• Mounyr Balouiri Laboratory of Medicinal and Aromatic Plants and Natural Substances, National Institute of Medicinal and Aromatic Plants – Taounate, Sidi Mohamed Ben Abdellah University, Fez, Morocco; Laboratory of Microbial Biotechnology,. Faculty of Science and Techniques, Sidi Mohamed Ben Abdellah University, Fez, Morocco
  • Samira Bouhdid
  • El Houssaine Harki
  • Moulay Sadiki
  • Wessal Ouedrhiri



Objective: This study aimed to investigate the antifungal activity of a microbial strain isolated from a non-exploited habitat (southeast region of
Morocco) against Candida albicans.
Methods: The antifungal producing strains were isolated from the rhizosphere of Calotropis procera Ait. The isolate (Cp-LMA-9) was selected based
on the primary antimicrobial screening. The optimization of the culture media for an antifungal production, and the extraction of bioactive metabolites
were performed. The bioautography and a partial characterization of these metabolites were also done.
Results: Based on the molecular identification, it was identified as Bacillus spp. with a similarity percentage of 98% with Bacillus tequilensis and
Bacillus subtilis. The maximum antifungal production against C. albicans was observed with malt extract-yeast extract-agar by solid-state fermentation
and malt extract-yeast extract-broth under stationary conditions. Antifungal fraction was extracted successfully from the solid-state fermentation by
acetone and methanol. It maintained the anti-Candida activity after heat treatment (autoclaving at 121°C for 15 minutes and boiling for 30 minutes),
within a pH range of 2-10, and after treatment with proteolytic enzymes.
Conclusion: The bacterium isolated and selected exhibit a remarkable antifungal effect against C. albicans. The resistance of the antifungal metabolites
extracted to heat, alkaline, and acidic conditions and to proteolitic enzymes making theme a promising antifungal compounds for the control of
Candida infections.

Keywords: Antifungal activity, Bacillus spp, C. albicans, Rhizosphere, Molecular identification.


Download data is not yet available.


Calderone RA, Fonzi WA. Virulence factors of Candida albicans. Trends Microbiol 2001;9(7):327-35.

Papon N, Courdavault V, Clastre M, Bennett RJ. Emerging and emerged pathogenic Candida species: Beyond the Candida albicans paradigm. PLoS Pathog 2013;9(9):e1003550.

McCullough MJ, Ross BC, Reade PC. Candida albicans: A review of its history, taxonomy, epidemiology, virulence attributes, and methods of strain differentiation. Int J Oral Maxillofac Surg 1996;25(2):136-44.

Mulu A, Kassu A, Anagaw B, Moges B, Gelaw A, Alemayehu M, et al. Frequent detection of ‘azole’ resistant Candida species among late presenting AIDS patients in northwest Ethiopia. BMC Infect Dis 2013;13:82.

Arendrup MC, Dzajic E, Jensen RH, Johansen HK, Kjaeldgaard P, Knudsen JD, et al. Epidemiological changes with potential implication for antifungal prescription recommendations for fungaemia: Data from a nationwide fungaemia surveillance programme. Clin Microbiol Infect 2013;19(8):E343-53.

Carlet J, Rambaud C, Pulcini C. WAAR (World Alliance against Antibiotic Resistance): Safeguarding antibiotics. Med Mal Infect 2012;42(9):446-51.

Hsieh T. Antifungal effects of bacilysin and fengymycin from bacillus suhtilis F-29-3 a comparison with activities of other bacillus antihiotics. J Phytopathol 1986;213:204-13.

Ahimou F, Jacques P, Deleu M. Surfactin and iturin A effects on Bacillus subtilis surface hydrophobicity. Enzyme Microb Technol 2000;27(10):749-54.

Tamehiro N, Okamoto-Hosoya Y, Okamoto S, Ubukata M, Hamada M, Naganawa H, et al. Bacilysocin, a novel phospholipid antibiotic produced by Bacillus subtilis 168. Antimicrob Agents Chemother 2002;46(2):315-20.

Sansinenea E, Ortiz A. Secondary metabolites of soil Bacillus spp. Biotechnol Lett 2011;33(8):1523-38.

Wu S, Jia S, Sun D, Chen M, Chen X, Zhong J, et al. Purification and characterization of two novel antimicrobial peptides subpeptin JM4-A and subpeptin JM4-B produced by Bacillus subtilis JM4. Curr Microbiol 2005;51(5):292-6.

Tan Z, Lin B, Zhang R. A novel antifungal protein of Bacillus subtilis B25. Springerplus 2013;2:543.

Chitarra GS, Breeuwer P, Nout MJ, van Aelst AC, Rombouts FM, Abee T. An antifungal compound produced by Bacillus subtilis YM 10‑20 inhibits germination of Penicillium roqueforti conidiospores. J Appl Microbiol 2003;94(2):159-66.

