Antifungal Activities of Azardirachta indica Crude Extracts and Fractions against Strain 1161, P37005 and RM1000

Main Article Content

A. D. M. Owoyale
M. Galadimma
S. Y. Daniyan
N. Adabara

Abstract

Azardirachta indica (neem) has served as an alternative medicine in the treatment of some common illnesses that have being associated to man. Azardirachta indica have proven effectively against certain fungi species that can infect human beings and cause disease. This study was aimed at determining the antifungal activities of Azardirachta indica crude extracts and fractions against certain Candida albicans strains: P37005, RM1000 and Strain1161. The reflux method was used for successive extraction of Azardirachta indica leaves which was done by three solvents namely: n- hexane, ethyl acetate and methanol. The leaves of Azardirachta indica was assayed for its phytochemicals components which was determined quantitatively. However, the various strains were subjected to the crude extracts of Azardirachta indica at a varying concentration of 40, 60, 80 and 100 mg/ml. The activity of n- hexane and methanol crude extracts had higher zones of inhibition than that of ethyl acetate crude extracts. The n- hexane crude extract showed a minimum inhibitory concentrations (MIC) and minimum fungicidal concentration (MFC) value of 12.5 mg/ml and 50 mg/ml respectively against the tested strain of P37005(Isolate B4). The value of 6.25 mg/ml and 50 mg/ml were also observed for the n- hexane crude extract against strain RM 1000(Isolate B2). Whereas, the methanol crude extract presented a value of 6.25 mg/ml and 100mg/ml respectively against strain RM1000(Isolate B2). More so, the antifungal activities of the fractions were determined at a concentration of 5mg/ml and 10 mg/ml which showed inhibition for all the strains. The n-hexane fraction F3 of Azardirachta indica had an MIC and MFC value of 12.5 mg/ml and 25 mg/ml against P37005(IsolateB4). The value for the methanol fractions(F6) of Azardirachta indica against strain 1161(Isolate S5) was 3.125 mg/ml and 6.25 mg/ml. It is obvious from this study that the antifungal activities of the crudes and fractions used were less in activity compared to the standard antibiotics(fluconazole).

Keywords:
Azardirachta indica, fractions, bioactive, inhibition strains

Article Details

How to Cite
Owoyale, A. D. M., Galadimma, M., Daniyan, S. Y., & Adabara, N. (2020). Antifungal Activities of Azardirachta indica Crude Extracts and Fractions against Strain 1161, P37005 and RM1000. Microbiology Research Journal International, 30(10), 63-78. https://doi.org/10.9734/mrji/2020/v30i1030274
Section
Original Research Article

References

Ahana N. The Medicinal Value of Azadirachta indica. Hindu Press, India; 2005.

Biswas K, Chattopadhyay I, Banerjee RK. Biological activities and Medical property of neem (Azardirachta indica) U. Current Science. 2002;82:1336-1345.

Chattopadhyay RR, Chattopadhyay RN, Maitra SK. Possible mechanism of anti -inflammatory activity of Azardirachta indica leaf extract. Indian Journal of Pharmacology. 1994;26:141-143.

Nat VD, Klerx J, Dijk HV, De Silva KT, Labadie RP. Immunomodulatory activities of an acqueous extract of Azardirachata indica stem bark. Journal of Ethno Pharmacology. 1987;27:143-145.

Sengupta P, Chowdhuri SN, Khastagir HN. Terpenoids and related compounds-1 constituents of the trunk bark of Melia azardirachta Linn and the structure of the ketophenol, nimbol. Tetra Hedron. 1960; 10-45-45.

Shumutterer H. The neem tree Azadirachta indica A. Juss. and other meliaceous plants. 2nd edition. International print-o-pac limited. Munbai. 2002;893.

Erdogan A, Rao SS. Small intestinal fungal overgrowth. Curr Gastroenterol Rep. 2015;17(4):16.

Bharvan PS, Rajkumar R, Radhakrishnan S, Seenivasan C, Kannan S. Culture and identification of Candida albicans from vaginal ulcer and separation of Enolase on SDS- PAGE. International Journal of Biology. 2010;2:84-93.

Koehler AP, Chu KC, Houang ET, Cheng AF. Simple, reliable and cost effective yeast identification scheme for the clinical laboratory. Journal of Clinical Microbiology. 1999;37:422–426

Cheesbrough M. Laboratory Manual. In: District Laboratory Practice in Tropical Countries. UR United Kingdom: Cambridge University Press. 2010;146-157.

