Bioactive Components Identification and Characterization of the Mechanisms of Action of Selected Medicinal Plant Extracts Against Salmonella enterica Serovar Typhi
Main Article Content
Abstract
The rise in antimicrobial resistant bacteria has necessitated the search for alternatives therapy, this study evaluates the antimicrobial potency of five medicinal plants against MDR S. Typhi. The leaves of Azadirachta indica, Bambusa vulgaris, Cymbopogon citratus, Moringa oleifera, Carica papaya, were collected, shed-dry, powdered, and soaked in ethanol for 72 hours, dried to generate the crude extract. The potency was tested on MDR S. Typhi isolated from Alheri laboratory. GCMS and Fourier Transform Infrared (FTIR) were performed to identify plant’s active components and functional group(s). Scanning Electron Microscope (SEM) and UV spectrophotometer were used to assay effects on bacterial membrane. MDR S. Typhi was isolated, confirmed through serological and molecular analysis. Both the invA and tviA genes were identified. Azadirachta indica showed the highest activity with a zone of inhibition of 19.6 ± 0.8 mm at 100 mg/mL. The MIC and MBC of A. indica was 50 mg/mL and 100 mg/mL, respectively. GCMS analysis identified cis-13-octadecenoic (19.63%), n-hexadecanoic (17.05%), octadecanoic (6.99%), trans-13-octadecenoic (3.74%) acids, as the major components. FTIR analysis confirmed the presence of hydroxyl, carbonyl, amine, ether, ester, and aromatic functional groups associated with antimicrobial phytochemicals such as phenols, flavonoids, alkaloids, and terpenoids. SEM analysis of treated bacterial cells showed morphological cell deformation. UV spectrophotometric analysis at 260 nm and 280 nm wavelength showed increased absorbance in treated cells, suggesting leakage of nucleic acids and proteins. Azadirachta indica possesses significant antibacterial activity against MDR S. Typhi, primarily through disruption of bacterial membrane integrity and leakage of intracellular contents.
Article Details

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
References
1. Abdalla WE, Abdallah EM. Antibacterial activity of ginger (Zingiber officinale Rosc.) rhizome: a mini review. Int J Pharmacogn Chinese Med. 2018;2(4):000142. https://doi.org/10.23880/ipcm-16000142
2. Chaachouay N, Zidane L. Plant-derived natural products: a source for drug discovery and development. Drugs Drug Candidates. 2024;3:184–207. https://www.scirp.org/reference/referencespapers?referenceid=3828824
3. Christenhusz MJM, Byng JW. The number of known plant species in the world and its annual increase. Phytotaxa. 2016;261(3):201–217. http://doi:10.11646/phytotaxa.261.3.1
4. Ssenku J.E., Okurut S.A., Namuli A., Kudamba A., Tugume P., Matovu P., Wasige G., Kafeero H.M., Walusansa A. Medicinal plant use, conservation, and associated traditional knowledge in rural communities in Eastern Uganda. Trop Med Health. 2022;50(1). http://doi:10.1186/s41182-022-00428-1
5. World Health Organization. Prioritization of pathogens to guide discovery, research and development of new antibiotics for drug-resistant bacterial infections, including tuberculosis. Geneva: WHO; 2017. (WHO/EMP/IAU/2017.12). https://www.who.int/publications/i/item/WHO-EMP-IAU-2017.12
6. Saganuwan SA. Competitive biodiversity of human and vascular plant species: implications for pharmaceutical industries, health and world economy part-2. Eur J Med Plants. 2016;16(1):1–29. https://doi.org/10.9734/EJMP/2016/27325
