Antifungal Activity of Juniperus Oxycedrus Tar; Growing Wild in North-West of Algeria B.Terfaya1*, A.Makhloufi1, A
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Applied Biology in Saharan Areas Vol. 1, N. 1, p. 33-36, March 2017 ISSN: 2571- 9823 Antifungal Activity of Juniperus oxycedrus Tar; Growing Wild in North-west of Algeria B.Terfaya1*, A.Makhloufi1, A. Mekboul1, L.Benlarbi1, D. Abdelouahed 2 1 Laboratory of valorization of vegetal Resource and Food Security in Semi Arid Areas, South West of Algeria, BP 417, University Tahri Mohamed , Bechar, Algeria. 2 Laboratory of Food, Biomedical and Environmental Microbiology, Aboubakr Belkaid University, Tlemcen, 13000 - Algeria. * Corresponding author: bouzybiochimie @yahoo.fr Abstract – Biological activities of medicinal plants have been recognized for a long time. The present study focused on the antifungal activity (In vitro) of Juniperus oxycedrus tar. The physicochemical results of vegetable tar showed a density value of 1.15, a refraction index about 1.5115, an acid pH (3.25) and the dries matter ratio of 34, 78%. The antifungal activity of studied tar against six strains of Fusarium oxysporum f.sp albedinis (FOA). (S1, S2, S3, S4, S5, S6), showed that Juniperus oxycedrus tar has a great antifungal activity against all the investigated strains. The antifungal activity results revealed that the fungi growths were completely inhibited by the tar. Whereas; (S3) was the most sensitive strain with minimal inhibition concentration value 0.003 mg/ml. Keywords: Tar, Juniperus oxycedrus, Antifungal activity, North-west Algeria. 1. Introduction J.oxycedrus was used since the antiquity to Essential oils and extracts from produce oil which is rich on sesquiterpene, aromatic plants have long been used for a named „cade oil‟, used in dermatology and wide variety of medicinal and domestic cosmetology. Otherwise, several studies purposes. Antimicrobial properties reported on the chemical composition of medicinal plants such as Matricaria solvent extracts from wood, berries and pubescens (Desf.), Rosmarinus officinalis leaves of J. oxycedrus, in which L and Artimisia herba alba against food sesquiterpenes, diterpene acids and related microorganisms as well as their polyphenolic compounds have been applications in food system have been isolated and characterized. investigated and reviewed (Gachkar & The use of tar is reserved mainly for al.,2007; Atik bekkara & al.,2007; patients with chronic stable, scalp Chanthaphon & al.,2008; ; Makhloufi & psoriasis, atopic dermatitis, and seborrhea al., 2011; Makhloufi & al., 2012). dermatitis. J.oxycedrus has been the Juniperus oxycedrus is a shrub belonging subject of recent research. In the to Cupressaceae family, in the pharmaceutical and food industries, it has mediterranean area, including Algeria excellent antioxidant and antimicrobial which lies on the arid slopes and hills properties, due to some compounds (Dellile, 2007). It is more frequent in the (carnosol, carnosic acid, ursolic acid, forest of northern Algeria. Available online at www.univ-bechar.dz/absa 33 Applied Biology in Saharan Areas Vol. 1, N. 1, p. 33-36, March 2017 ISSN: 2571- 9823 betulinic acid, and the rosmaridiphenol albedinis(5) FOA(5), Fusarium oxysporum rosmanol) (Thoresen & al., 2003). f. sp. albedinis(6) FOA(6) were isolated from infected date palm from Mazer area 2. Materials and Methods in Saoura region, south-west of Algeria. J.oxycedrus specimens were collected Fungal spores were prepared by from Tlemcen forest, north-west Algeria, growing mold on Potato Dextrose Agar during January and February 2014. These (PDA) at 27°C for 7 days, and spores were biomasses were dried for fifteen days in suspended in sterile 1% tween-80. Spore the dark at ambient laboratory temperature count was performed by using (20-28°C). hemacytometer, and adjusted to obtain 105 to 106 spores/ml with Potato Dextrose 2.1 Distillation of Tar Broth, (Ouraïni & al., 2007; Makhloufi & Tar extraction was realized in the al., 2011). research laboratory “VRVSA” of Tahri Mohamed university of Bechar, the wood 2.3.2 Dilution Method (MIC) of the aerial parts were cut before the Antifungal tests were performed operation, and then, the cut wood was according to the method reported by submitted to distillation for 3 to 4 hours Hassikou & al., (2002). The antifungal using a special dispositive. The distilled tar activity (Assay for radial growth was stored at ambient temperature till it inhibition), Petri dishes containing PDAa was used. media and tar at different concentrations were inoculated with fungus suspension 2.2 Physico-chemical Characterization obtained from pure culture, and they were of J.oxycedrus Tar incubated at 25°C for 7 days. The diameter of the fungus colony was obtained by Methods conform to AFNOR (1986) calculating the average of two have determined the specific density at perpendicular diameters. 