Investigating the Antibacterial Effects of Zataria multiflora Methanolic Extract on Standard Pathogenic Bacteria in Laboratory Conditions

Abstract

Alternative treatments are becoming more and more necessary due to the increasing growth in medication resistance to synthetic antibiotics. The purpose of this study was to examine the antibacterial capabilities of an extract from Zataria multiflora (Z. multiflora) against a variety of pathogenic bacteria. Z. multiflora plants were collected from the natural resources of Isfahan province and verified as Z. multiflora by botanists from the Agricultural Jihad Organization. Methanolic extract of Z. multiflora was prepared, and its effects were tested at concentrations of 50 mg/ml, 100 mg/ml, 200 mg/ml, and 400 mg/ml using the well diffusion and disk diffusion methods on Staphylococcus aureus (S. aureus), Bacillus cereus (B. cereus), Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa). Minimum inhibitory concentration of bacterial growth and minimum bacterial pathogen concentration (MIC/MBC) were determined through dilution in tubes. The outcomes showed that both gram-positive and gram-negative bacteria were significantly inhibited by the methanolic extract of Z. multiflora. The MBC/MIC analysis revealed that S. aureus was the most sensitive, while E. coli was the least sensitive to the Z. multiflora extract. The study shows the potential of the methanolic extract from Z. multiflora against various pathogenic bacteria, including Gram-positive and Gram-negative ones. It suggests it could be a viable alternative to synthetic antibiotics, especially in the face of antibiotic resistance. Further research is needed to understand its mechanisms, optimize extract preparation, and explore its clinical applications..

Keywords

• Medicinal plants
• Antibacterial effects
• Pathogenic bacteria
• Extract

References

1. Al-Snafi, A. E., Salman Khadem, H., Al-Saedy, H. A., Alqahtani, A. M., El-Saber Batiha, G., & Jafari-Sales, A. (2021). A review on Medicago sativa: A potential medicinal plant. International Journal of Biological and Pharmaceutical Sciences Archive, 1 (2), 22-33. https://doi.org/10.30574/ijbpsa.2021.1.2.0302
2. Alizadeh, H., Jafari, B., & Babai, T. (2012). The study of antibacterial effect of Capsella bursapastoris on some of gram positive and gram negative bacteria. Journal of Basic and Applied Scientific Research, 2 (7), 6940-6945.
3. Andacheh, F., Moslehishad, M., Shojaee-Aliabadi, S., & Jannatyha, N. (2020). Production and characterization of antimicrobial carboxymethyl cellulose (cmc) films containing essentiol oils of Satureja khuzistanica, Zataria multiflora, Allium sativum and Bunium persicum. Journal of Medicinal plants and By-Products, 9 (Special), 35-45. htpp://doi.org/10.22092/jmpb.2020.121749
4. Atapour, M., Zahedi, M. J., Mehrabani, M., Safavi, M., Keyvanfard, V., Foroughi, A., Siavoshi, F., & Foroumadi, A. (2009). In vitro susceptibility of the gram-negative bacterium Helicobacter pylori to extracts of Iranian medicinal plants. Pharmaceutical Biology, 47 (1), 77-80. htpp://doi.org/10.1080/13880200802434401
5. Azizkhani, M., Misaghi, A., Akhondzadeh Basti, A., Gandomi Nasrabadi, H., & Hosseini, H. (2012). Effect of Zataria multiflora Boiss. essential oil on growth and enterotoxin e production of Staphylocuccus aureus ATCC 29213. Journal of Medicinal Plants, 11 (44), 185-192.
6. Cockerill, F. R. (2013). Performance Standards for Antimicrobial Susceptibility Testing: Twenty-Third Informational Supplement. Wayne, Pennsylvania: Clinical and Laboratory Standards Institute.
7. Cowan, M. M. (1999). Plant Products as antimicrobial agents. Clinical Microbiology Reviews, 12 (4), 564-582.
8. Dadashi, M., Hashemi, A., Eslami, G., Fallah, F., Goudarzi, H., Erfanimanesh, S., & Taherpour, A. (2016). Evaluation of antibacterial effects of Zataria multiflora Boiss. extracts against ESBL-producing Klebsiella pneumoniae strains. Avicenna Journal of Phytomedicine, 6 (3), 336-343.

