Investigation of the antimicrobial activities of solvent extracts of two endemic species from Turkey: Campanula tomentosa Lam. and Verbascum mykales Bornm.

Year 2021, Volume: 51 Issue: 3, 348 - 356, 29.12.2021

Esin Poyrazoğlu , Esra Barışık

Abstract

Background and Aims: Campanula tomentosa Lam. and Verbascum mykales Bornm. are endemic species in Turkey. Extracts of these plants contain important natural compounds such as flavonoids, saponins and tannins. This study investigates the antimicrobial effects of leaf extracts of C. tomentosa and V. mykales against some bacteria and yeasts. Materials and Methods: Leaves of plant samples were air-dried and ground into powder. Five solvents (ethyl acetate, methanol, acetone, chloroform, boiled water) were used for extraction. Experiments were conducted using these crude extracts on seventeen bacteria, three yeasts and three microfungi. The agar well diffusion method was used for the antimicrobial activities of the extracts. In addition, minimum inhibitory concentrations, minimum bacteriocidal concentration, minimal fungicidal concentrations were carried out. Results: The ethyl acetate and methanol extracts of C. tomentosa and V. mykales were found to be highly effective against the tested microorganisms. According to the MIC values, the ethyl acetate extracts of C. tomentosa and V. mykales had a strong effect (4-8 μg/mL) against Escherichia coli ATCC 35218, Micrococcus luteus ATCC 9341, Streptococcus pneumonia ATCC 27336, Pseudomonas aeruginosa ATCC 35032, Mycobacterium smegmatis ATCC 607, Proteus vulgaris ATCC 33420, Enterococcus faecalis ATCC 29212, Bacillus cereus ATCC 11778, and Bacillus subtilis ATCC 6633. The ethyl acetate extract of C. tomentosa had a moderate effect (64 μg/mL) against Candida albicans ATCC 10231 and Saccharomyces cerevisiae ATCC 9763, and the ethyl acetate extract of V. mykales had a moderate effect (64 μg/mL) against Aspergillus flavus ATCC 9807 and Aspergillus niger ATCC 16404. However, the boiled water extract of C. tomentosa and V. mykales had no effect on the tested microorganisms. Conclusion: C. tomentosa and V. mykales used in the study are endemic plants and their antimicrobial activities are being investigated for the first time. The ethyl acetate extract of both plants was found to be most effective against the Gram (+) and Gram (-) bacteria. However, all extracts of both plants were found to have fewer antimicrobial effects against used yeasts and microfungi. This study demonstrates that plant extracts are more effective against prokaryotic microorganisms than eukaryotes.

Keywords

Campanula tomentosa and Verbascum mykales, antimicrobial activity, agar well diffusion, MIC/MBC/MFC

Supporting Institution

This study was funded by Scientific Research Projects Coordination Unit of Aydin Adnan Menderes University

Project Number

FEF–17039

Thanks

This work was carried out by Aydin Adnan Menderes University Biology Department, Microbiology Laboratory. We thank Assoc. Prof. Dr. Özkan EREN for tlıe support provided to us in the collection and identification of plants.

