Comparative Effects of Crude Libyan Propolis and Commercial Propolis Extracts on Aspergillus Ochraceus, Aspergillus Niger, and Penicillium spp. in Vitro

Rokaya Amara (1) , Maisoun Khashur (1) , Zanib Alabozidy (2) , Ahlam Rhuoma (1)
(1) Libyan Biotechnology Research Center, Tajoura, Tripoli, Libya,
(2) Faculty of Agriculture, University of Tripoli, Tripoli, Libya

Abstract

Propolis, a resinous substance collected by honeybees, exhibits antimicrobial properties attributed to its flavonoid and phenolic content, which vary significantly with geographical origin. This study evaluated and compared the antifungal efficacy of crude Libyan propolis (CLP) and commercial propolis extract (CPE) against Aspergillus niger, Aspergillus ochraceus, and Penicillium spp. using mycelial growth inhibition on agar (10, 25, and 50 mg mL⁻¹) and biomass dry-weight quantification in broth (1, 5, and 10 mg mL⁻¹), with all experiments performed in triplicate and data analysed by three-way ANOVA. Both propolis types significantly inhibited fungal growth compared with controls (p < 0.001). CLP showed superior activity against A. niger (64.7% inhibition at 25 mg mL⁻¹) compared with CPE, which achieved comparable inhibition (45.1%) only at 50 mg mL⁻¹, a twofold higher concentration. Against A. ochraceus, CLP exhibited maximal inhibition at 10 mg mL⁻¹ (76.4%), while CPE provided stable inhibition (58.8–67.0%) across concentrations. In biomass assays, Penicillium spp. was most sensitive to CPE at 10 mg mL⁻¹ (1.42 g), and a hormetic effect was observed at 1 mg mL⁻¹ CPE, where biomass exceeded controls by up to 86%. These findings demonstrate that both CLP and CPE possess species-specific antifungal activity. CLP was particularly effective against A. niger, whereas CPE showed greater activity against Penicillium spp. The observed biphasic and hormetic responses underscore the importance of dose optimisation. Libyan propolis therefore represents a promising natural antifungal agent for food preservation.

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References

[1] Simone-Finstrom, M., & Spivak, M. (2010). Propolis and bee health: The natural history and significance of resin use by honey bees. Apidologie, 41(3), 295–311. https://doi.org/10.1051/apido/2010016

[2] Zulhendri, F., Chandrasekaran, K., Kowacz, M., Ravalia, M., Kripal, K., Fearnley, J., & Perera, C. O. (2021). Antiviral, antibacterial, antifungal, and antiparasitic properties of propolis: A review. Foods, 10(6), 1360. https://doi.org/10.3390/foods10061360

[3] Wagh, V. D. (2013). Propolis: A wonder bees product and its pharmacological potentials. Advances in Pharmacological Sciences, 2013, 308249. https://doi.org/10.1155/2013/308249

[4] Bankova, V., Popova, M., & Trusheva, B. (2016). New emerging fields of application of propolis. Macedonian Journal of Chemistry and Chemical Engineering, 35(1), 1–11. https://doi.org/10.20450/mjcce.2016.864

[5] Anjum, S. I., Ullah, A., Khan, K. A., Attaullah, M., Khan, H., Ali, H., Bashir, M. A., Tahir, M., Ansari, M. J., Ghramh, H. A., & Adgaba, N. (2019). Composition and functional properties of propolis (bee glue): A review. Saudi Journal of Biological Sciences, 26(7), 1695–1703. https://doi.org/10.1016/j.sjbs.2018.08.013

[6] Siheri, W., Zhang, T., Ebiloma, G. U., Biddau, M., Woods, N., Hussain, M. Y., Clements, C. J., Fearnley, J., Edrada Ebel, R., Paget, T., Müller, S., Carter, K. C., Ferro, V. A., De Koning, H. P., & Watson, D. G. (2016). Chemical and antimicrobial profiling of propolis from different regions within Libya. PLoS ONE, 11(5), e0155355. https://doi.org/10.1371/journal.pone.0155355

[7] Siheri, W., Almutairi, S., & Watson, D. G. (2019). Propolis from Libya: Chemical composition and biological activities. In V. Bankova & M. Popova (Eds.), Propolis: Chemistry, biology and applications (pp. 45–62). Springer.

[8] Ali. M Alhaliba , Alauldin. O Alturshania, Adel. M Mlitan (2025). Chemical composition and antibacterial activity of Libyan propolis. Lebda Medical Journal; [LMJ]. https://lebmedj.elmergib.edu.ly/index.php/LMJ/article/view/259

[9] Przybyłek, I., & Karpiński, T. M. (2019). Antibacterial properties of propolis. Molecules, 24(11), 2047. https://doi.org/10.3390/molecules24112047

[10] Elsharkawy, E. R., & Aljabre, S. H. M. (2023). Antibacterial and antifungal activity of the propolis growing in the basin of Mediterranean city Misurata, Libya. The Pharmaceutical and Chemical Journal, 10(4), 1–8.

