Journal of Ethnopharmacology and Toxicology | Volume 4 Issue 2 | Pages: 22-30 | Doi : 10.37446/jet/rsa/4.2.2026.22-30

Research Article

OPEN ACCESS | Published on : 25-Aug-2026

Inhibitory activity of propolis and nano-propolis against isolates of Staphylococcus aureus and Escherichia coli

• Mohammed A. Aboktifa
• Department of physiology, biochemistry and pharmacology, College of Veterinary Medicine, Al-Qasim Green University, Babylon 51013 Iraq.

Alaa Abdelkadhim Jawad
• Department of microbiology and parasitology/ College of Veterinary Medicine, University of Al-Qadisiyah, Al-Diwaniyah, Iraq.

Dhama Al-Sallami
• Department of Physiology, Pharmacology and Biochemistry/ College of Veterinary Medicine, University of Al-Qadisiyah, Al-Diwaniyah, Iraq.

Abstract

Background: Propolis, a plant-derived material, is widely used to enhance antibiotic activity against common pathogenic bacteria which causes significant global economic losses. This research was designed to detect the role of propolis and nano-propolis as natural antibiotic.

Methods: Fifteen isolates of S. aureus and 15 of E. coli were cultured on plates of Nutrient Agar, incubated at 37 °C, then stored at -20 °C. Bacterial sensitivity of the 15 isolates to different important antibiotics were performed separately for each type of bacteria. Also, susceptibility test by Kirby Bauer disc diffusion method was utilized with three concentrations (125,250,375 μg/ml) of propolis and nano-propolis. Zones diameters of bacterial growth inhibition for the tested isolates were measured after 24 h of incubation. In the same time MIC and MBC for both materials was determined.

Results: Results of bacterial sensitivity test show that 13 bacterial isolates of S. aureus were highly resistant to Benzylpenicillin (86.6%) and 12 isolates of S. aureus were resistant to Oxacillin (80%). While, 13 isolates of S. aureus were sensitive to Linezolid and Teicoplanin (86.66%). In addition, E. coli showed that 13 isolates were resistant to ampicillin (86.66) and 10 out of 15 isolates were sensitive to Piperacillin (66.66%). Furthermore, the inhibition zone of S. aureus was higher at 375 µg/ml as 18.36 mm and 11.87 mm with nano-propolis and propolis respectively. Moreover, E. coli cultures showed higher inhibition zone at 375 µg/ml as 11.78 mm with propolis and 12.56 mm with nano-propolis.

Conclusion: This study indicates that propolis and nano-propolis inhibited bacterial growth and may potentiate antibiotic effect against gram positive and gram-negative bacteria. Suggesting a good strategy against pathogens. However, nano-propolis is a promising candidate as plant derived material to decrease MDR microbes.

Keywords

Propolis, nano propolis, antibiotic, MDR, S. aureus and E. coli

References

• Ab Rahman, J. B. (2015). Brief guidelines for methods and statistics in medical research. Springer.

  Adul-Rasool, A. A., Athair, D. M., Zaidan, H. K., Rheima, A. M., Al-Sharify, Z. T., & Mohammed, S. H. (2024). 0, 1, 2, 3D nanostructures, types of bulk nanostructured materials, and drug nanocrystals: an overview. Cancer Treatment and Research Communications, 40, 100834.

  Agarwal, G., Vemanaradhya, G. G., & Mehta, D. S. (2012). Evaluation of chemical composition and efficacy of Chinese propolis extract on Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans: An in vitro study. Contemporary Clinical Dentistry, 3(3), 256-261.

  Alara, O. R., Abdurahman, N. H., & Ukaegbu, C. I. (2021). Extraction of phenolic compounds: A review. Current research in food science, 4, 200-214.

   

  Alawadi, Z. I., Alsultan, A., Alsallami, D., Alnomasy, S. F., Alqasmi, M., Almufarriji, F. M., ... & Alenazy, R. (2024). Designing a multiepitope mRNA-based vaccine against enterotoxigenic Escherichia coli infection in calves: immunoinformatics and molecular modeling approach. Open Veterinary Journal, 14(6), 1417.