References

• • Abdallah, L., & Omar, G. (2019). Antibacterial effect of some wild medicinal plants in Paletsine against multi drug resistant Escherichia coli clinical isolate. Brazilian Journal of Biological Sciences, 6(12), 103-113.
• • Alhage, J., Elbitar, H., Taha, S., & Benvegnu, T. (2018). Invitro assessment of antioxidant, antimicrobial, cytotoxic, antiinflammatory and antidiabetic activities of Campanula retrorsa crude extracts. Pharmacognosy Research, 10, 39-403.
• • Amirnia, R., Khoshnoud, H., Alahyary, P., Ghiyasi, M., Tajbakhsh, M., & Valizadegan, O. (2011). Fresenius Environmental Bulletin, 20(3a), 690-693.
• • Baytop, T. (1999). Türkiye’de bitkiler ile tedavi, geçmişte ve bugün. Nobel Tıp Kitabevleri, II. Baskı, (pp. 480), İstanbul, Türkiye.
• • Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71-79.
• • Benli, M., Bingol, U., Geven, F., Guney, K., & Yigit, N. (2008). An investigation on the antimicrobial activity of some endemic plant species from Turkey. African Journal of Biotechnology, 7(1), 1-5.
• • Bıyık, H. H., Onur, M., Törün, B., & Çoban, E. P. (2018). Antibacterial and anticandidal effects of the leaf extracts of Persea americana Mill. Annals of Phytomedicine, 7(2), 88-93.
• • Borchardt, J. R., Wyse, D. L., Sheaffer, C. C., Kauppi, K. L., Fulcher, R. G., Ehlke, N. J., Biesboer, D. D., & Bey, R. F. (2008). Antimicrobial activity of native and naturalized plants of Minnesota and Wisconsin. Journal of Medicinal Plant Research, 2(5), 98-110.
• • Buruk, K., Sokmen, A., Aydin, F., & Erturk, M. (2006). Antimicrobial activity of some endemic plants growing in the Eastern Black Sea Region, Turkey. Fitoterapia, 77: 388-391.
• • Clinical and Laboratory Standards Institute. (2004). Method for antifungal disk diffusion susceptibility testing of yeasts, approved guideline. CLSI document M44-A. Wayne, PA, USA.
• • Clinical and Laboratory Standards Institute. (2009). M07–A8: Methods for dilution antimicrobial susceptibility testing for bacteria that grow aerobically: Approved Standard (8thed). (pp. 1-65). CLSI, Wayne, PA, USA.
• • Clinical and Laboratory Standards Institute. (2013). Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard. CLSI document VET01–A4(4thed). CLSI, Wayne, PA, USA.
• • Clinical and Laboratory Standards Institute. (2015). M02-A12: Performance standards for antimicrobial disk susceptibility tests: Approved standard. (12thed). (pp. 1-73). CLSI, Wayne, PA, USA.
• • Collins, C. H., Lyne, P. M., Grange, J. M., & Falkinham, J. O. (2004). Collins and Lyne’s microbiological methods (8thed). (pp. 456). London: Butterworths.
• • Coşkun, M. I., Gemici, M., &Yildirim, H. (2017). A study on endogenous hormone levels and phenolic profiles in embryogenic and non- embryogenic calli of endemic plant Campanula tomentosa L. Asian Journal of Biology, 4(2), 1-7.
• • Çoban, E. P., Bıyık, H. H., Törün, B., & Yaman, F. (2017). Evaluation the antimicrobial effects of Pistacia terebinthus L. and Papaver rhoeas L. extracts against some pathogen microorganisms. Indian Journal of Pharmaceutical Education and Research, 51(3), 377-380.
• • Çoban, E. P., Erçin, Z., Törün, B., & Bıyık, H. H. (2018). Antimicrobial effects of the stem extracts of Apium graveolens Mill. Annals of Phytomedicine, 7(2), 70-75.
• • Dar, R. A., Shahnawaz, M., & Qazi, P. H. (2016). General overview of medicinal plants: A review. Journal of Phytopharmacy, 6(6), 349-351. • Dulger, B., Kirmizi, S., Arslan, H., Güleryüz, G. (2002). Antimicrobial activity of three endemic Verbascum species. Pharmaceutical Biology, 40(8): 587-589.
• • Dulger, B., & Dulger, G. (2018). Antibacterial activity of Verbascum antinori. Konuralp Tıp Dergisi, 10(3), 395-398.
• • Dülger, B., & Hacıoğlu, N. (2008). Antimicrobial activity of some endemic Verbascum and Scrophularia species from Turkey. Asian Journal of Chemistry, 20(5), 3779-3785. • EUCAST (2019). Disk diffusion method for antimicrobial susceptibility testing (Version 7.0).
