Comparative Evaluation of Two Commercial Propolis Extracts as Plant Defense Activator and Antimicrobial Agent Against Pseudomonas syringae pv. tomato (Pst) strain DC3000
Year 2021,
Volume: 8 Issue: 2, 213 - 219, 30.06.2021
Melda Ateş
,
Nazlı Özkurt
,
Yasemin Bektaş
Abstract
Propolis is a natural bee product that protects the beehives from internal and external factors. It is a natural-complex compound with a wide potential use as an antimicrobial, anti-fungal, and anti-inflammatory effect. The role of propolis as a plant protection agent and an alternative to pesticides is an unexplored area. Here, two different commercially available propolis extracts were evaluated for antibacterial effects on Pseudomonas syringae pv. tomato (Pst) strain DC3000. Also, the activities of defense response genes WRKY70 and CaBP22 in Arabidopsis thaliana under propolis application were compared. According to the results, each propolis extract and dose had a different effect on gene expressions as well as antibacterial activity. One of the commercial brands had a significant effect at all doses while another brand’s propolis extract had its activity at only 1% concentration. Propolis reduced bacterial growth up to 93% with a 2% concentration. For the first time, propolis is also evaluated for its capacity as a plant defense activator agent and it induced WRKY70 and CaBP22 gene expression. The differences in gene expression and bacterial growth inhibition levels suggest the importance of the origins of propolis, such as plant species and regions it collected. While preliminary in nature, these results suggest a significant potential of propolis in plant protection in commercial and organic agriculture.
Supporting Institution
TUBİTAK-2209-National/International Research Projects Fellowship Programme for Undergraduate Students 2017/2
Project Number
1919B011702852.
References
- Abd-El-Rhman, A.M., 2009. Antagonism of Aeromonas hydrophila by propolis and its effect on the performance of Nile tilapia, Oreochromis niloticus. Fish and Shellfish Immunology, 27(3): 454-459.
- Abo-Elyousr, K.A.M., Seleim, M.E.A., El-Sharkawy, R.M., Khalil Bagy, H.M.M., 2017. Effectiveness of Egyptian propolis on control of tomato bacterial wilt caused by Ralstonia solanacearum. Journal of Plant Diseases and Protection, 124(5): 467-472.
- Ali, A.M., Kunugi, H., 2021. Propolis, bee honey, and their components protect against coronavirus disease 2019 (COVID-19): A review of in silico, in vitro, and clinical studies. Molecules, 26(5): 1232.
- Aparicio-García, P.F., Ventura-Aguilar, R.I., Del Río-García, J.C., Hernández-López, M., Guillén-Sánchez, D., Salazar-Piña, D.A., Ramos-García, M.D.L., Bautista-Baños, S., 2021. Edible chitosan/propolis coatings and their effect on ripening, development of Aspergillus flavus, and sensory quality in fig fruit, during controlled storage. Plants, 10(1): 112.
- Awale, S., Li, F., Onozuka, H., Esumi, H., Tezuka, Y., Kadota, S., 2008. Constituents of Brazilian red propolis and their preferential cytotoxic activity against human pancreatic PANC-1 cancer cell line in nutrient-deprived condition. Bioorganic & Medicinal Chemistry, 16(1): 181-189.
- Basim, E., Basim, H., Özcan, M., 2006. Antibacterial activities of Turkish pollen and propolis extracts against plant bacterial pathogens. Journal of Food Engineering, 77(4): 992-996.
- Bektas, Y., Eulgem, T., 2015. Synthetic plant defense elicitors. Frontiers in Plant Science, 5(804): 1-17.
- Bohinc, T., Vučajnk, F., Trdan, S., 2019. The efficacy of environmentally acceptable products for the control of major potato pests and diseases. Zemdirbyste-Agriculture, 106(2): 135-142.
- Botteon, C.E.A., Silva, L.B., Ccana-Ccapatinta, G.V., Silva, T.S., Ambrosio, S.R., Veneziani, R.C.S., Bastos, J.K., Marcato, P.D., 2021. Biosynthesis and characterization of gold nanoparticles using Brazilian red propolis and evaluation of its antimicrobial and anticancer activities. Scientific Reports, 11(1): 1-16.
- Burdock, G.A., 1998. Review of the biological properties and toxicity of bee propolis (Propolis). Food and Chemical Toxicology, 36(4): 347-363.