Tendulkar SR, Saikumari YK, Patel V, Raghotama S, Munshi TK, Balaram P, et al. Isolation, purification and characterization of an antifungal molecule produced by Bacillus licheniformis BC98, and its effect on phytopathogen Magnaporthe grisea. J Appl Microbiol 2007;103(6):2331-9.

Konishi M, Nishio M, Saitoh K, Miyaki T, Oki T, Kawaguchi H. Cispentacin, a new antifungal antibiotic. I. Production, isolation, physico-chemical properties and structure. J Antibiot (Tokyo) 1989;42(12):1749-55.

Intana W, Yenjit P, Suwanno T, Sattasakulchai S, Suwanno M, Chamswarng C. Efficacy of antifungal metabolites of Bacillus spp. for controlling tomato damping-off caused by Pythium aphanidermatum. Walaikak J Sci Technol 2008;5(1):29-38.

Kumar A, Saini P, Shrivastava JN. Production of peptide antifungal antibiotic and biocontrol activity of Bacillus subtilis. Indian J Exp Biol 2009;47(1):57-62.

Ren JJ, Shi GL, Wang XQ, Liu JG, Wang YN. Identification and characterization of a novel Bacillus subtilis strain with potent antifungal activity of a flagellin-like protein. World J Microbiol Biotechnol 2013;29(12):2343-52.

Khalil R, Djadouni F, Elbahloul Y, Omar S. The influence of cultural and physical conditions on the antimicrobial activity of bacteriocinproduced by a newly isolated Bacillus megaterium 22 strain. Afr J Food Sci 2009;3:11-22.

Pochon J, Tardieux P. Techniques d’analyse en microbiologie du sol. Edition de. Saint-Mande; 1962 Available from: [Last cited on 2013 Dec 19].

Ganesh Kumar C, Mongolla P, Joseph J, Nageswar YV, Kamal A. Antimicrobial activity from the extracts of fungal isolates of soil and dung samples from Kaziranga National Park, Assam, India. J Med Mycol 2010;20(4):283-9. Available from: [Last cited on 2012 Nov 29].

Anibou M, Chait A, Zyad A, Taourirt M, Ouhdouch Y, Benherref A. Actinomycetes from Moroccan habitats: Isolation and screening for cytotoxic activities. World J Microbiol Biotechnol 2008;24(10):2019‑25. Available from:‑008-9705-7. [Last cited on 2012 Feb 05].

Ibrahim D, Lee CC, Sheh-Hong L. Antimicrobial activity of endophytic fungi isolated from Swietenia macrophylla leaves. Nat Prod Commun 2014;9(2):247-50.

Gajbhiye M, Kesharwani J, Ingle A, Gade A, Rai M. Fungus-mediated synthesis of silver nanoparticles and their activity against pathogenic fungi in combination with fluconazole. Nanomedicine 2009;5(4):382‑6.

Mostakim M, El Abed S, Iraqui M, Benbrahim KF, Houari A, Gounni AS, et al. Biocontrol potential of a Bacillus subtilis strain against Bactrocera oleae. Ann Microbiol 2011;62(1):211-6. Available from: [Last cited on 2014 Jan 01].

Logan NA, Berkeley RC. Identification of Bacillus strains using the API system. J Gen Microbiol 1984;130(7):1871-82.

Schleifer KH. Phylum XIII. Firmicutes Gibbons and Murray 1978, 5 (Firmacutes [sic] Gibbons and Murray 1978, 5). In: De Vos P, Garrity G, Jones D, Krieg N, Ludwig W, Rainey F, et al., editors. Bergey’s Manual® of Systematic Bacteriology SE - 3. New York: Springer; 2009. p. 19-1317.

Choma IM, Grzelak EM. Bioautography detection in thin-layer chromatography. J Chromatogr A 2011;1218(19):2684-91.

Gatson JW, Benz BF, Chandrasekaran C, Satomi M, Venkateswaran K, Hart ME. Bacillus tequilensis sp. nov. isolated from a 2000-year-old Mexican shaft-tomb, is closely related to Bacillus subtilis. Int J Syst Evol Microbiol 2006;56:1475-84.

Chattopadhyay D, Sen SK. Optimisation of cultural conditions for antifungal antibiotic accumulation by Streptomyces rochei G164. Hindustan Antibiot Bull 1997;39(1-4):64-71



How to Cite

Balouiri, M., S. Bouhdid, E. H. Harki, M. Sadiki, and W. Ouedrhiri. “ANTIFUNGAL ACTIVITY OF BACILLUS SPP. ISOLATED FROM CALOTROPIS PROCERA AIT. RHIZOSPHERE AGAINST CANDIDA ALBICANS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 8, no. 2, Mar. 2015, pp. 213-7,



Original Article(s)