In Vitro Antibacterial Activity, Preliminary Phytochemical Screening Profile, and in Vivo Toxicity of Seven Traditional Medicinal Plants in Ethiopia
Abstract
The threat of antibiotic-resistance calls for novel antibacterial agents. This study was aimed at screening medicinal plants for their antibacterial properties, phytochemical content and safety. Leaves of Allophylus abyssinicus (Hochst.) Radlk., Dicliptera laxata C.B.Clarke, Ligustrum vulgare L., Solanecio gigas (Vatke) c. Jeffrey and Gymnanthemum myrianthum (Hook.f.) H.Rob.; leaf and stem-bark of Olinia rochetiana A. Juss. and the seed of Cucurbita pepo L. were used. Chloroform and ethanol were used to extract G. myrianthum, D. laxata and O. rochetiana; ethyl acetate and methanol for the rest, and water for all. The extracts were tested against clinical/standard strains of Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi and Staphylococcus aureus by the agar-diffusion method. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were determined. Acute toxicity to mice was checked and preliminary phytochemical screening was done. Thirteen extracts, out of 24, were active (inhibition zone >7 mm) at differing levels (9.67±0.33-25.66±0.57 mm) against at least one bacterial strain. The MICs and MBCs were 1.95-15.6 mg/mL and 7.8-125 mg/mL respectively. The aqueous extract of S. gigas, methanol extracts of L. vulgare and A. abyssinicus, and ethanol extract of O. rochetiana leaf were the most active (MIC 1.95mg/ml) against S. aureus. Ethyl acetate extracts of A. abyssinicus, L. vulgare and S. gigas; aqueous of C. pepo, O. rochetiana and G. myrianthum; and all D. laxata had no antibacterial activity. P. aeruginosa was the least susceptible to any extract, although the methanol and aqueous extracts of S. gigas performed better against it. Preliminary phytochemical screening of selected extracts for phenols, flavonoids, tannins, steroids, terpenoids, steroidal glycosides, alkaloids, saponins, resins and glycosides showed positivity at least for four of these phytochemicals with glycoside and terpenoids in nearly all extracts and resin in none. The plants were not toxic to mice at 2000 mg/kg. Further consideration of S. gigas, L. vulgare, A. abyssinicus and O. rochetiana is recommended in light of their promising potential and safety.
Fauci AS, Morens DM. The perpetual challenge of infectious diseases. N Engl J Med 2012;366:454-461.
Fletcher SM, Mclaws ML, Ellis JT. Prevalence of gastrointestinal pathogens in developed and developing countries: systematic review and meta-analysis. J Pub Health Research 2013;2:42-53.
Amani J, Mirhosseini SA, Fooladi AA. A review approaches to identify enteric bacterial pathogens. Jundishapur J Microbiol 2015;8:e17473.
Rojas-Lopez M, Monterio R, Pizza M, Desvaux M, Rosini R. Intestinal pathogenic Escherichia coli: insights for vaccine development. Frontier Microbiol 2018;9:440.
Als D, Radhakrishnan A, Arora P, Gaffey MF, Campisi S, et al. Global trends in typhoidal salmonellosis: A systematic review. Am J Trop Med Hyg 2018;99:10-19.
Zhoua Z, McCann A, Weill FX, Blin C, Nair S, et al. Transient darwinian selection in salmonella enteric serovar paratyphi a during 450 years of global spread of enteric fever. PNAS 2014;111:12199-12204.
Khan HA, Ahmad A, Mehboob. Nosocomial infection and their control strategy. Asian Pac Trop Biomed 2017;5:509-514.
Vitkauskiene A, Skrodeniene E, Dambrauskiene A, Macas A, Sakalauskas R. Pseudomonas aeruginosa bacteremia: resistance to antibiotics, risk factors, and patient mortality. Medicina (Kaunas) 2010;46:490-495.
Tong SY, Davis JS, Eichenberger E, Holland T, Fowler VG. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev 2015;28:603-661.
Tadesse DA, Zhao S, Tong E, Ayers S, Singh A, et al. Antimicrobial drug resistance in Escherichia coli from humans and food animals, United States, 1950-2002. Emerg Infect Dis 2012;8:741-749.