- Cai, Y., Cao, X., Aballay, A., Nishikawa, Y., Davies, J.E., 2014. Whole-animal chemical screen identifies colistin as a new immunomodulator that targets conserved pathways. mBio, 5(4): 1-11.
- Cohen, Y., Vaknin, M., Mauch-Mani, B., 2016. BABA-induced resistance: milestones along a 55-year journey. Phytoparasitica, 44(4): 513-538.
- Da Silva Barboza, A., Aitken-Saavedra, J.P., Ferreira, M.L., Fábio Aranha, A.M., Lund, R.G., 2021. Are propolis extracts potential pharmacological agents in human oral health? - A scoping review and technology prospecting. Journal of Ethnopharmacology, 271: 113846.
- Er, Y., 2021. In vitro and in vivo antimicrobial activity of propolis extracts against various plant pathogens. Journal of Plant Diseases and Protection, 128: 693-701.
- Farace, G., Fernandez, O., Jacquens, L., Coutte, F., Krier, F., Jacques, P., Clément, C., Barka, E.A., Jacquard, C., Dorey, S., 2015. Cyclic lipopeptides from Bacillus subtilis activate distinct patterns of defence responses in grapevine. Molecular Plant Pathology, 16(2): 177-187.
- Freires, I.A., Queiroz, V., Furletti, V.F., Ikegaki, M., de Alencar, S.M., Duarte, M.C.T., Rosalen, P.L., 2016. Chemical composition and antifungal potential of Brazilian propolis against Candida spp. Journal of Medical Mycology, 26(2): 122-132.
- Gallez, L., Kiehr, M., Fernández, L., Delhey, R., Stikar, D., 2015. Antifungal activity in vitro of propolis solutions from Argentina against two plant pathogenic fungi: Didymella bryoniae and Rhizotocnia solani. Journal of Apicultural Research, 53(4): 438-440.
- Halder, V., Suliman, M.N.S., Kaschani, F., Kaiser, M., 2019. Plant chemical genetics reveals colistin sulphate as a SA and NPR1-independent PR1 inducer functioning via a p38-like kinase pathway. Scientific Reports, 9(1): 11196.
- Henry, G., Deleu, M., Jourdan, E., Thonart, P., Ongena, M., 2011. The bacterial lipopeptide surfactin targets the lipid fraction of the plant plasma membrane to trigger immune-related defence responses. Cellular Microbiology, 13(11): 1824-1837.
- Huang, S., Zhang, C P., Wang, K., Li, G.Q., Hu, F.L., 2014. Recent advances in the chemical composition of propolis. Molecules, 19(12): 19610-19632.
- Ishihara, M., Naoi, K., Hashita, M., Itoh, Y., Suzui, M., 2009. Growth inhibitory activity of ethanol extracts of Chinese and Brazilian propolis in four human colon carcinoma cell lines. Oncology reports, 22(2): 349-354.
- Jug, M., Končić, M.Z., Kosalec, I., 2014. Modulation of antioxidant, chelating and antimicrobial activity of poplar chemo-type propolis by extraction procures. LWT-Food Science and Technology, 57(2): 530-537.
- Knoth, C., Salus, M.S., Girke, T., Eulgem, T., 2009. The synthetic elicitor 3,5-dichloroanthranilic acid induces NPR1-dependent and NPR1-independent mechanisms of disease resistance in Arabidopsis. Plant Physiology, 150(1): 333-347.
- Kolayli, S., Palabiyik, I., Atik, D.S., Keskin, M., Bozdeveci, A., Karaoglu, S.A., 2020. Comparison of antibacterial and antifungal effects of different varieties of honey and propolis samples. Acta Alimentaria, 49(4): 515-523.
- Lima Cavendish, R., de Souza Santos, J., Belo Neto, R., Oliveira Paixao, A., Valeria Oliveira, J., Divino de Araujo, E., Berretta, E., Silva, A.A., Maria Thomazzi, S., Cordeiro Cardoso, J., Zanardo Gomes, M., 2015. Antinociceptive and anti-inflammatory effects of Brazilian red propolis extract and formononetin in rodents. Journal of Ethnopharmacology, 173: 127-133.
- Martinello, M., Mutinelli, F., 2021. Antioxidant activity in bee products: A review. Antioxidants, 10(1): 71.