John Thorne C, Heidi L, Lawrence CP. Manual of Medical Mycology. Cambridge, Mass., USA: Blackwell Scientific, C1995; 2010.

Promega Corporation. Wizard (R) Genomic DNA Purification Kit Technical Manual TM050; 2014.

Available:https://www.promega.com/-/media/files/resources/protocols/technical-manuals/0/wizard-genomic-dna-purification-kit-protocol.pdf. (Accessed: 19.12.2016).

Hymete ATH, Iversen J, Rohloff B, Erko E. Screening of Echinops ellenbeckii and Echinops longisetus for biological activities and chemical constituents. Phytomedicine. 2004;01-03.

Chang C, Yang M, Wen H, Chern J. Estimation of total flavonoid contents in propolis by two complementary colorimetric methods. Journal of Food Drug Analysis. 2002;10:178-182.

Singleton VL, Orthofer R, Lamuela- Raventos RM. Analysis of total phenols and other oxidants substrates and antioxidants by means of Folin -Ciocalteu reagent. Method in Enzymology. 1999; 299:152-178.

Oloyed OI. Chemical profile of unripe pulp of Carica pagaya. Pakistan Journal of Nutrition; 2005.

Emmanuel EO, Helmina O, Egwin CE. Phytochemical constituents of seeds of ripe and unripe Blighia Sapida (K Koening) and physicochemical properties of the seed Oil. International Journal of Pharmaceutical Science Invention. 2014; 3(9):31-40.

Wheeler EL, Ferrel RE. Western Regional Research Laboratory, Agricultural Research Servive, U.S. Department of Agriculture, Albany, California 94710; 1971.

Oke OL. Chemical studies on some Nigeria Food stuffs. Journal of West African Science Association. 1996;11:42-48.

McFarland J. Nephelometer. Journal of the American Medical Association. 1997;14:1176-1178.

Magaldi S, Mata S, Hartung C. ‘In vitro’ susceptibility of 137 Candida sp. isolates from HIV positive patients to several antifungal drugs. Mycopathologia. 2002; 149:63–68.

Ewansiha JU, Garba SA, Musa G, Daniyan SY, Busari, Innalegwu DA, Doughari JH. Preliminary phytochemical and antimicrobial activity of citrus x limon (L) Burm. f. (lemon) leaf extract against some pathogenic microorganisms. Journal of Applied life Science International. 2016; 6(4):1-10.

Fair JD, Kosmos CM. Thin Layer Chromatograhy. Journal on Chromatography. 2008;1211(1-2):49-54.

Edeoga HO, Okwu DE, Mbaebie BO. Phytochemical constituents of some Nigerian medicinal plants. African Journal of Biotechnology. 2005;4:685-688.

National Centre for Biotechnology and Information (NCBI). Sequence BLAST; 1988

Available:www.ncbi.nlm.nih.gov

Mahmoud DA, Hassanein NM, Youssef KA, Abouzeid MA. Antifungal activity of neem leaf extracts and the nimonol against some important human pathogen. Brazilian Journal of Microbiology. 2011;42(3):1007-1016.

Oke F, Aslim B. Biological potentials and cytotoxicity of various extracts from endemic Origanum minutiflorum O. Schwartz & P.H. Davis. Food and Chemical Toxicology. 2010;48:1728-1733.

Costa AF. Pharmacognosy: experimental pharmacognosy. 3rd. edition. Lisboa: Calouste Gulbenkian. 2001;3(303):308-309.

Prescott LM, Harley JP, Klein DA. Microbiology (International edition). 5th Edition. New York: McGraw-Hill Companies, Incorporation. 2002;811.

Simhadri VS, Nagesh NA, Muniappan M, Kannan I, Viswanathan S, Jayachandra K. Antifungal activity of various extracts of Azardirachta indica leaf –an invitro study . International Journal of Chem Tech Research. 2007;10(15):305-31.

Al-Humaid AI, Mousa RA, El-mergawi, Abdel-Salam AM. Chemical composition and antioxidant activity of dates and dates- Camel-Milk mixtures as a protective meal against lipid peroxidation in rats. American Journal of Food Technology. 2010;5:22-30.

Jones GA, McAllister TA, Muir AD, Cheng KJ. Effects of Safonin (Onorbrychis viciifolia scop) condensed tannins on growth and proteolysis by four strains of ruminal bacteria. Appl. Environ Microbiol. 1994;60:1374-1378.