European Journal of Clinical and Biomedical Sciences

Submit a Manuscript

Publishing with us to make your research visible to the widest possible audience.

Propose a Special Issue

Building a community of authors and readers to discuss the latest research and develop new ideas.

Combined Antibacterial Effect of Croton macrostachyus, Calpurina aurea and Ocimum gratissimum Against Selected Clinical and Standard Pathogenic Bacteria

Background: The use of medicinal plants as treatment options of human and animal diseases can be traced back in human history, and about ten percent of identified medicinal plants serve a pharmaceutical role because they have active chemical constituents such phenolic acids, flavonoids, tannins and lignin. Ethiopia is a place rich in medicinal plants, though most studies in the region have only considered the individual effects of their extracts while under-exploring their combined effects. Objective: The objective of this research was to assess the synergistic antibacterial activity of crude extracts of leaves of Croton macrostachyus, Calpurnia aurea and Ocimum gratissimum collected from Bahir Dar town against standard and clinical isolates of Staphylococcus aureus, Escherichia coli and Salmonella typhi by using methanol, acetone and chloroform as solvents. Methods: Extracts were prepared at a plant-to-solvent ratio of 50 g to 500 mL and then set to a concentration of 50 mg/mL by dissolving 100 mg of crude extract in 2 ml of 10% dimethyl sulfoxide in small cups, from which 100 µL was used for antibacterial assays using the disc diffusion method. Minimum inhibitory and bactericidal concentration assays were assessed with the broth microdilution and overnight bacterial culture preparation techniques, respectively. The fractional inhibitory concentration index was used for synergistic activity analysis. Results: Combinations of extracts showed relatively better effects against most test bacteria with inhibition zones reaching up to 23.00 ± 1.00 mm (Salmonella typhi) despite limited activity on both standard and clinical isolates of Escherichia coli. The lowermost minimum inhibitory and bactericidal concentrations were 3.125 mg/mL and 6.25 mg/mL, respectively, and a few synergistic and many additive effects were recorded for different forms of combinations on different bacterial isolates. Conclusion: The combined use of extracts is relatively promising, though further work is required to clearly set the safety margins of combinations used in vivo, as this is the first report on all settings used here. The findings of this study provide scientific evidence for communities, pharmaceutical industries, and other concerned bodies regarding alternative formulations of phytochemicals for the relief of different physiological deviations, with the combined use of plants showing better performance.

Antibacterial Effect, Calpurina Aurea, Croton Macrostachyus, Medicinal Plant, Ocimum Gratissimum

Ayichew Teshale, Nega Berhane, Wagaw Sendeku. (2023). Combined Antibacterial Effect of Croton macrostachyus, Calpurina aurea and Ocimum gratissimum Against Selected Clinical and Standard Pathogenic Bacteria. European Journal of Clinical and Biomedical Sciences, 9(3), 38-46.

Copyright © 2023 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License ( which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