9. Davazdahemami, A., & Behbahani, M. (2022). Investigating in silico and in vitro anti-bacterial activity of eight monofloral iranian honey types. Research in Molecular Medicine, 10(2), 133-142. https://doi.org/10.32598/rmm.10.2.1195
10. Deans, S. G., Simpson, E., Noble, R. C., MacPherson, A., & Penzes, L. (1993). Natural antioxidants from Thymus vulgaris (Thyme) volatile oil: the beneficial effects upon mammalian lipid metabolism. Acta Horticulturae, 342, 237-241.
11. Fallah, F., Taherpour, A., Borhan, R. S., Hashemi, A., Habibi, M., & Sajadi Nia, R. (2013). Evaluation of Zataria multiflora Boiss. and Carum copticum antibacterial activity on IMP-type metallo-beta-lactamase-producing Pseudomonas aeruginosa. Annals of Burns and Fire Disasters, 26 (4), 193-198.
12. Fazeli, M. R., Amin, G., Attari, M. M. A., Ashtiani, H., Jamalifar, H., & Samadi, N. (2007). Antimicrobial activities of Iranian sumac and avishan-e shirazi (Zataria multiflora) against some food-borne bacteria. Food Control, 18 (6), 646-649. https://doi.org/10.1016/j.foodcont.2006.03.002
13. Foladvand, Z., Kiani, Z., Javadian, F., Hesaraki, M., Nasiri, A., & Sepehri, Z. (1970). Evaluation of the effect of antimicrobial activity of ethanol extract of Myrtus communis, Zataria multiflora Boiss. and Allium sativum on biofilm formation by Staphylococcus aureus. Journal of Sabzevar University of Medical Sciences, 21 (6), 1019-1027.
14. Ghannadi, A., Sajjadi, S.-E., Abedi, D., Yousefi, J., & Daraei-Ardekani, R. (2004). The in vitro activity of seven Iranian plants of the Lamiaceae family against Helicobacter pylori. Nigerian Journal of Natural Products and Medicine, 8 (1), 40-42. https://doi.org/10.4314/njnpm.v8i1.11812
15. Golkar, P., Mosavat, N., & Jalali, S. A. H. (2020). Essential oils, chemical constituents, antioxidant, antibacterial and in vitro cytotoxic activity of different Thymus species and Zataria multiflora collected from Iran. South African Journal of Botany, 130, 250-258. https://doi.org/10.1016/j.sajb.2019.12.005
16. Goudarzi, M., Sattari, M., Najar Piraieh, S., Goudarzi, G., & Bigdeli, M. (2006). Antibacterial effects of aqueous and alcoholic extracts of Thyme on enterohemorrhagic Escherichia coli. Yafteh, 8 (3), 63-69.
17. Hatami Pirghibi, T., Chehri, K., Abiri, R., & Karimi, I. (2021). Effect of hydro-alcoholic extracts of Rosmarinus officinalis L., Mentha piperita L., and Zataria multiflora Boiss. on biofilm formation of Pseudomonas. Research in Medicine, 45 (3), 11-17.
18. Hornok, L. (1988). Effect of environmental factors on the production of some essential oil plants. Developments in Food Science, 18, 129-140.
19. Izadiyan, P., Salehi, A., Moaddeli, A., Zarenezhad, M., & Izadiyan, M. (2022). How quantity of bioactive compounds of Zataria multiflora differ using traditional or modern extraction methods. Journal of Medicinal plants and By-Products, 11 (Special), 107-115. https://doi.org/10.22092/jmpb.2021.353668.1339
20. Jafari-Sales, A., & Hossein-Nezhad, P. (2020). Antimicrobial effects of Rosmarinus officinalis methanolic extract on Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa in laboratory conditions. Journal of Medicinal and Chemical Sciences, 3 (2), 103-108. https://doi.org/10.26655/jmchemsci.2020.2.2
21. Jafarı-Sales, A., Jafari, B., Khaneshpour, H., & Pashazadeh, M. (2020). Antibacterial effect of methanolic extract of Rosa damascena on standard bacteria Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa in vitro. International Journal of Nature and Life Sciences, 4 (1), 40-46.