  Almuhayawi, M. S. (2020). Propolis as a novel antibacterial agent. Saudi journal of biological sciences, 27(11), 3079-3086.

  Al-Mutairi, R., Tovmasyan, A., Batinic-Haberle, I., & Benov, L. (2018). Sublethal photodynamic treatment does not lead to development of resistance. Frontiers in Microbiology, 9, 1699.

  Alsultan, A., & Alsallami, D. (2022). Efflux-mediated bile resistance in Gram-positive pathogens. Journal of Pure and Applied Microbiology, 16(1), 10-18.

  Alsultan, A., Al-Sallami, D., & Alsaadi, M. (2023). Antibiotic resistance genes in farm animal slaughterhouse wastes in Al-Dewanyiah province, Iraq: https://doi. org/10.12982/VIS. 2023.041. Veterinary Integrative Sciences, 21(2), 577-586.

  Al-Ubaidy, Y., & Alsultan, A. (2023). Using of integrons as biomarker to assess dissemination and diversity of antimicrobial resistance genes in farm animal manure. Journal of Pure and Applied Microbiology, 17(3), 1708-1714.

  Bankova, V., Popova, M., & Trusheva, B. (2014). Propolis volatile compounds: chemical diversity and biological activity: a review. Chemistry Central Journal, 8(1), 28.

  Blanco, P., Hernando-Amado, S., Reales-Calderon, J. A., Corona, F., Lira, F., Alcalde-Rico, M., ... & Martinez, J. L. (2016). Bacterial multidrug efflux pumps: much more than antibiotic resistance determinants. Microorganisms, 4(1), 14.

  Chandrakala, V., Aruna, V., & Angajala, G. (2022). Review on metal nanoparticles as nanocarriers: current challenges and perspectives in drug delivery systems. Emergent Materials, 5(6), 1593-1615.

  Darwish, R. M., Abu Fares, R. J., Abu Zarga, M. H., & Nazer, I. K. (2010). Antibacterial effect of Jordanian propolis and isolated flavonoids against human pathogenic bacteria. African Journal of Biotechnology, 9(36), 5966–5974.

  Davies, J., & Davies, D. (2010). Origins and evolution of antibiotic resistance. Microbiology and molecular biology reviews, 74(3), 417-433.

  de Souza Ferreira, S. B., de Assis Dias, B. R., Obregón, C. S., Gomes, C. C., de Araújo Pereira, R. R., Ribeiro Godoy, J. S., … Bruschi, M. L. (2014). Microparticles containing propolis and metronidazole: in vitro characterization, release study and antimicrobial activity against periodontal pathogens. Pharmaceutical Development and Technology, 19(2), 173–180.

  Guo, Y., Song, G., Sun, M., Wang, J., & Wang, Y. (2020). Prevalence and therapies of antibiotic-resistance in Staphylococcus aureus. Frontiers in cellular and infection microbiology, 10, 107.

  Hong, J., Hu, J., & Ke, F. (2016). Experimental induction of bacterial resistance to the antimicrobial peptide tachyplesin I and investigation of the resistance mechanisms. Antimicrobial agents and chemotherapy, 60(10), 6067-6075.

  Humphreys, H. (2012). Staphylococcus aureus: the enduring pathogen in surgery. the surgeon, 10(6), 357-360.

  Khudeir, M. A., Al-sallami, D., & Alsultan, A. (2024). Draft genome sequence of multidrug resistant Proteus mirabilis strain MAD23: https://doi. org/10.12982/VIS. 2024.008. Veterinary Integrative Sciences, 22(1), 93-109.

  Kwong, L. H., Ercumen, A., Pickering, A. J., Arsenault, J. E., Islam, M., Parvez, S. M., ... & Luby, S. P. (2020). Ingestion of fecal bacteria along multiple pathways by young children in rural Bangladesh participating in a cluster-randomized trial of water, sanitation, and hygiene interventions (WASH benefits). Environmental science & technology, 54(21), 13828-13838.

  Livermore, D. M. (2003). Linezolid in vitro: mechanism and antibacterial spectrum. Journal of Antimicrobial Chemotherapy, 51(suppl_2), ii9-ii16.