1. Firenzuoli, F.; Gori, L. Herbal medicine today: Clinical and research issues. Evid. Based Complement. Altern. Med. 2007, 4 (Suppl. 1), 37–40.
2. Sahoo, S. K.; Labhasetwar, V. Nanotech approaches to drug delivery and imaging. Drug Discov. Today 2003, 8, 1112–1120.
3. Rasool Hassan, B. A. Medicinal Plants (Importance and Uses). Pharm. Anal. Acta 2012, 3, 2153–2435.
4. Jamshidi-Kia, F.; Lorigooini, Z.; Amini-Khoei, H. Medicinal plants: Past history and future perspective. J. Herbmed Pharmacol. 2018, 7, 1–7.
5. Halberstein, R. A. Medicinal Plants: Historical and Cross-Cultural Usage Patterns. Ann. Epidemiol. 2005, 15, 686–699.
6. Huang, H. Plant diversity and conservation in China: Planning a strategic bioresource for a sustainable future. Bot. J. Linn. Soc. 2011, 166, 282–300.
7. Rafieian-Kopaei, M. Medicinal plants and the human needs. J. Herbmed Pharmacol. 2012, 1, 1–12.
8. Verma, S. P. S. S.; Singh, S. Current and future status of herbal medicines. Veter. World 2008, 2, 347–350.
9. Phillipson, J. Phytochemistry and medicinal plants. Phytochemistry 2001, 56, 237–243.
10. World Health Organization. The world health report 2002: reducing risks, promoting healthy life. World Health Organization, 2002.
11. Cowan, M. M. Plant Products as Antimicrobial Agents. Clin. Microbiol. Rev. 1999, 12, 564–582.
12. Auwal, M.; Mairiga, I.; Shuaibu, A.; Ibrahim, A.; Gulani, I.; Wampana, B. Preliminary phytochemical and in vitro antibacterial evaluation of the crude pericarp extract of Hyphaene thebaica (doumpalm). J. Med. Plant Herb. Ther. Res. 2013, 1, 1–7.
13. Jack, D. B. One hundred years of aspirin. Lancet 1997, 350, 437–439.
14. Alviano, D. S.; Alviano, C. S. Plant Extracts: Search for New Alternatives to Treat Microbial Diseases. Curr. Pharm. Biotechnol. 2009, 10, 106–121.
15. Gardam, M. A. Is methicillin-resistant Staphylococcus aureus an emerging community pathogen? A review of the literature. The Canadian journal of infectious diseases. Can. J. Infect. Dis. Med. Microbiol. 2000, 11, 202–211.
16. Woldeab, B.; Regassa, R.; Alemu, T.; Megersa, M. Medicinal Plants Used for Treatment of Diarrhoeal Related Diseases in Ethiopia. Evid.-Based Complement. Altern. Med. 2018, 2018, 1–20.
17. Bekele, E. Study on Actual Situation of Medicinal Plants in Ethiopia; JAICAF: Tokyo, Japan, 2008.
18. Balunas, M. J.; Kinghorn, A. D. Drug discovery from medicinal plants. Life Sci. 2005, 78, 431–441.
19. Teklehaymanot, T.; Giday, M. Ethnobotanical study of medicinal plants used by people in Zegie Peninsula, Northwestern Ethiopia. J. Ethnobiol. Ethnomedicine 2007, 3, 12–12.
20. Deo, S.; Utane, R.; Khubalkar, R.; Thombre, S. Extraction and isolation, synthesis, physiological activity of 1-phenyl naphthalene and its derivatives: A review. Pharma Innov. 2017, 6 (Pt A), 21–30.
21. Dog, T. L. Smart Talk on Supplements and Botanicals: Flor de Jamaica—Herbal Diuretics—Hawthorn. Altern. Complement. Ther. 2009, 15, 214–215.
22. Eloff, J. N. Which extractant should be used for the screening and isolation of antimicrobial components from plants? J. Ethnopharmacol. 1998, 60, 1–8.
23. Minale, A. S.; Alemu, K. Mapping malaria risk using geographic information systems and remote sensing: The case of Bahir Dar City, Ethiopia. Geospat. Health 2018, 13, 660.
24. Central Statistical Agency. Census on Ethiopian Population Growth; Central Statistical Agency: Addis Ababa, Ethiopia, 2013; Volume 1, pp. 39–41.
25. Edwards, S.; Tadesse, M.; Demissew, S.; Hedberg, I. (Eds.) Flora of Ethiopia and Eritrea. Magnoliaceae to Flacourtiaceae; The National Herbarium: Addis Ababa, Ethiopia; Department of Systematic Botany: Uppsala, Sweden, 2000; Volume 2, Issue Part 1.
26. Adam, S.; Salih, S.; Abdelgadir, W. In vitro Antimicrobial Assessment of Lepidium sativum L. Seeds Extracts. Asian J. Med. Sci. 2011, 3, 261–266.
27. Obeidat, M.; Shatnawi, M.; Al-Alawi, M.; Al-Zu`bi, E.; Al-Dmoor, H.; Al-Qudah, M.; El-Qudah, J.; Otri, I. Antimicrobial Activity of Crude Extracts of Some Plant Leaves. Res. J. Microbiol. 2012, 7, 59–67.
28. Ericsson, H.; Hogman, C.; Wickman, K. A paper disk method for determination of bacterial sensitivity to chemotherapeutic and antibiotic agents. Scand. J. Clin. Lab. Investig. 1954, 6 (Suppl. 11), 23–36.
29. Taye, B.; Giday, M.; Animut, A.; Seid, J. Antibacterial activities of selected medicinal plants in traditional treatment of human wounds in Ethiopia. Asian Pac. J. Trop. Biomed. 2011, 1, 370–375.
30. Balows, A.; Hausler, W. J., Jr.; Herrmann, K. L.; Isenberg, H. D. Manual of Clinical Microbiology. Rev. Inst. De Med. Trop. Sao Paulo 1991, 33, 434.
31. Mueller, M. S.; Mechler, E. Medicinal Plants in Tropical Countries: Traditional Use-Experience-Facts; Thieme: Stuttgart, Germany; New York, NY, USA, 2005.
32. Stefanović, O. D. Synergistic Activity of Antibiotics and Bioactive Plant Extracts: A Study Against Gram-Positive and Gram-Negative Bacteria. In Bacterial Pathogenesis and Antibacterial Control; Sahra, S., Ed.; Intech Open: London, UK, 2017.
33. Satish, K. P.; Moellering, R. C.; Eliopoulos, G. M. Antimicrobial Combinations, In Antibiotics in Laboratory Medicine, 5th ed.; Lorian, V., Ed.; Lippincott Williams &Wilkins: Philadelphia, PA, USA, 2005; pp. 290–365. ISBN 0-7817-4983-2.
34. Firomsa, W.; Tamirat, A.; Tekle, B.; Tesfaye, B.; Shafi, H.; Suresh, A. Bactericidal action of Croton macrostachyus leaf extract against common human pathogenic bacteria. J. Med. Plants Stud. 2018, 6, 33–36.
35. Sendeku W, Alefew B, Mengiste D, Seifu K, Girma S, Wondimu E, Bekuma G, Verma D, Berhane N. Antibacterial activity of Croton macrostachyus against some selected pathogenic bacteria. Biotechnology International. 2015; 8 (1): 11-20.
36. Umer, S.; Tekewe, A.; Kebede, N. Antidiarrhoeal and antimicrobial activity of Calpurnia aurea leaf extract. BMC Complement. Altern. Med. 2013, 13, 21.
37. Birhan, M.; Tessema, T.; Kenubih, A.; Yayeh, M. In Vitro Antimicrobial Evaluation of C aurea. Asian J Med Pharm Res. 2018, 8, 33–43.
38. Amabye, T. G.; Mussa, S. In Vitro Antimicrobial Efficacy of Fractions from Demakese (Ocimum lamifolium) Leaves Extract from Mekelle Tigray, Ethiopia. Nat. Prod. Chem. Res. 2015, 3, 1000196.
39. Melese, A.; Dobo, B.; Mikru, A. Antibacterial activities of Calpurnia aurea and Ocimum lamiifolium extracts against selected gram positive and gram-negative bacteria. Ethiop. J. Sci. Technol. 2019, 12, 203–220.
40. Jasuja, N. D.; Choudhary, J.; Sharama, P.; Sharma, N.; Joshi, S. C. A Review on Bioactive Compounds and Medicinal Uses of Commiphora mukul. J. Plant Sci. 2012, 7, 113–137.