22. Jafari-Sales, A., Jafari, B., Sayyahi, J., & Zohoori-Bonab, T. (2015). Evaluation of antibacterial activity of ethanolic extract of Malva neglecta and Althaea officinalis L. on antibiotic-resistant strains of Staphylococcus aureus. Journal of Biology and Today's World, 4 (2), 58-62. https://doi.org/10.15412/J.JBTW. 01040205
23. Jafari-Sales, A., Meshinchi, P., & Shabkhosh, A. (2021). In vitro antibacterial activity of ethanolic extract of Aloe vera and silver nanoparticles on standard strains of some pathogenic bacteria. Journal of Clinical and Basic Research, 5 (4), 22-30. https://doi.org/10.29252/Jcbr.5.4.22
24. Jafarı-Sales, A., & Pashazadeh, M. (2020). Antibacterial effect of methanolic extract of saffron petal (Crocus sativus L.) on some standard gram positive and gram negative pathogenic bacteria In vitro. Current Perspectives on Medicinal and Aromatic Plants, 3 (1), 1-7. https://doi.org/10.38093/cupmap.692879
25. Jafari, B., Jafari-Sales, A., Khaneshpour‬‏, H., Fatemi, S., Pashazadeh, M., Al-Snafi, A. E., & Shariat, A. (2020). Antibacterial effects of Thymus vulgaris, Mentha pulegium, Crocus sativus and Salvia officinalis on pathogenic bacteria: A brief review study based on gram-positive and gram-negative bacteria. Jorjani Biomedicine Journal, 8 (3), 58-74. https://doi.org/10.29252/jorjanibiomedj.8.3.58
26. Jafari Sales, A., & Shariat, A. (2020). Synergistic Effects of silver nanoparticles with ethanolic extract of Eucalyptus globules on standard pathogenic bacteria in vitro. Tabari Biomedical Student Research Journal, 2 (3), 13-21. https://doi.org/10.18502/tbsrj.v2i3.4528
27. Jebelli Javan, A. (2016). Combinational effects of Trachyspermum ammi and Zataria multiflora Boiss. essential oils on some pathogenic food-borne bacteria. Koomesh, 17 (2), 374-383.
28. Karbasizade, V., Dehghan, P., Sichani, M. M., Shahanipoor, K., Jafari, R., & Yousefian, R. (2017). Evaluation of three plant extracts against biofilm formation and expression of quorum sensing regulated virulence factors in Pseudomonas aeruginosa. Pakistan Journal of Pharmaceutical Sciences, 30, 2 (Suppl.), 585-589.
29. Mahmoudi, S., Nasiri, R., & Jafari Sales, A. (2019). In-vitro antibacterial effects of methanolic extract of peppermint (Mentha Piperita Lamiaceae) on standard Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa strain. Jorjani Biomedicine Journal, 7 (4), 4-10. https://doi.org/10.29252/jorjanibiomedj.7.4.4
30. Meskini, M., Khaledi, A., & Esmaeili, D. (2019). Inhibitory Effects of a Herbal Ointment against Pseudomonas aeruginosa. Medical Laboratory Journal, 13 (1), 1-5. https://doi.org/10.29252/mlj.13.1.1
31. Mobaiyen, H., Jafari Sales, A., & Sayyahi, J. (2015). Evaluating antimicrobial effects of Centaurea plant’s essential oil on pathogenic bacteria: Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli Isolated from clinical specimens. Journal of Advanced Biomedical Sciences, 5 (4), 479-487.
32. Mosadegh, M., & Naghibi, F. (2002). Iran Traditional Medicine: Past & Present. Traditional Medicine and Materia Medica Research Center, 1, 2-20.