European Centre for Disease Prevention and Control. Antimicrobial resistance surveillance in Europe Annual report of the European Antimicrobial Resistance Surveillance Network, 2011. https://www.ecdc.europa.eu/en/publications-data/antimicrobial-resistance-surveillance-europe-2011
Ayukekbong JA, Ntemgwa M, Atabe AN. The threat of antimicrobial resistance in developing countries: causes and control strategies. BMC Antimicrob Resist Infect Control 2017;6:47.
WHO. Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics. World Health Organization, Geneva, Switzerland 2017. https://www.who.int/medicines/publications/global-priority-list-antibiotic-resistant-bacteria/en/
Fabricant DS, Farnsworth NR. The value of plants used in traditional medicine for drug discovery. Rev Environ Health 2001;109:69-75.
Yesuf A, Asres K. Wound healing and anti-inflammatory properties of Allophylus abyssinicus (Hochst.) Radlk. Phytopharm 2013;4:442-453.
Czerwinska ME, Granica S, Kiss AK. Effects of an aqueous extract from leaves of Ligustrum vulgare on mediators of inflammation in a human neutrophils model. Planta Med 2013;79:924-932.
Nagy M, Krizková L, Mucaji P, Kontseková Z, Sersen F, et al. Antimutagenic activity and radical scavenging activity of water infusions and phenolics from ligustrum plants leaves. Molecules 2009;14:509-518.
Kiss AK, Mank M, Melzig MF. Dual inhibition of metallopeptidases ACE and NEP by extracts, and iridoids from Ligustrum vulgare L. J Ethnopharm 2008;120:220-225.
Brima EI. Toxic elements in different medicinal plants and the impact on human health. Int J Environ Res Pub Health 2017;14:1209.
Ernst E. Toxic heavy metals and undeclared drugs in Asian herbal medicines. Trends Pharm Sci 2002;23:136-139.
Robber, Huxtable RJ. The myth of beneficent nature: The risks of herbal preparations. Ann Intern Med 1992;117:165-166.
Harborne JB. Phytochemical Methods. A Guide to Modern Techniques of Plant Analysis. 3rd ed. Chapman and Hall Ltd, London 1973; pp 5-107.
CLSI. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard. 10th ed. Clinical and Laboratory Standards Institute USA 2015.
McFarland J. The nephelometer: an instrument for estimating the number of bacteria in suspensions for calculating the opsonic index and vaccines. J Am Med Assoc 1907;49:1176.
WHO. General Guidelines for Methodologies on Research and Evaluation of Traditional Medicine WHO/EDM/TRM/2000.1, Geneva. Switzerland 2000.
Andrews JM. Determination of minimum inhibitory concentrations. J Antimicrobial Chemother 2001;48:5-16.
Rios JL, Racio MC, Villar A. Screening methods for natural products with antimicrobial activity: A review of the literature. J Ethnopharmacol 1988;127-149.
Committee for the Update of the Guide for the Care and Use of Laboratory Animals. Institute for Laboratory Animal Research. Division on Earth and Life Studies. Guide for the Care and Use of Laboratory Animals. Eighth edition. The National Academies Press. Washington DC 2011. www.national-academies.org
OECD. Organization for Economic Cooperation and Development. Guideline for testing of chemicals. 2001.
Kelly AS, Nehrenberg LD, Hua K, Garland T, Pomp D. Exercise, weight loss, and changes in body composition in mice: phenotypic relationships and genetic architecture. Physiol Genomics 2011;43:199-212.
https://www.fitwatch.com/calculator/weight-loss-percentage
Bull BS, Koepke JA, Simson E, Van-Assendelft OW. Procedure for determining packed cell volume by the micro hematocrit method. Approved standard. 3rd ed. Clinical and Laboratory Standards Institute 2000.
Anibijuwon II, Oladejo BO, Adetitun DO, Kolawole OM. Antimicrobial activities of Vernonia amygdalina against oral microbes. G J Pharmacol 2012;6:178-185.
Orebo T. Phytochemical investigation and antimicrobial activities of extracts of vernonia auriculifera heir leaves [MSc thesis]. Haromaya University, Haromaya, Ethiopia 2015.
Borges A, Abreu AC, Dias C, Saavedra MJ, Borges F, et al. New perspectives on the use of phytochemicals as an emergent strategy to control bacterial infections including biofilms. Molecules 2016;21:877.