• • Fares, S. S. F. (2018). Antimicrobial activities and phytochemical analysis of Verbascum fruticulosum (Master DegreeThesis). An- Najah National University Faculty of Graduate Studies.
• • Faydaoğlu, E., & Sürücüoğlu, M. S. (2011). Geçmişten günümüze tıbbi ve aromatik bitkilerin kullanılması ve ekonomik önemi. Kastamonu Üniversitesi Orman Fakültesi Dergisi, 11(1), 52-67.
• • Faydaoğlu, E., & Sürücüoğlu, M. S. (2013). Tıbbi ve aromatik bitkilerin antimikrobiyal, antioksidan aktiviteleri ve kullanım olanakları. Erzincan Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 6(2), 233-265.
• • Göse, M. & Hacıoğlu Doğru (2021). Bioactive compounds, antimicrobial and antibiofilm activity of two Verbascum species. KSU Journal of Agriculture and Nature, 24(3): 479-487.
• • Guarino, C. (2002). Antimicrobial activity of Verbascum macrurum Ten. (Scrophulariaceae). Bollettino Chimico Farmaceutico, 141(3), 238-242.
• • Herkul, M.M, & Köroğlu, A. (2019). A review on the ethnobotanical use, chemical content and biological activity of the Campanulaceae family. Türk Farmakope Dergisi, 4(3), 70–84.
• • Ismaiel, A. A., & Tharwat, N. A. (2014). Antifungal activity of silverion on ultra structure and production of aflatoxin B1 and patulin by two mycotoxigenic strains, Aspergillus flavus OC1 and Penicillium vulpinum CM1. Journal de Mycologie Médicale, 24,193-204.
• • Jorgensen, J. H., & Ferraro, M. J. (2009). Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Journal of Medical Microbiology, 49, 1749-1755.
• • Kahraman, Ç., Ekizoğlu, M., Kart, D., Akdemir, Z. Ş., & Tatlı, I. I. (2011). Antimicrobial activity of some Verbascum species growing in Turkey. FABAD Journal of Pharmaceutical Sciences, 36, 11-15.
• • Kahraman, Ç., Tatlı, İ. İ., Kart, D., Ekizoğlu, M., & Akdemir, Z. Ş. (2018). Structure elucidation and antimicrobial activities of secondary metabolites from the flowery parts of Verbascum mucronatum Lam. Turkish Journal of Pharmaceutical Sciences, 15(3), 231-237.
• • Karalija, E., Parić, A., Dahija, S., Bešta-Gajević, R., & Zeljković, S. Ć. (2018). Phenolic compounds and bioactive properties of Verbascum glabratum subsp. Bosnense (K. Malý) Murb., an endemic plant species. Natural Product Research, 32, 1-5.
• • Keskin, C. (2018). Medicinal plants and their traditional uses. Journal of Advances in Plant Biology, 1(2), 1-12.
• • Khafagi, I. K. (2001). Screening in vitro cultures of some Sinai medicinal plants for their antibiotic activity. Journal Microbiology, 34: 613-627.
• • Küçük, S., Özdemir, F., İşcan, G., & İncesu, Z. (2016). Determination of cytotoxic and anticandidal activities of three Verbascum L. species fromTurkey: V. cheiranthifolium Boiss. var. asperulum (Boiss.) Murb. Monorg. V. pycnostachyum Boiss. & Heldr and V. orgyale Boiss. & Heldr. Turkish Journal of Pharmaceutical Sciences, 13(3), 318-322.
• • Moosavi, S. R., Ardekani, M. R. S., Vazirian, M., & Lamardi, S. N. S. (2018). Campanula latifola, giant bellflower; ethno-botany, phytochemical and antioxidant evaluation. Traditional and Integrative Medicine, 3(3), 113-119.
• • Muhammad, A., Shandana, A., Khushboo, S., & Rahila, A. (2019). Review on antibacterial activity of Himalayan medicinal plants traditionally used to treat pneumonia and tuberculosis. Journal of Pharmacy and Pharmacology, 71, 1599-1625.
• • Nofouzi, K., Mahmudi, R., Tahapour, K., Amini, E., & Yousefi, K. (2016). Verbascum speciosum methanolic extract: Phytochemical components and antibacterial properties. Journal of Essential Oil Bearing Plants, 19(2), 499-505.
• • Nofouzi, K. (2015). Study on the antioxidant activity and in vitro antifungal activity of Verbascum speciosum methanolic extract. Journal of Mycology Research, 2(2), 97-103.
• • Noori, M., Malayeri, B., Moosaei, M., Pakzad, R., & Piriye, M.H. (2012). Effects of heavy metals on the antibacterial properties of Verbascum speciosum Schard. Revista Científica UDO Agrícola, 12(2), 463-471.