33. Motevasel, M., Okhovat, M. A., Zomorodian, K., & Farshad, S. (2014). Antibacterial effect of Zataria multiflora extract on MRSA. Iranian South Medical Journal, 17 (5), 900-906.
34. Neef, H., Declercq, P., & Laekeman, G. (1995). Hypoglycaemic activity of selected European plants. Phytotherapy Research, 9 (1), 45-48. https://doi.org/10.1002/ptr.2650090111
35. Ojagh, M., Mohammadi, N., Babakhani Lashkan, A., & Mohammadi, E. (2016). Evaluation of antibacterial activity of aqueous and ethanolic extracts of Dorema aucheri, Zataria multiflora Boiss. and Ferulago angulata against certain pathogenic microbes. Journal of Medicinal Plants Biotechnology, 2 (4), 1-9.
36. Omidpanah, S., Vazirian, M., Hadjikhondi, A., Nabavi, S., & Manayi, A. (2016). Evaluation of antibacterial activity of some medicinal plants against isolated Escherichia coli from diseased laying hens. Progress in Nutrition, 18 (4), 429-435.
37. World Health Organization. (1978). The Promotion and Development of Traditional Medicine: Report af A WHO Meeting (Held in Geneva from 28 November to 2 December 1977). In, Geneva, Switzerland: World Health Organization.
38. World Health Organization. (2002). WHO Traditional Medicine Strategy 2002-2005. In, Geneva, Switzerland: World Health Organization.
39. Owlia, P., Saderi, H., Matloob, F., & Rezaee, M. B. (2006). Antimicrobial effect of Zataria multiflora Boiss. extract and oxacillin against Staphylococcus aureus. Iranian Journal of Medicinal and Aromatic Plants Research, 22 (1), 22-26. https://doi.org/10.22092/ijmapr.2006.114996
40. Pirbalouti, A. G., Hamedi, B., Poor, F. M., Rahimi, E., & Nejhad, R. N. (2011). Inhibitory activity of Iranian endemic medicinal plants against Vibrio parahaemolyticus and Vibrio harveyi. Journal of Medicinal Plants Research, 5 (32), 7049-7053. https://doi.org/10.5897/JMPR11.1256
41. Pourhosseini, S. H., Ahadi, H., Aliahmadi, A., & Mirjalili, M. H. (2020). Chemical composition and antibacterial activity of the carvacrol-rich essential oils of Zataria multiflora Boiss. (Lamiaceae) from southern natural habitats of Iran. Journal of Essential Oil Bearing Plants, 23 (4), 779-787. https://doi.org/10.1080/0972060X.2020.1824688
42. Ramezanpour, S., Ardestani, F., & Asadollahzadeh, M. J. (2016). Combination effects of Zataria multiflora, Laurus nobilis and Chamaemelum nobile essences on pathogenic E. coli and determination of optimum formulation using fraction and factorial statistical method. Iranian Journal of Medical Microbiology, 10 (2), 53-62.
43. Rezvannejad, E., Nasirifar, E., Lotfi, S., & Abdolinasab, M. (2019). Study and comparison of antibacterial activities of extracts of Zataria multiflora and Teucrium polium on Penibacillus alvei. Journal of the Hellenic Veterinary Medical Society, 70 (1), 1421-1428.
44. Sabzikar, A., Hosseinihashemi, S. K., Shirmohammadli, Y., & Jalaligoldeh, A. (2020). Chemical composition and antimicrobial activity of extracts from thyme and rosemary against Staphylococcus aureus and Candida albicans. BioResources, 15 (4), 9656-9671.
45. Sadeghi, H., Robati, Z., & Saharkhiz, M. J. (2015). Variability in Zataria multiflora Bioss. essential oil of twelve populations from Fars province, Iran. Industrial Crops and Products, 67, 221-226. https://doi.org/10.1016/j.indcrop.2015.01.021
46. Safari, R., Adel, M., Monji, H., Riyahi Cholicheh, H., & Nematolahi, A. (2015). Evaluation of antibacterial effect of some of the endemic herbal essential oils on Streptococcus iniae in invitro. Journal of Aquatic Ecology, 4 (4), 40-33.