Chung KT, Wong YT, Huang YW, Lin Y. Tannins and human health: a review. Crit Rev Food Sci Nutr 1998;38:421-464.
Haslaam E. Natural polyphenols (vegetable tannins) as drugs: possible modes of action. J Nat Prod 1996;59:205-215.
Wang X, Bunkers G. Potent heterologous antifungal proteins from cheese weed (Malvapar viflora). Biochem Biophys Res Commn 2000;279:669-673.
Kazmi M, Malik A, Hameed S, Akhtar N, Noor AS. An anthraquinone derivative from Cassia italic. Phytochemistry 1994;36:761-763.
Ulubelen A, Topcu G, Eriş C, Sönmez U, Kartal M, et al. Terpenoids from salvia sclerea. Phytochemistry 1994;36:971-974.
Chonoko UG, Rufai AB. Phytochemical screening and antibacterial activity of cucurbita pepo (pumpkin) against Staphylococcus aureus and salmonella typhi. Bajop S 2011;4:145-147.
Umadevi P, Murugan S, Suganthi S, Subakanmani S. Evaluation of antidepressant-like activity of Cucurbita pepo seed extracts in rats. Int J Curr Pharm Res 2011;3:108-113.
Asres K, Sporer F, Wink M. Identification and quantification of hepatotoxic pyrrolizidine alkaloids in the Ethiopian medicinal plant solanecio gigas (asteraceae). Pharmazie 2007;62:709-713.
Bello OA, Ayanda OI, Aworunse OS, Olukanmi BI, Soladoye MO, et al. Solanecio biafrae: an underutilized nutraceutically-important African indigenous veritable. Phcog Rev 2018;12:128-132.
Tadege H, Mohamad E, Asres K, Gebre-Mariam T. Antimicrobial activities of some selected traditional ethiopian medicinal plants used in the treatment of skin disorders. J Ethnopharmacol 2005;100:168-175.
Amuka O, Mulie J, Jeruto P. Intrinsic biological activities of crude methanol extract of O. rochetiana A. Juss. Eur J Pharm Sci 2015;2:83-89.
Chavan RB, Gaikwad DK. The ethnobotany, photochemistry and biological properties of Allophylus species used in traditional medicine. W J Pharm Pharm Sci 2016;5:664-682.
Mishra MP, Padhy RN. In vitro antibacterial efficacy of 21 Indian timber-yielding plants against multidrug-resistant bacteria causing urinary tract infection. Osong Public Health Res Perspect 2013;4:347-357.
Mulu W, Abera B, Yimer M, Hailu T, Ayele H, et al. Bacterial agents and antibiotic resistance profiles of infections from different sites that occurred among patients at debre markos referral hospital, Ethiopia: a cross-sectional study. BMC Res Notes 2017;10:254.
Ibrahim SE, Yuan Q, Ahmed IF, Khalil O, Osman EB. In vitro antimicrobial activity of summer squash (Cucurbita pepo L.). University of Africa Journal of Science 2010;1:77-89.
Abd EI-Aziz AB, Abd EI-Kalek HH. Antimicrobial proteins and oil seeds from pumpkin (Cucurbita moschata). Nat Sci 2011;9:105-119.
Barbieri R, Coppo E, Marchese A, Daglia M, Sobarzo-Sánchez E, et al. Phytochemicals for human disease: An update on plant-derived compounds antibacterial activity. Microbiol Res 2017;196:44-46.
Saxena J, Nema R, Singh D, Gupta A. Phytochemistry of medicinal plants. J Pharmacogn and Photochem 2013;1:168-182.
Muguweru FG, Nyamai DW, Arika MW, Ngugi MP, Gathumbi PK, et al. Antimicrobial activity of aqueous extracts of maytemus putterlickoides, senna spectabilis and olinia usambarensis on selected diarrhea causing bacteria. J Bacteriol Parasitol 2016;7:2155-2159.
Bode AM, Dong Z. Toxic phytochemicals and their potential risks for human cancer. Cancer Prev Res 2015;8:1-8.
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| Issue | Vol 6, No 4, 2021 | |
| Section | Research Article(s) | |
| DOI | https://doi.org/10.18502/tim.v6i4.8273 | |
| Keywords | ||
| Antibiotic-resistance Antibacterial agent Minimum inhibitory concentration (MIC) Minimal bactericidal concentration (MBC) Agar-diffusion | ||
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