Please enter verification code
In-Vitro Antimicrobial Activities and Phytochemical Screening of Calotropis Procera (Ait) and Vernonia Amygdalina (Del.) Extracts Against Some Medically Important Pathogenic Bacteria
American Journal of Bioscience and Bioengineering
Volume 6, Issue 6, December 2018, Pages: 42-55
Received: Jan. 10, 2019; Accepted: Feb. 27, 2019; Published: Mar. 22, 2019
Views 739      Downloads 135
Yirgashewa Asfere, Department of Biotechnology, Wolkite University, Gubrea, Ethiopia
Ameha Kebede, Department of Biology, Haramaya University, Haramaya, Ethiopia
Manikandan Muthuswamy, Department of Biology, Haramaya University, Haramaya, Ethiopia
Article Tools
Follow on us
The aim of this study is to evaluate phytochemical screening and antibacterial activities of crude extracts obtained from different parts of Calotropis procera and Vernonia amygdalina against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. The stems, roots and leaves of the selected plant species were shade dried and ground to powders and the bioactive components were extracted using ethanol (99.5%), methanol (99.8%), hexane (99.8%) and distilled water. The antibacterial activities of the resulting extracts against the three selected pathogens were evaluated using paper disc method and inhibitory zones were recorded in millimeters at five different concentrations (20, 30, 40, 50 and 60 mg/ml). Agar dilution method is used to determine minimum inhibitory concentrations (MICs) of the plant extracts against selected pathogens. Chloramphenicol and DMSO used as positive and negative controls, respectively. The bioassay results revealed that the crude extracts of ethanol, methanol, hexane and water had antibacterial activities on all three bacterial species at all concentration except at 20 mg/ml of all solvent extracts. Methanol and ethanol extracts had the highest growth inhibitory effects as compared with those of the aqueous and hexane crude extracts. However, the four solvent crude extracts had less antibacterial activities than chloramphenicol. S. aureus found to be the most susceptible pathogen to the crude ethanol (99.5%) and methanol (99.8) extracts of the leaves of Vernonia amygdalina (22 mg/ml) and ethanol extract of the leaves of Calotropis procera (22 mg/ml). Whereas Pseudomonas auruginosa was the least susceptible bacterium to crude ethanol extract (99.5%) of the root of Calotropis procera at 28 mg/ml and crude water, extract of the root of Vernonia amygdalina at 28 mg/ml. The growth inhibitory activities of the crude extracts were found to be significantly different for the four concentrations (30, 40, 50 and 60mg/ml) in both plant parts (p < 0.05). Phytochemical screening were done and the following bioactive components are detected such as Tannins, Phenolics, Resins, Amino acids, Flavonoids, Saponins, Reducing sugar, Glycosides, Steroids, Triterpenoids, Anthocyanidins, Sterol and Volatile Oil. In conclusion, this study did not only show the antibacterial activities of Calotropis procera and Vernonia amygdalina, but also offered a scientific validation for its traditional use against some diseases. There is a need for conducting more studies to identify and characterize the medicinal properties in the tested plant, which may serve as novel compounds for the development of new and more effective antimicrobial drugs.
Phytochemical Screening, Paper disc Diffusion, Antibacterial Activities, Minimum Inhibitory Concentration, C. Procera, V. Amaygdalina
To cite this article
Yirgashewa Asfere, Ameha Kebede, Manikandan Muthuswamy, In-Vitro Antimicrobial Activities and Phytochemical Screening of Calotropis Procera (Ait) and Vernonia Amygdalina (Del.) Extracts Against Some Medically Important Pathogenic Bacteria, American Journal of Bioscience and Bioengineering. Vol. 6, No. 6, 2018, pp. 42-55. doi: 10.11648/
Copyright © 2018 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.
Ghani, A. 2003. Medicinal Plants of Bangladesh. The Asiatic Society of Bangladesh, pp: 21-28.
Bibitha, B. J., V. K. Jisha, C. V, Salitha, S. Mohan and A. K. Valsa. 2002. Antibacterial activity of different plant extracts, short communication. Indian Journal of Microbiology. 42:361 363.