- Noutoshi, Y., Okazaki, M., Kida, T., Nishina, Y., Morishita, Y., Ogawa, T., Suzuki, H., Shibata, D., Jikumaru, Y., Hanada, A., Kamiya, Y., Shirasu, K., 2012. Novel plant immune-priming compounds identified via high-throughput chemical screening target salicylic acid glucosyltransferases in Arabidopsis. Plant Cell, 24(9): 3795-3804.
- Ordonez, R.M., Zampini, I.C., Moreno, M.I., Isla, M.I., 2011. Potential application of Northern Argentine propolis to control some phytopathogenic bacteria. Microbiological Research, 166(7): 578-584.
- Pascoal, A., Feás, X., Dias, T., Dias, L.G., Estevinho, L.M., 2014. The role of honey and propolis in the treatment of infected wounds. In: K. Kon, M. Rai (Eds.), Microbiology for Surgical Infections, Academic Press, Amsterdam, pp. 221-234.
- Reddy, P.P., 2013. Plant Defence Activators. Springer, New Delhi.
- Regueira, M.S.N., Tintino, S.R., Da Silva, A.R.P., Costa, M.D.S., Boligon, A.A., Matias, E.F.F., De Queiroz Balbino, V., Menezes, I.R.A., Melo Coutinho, H.D., 2017. Seasonal variation of Brazilian red propolis: Antibacterial activity, synergistic effect and phytochemical screening. Food and Chemical Toxicology, 107(Pt B): 572-580.
- Rivera-Yañez, N., Rivera-Yañez, C.R., Pozo-Molina, G., Méndez-Catalá, C.F., Méndez-Cruz, A.R., Nieto-Yañez, O., 2020. Biomedical properties of propolis on diverse chronic diseases and its potential applications and health benefits. Nutrients, 13(1): 78.
- Salatino, A., Teixeira, E.W., Negri, G., Message, D., 2005. Origin and chemical variation of Brazilian propolis. Evidence-Based Complementary and Alternative Medicine, 2(1): 33-38.
- Scorza, C.A., Gonçalves, V.C., Scorza, F.A., Fiorini, A.C., De Almeida, A.C.G., Fonseca, M.C.M., Finsterer, J., 2020. Propolis and coronavirus disease 2019 (COVID-19): Lessons from nature. Complementary Therapies in Clinical Practice, 41: 101227.
- Sforcin, J.M., Bankova, V., 2011. Propolis: is there a potential for the development of new drugs? Journal of Ethnopharmacology, 133(2): 253-260.
- Silva-Castro, I., Diez, J., Martín-Ramos, P., Pinto, G., Alves, A., Martín-Gil, J., Martín-García, J., 2018.
Application of bioactive coatings based on chitosan and propolis for Pinus spp. protection against Fusarium circinatum. Forests, 9(11): 685.
- Valente, M.J., Baltazar, A.F., Henrique, R., Estevinho, L., Carvalho, M., 2011. Biological activities of Portuguese propolis: protection against free radical-induced erythrocyte damage and inhibition of human renal cancer cell growth in vitro. Food and Chemical Toxicology, 49(1): 86-92.
- Wagh, V.D., 2013. Propolis: a wonder bees product and its pharmacological potentials. Advances in Pharmacological and Pharmaceutical Sciences, 2013: 308249.
- Xin, X.F., He, S.Y., 2013. Pseudomonas syringae pv. tomato DC3000: a model pathogen for probing disease susceptibility and hormone signaling in plants. Annual Review of Phytopathology, 51: 473-498.
- Xu, J., Zhang, Y., 2020. Traditional Chinese Medicine treatment of COVID-19. Complementary Therapies in Clinical Practice, 39: 101165.
- Yong, H., Liu, J., 2021. Active packaging films and edible coatings based on polyphenol‐rich propolis extract: A review. Comprehensive Reviews in Food Science and Food Safety, 20(2): 2106-2145.
- Zabaiou, N., Fouache, A., Trousson, A., Baron, S., Zellagui, A., Lahouel, M., Lobaccaro, J.A. 2017. Biological properties of propolis extracts: Something new from an ancient product. Chemistry and Physics of Lipids, 207(Pt B): 214-222.
- Zampini, I.C., Salas, A.L., Maldonado, L.M., Simirgiotis, M.J., Isla, M.I., 2021. Propolis from the Monte region in Argentina: a potential phytotherapic and food functional ingredient. Metabolites, 11(2): 76.