  Lopes, J. G., & Sourjik, V. (2018). Chemotaxis of Escherichia coli to major hormones and polyamines present in human gut. The ISME journal, 12(11), 2736-2747.

  Marek, A., Dec, M., Urban-Chmiel, R., Stępień-Pyśniak, D., Wieczorek, K., Nowaczek, A., & Osek, J. (2022). Antibiotic Resistance in Bacteria-A Review. Antibiotics, 11, 1079-1079.35.     

  Mirsepasi-Lauridsen, H. C., Vallance, B. A., Krogfelt, K. A., & Petersen, A. M. (2019). Escherichia coli pathobionts associated with inflammatory bowel disease. Clinical microbiology reviews, 32(2), 10-1128.

  Moosavy, M. H., Hallaj Salahipor, M., Mostafavi, E., & Khatibi, S. A. (2018). Risk factors for human brucellosis in Mianeh, Iran. Journal of Zoonotic Diseases, 3(1), 10-21.

  Mosolygó, T., Kincses, A., Csonka, A., Tönki, Á. S., Witek, K., Sanmartín, C., ... & Rutledge, P. J. (2019). Selenocompounds as Novel Antibacterial Agents and Bacterial Efflux Pump Inhibitors. Molecules, 24(8), Article 1487.

  Nandre, V. S., Bagade, A. V., Kasote, D. M., Lee, J. H., Kodam, K. M., Kulkarni, M. V., & Ahmad, A. (2021). Antibacterial activity of Indian propolis and its lead compounds against multi-drug resistant clinical isolates. Journal of Herbal Medicine, 29, 100479.

  Puvača, N., & de Llanos Frutos, R. (2021). Antimicrobial resistance in Escherichia coli strains isolated from humans and pet animals. Antibiotics, 10(1), 69.

  Rao, M., Padyana, S., Dipin, K. M., Kumar, S., Nayak, B., & Varela, M. F. (2018). Antimicrobial compounds of plant origin as efflux pump inhibitors: new avenues for controlling multidrug resistant pathogens. Journal of Antimicrobial Agents, 4: 1000159.

  Ruvalcaba-Gómez, J. M., Villagrán, Z., Valdez-Alarcón, J. J., Martínez-Núñez, M., Gomez-Godínez, L. J., Ruesga-Gutiérrez, E., ... & Villarruel-López, A. (2022). Non-antibiotics strategies to control Salmonella infection in poultry. Animals, 12(1), 102.

  Salama, S. A., Essam, D., Tagyan, A. I., Farghali, A. A., Khalil, E. M., Abdelaleim, Y. F., ... & Mahmoud, R. (2024). Novel composite of nano zinc oxide and nano propolis as antibiotic for antibiotic-resistant bacteria: a promising approach. Scientific Reports, 14(1), 20894.

  Shabbir, A., Rashid, M., & Tipu, H. N. (2016). Propolis, a hope for the future in treating resistant periodontal pathogens. Cureus, 8(7).

  Sharma, A., Gupta, V. K., & Pathania, R. (2019). Efflux pump inhibitors for bacterial pathogens: From bench to bedside. Indian Journal of Medical Research, 149(2), 129-145.

  Sugrue, I., Tobin, C., Ross, R. P., Stanton, C., & Hill, C. (2019). Foodborne pathogens and zoonotic diseases. In Raw milk (pp. 259-272). Academic Press.

  Varghese, M., & Balachandran, M. (2021). Antibacterial efficiency of carbon dots against Gram-positive and Gram-negative bacteria: A review. Journal of Environmental Chemical Engineering, 9(6), 106821.

  Yaseen, M. M., Mohammed Merah, M. H., & GHAZI, A. M. (2023). GC-mass characterization of tomato ethyl acetate extract and its antibacterial and antioxidant properties. Nativa, 11(4).

  Zhang, W., Margarita, G. E., Wu, D., Yuan, W., Sha, Y., Qi, S., ... & Wu, L. (2022). Antibacterial activity of Chinese red propolis against Staphylococcus aureus and MRSA. Molecules, 27(5), 1693.