47. Sales, A. J. (2014). Evaluation of antibacterial activity of ethanol extract of Lavandula stoechas L. plant on antibiotic-resistant strains of Staphylococcus aureus. Journal of Current Research in Science, 2 (6), 641-645.
48. Sayyahi, J., Mobaiyen, H., Jafari, B., & Jafari-Sales, A. (2019). Antibacterial effects of methanolic extracts of Reum ribes L. and Hyssopus officinalis on some standard pathogenic bacteria. Jorjani Biomedicine Journal, 7 (3), 34-44. https://doi.org/10.29252/jorjanibiomedj.7.3.34
49. Sayyahi, J., Mobayen, H., Jafari, B., & Jafari-Sales, A. (2021). Antibacterial effects of ethanolic extracts of Ziziphus jujuba, Medicago sativa, Reum ribes and Hyssopus officinalis on some standard gram-positive and gram-negative bacteria in vitro. Armaghane Danesh, 26 (3), 338-350. https://doi.org/10.52547/armaghanj.26.3.338
50. Shafaroudi, A. M., Gorji, N. E., Nasiri, P., Javidnia, J., & Saravi, M. E. (2022). Antifungal properties of Zataria multiflora on Candida species: A systematic review. Journal of Evidence-Based Integrative Medicine, 27, 1-15. https://doi.org/10.1177/2515690X221132272
51. Shariff, Z. U. (2001). Modern Herbal Therapy for Common Ailments. Ibadan, Nigeria: Spectrum Books.
52. Sharififar, F., Moshafi, M. H., Mansouri, S., Khodashenas, M., & Khoshnoodi, M. (2007). In vitro evalution of antioxidant and antibacterial activities of the essential oil and methanol extract of endemic Zataria multiflora Boiss. Food Control, 18 (7), 800-805. https://doi.org/10.1016/j.foodcont.2006.04.002
53. Sheikholeslami, S., Mousavi, S. E., Ahmadi Ashtiani, H. R., Hosseini Doust, S. R., & Mahdi Rezayat, S. (2016). Antibacterial activity of silver nanoparticles and their combination with Zataria multiflora essential oil and methanol extract. Jundishapur Journal of Microbiology, 9 (10), e36070. https://doi.org/10.5812/jjm.36070
54. Skaltsa, H., Demetzos, C., Lazari, D., & Soković, M. (2003). Essential oil analysis and antimicrobial activity of eight Stachys from Greece. Phytochemistry, 64 (3), 743-752. https://doi.org/10.1016/S0031-9422(03)00386-8
55. Skaltsa, H., Lazari, D. M., Chinou, I. B., & Loukis, A. E. (1999). Composition and antibacterial activity of the essential oils of Stachys candida and S. chrysantha from southern Greece. Planta medica, 65 (3), 255-256. https://doi.org/10.1055/s-2006-960471
56. Taherpour, A., Hashemi, A., Fallah, F., Erfani-Manesh, S., & Taki, E. (2015). Evaluation of Zenian and Avishan-e Shirazi antibacterial activity against Vibrio cholerae strains. Journal of Medical Bacteriology, 3 (1-2), 17-21.
57. Weinstein, R. A. (2001). Controlling antimicrobial resistance in hospitals: infection control and use of antibiotics. Emerging Infectious Diseases 7 (2), 188-192. http://doi.org/10.3201/eid0702.010206
58. Yadegar, A., Sattari, M., Bigdeli, M., & Bakhtiari, F. (2010). Evaluation and comparison of antibacterial effects of alcoholic extracts of Zataria multiflora Boiss. leaves, flowers and root on methicillin-resistant Staphylococcus aureus. Journal of Medicinal Plants, 9 (33), 58-65.
59. Zare Bidaki, M., Arab, M., Khazaei, M., Afkar, E., & Zardast, M. (2015). Anti-bacterial effect of Zataria multiflora Boiss. essential oil on eight gastrointestinal pathogenic species. Quarterly of the Horizon of Medical Sciences, 21 (3), 155-161. http://doi.org/10.18869/acadpub.hms.21.3.155