[11] Amara, R. O., Khashur, M. M., Alabozidy, Z. A., & Rhuoma, A. A. (2025). Effectiveness of Libyan propolis against mold isolated from spoiled bread. Journal of Research in Agriculture and Food Sciences, 2(3), 232–240.

[12] EFSA Panel on Contaminants in the Food Chain (CONTAM), Schrenk, D., Bodin, L., Chipman, J. K., del Mazo, J., Grasl-Kraupp, B., Hogstrand, C., Hoogenboom, L. R., Leblanc, J. C., Nebbia, C. S., Nielsen, E., Ntzani, E., Petersen, A., Sand, S., Schwerdtle, T., Vleminckx, C., Mark, R., Wallace, H., & Alexander, J. (2020). Risk assessment of ochratoxin A in food. EFSA Journal, 18(5), e06113. https://doi.org/10.2903/j.efsa.2020.6113

[13] Saranraj, P., & Geetha, M. (2012). Microbial spoilage of bakery products and its control by preservatives. International Journal of Pharmaceutical and Biological Archives, 3(1), 38–48.

[14] Segueni, N., Boutaghane, N., Asma, S. T., Tas, N., Acaroz, U., Arslan-Acaroz, D., & Akkal, S. (2023). Review on propolis applications in food preservation and active packaging. Plants, 12(8), 1654. https://doi.org/10.3390/plants12081654

[15]Mesías, F. J., Martín, A., & Hernández, A. (2021). Consumers' growing appetite for natural foods: Perceptions towards the use of natural preservatives in fresh fruit. Food Research International, 150, 110749. https://doi.org/10.1016/j.foodres.2021.110749

[16] Wo´zniak, M.; Mrówczy´ nska, L.; Kwa´sniewska-Sip, P.;Wa´skiewicz, A.; Nowak, P.; Ratajczak, I. Effect of the solvent on propolis phenolic profile and its antifungal, antioxidant, and in vitro cytoprotective activity in human erythrocytes under oxidative stress.Molecules 2020, 25, 4266. [CrossRef]

[17] Kasote, D., Bankova, V., & Viljoen, A. M. (2022). Propolis: Chemical diversity and challenges in quality control. Phytochemistry Reviews, 21(6), 1887–1911. https://doi.org/10.1007/s11101-022-09816-1

[18] Pandey, D. K., Tripathi, N. N., Tripathi, R. D., & Dixit, S. N. (1982). Fungitoxic and phytotoxic properties of the essential oil of Hyptis suaveolens. Zeitschrift für Pflanzenkrankheiten und Pflanzenschutz, 89(6), 344–349.

[19] Eloff, J. N. Quantifying the antifungal activity of plant extracts against fungal growth in liquid culture. South African Journal of Botany,2019.126,234-240.

[20] Biswas, D., et al. Standardization of biomass quantification methods for antifungal susceptibility testing. Journal of Microbiological Methods,2020. 178, 106072.

[21] Logan A, Wolfe A, Williamson JC. Antifungal Resistance and the Role of New Therapeutic Agents. Curr Infect Dis Rep. 2022;24(9):105-116. doi: 10.1007/s11908-022-00782-5. Epub 2022 Jul 5. PMID: 35812838; PMCID: PMC9255453.

[22] Djenontin E, Lavergne RA, Morio F, Dannaoui E. Antifungal Resistance in Non-fumigatus Aspergillus Species. Mycoses. 2025 Apr;68(4):e70051. doi: 10.1111/myc.70051. PMID: 40219727; PMCID: PMC11992613.

[23] Heuer C, Leonard H, Nitzan N, Lavy-Alperovitch A, Massad-Ivanir N, Scheper T, Segal E. Antifungal Susceptibility Testing of Aspergillus niger on Silicon Microwells by Intensity-Based Reflectometric Interference Spectroscopy. ACS Infect Dis. 2020 Oct 9;6(10):2560-2566. doi: 10.1021/acsinfecdis.0c00234. Epub 2020 Sep 21. PMID: 32930571; PMCID: PMC7584364.

[24] Lobo-Vega R, Megías D, Río-Ropero A, Mato-López Á, Ashraph K, Manosalva J, Alcázar Fuoli L, Alastruey-Izquierdo A, Mellado E, Amich J. Establishing thresholds for azole tolerance and persistence in Aspergillus fumigatus to study their impact on voriconazole treatment in vivo. bioRxiv 2025.12.26.693403. doi: 10.64898/2025.12.26.693403.