Comparative Evaluation of Two Commercial Propolis Extracts as Plant Defense Activator and Antimicrobial Agent Against Pseudomonas syringae pv. tomato (Pst) strain DC3000
Year 2021,
Volume: 8 Issue: 2, 213 - 219, 30.06.2021
Melda Ateş
,
Nazlı Özkurt
,
Yasemin Bektaş
Abstract
Propolis is a natural bee product that protects the beehives from internal and external factors. It is a natural-complex compound with a wide potential use as an antimicrobial, anti-fungal, and anti-inflammatory effect. The role of propolis as a plant protection agent and an alternative to pesticides is an unexplored area. Here, two different commercially available propolis extracts were evaluated for antibacterial effects on Pseudomonas syringae pv. tomato (Pst) strain DC3000. Also, the activities of defense response genes WRKY70 and CaBP22 in Arabidopsis thaliana under propolis application were compared. According to the results, each propolis extract and dose had a different effect on gene expressions as well as antibacterial activity. One of the commercial brands had a significant effect at all doses while another brand’s propolis extract had its activity at only 1% concentration. Propolis reduced bacterial growth up to 93% with a 2% concentration. For the first time, propolis is also evaluated for its capacity as a plant defense activator agent and it induced WRKY70 and CaBP22 gene expression. The differences in gene expression and bacterial growth inhibition levels suggest the importance of the origins of propolis, such as plant species and regions it collected. While preliminary in nature, these results suggest a significant potential of propolis in plant protection in commercial and organic agriculture.
Project Number
1919B011702852.
References
- Abd-El-Rhman, A.M., 2009. Antagonism of Aeromonas hydrophila by propolis and its effect on the performance of Nile tilapia, Oreochromis niloticus. Fish and Shellfish Immunology, 27(3): 454-459.
- Abo-Elyousr, K.A.M., Seleim, M.E.A., El-Sharkawy, R.M., Khalil Bagy, H.M.M., 2017. Effectiveness of Egyptian propolis on control of tomato bacterial wilt caused by Ralstonia solanacearum. Journal of Plant Diseases and Protection, 124(5): 467-472.
- Ali, A.M., Kunugi, H., 2021. Propolis, bee honey, and their components protect against coronavirus disease 2019 (COVID-19): A review of in silico, in vitro, and clinical studies. Molecules, 26(5): 1232.
- Aparicio-García, P.F., Ventura-Aguilar, R.I., Del Río-García, J.C., Hernández-López, M., Guillén-Sánchez, D., Salazar-Piña, D.A., Ramos-García, M.D.L., Bautista-Baños, S., 2021. Edible chitosan/propolis coatings and their effect on ripening, development of Aspergillus flavus, and sensory quality in fig fruit, during controlled storage. Plants, 10(1): 112.
- Awale, S., Li, F., Onozuka, H., Esumi, H., Tezuka, Y., Kadota, S., 2008. Constituents of Brazilian red propolis and their preferential cytotoxic activity against human pancreatic PANC-1 cancer cell line in nutrient-deprived condition. Bioorganic & Medicinal Chemistry, 16(1): 181-189.
- Basim, E., Basim, H., Özcan, M., 2006. Antibacterial activities of Turkish pollen and propolis extracts against plant bacterial pathogens. Journal of Food Engineering, 77(4): 992-996.
- Bektas, Y., Eulgem, T., 2015. Synthetic plant defense elicitors. Frontiers in Plant Science, 5(804): 1-17.
- Bohinc, T., Vučajnk, F., Trdan, S., 2019. The efficacy of environmentally acceptable products for the control of major potato pests and diseases. Zemdirbyste-Agriculture, 106(2): 135-142.
- Botteon, C.E.A., Silva, L.B., Ccana-Ccapatinta, G.V., Silva, T.S., Ambrosio, S.R., Veneziani, R.C.S., Bastos, J.K., Marcato, P.D., 2021. Biosynthesis and characterization of gold nanoparticles using Brazilian red propolis and evaluation of its antimicrobial and anticancer activities. Scientific Reports, 11(1): 1-16.
- Burdock, G.A., 1998. Review of the biological properties and toxicity of bee propolis (Propolis). Food and Chemical Toxicology, 36(4): 347-363.