• • Okoye, E. L., Uba, B. O., Dike, U. C., & Eziefule, C. J. (2020). Growth rate and antifungal activities of acetone extracts of Ocimum gratissimum (ScentLeaf ) and Alliumsativum (Garlic) on cassava and banana peels formulated media. Journal of Advances in Microbiology, 20(4), 19-29.
• • Oyeka, E. E., Asegbeloyin, J. N., Babahan, I., Eboma, B., Okpareke, O., Lane, J., Ibezim, A, Bıyık, H.H, Törün, B, & Izuogu, D. C. (2018). Synthesis, crystal structure, computation analysis and biological properties of 1-(4-chlorobenzoyl)-3-[2-(2-{2-[3-(4-chlorobenzoyl)- thioureido]-ethoxy} ethoxy) ethyl]-thiourea and its Ni(II) and Cu(II) complexes. Journal of Molecular Structure, 1168, 153-164.
• • Ozhatay, N., Kultur, Ş., & Aslan, Ş. (2009). Check-list of additional taxa to the supplement Flora of Turkey IV. Turkish Journal of Botany, 33, 191-226.
• • Politeo, O., Skocibusic, M., Burcu, F., Maravic, A., Carev, I., Ruscic, M., & Milos, M. (2013). Campanula portenschlagiana Roem.et Schult.: Chemical and antimicrobial activities. Chemistry and Biodiversity, 10: 1072-1080.
• • Prakash, V., Rana, S., & Sagar, A. (2016). Studies on antibacterial activity of Verbascum thapsus. Journal of Medicinal Plants Studies, 4(3), 101-103.
• • Sepahi, S., Ghorani-Azam, A., Sepahi, S., Asoodeh, A., & Rostami, S. (2014). In vitro study to evaluate antibacterial and non-haemolytic activities of four Iranian medicinal plants. West Indian Medical Journal, 63(4), 289-293.
• • Sinek, K., İskender, N, Y., Yaylı, B., Karaoğlu, S. A., & Yaylı N. (2012). Antimicrobial activity and chemical composition of the essential oil from Campanula glomerata L. subsp. Hispida (Witasek) Hayek. Asian Journal of Chemistry, 24(5), 1931-1934.
• • Stojkovi´c, D., Dias, M. I., Drakuli´c, D., Barros, L., Stevanovi´c, M., Ferreira, I. C. F. R., &Sokovi´c, M., D. (2020). Methanolic extract of the herb Ononis spinosa L. is an antifungal agent with no cytotoxicity to primary human cells. Pharmaceuticals, 13(78), 1-13.
• • Şen, B., Döşler, S., Meriçli, A. H. (2012). The antimicrobial activity screening of three Verbascum species in Marmara region. Planta Medica, 78, PI310.
• • Tatli, İ.İ., Schuhly, W., Kunert, O., Bedir, E., & Akdemir Z. S. (2008). Secondary metabolites from the aerial parts of Verbascum dudleyanum and their biological activities. Chemistry of Natural Compounds, 44(3), 292-295.
• • Tatli, İ., & Akdemir. Z. S. (2005). Antimicrobial and antimalarial activities of secondary metabolites from some Turkish Verbascum species. FABAD Journal of Pharmaceutical Sciences, 30(2), 84–92.
• • Temel, M. Tınmaz, B. Öztürk, M. & Gündüz, O. (2018). Dünyada ve Türkiye’de tıbbi-aromatik bitkilerin üretimi ve ticareti. KSU Journal of Agriculture and Nature, 21(Special Issue): 198-214.
• • Tosun, G. Kahrıman, N. Çoskun Çelebi, K. Genç, H. Alpay Karaoglu, S. & Yaylı, N. (2011). Chemical composition and biological activity of the essential oil of Campanula olympica Boiss. Asian Journal of Chemistry, 23(6), 2389-2391.
• • Törün, B., Çoban, E. P., Bıyık, H. H., & Barışık, E. (2017). Antimicrobial activity of Echinophora tenuifolia L. and Raphanus sativus L. extracts. Indian Journal of Pharmaceutical Education and Research, 51(3), 373-376.
• • Turker, H., & Yıldırım, A. B. (2015). Screening for antibacterial activity of some Turkish plants against fish pathogens: a possible alternative in thetreatment of bacterial infections. Biotechnology and Biotechnological Equipment, 29(2), 281-288.
• • Usta, C., Yildirim, B. A., & Turker, U. A. (2014). Antibacterial and antitumor activities of some plants grown in Turkey. Biotechnology and Biotechnological Equipment, 28(2), 306-315.
• • Yaldız, G., & Çamlıca, M. (2018). Türkiye’de tıbbi ve aromatik bitkilerin üretimi ve ticareti. Bahçe, 47: (Özel Sayı): 224-229.
• • Zamri, N. A. F. M., Bakar, R. A., Noor, S. S. M., & Fuad, W. E. M. (2020). Antimicrobial effects of Peperomia pellucida (Ketumpang Air) against multi drug resistant and foodborne organisms including toxicity study in Sprague Dawley rats. Asian Journal of Medicine and Biomedicine, eISNN: 2600–8173.