[25] Calabrese EJ. Preconditioning is hormesis part I: Documentation, dose-response features and mechanistic foundations. Pharmacol Res. 2016 Aug; 110:242-264. doi: 10.1016/j.phrs.2015.12.021. Epub 2016 Jan 3. PMID: 26757428.

[26] López-Malo, A., Alzamora, S. M., & Palou, E. (2002). Aspergillus flavus dose–response curves to selected natural and synthetic antimicrobials. International Journal of Food Microbiology, 73(2-3), 213–218. https://doi.org/10.1016/S0168-1605(01)00639-0

[27] Barboráková, Z., Tančinová, D., Medo, J., Jakabová, S., Häubl, G., Jaunecker, G., Mašková, Z., & Labuda, R. (2025). Strain-Dependent Variability in Ochratoxin A Production by Aspergillus spp. Under Different In Vitro Cultivation Conditions. Microorganisms, 13(12), 2850. https://doi.org/10.3390/microorganisms13122850

[28] Berretta AA, Nascimento AP, Bueno PC, Vaz MM, Marchetti JM. Propolis standardized extract (EPP-AF®), an innovative chemically and biologically reproducible pharmaceutical compound for treating wounds. Int J Biol Sci. 2012;8(4):512-21. doi: 10.7150/ijbs.3641. Epub 2012 Mar 21. PMID: 22457606; PMCID: PMC3314192.

[29] Long N, Li F. Antifungal Mechanism of Natural Products Derived from Plants: A Review. Natural Product Communications. 2024;19(8). doi:10.1177/1934578X241271747

[30] Türkekul İ, Gülmez Y. Propolis: An enrichment material for mycelium development of oyster mushroom (Pleurotus ostreatus). Natural Resources. 2016;7(03):119–123.

[31] Dharma KS, Suryanti S, Widiastuti A. Hormesis in pathogenic and biocontrol fungi: From inhibition to stimulation. AGRIS. 2024.

[32] Garzon CD, Flores FJ. Detection and assessment of chemical hormesis on the radial growth in vitro of oomycetes and fungal plant pathogens. Dose-Response. 2013;11(3):361–373.

[33] Baur JA, et al. What is xenohormesis? American Journal of Pharmacology and Toxicology. 2008;3(1):152–159.

[34] Howitz KT, Sinclair DA. Xenohormesis: Sensing the chemical cues of other species. Cell. 2008;133(3):387–391.

[35] Calabrese EJ, Baldwin LA. Hormesis: A general biological phenomenon. Human and Experimental Toxicology. 2002;21(2):91–97.

[36] Garzon CD, Flores FJ. Hormesis in plant pathogens: A review of low dose stimulation of fungal and oomycete growth. Plant Disease. 2013;97(5):602–612.

[37] Wadhwa K, Kapoor N, Kaur H, Abu-Seer EA, Tariq M, Siddiqui S, Yadav VK, Niazi P, Kumar P, Alghamdi S. A Comprehensive Review of the Diversity of Fungal Secondary Metabolites and Their Emerging Applications in Healthcare and Environment. Mycobiology. 2024 Dec 3;52(6):335-387. doi: 10.1080/12298093.2024.2416736. PMID: 39845176; PMCID: PMC11749308

[38] Qin, Y., Xia, Y. Melanin in fungi: advances in structure, biosynthesis, regulation, and metabolic engineering. Microb Cell Fact 23, 334 (2024). https://doi.org/10.1186/s12934-024-02614-8

[39] Houbraken J, Samson RA. Phylogeny of Penicillium and the segregation of Trichocomaceae into three families. Stud Mycol. 2011 Nov 15;70(1):1-51. doi: 10.3114/sim.2011.70.01. PMID: 22308045; PMCID: PMC3233907.

Authors

Rokaya Amara
[email protected] (Primary Contact)
Maisoun Khashur
Zanib Alabozidy
Ahlam Rhuoma
Comparative Effects of Crude Libyan Propolis and Commercial Propolis Extracts on Aspergillus Ochraceus, Aspergillus Niger, and Penicillium spp. in Vitro. (2026). Journal of Pure & Applied Sciences , 25(2), 1-6. https://doi.org/10.51984/ygn5by41

Article Details

How to Cite

Comparative Effects of Crude Libyan Propolis and Commercial Propolis Extracts on Aspergillus Ochraceus, Aspergillus Niger, and Penicillium spp. in Vitro. (2026). Journal of Pure & Applied Sciences , 25(2), 1-6. https://doi.org/10.51984/ygn5by41

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