- Cai, Y., Cao, X., Aballay, A., Nishikawa, Y., Davies, J.E., 2014. Whole-animal chemical screen identifies colistin as a new immunomodulator that targets conserved pathways. mBio, 5(4): 1-11.
- Cohen, Y., Vaknin, M., Mauch-Mani, B., 2016. BABA-induced resistance: milestones along a 55-year journey. Phytoparasitica, 44(4): 513-538.
- Da Silva Barboza, A., Aitken-Saavedra, J.P., Ferreira, M.L., Fábio Aranha, A.M., Lund, R.G., 2021. Are propolis extracts potential pharmacological agents in human oral health? - A scoping review and technology prospecting. Journal of Ethnopharmacology, 271: 113846.
- Er, Y., 2021. In vitro and in vivo antimicrobial activity of propolis extracts against various plant pathogens. Journal of Plant Diseases and Protection, 128: 693-701.
- Farace, G., Fernandez, O., Jacquens, L., Coutte, F., Krier, F., Jacques, P., Clément, C., Barka, E.A., Jacquard, C., Dorey, S., 2015. Cyclic lipopeptides from Bacillus subtilis activate distinct patterns of defence responses in grapevine. Molecular Plant Pathology, 16(2): 177-187.
- Freires, I.A., Queiroz, V., Furletti, V.F., Ikegaki, M., de Alencar, S.M., Duarte, M.C.T., Rosalen, P.L., 2016. Chemical composition and antifungal potential of Brazilian propolis against Candida spp. Journal of Medical Mycology, 26(2): 122-132.
- Gallez, L., Kiehr, M., Fernández, L., Delhey, R., Stikar, D., 2015. Antifungal activity in vitro of propolis solutions from Argentina against two plant pathogenic fungi: Didymella bryoniae and Rhizotocnia solani. Journal of Apicultural Research, 53(4): 438-440.
- Halder, V., Suliman, M.N.S., Kaschani, F., Kaiser, M., 2019. Plant chemical genetics reveals colistin sulphate as a SA and NPR1-independent PR1 inducer functioning via a p38-like kinase pathway. Scientific Reports, 9(1): 11196.
- Henry, G., Deleu, M., Jourdan, E., Thonart, P., Ongena, M., 2011. The bacterial lipopeptide surfactin targets the lipid fraction of the plant plasma membrane to trigger immune-related defence responses. Cellular Microbiology, 13(11): 1824-1837.
- Huang, S., Zhang, C P., Wang, K., Li, G.Q., Hu, F.L., 2014. Recent advances in the chemical composition of propolis. Molecules, 19(12): 19610-19632.
- Ishihara, M., Naoi, K., Hashita, M., Itoh, Y., Suzui, M., 2009. Growth inhibitory activity of ethanol extracts of Chinese and Brazilian propolis in four human colon carcinoma cell lines. Oncology reports, 22(2): 349-354.
- Jug, M., Končić, M.Z., Kosalec, I., 2014. Modulation of antioxidant, chelating and antimicrobial activity of poplar chemo-type propolis by extraction procures. LWT-Food Science and Technology, 57(2): 530-537.
- Knoth, C., Salus, M.S., Girke, T., Eulgem, T., 2009. The synthetic elicitor 3,5-dichloroanthranilic acid induces NPR1-dependent and NPR1-independent mechanisms of disease resistance in Arabidopsis. Plant Physiology, 150(1): 333-347.
- Kolayli, S., Palabiyik, I., Atik, D.S., Keskin, M., Bozdeveci, A., Karaoglu, S.A., 2020. Comparison of antibacterial and antifungal effects of different varieties of honey and propolis samples. Acta Alimentaria, 49(4): 515-523.
- Lima Cavendish, R., de Souza Santos, J., Belo Neto, R., Oliveira Paixao, A., Valeria Oliveira, J., Divino de Araujo, E., Berretta, E., Silva, A.A., Maria Thomazzi, S., Cordeiro Cardoso, J., Zanardo Gomes, M., 2015. Antinociceptive and anti-inflammatory effects of Brazilian red propolis extract and formononetin in rodents. Journal of Ethnopharmacology, 173: 127-133.
- Martinello, M., Mutinelli, F., 2021. Antioxidant activity in bee products: A review. Antioxidants, 10(1): 71.