Ojo, O. O, I. O. Tella and O. O. Ademola Aremu. 2005. Effects of Azadiractha indica, Tamarindus indica and Euca-lyptus camaldulensis on Paracetamol Induced Lipid Peroxida-tion in Rats, Journal of Sustainable Development Agricultural Environment. 1:755-760.
Endashaw, B. 2007. Study on Actual Situation of Medicinal Plants in Ethiopia. Japan Association for International Collaboration of Agriculture and Forestry, 2: 1-9.
Yelne MB, Sharma PC, Dennis TJ. 2000. Database on Medicinal Plants Used in Ayurveda, Central Council For Research in Ayurveda and Siddha, Delhi; Vol. 2, 69-73.
Grace, BS. 2009. Calotropis Procera (Aiton) W. T. Aiton, In the Biology of Australian Weeds, Volume 3 (Ed. FD Panetta), RG and FJ Richardson, Melbourne.
Himalaya, 2002. Herbal Monograph, Himalaya Herbal Health care.
Ibrahim, NDG, EM. Abdurahman, G. Ibrahim. 2000. Histological Studies of the Effects of Chronic Feeding of Vernonia amygdalina Del Leaves on Rats. Nig. J. Surg. Res. 2: 68-74.
Egedigwe CA. 2010. Effect of dietary incorporation of Vernonia amygdalina and Vernonia colorata on blood lipid profile and relative organ weights in albino rats (Thesis). Department of Biochemistry, MOUAU and Nigeria.
Argheore, E. M., H. P. S. Makkar, K. Becker. 2000. Feed Value of Some Browse Plants From the Central Zone of Delta State Nigeria. Tropical Science 38(2), pp. 97 – 104.
Singh, K. N. and R. Kaushal. 2007. Comprehensive Notes on Commercial Utilization, Characteristics and Status of Steroid Yielding Plants in India. Ethno botanical Leaflets 11: 45-51.
Cheesbrough, M. 2002. “District Laboratory practice in Tropical countries”, Cambridge University Press London 2: 137-140.
Newtons, S. M., C. Lau, S. S. Gurcha, G. S. Besra and C. W. Wright. 2002. The evolution of forty- three plant species for invitro antimicrobial activities; isolation active constituents’ from psoraleacorylifolia and sanguinaria Canadensis. Journal of ethno pharmacology.79 1:57-67.
Amit, P., S. Agrawal, A. K. Bhatia, A. Saxena. 2015. “In-vitro Assesment of Antibacterial Activity of Calotropis Procera and Coriandrum Sativum against Various Pathogens”. Int. J. Of Pharm. Res. & All. Sci.; 4(1):33-44.
Dewanjee, S., M. Kundu, A. Maiti, R. Mjumdar, A. Majumdar and S. C. Mandal. 2007. In-vitro Evaluation of Antimicrobial Activity of Crude Extract from Plants Diospyros peregrine, Coccinia grandis and Swietenia macrophylla. Tropical Journal of Pharmaceutical research. 6(3):773-778.
De Flora S, Izzotti A, 2007. Mutagenesis and Cardiovascular diseases: molecular mechanisms, risk factors, and protective factors. Mut. Res., 621: 5-17.
Ronald, M. A. 1995. Microorganisms in our world. MosbyYear Book, Inc.St.louis.765pp.
Arekemase. 2013. Assessment of Bitter Leaf (Vernonia amygdalina) on Some Selected Pathogenic Microorganisms (5) 2360-2365.
Estimone, C. O., M. U. Adikwu and J. M. Okunta. 1998. Preliminary antimicrobial screening of the ethanolic extract from the lichen Usneasubfloridans L. Journal of Pharmaceutical Research and Development. 32: 99-101.
Obeidat. M, Shatnawi. M, Al-alawi. M, Al-Zu`bi. E. Al-Dmoor, H. AlQudah. M, El-Qudah and I. Otri. 2012. Antimicrobial Activity of Crude Extracts of Some Plant Leaves. Research Journal of Microbiology, Vol.7:59-67.
Hailu, T., M. Endris, A. Kaleab, G. Tsige. 2005. Antimicrobial Activities of some Selecte Traditional Ethiopian Medicinal Plants Used in the Treatment of Skin Disorders. Journal of Ethnopharmacology 100.168–175.
Baeur RW, MDK Kirby, JC Sherris, M Turk 1996. Antibiotic Susceptibility Testing by Standard Single Disc Diffusion Method. Am. Clin. Pathol. 45:493-496.