7. O’Neill J. Tackling drug-resistant infections globally: final report and recommendations. Arch Pharm Pract. 2016;7(3). http://doi:10.4103/2045-080x.186181
8. Zenoh DA, Josephus B, Halley N, Endurance O, Chukwuemeka H, Akumbo G. Evaluation of antimicrobial properties of five medicinal plants used against bacterial infections in Jalingo, Nigeria. Afr J Clin Exp Microbiol. 2024;25(2):219–226. http://doi:10.4314/ajcem.v25i2.13
9. Wong W, Reed C, Holt KE. The molecular epidemiology of quinolone-resistant Salmonella typhi in Asia. Lancet Infect Dis. 2015;15(10):1101–1108. https://doi.org/10.1038/ng.3281
10. Wain J, Gregson DJ, Aarestrup FM. Dihydrofolate reductase mutations and resistance to trimethoprim in Salmonella typhi. Int J Antimicrob Agents. 2016;48(5):465–468. https://doi.org/10.1016/j.ijantimicag.2016.06.005
11. Pant P, Pandey S, Dall’Acqua S. The influence of environmental conditions on secondary metabolites in medicinal plants: a literature review. Chem Biodivers. 2021;18(11):e2100345. https://doi.org/10.1002/cbdv.202100345
12. Vázquez-León L.A., Páramo-Calderón D.E., Robles-Olvera V.J., Valdés-Rodríguez O.A., Pérez-Vázquez A., García-Alvarado M.A., Rodríguez-Jimenes G.C. Variation in bioactive compounds and antiradical activity of Moringa oleifera leaves: influence of climatic factors, tree age, and soil parameters. Eur Food Res Technol. 2017; 243:1593–1608. https://doi.org/10.1007/s00217-017-2868-4
13. Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing. 33rd ed. CLSI supplement M100. 2023. https://clsi.org/standards/products/microbiology/documents/m100/
14. Cheesbrough M. District laboratory practice in tropical countries. 2nd ed. Cambridge: Cambridge University Press; 2006. https://doi.org/10.1017/CBO9780511543470
15. Forbes BA, Sahm DF, Weissfeld AS. Bailey & Scott’s diagnostic microbiology. 14th ed. Philadelphia: Elsevier; 2016. https://shop.elsevier.com/books/bailey-and-scotts-diagnostic-microbiology/tille/978-0-323-35482-0
16. Crump JA, Mintz ED. Global trends in typhoid and paratyphoid fever. Clin Infect Dis. 2010;50(2):241–246. http://doi:10.1086/649541
17. Ranjbar, R., Mortazavi, S. M., Tavana, A. M., Meysam Sarshar, Najafi, A., & Zanjani, R. S. Simultaneous Molecular Detection of Salmonella enterica Serovars typhi, Enteritidis, Infantis, and Typhimurium. Iranian J Public Health, (2017);46(1), 103–111. https://pmc.ncbi.nlm.nih.gov/articles/PMC5401918/?utm
18. Kumar S, Balakrishna K, Batra HV. Detection of Salmonella enterica serovar Typhi by selective amplification of invA, viaB, fliC-d and prt genes by PCR in multiplex format. Lett Appl Microbiol. 2006;42(2):149–154. http://doi:10.1111/j.1472-765x.2005.01813.x
19. Parkhill J., Dougan G., James K.D., Thomson N.R., Pickard D., Wain J., Churcher C., Mungall K.L., Bentley S.D., Holden M.T.G., Sebaihia M., Baker S., Basham D., Brooks K., Chillingworth T., Connerton P., Cronin A., Davis P., Davies R.M., Dowd L., White N., Farrar J., Feltwell T., Hamlin N., Haque A., Hien T.T., Holroyd S., Jagels K., Krogh A., Larsen T.S., Leather S., Moule S., Ó’Gaora P., Parry C., Quail M., Rutherford K., Simmonds M., Skelton J., Stevens K., Whitehead S., Barrell B.G. Complete genome sequence of a multiple drug-resistant Salmonella enterica serovar typhi CT18. Nature. 2001;413(6858):848–852. http://doi:10.1038/35101607
