significantly reduce the C. glabrata biofilms in a dose- dependent manner.
Hungary	Papp et al. (2021)	Candida albicans ATCC 44829	Ethanolic extract/NA	Broth microdilution (CLSI)	The extracts showed an MIC80 and IC50 in the range of 100–200 µg/mL and 72–134 µg/mL respectively.
Iran	Moghim et al. (2021)	Reference strain of Candida albicans	Ethanolic extract/Brown	Broth microdilution (CLSI)	The mean of MIC, MIC 50, and MFC of Iranian propolis extract on C. albicans were, respectively, 0.030 ± 0.015, 0.0618 ± 0.027, and 0.0833 ± 0.0599 mg/mL.
Brazil	Sokolonski et al. (2021)	Clinical oral isolates of C. albicans, C. dubliniensis and C. tropicalis, and reference strains of Candida albicans	Ethanolic extracts/ green and red	Broth microdilution (CLSI)	Green propolis extract inhibited Candida species by 50% at 0.125 mg/mL and by 95% at 8 mg/mL, whereas red propolis extract, obtained through ultrasound pretreatment, inhibited Candida growth at a concentration of 0.015 mg/mL. Additionally, the ethanolic extract of red propolis inhibited biofilm formation.
Serbia	Tambur et al. (2021)	Actinomyces odontolyticus ATCC 17929, Streptococcus mitis ATCC 6249, Streptococcus. sanguis ATCC 10556, Eikenella corrodens ATCC 23834, Fusobacterium nucleatum ATCC 25586, Lactobacillus acidophilus ATCC 4356, Streptococcus mutans ATCC 25175, Porphyromonas gingivalis ATCC 33277	Propolis solutions dissolved in benzene, diethyl ether, methyl chloride, and in acetone/ Brown	Agar dilution	Propolis solutions dissolved in benzene, diethyl ether and methyl chloride, demonstrated equal effectiveness against all investigated oral bacteria (MIC=12.5 μg/mL). Propolis solution dissolved in acetone displayed MIC of 6.3 μg/mL only for Lactobacillus acidophilus.
Colombia	Salamanca-Grosso et al. (2022)	Clinical isolates of Staphylococcus aureus and Escherichia coli	Ethanolic extract /Brown	Disk diffusion.	Propolis samples from Nariño showed greater effectiveness against Gram positive bacteria.
Mexico	Rivera-Yañez et al. (2022)	Clinical isolates of C. albicans, C. krusei and C. glabrata	Methanolic extract/ Brown	Disk diffusion	C. glabrata was the most sensitive to propolis, as it exhibited 21.43 ± 1.30 mm inhibition halos. In contrast, C. krusei presented the smallest inhibition halos (7.60 ± 0.10 mm).
Brazil	Campos et al. (2023)	Staphylococcus aureusATCC 6538, clinical isolates MRSA, Enterococcus faecalisATCC 43300, clinical isolates E. faecalis vancomycin-resistant, Escherichia coliATCC 29998, clinical isolates cephalosporin-resistant, Pseudomonas aeruginosa ATCC 15442, clinical isolates P. aeruginosa imipenem-resistant, Cryptococcus neoformans ATCC 32264, clinical isolates C. neoformans amphotericin B-resistant, Candida albicans ATCC 10231 and clinical isolates C. albicans amphotericin B-resistant	Ethanolic extract/ Brown	Broth microdilution	The extracts showed bactericidal and fungicidal activity against reference strains and hospital origin resistant isolates.
Ghana	Sa-Eed et al. (2023)	S. aureus ATCC 25923, E. coli NCTC 13351, P. aeruginosa ATCC 27853, along with one clinical isolate of each microorganism	Crude propolis and fractions/ Brown	Macrobroth dilution (CLSI), and agar diffusion	The crude propolis extracts and fractions from the various regions showed different degrees and patterns of antimicrobial activity against the tested bacterial isolates. The mean MIC range of the most active fractions was greatest for S. aureus.