Brain, K. R. and Tuner, T. D. (1975): The practical evaluation of phytopharmaceuticals. Wright Scientectica Publishers, Bristol. Pp57-58.
Cuilel, I. (1994): Methodology for the analysis of vegetables and drugs. Chemical Industry Division, NNIDO Romania. Pp24-67.
Sofowora, A. (1993): Medicinal plants and traditional medicine in Africa. Spectrum books limited, Ibadan, Nigeria. Pp220-224.
Kesatebrhan H. A. 2013. Antimicrobial Activity and Phytochemical Screening of Crude Extracts of Medicinal Plants Grown in Eastern Ethiopia Int J Pharm Bio Sci 2013 Oct; 4(4): (P) 326 – 333.
Cowan, M. M.1999. Plant Products as Antimicrobial Agents. Clin. Microbial Rev., 12(4):564-582.
Wynn, S. G. and Fougere, B. 2006. Veterinary Herbal Medicine. Elsevier Health Sciences.
Preethi, RM, V. V, Devanathan, M. Loganathan. 2010. Antimicrobial and Antioxidant Efficacy of Some Medicinal Plants against Food Borne Pathogens. Advances in Biological Research., 4:122- 125.
Cvetnic and K. S. Vladmir. 2004. Antimicrobial activity of grapefruit seed and pulp ethanol extract. Acta Pharmaceutical, 54(3):243-250.
Kawo, AH, A, Mustapha, BA, Abdullahi, LD, Rogo ZA., Gaiya and AS. Kumurya. 2009. Phytochemical properties and antibacterial activities of the leaf and latex extracts.
Kareem SO, I., Akpan, OP. Ojo. 2008. Antimicrobial Activities of Calotropis procera (Ait.) R. Br. Flowers.
Bhaskar VH. Antimicrobial Activity of Calotropis procera Seeds. Asian journal of chemistry, 2000, 21(7): 5788-5790.
Calixto, J. B. 2000. Efficacy, safety, quality control, marketing and regulatory guidelines for herbal medicines (Phytotherapeutic agents). Brazil Journal Medical Biologica Research, 33(2), 19–189.
Okigbo, R. N., O. D., Omodamiro. 2006. Antimicrobial effects of leaf extracts of pigeon pea (Cajanuscajan (L.) Mill sp) on some human pathogens. Journal of Herbs Spices of Medicinal Plants, 12 (1 & 2), 117–127 on Selected Pathogenic Microorganisms. African Journal of Biomedical Research.
Ijeh, II and A. T. Adedokun. 2006. Effects of administration of ethanolic extract of Vernonia amygdalina Del. on kidney function of experimental 69: 793-825. Rabbit model. Res. J. Biotech., 1: 34-35.
Okigbo, R. N., D. I., IGWE. 2007. The antimicrobial effects of Piper guineense ‘uziza’ and Phyllantusamarus ‘ebe- benizo’ on Candida albicans and Streptococcus faecalis. Acta Microbiologicaet Immunologica Hungarica, 54(4), 353–366.
Gotep, J. G., G. O. Agada, D. S. Gbise and S. Chollom. 2010. Antibacterial activity of ethanolic extract of Acalypha wilkesiana leaves growing in Jos, Plateau State, Nigeria. Malaysian Journal of Microbiology, 6(2): 69-74.
Lu, Y., Joerger, R. and Wu, C. 2011. Study of the chemical composition and antimicrobial activities of ethanolic extracts from roots of scutellaria baicalensis Georgi. Journal of Agriculture and food chemistry, 59(20): 10934-10942. DOI:10.1021/jf202741x.
Meher, B. R., S. Mahar, B. G. Rath and S. K. Sahoo. 2013. Antimicrobial activities of ethanolic extracts of leaves of sphaeranthus inducus. Der Pharmacia Lettre, 5(1):8-10.
Wojtyczka, R. D., M., Kepa, D., Idizik, R., Kubina, A., Kabala-Dzik, A. Dziedzic, and T. J. Wasik. 2013. In Vitro Antimicrobial Activity of Ethanolic Extract of Polish Propolis against Biofilm Forming Staphylococcus epidermidis Strains. Evidence-Based Complementary and Alternative Medicine, 2013: 1-12.