- Noutoshi, Y., Okazaki, M., Kida, T., Nishina, Y., Morishita, Y., Ogawa, T., Suzuki, H., Shibata, D., Jikumaru, Y., Hanada, A., Kamiya, Y., Shirasu, K., 2012. Novel plant immune-priming compounds identified via high-throughput chemical screening target salicylic acid glucosyltransferases in Arabidopsis. Plant Cell, 24(9): 3795-3804.
- Ordonez, R.M., Zampini, I.C., Moreno, M.I., Isla, M.I., 2011. Potential application of Northern Argentine propolis to control some phytopathogenic bacteria. Microbiological Research, 166(7): 578-584.
- Pascoal, A., Feás, X., Dias, T., Dias, L.G., Estevinho, L.M., 2014. The role of honey and propolis in the treatment of infected wounds. In: K. Kon, M. Rai (Eds.), Microbiology for Surgical Infections, Academic Press, Amsterdam, pp. 221-234.
- Reddy, P.P., 2013. Plant Defence Activators. Springer, New Delhi.
- Regueira, M.S.N., Tintino, S.R., Da Silva, A.R.P., Costa, M.D.S., Boligon, A.A., Matias, E.F.F., De Queiroz Balbino, V., Menezes, I.R.A., Melo Coutinho, H.D., 2017. Seasonal variation of Brazilian red propolis: Antibacterial activity, synergistic effect and phytochemical screening. Food and Chemical Toxicology, 107(Pt B): 572-580.
- Rivera-Yañez, N., Rivera-Yañez, C.R., Pozo-Molina, G., Méndez-Catalá, C.F., Méndez-Cruz, A.R., Nieto-Yañez, O., 2020. Biomedical properties of propolis on diverse chronic diseases and its potential applications and health benefits. Nutrients, 13(1): 78.
- Salatino, A., Teixeira, E.W., Negri, G., Message, D., 2005. Origin and chemical variation of Brazilian propolis. Evidence-Based Complementary and Alternative Medicine, 2(1): 33-38.
- Scorza, C.A., Gonçalves, V.C., Scorza, F.A., Fiorini, A.C., De Almeida, A.C.G., Fonseca, M.C.M., Finsterer, J., 2020. Propolis and coronavirus disease 2019 (COVID-19): Lessons from nature. Complementary Therapies in Clinical Practice, 41: 101227.
- Sforcin, J.M., Bankova, V., 2011. Propolis: is there a potential for the development of new drugs? Journal of Ethnopharmacology, 133(2): 253-260.
- Silva-Castro, I., Diez, J., Martín-Ramos, P., Pinto, G., Alves, A., Martín-Gil, J., Martín-García, J., 2018.
Application of bioactive coatings based on chitosan and propolis for Pinus spp. protection against Fusarium circinatum. Forests, 9(11): 685.
- Valente, M.J., Baltazar, A.F., Henrique, R., Estevinho, L., Carvalho, M., 2011. Biological activities of Portuguese propolis: protection against free radical-induced erythrocyte damage and inhibition of human renal cancer cell growth in vitro. Food and Chemical Toxicology, 49(1): 86-92.
- Wagh, V.D., 2013. Propolis: a wonder bees product and its pharmacological potentials. Advances in Pharmacological and Pharmaceutical Sciences, 2013: 308249.
- Xin, X.F., He, S.Y., 2013. Pseudomonas syringae pv. tomato DC3000: a model pathogen for probing disease susceptibility and hormone signaling in plants. Annual Review of Phytopathology, 51: 473-498.
- Xu, J., Zhang, Y., 2020. Traditional Chinese Medicine treatment of COVID-19. Complementary Therapies in Clinical Practice, 39: 101165.
- Yong, H., Liu, J., 2021. Active packaging films and edible coatings based on polyphenol‐rich propolis extract: A review. Comprehensive Reviews in Food Science and Food Safety, 20(2): 2106-2145.
- Zabaiou, N., Fouache, A., Trousson, A., Baron, S., Zellagui, A., Lahouel, M., Lobaccaro, J.A. 2017. Biological properties of propolis extracts: Something new from an ancient product. Chemistry and Physics of Lipids, 207(Pt B): 214-222.
- Zampini, I.C., Salas, A.L., Maldonado, L.M., Simirgiotis, M.J., Isla, M.I., 2021. Propolis from the Monte region in Argentina: a potential phytotherapic and food functional ingredient. Metabolites, 11(2): 76.