20. Marchello, C. S., Carr, S. D., & Crump, J. A. (2020). A Systematic Review on Antimicrobial Resistance among Salmonella typhi Worldwide. Am J Trop Med Hyg. 103(6), 2518–2527. https://doi.org/10.4269/ajtmh.20-0258
21. Klemm E.J., Shakoor S., Page A.J., Qamar F.N., Judge K., Saeed D.K., Wong V.K., Dallman T.J., Nair S., Baker S., Shaheen G., Qureshi S., Yousafzai M.T., Saleem M.K., Hasan Z., Dougan G., Hasan R. Emergence of an extensively drug-resistant Salmonella enterica serovar typhi clone. mBio. 2018;9(1):e00105-18. http://doi:10.1128/mbio.00105-18
22. World Health Organization. Antimicrobial resistance. Geneva: WHO; 2023. Available from: https://www.who.int/news-room/fact-sheets/detail/antimicrobial-resistance
23. Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev. 1999;12(4):564–582. https://doi.org/10.1128/CMR.12.4.564
24. Biswas K, Chattopadhyay I, Banerjee RK, Bandyopadhyay U. Biological activities and medicinal properties of neem (Azadirachta indica). Curr Sci. 2002;82(11):1336–1345. https://www.sciepub.com/reference/88916
25. Subapriya R, Nagini S. Medicinal properties of neem leaves: a review. Curr Med Chem Anti-Cancer Agents. 2005;5(2):149–156. http://doi:10.2174/1568011053174828
26. Do Q.D., Angkawijaya A.E., Tran-Nguyen P.L., Huynh L.H., Soetaredjo F.E., Ismadji S., Ju Y.-H. Effect of extraction solvent on total phenolic content, flavonoid content and antioxidant activity. J Food Drug Anal. 2014;22(3):296–302. http://doi:10.1016/j.jfda.2013.11.001
27. Akinyemi KO, Oladapo O, Okwara CE, Ibe CC, Fasure KA. Screening of crude extracts of medicinal plants for anti-MRSA activity. BMC Complement Altern Med. 2005; 5:6. http://doi:10.1186/1472-6882-5-6
28. Harborne JB. Phytochemical methods: a guide to modern techniques of plant analysis. 3rd ed. London: Chapman & Hall; 1998.
29. Desbois AP, Smith VJ. Antibacterial free fatty acids: activities and mechanisms of action. Appl Microbiol Biotechnol. 2009;85(6):1629–1642. http://doi:10.1007/s00253-009-2355-3
30. Kumar PP, Kumaravel S, Lalitha C. GC–MS study of Vitex negundo. Afr J Biochem Res. 2010;4(7):191–195. https://academicjournals.org/article/article1380110714_Kumar%20et%20al.pdf
31. Marchese A., Arciola C.R., Barbieri R., Silva A.S., Nabavi S.F., Sokeng A.J.T., Izadi M., Jafari N.J., Suntar I., Daglia M., Nabavi S.M. Update on monoterpenes as antimicrobial agents. Materials. 2017;10(8):947. http://doi:10.3390/ma10080947
32. Padmavathi AR, Abinaya B, Pandian SK. Antibiofilm activity of phenolic compound against Serratia marcescens. Biofouling. 2014;30(9):1111–1122. http://doi:10.1080/08927014.2014.972386
33. Adam M., Obi C., Zenoh D.A., Mshelia, M.B., Nuhu T., Alegbe S.D., Micheal N.Y. & Fasogbon, I.V. Antimicrobial Activity of Medicinal Plants as a Panacea for AntibioticResistance: A Systematic Review. Journal of Medicinal Herbs 2022:11-23. (DOI): 10.30495/MEDHERB.2022.698251
34. Pandey KB, Rizvi SI. Plant polyphenols as dietary antioxidants. Oxid Med Cell Longev. 2009;2(5):270–278. http://doi:10.4161/oxim.2.5.9498
35. Tiwari P., Kumar B., Kaur M., Kaur G., Kaur H. Phytochemical screening and extraction: a review. Int Pharm Sci. 2011;1(1):98–106. https://www.sciepub.com/reference/180569
36. Burt S. Essential oils: antibacterial properties and applications. Int J Food Microbiol. 2004;94(3):223–253. http://doi:10.1016/j.ijfoodmicro.2004.03.022
37. Ultee A, Bennik MHJ, Moezelaar R. Role of phenolic hydroxyl group of carvacrol against Bacillus cereus. Appl Environ Microbiol. 2002;68(4):1561–1568. http://doi:10.1128/AEM.68.4.1561-1568.2002