Igbinosa, O. O., E. O. Igbinosa and O. A. Aiyegoro. 2009. Antibacterial activity and phytochemical screening of stem bark extract from Jatropha curcas Linn. Afr. J. of pharmacy and pharmacology. Vol 32:058-062.
Oyewole, A. O., Audu, O. T. and Amupitan, J. O. (2004): A survey of chemical constituents and biological activities of some medicinal plants. Journal of Chemical Society of Nigeria 4:162165.
Rucker, G., Kehrbaum, S., Salakus, H., Lawong, B. and Goeltenboth, F. (1992): Acetylenic glucosides from aerial parts of Microglossa pyrifolia. Planta Medica 58:266-269.
Murakami, A. H., Ohigashi, S., Tanaka, A., Tatematsu, T and Koshimi, Z. W. (1993): Bitter cyanoglucosides from Laphira alata. Phytochemistry 32:1461-1466.
Trease, G. E. and Evans, W. C. (1989): Pharmacognosy. 1 st edition. Ahmadu Bello University press Ltd., Zaria, Nigeria. Pp188-198.
Afolayan, A. J. and Meyer, J. J. M. (1997): The antimicrobial activity of 3,5,7 trihydroxyflavone isolate from the shoot of Helichrysum aureonites. Journal of Ethnopharmacology 57:177-181.
Goodman A, Culman A, Geodman L (1980). Goodman and Gilmans Pharmacological Basic of Therapeutic Microbiology (6 th edition) MacMillian Publishing Co. Inc. New York pp.1080-1085.
Kaufman T, Kalderon N, Ullmann Y, Berger J (1988). J. Burn Care Rehabil. 9(2):156-159.
Dutta AC (1993). Botany for Degree Students. Fifth edition. Oxford University Press, Oxford pp.810-844.
Hodek, P., Trefil, P., Stiborova, M 2002. Flavonoids potent and versatile biologically active compounds interacting with cytochrom P450, chemical- Biol. Intern. 131:1-21.
Parekh, J., Chanda, S. 2007. Invitro antibacterial activities of crude methanol extract of wood Fordia Fruticosa Kurtz flower cythacease. Braz. J. Microbial 38:2.
Adegboye, M. F., Akinpelu, D. A and Okoli, A. I 2008. The bioactive and phytochemical properties of Garnicia Kola Hackel seed extract on some pathogens Afr. J. of Biotechnol. 721:3934-3938.
Adegboye, M. F., Akinpelu, D. A and Okoli, A. I 2008. The bioactive and phytochemical properties of Garnicia Kola Hackel seed extract on some pathogens Afr. J. of Biotechnol. 721:3934-3938.
Shimada, T 2006. Salivary protein as a defense against dietery tannins. J. chem. Ecol. 322:1149- 1163.
Dharmananda, S. 2003. Allnuts and the uses of tannins in Chinese medicine in proceedings of institutes for traditional medicine port. 3938 land Oregin. Afr. J. Biotechnol.
Li, H., Wang; Z., Liu, Y. 2003. Reviewed in the studies on tannins activities of cancer prevention and anticancer. Zhong-Yaoca. 266:444-448.
Okwu, D. E. 2001. Evaluation of the chemical composition of medicinal plant belonging to euphorbiacaec. Pak. Vet. S. 14:160.
Tripathi R. 2017. Antimicrobial investigation of some arid zone plants of Rajasthan. IJPSR; eight (11):4848-51.
Sharma SK, Sharma R. 2016. Essential oil constituents and Antimicrobial potency of Thuja orientalis grown in Rajasthan. Int. J. of Environment, Science and Technology; 2(2):1-7.
Verma M, Kumar A. 2017. Antimicrobial activity of some folk medicinal plants used in Rajasthan against selected pathogenic microorganisms. Int. J Curr. Pharm. Res.; 9(3):84-86.
Arora A, Matiya J. 2017. Quantitative ethno-medicinal studies of wound healing plants with reference to their hypoglycemic potential. Advances in Pharmacology and Toxicology 18(2):43-52.
Science Publishing Group
1 Rockefeller Plaza,
10th and 11th Floors,
New York, NY 10020
Tel: (001)347-983-5186