20°C; Refractive index (RI) was measured with a refractometer, at 20ºC. The methods used for the determination of acid index 3. Results and Discussion and ester are also used with the standards J.oxycedrus Tar is a dark brown, AFNOR (1986). viscous liquid with a smoky odor and acrid, slightly aromatic taste with the yield 2.3 Antifungal Activity (In vitro) of 1.20 % (v/w) on dry weight basis. It is soluble in the following solvents: very 2.3.1 Fungal Strains slightly soluble in water; soluble in ether, The antifungal activity was evaluated chloroform, amyl alcohol, ethyl acetate, by dilution methods against six selected alcohol; and partly soluble in petroleum fungi of Fusarium oxysporum f. sp. ether. The results of physico-chemical Albedinis(1) FOA(1), Fusarium oxysporum index of vegetable tar of J.oxycedrus are f. sp. Albedinis(2) FOA(2), Fusarium shown in Table 1. oxysporum f. sp. Albedinis(3) FOA(3), Table 1. Physico-chemical index of Fusarium oxysporum f. sp. albedinis(4) vegetable tar of J. oxycedrus FOA(4), Fusarium oxysporum f. sp. Available online at www.univ-bechar.dz/absa 34 Applied Biology in Saharan Areas Vol. 1, N. 1, p. 33-36, March 2017 ISSN: 2571- 9823 Specific Rate of 3.1 Antifungal Activity Refractive Acid density matter Dilution Method (MIC) index index at 20 °C dries The growth inhibition zones measured 1.5115 1.15 3.25 34,78% by dilution method (MIC) are presented in Table 2. Table 2. MIC of tar (mg/ml) on fungal strains Concentration 0.0006 0.001 0.002 0.003 0.005 0.006 0.013 0.026 0.039 (mg/ml) witness Strains S1 + + + + + + MIC - - - S 2 + + + + + + MIC - - - S 3 + + + + MIC - - - - - S 4 + + + + + + MIC - - - S 5 + + + + + MIC - - - - S 6 + + + + + MIC - - - - (-):Inhibition (+): Growth According to these In vitro results, the psychiatric disorder, cancer, fever, vegetable tar of Juniperus oxycedrus, have cephalic, angina, weight decrease, the great and a good inhibitory antifungal common cold, and hypotonia, without any activity against all the investigated fungi. scientific evidence to support these uses The concentration inhibition of the tar is despite such common use. The tar from the between the value of 0.003 and 0.006 mg/ dry leaves of Juniperus oxycedrus is ml. The diameters of growth inhibition obtained by steam distillation with a yield zone ranged from 15 to 50mm with the of 0.15%. (Skalli & al., 2014). highest inhibition zone values observed against the strain of Fusarium oxysporum f. In the presence of the vegetable tar, the sp. albedinis(3) FOA(3) (50mm). liberation of the polyphenol and the tannin However, the fungies studied did not show are realized because the Juniperus the same sensitivity against the vegetable oxycedrus tar has an antifungal activity, it tar, the same inhibited concentration with contains more than 50 compounds; S1, 2, 4 which is 0.006; 0.005 for S 5, 6 including α-pinéne 33.41%, 4.76% γ - and finally 0.003 for S3. cadinéne and germacrene D23.66% represent the major compounds. (Skalli & Juniperus oxycedrus is used in folk al., 2014). Ben khalifa A. & al (2003). medicine of North African countries. The 4. Conclusion tar is accessible to consumers without a Tar from J.oxycedrus has a good prescription, and it is known to be used biological activity. The strong antifungal externally for skin disorders in activity of J. oxycedrus against array of dermatology and hair care, essentially for filamentous fungi strains is an indication of scalp care, eczema, scale affections, loss of the broad spectrum antifungal potential of hair, and psoriasis. It is also used against the tar. This could make the tar a evil eye, abdominal pain and diarrhea, Available online at www.univ-bechar.dz/absa 35 Applied Biology in Saharan Areas Vol. 1, N. 1, p. 33-36, March 2017 ISSN: 2571- 9823 promising group of natural compounds for Characteristics of Cuminum, Cyminum, development of safer antimicrobial agents. and Rosmarinus Officinalis Essential Oils”. Food Chemistry, 102, 898904. 5. Acknowledgments -We thank the Laboratory of valorization Makhloufi, A., Moussaoui, A., Lazouni, of vegetal resources and food security in H.A., Hasnat, N., Abe El Ouahid D.E., semi arid area, south west of Algeria, (2011). Antifungal Activity of Tahri Mohamed university of Bechar, Rosmarinus Officinalis L. and its Impact Algeria, for providing facilities and on the Conservation of a Local Variety of materials. Dates. Journal of Medicinal Plants, Vol. 3(2), June 2001, pp. 1-6. 6. References Makhloufi, A., Moussaoui, A., Lazouni, Atik bekkara, F., Bousmaha, L., Taleb H.A., (2012). Antibacterial Activities of bendiab, S.A., Boti, J.B., Casanova, J., Essential Oil and Crude Extracts from (2007). Composition Chimique de L‟huile Matricaria Pubescens (Desf) Growing Essentielle de Rosmarinus Officinalis L: Wild in Béchar, South-west of Algeria. poussant à l‟état spontané et cultivé de la Journal of Medicinal Plants Research Vol. région de Tlemcen. J Biologie & Santé vol. 6(16), pp. 3124-3128. 7, n° 1, p. 6-11. Thoresen, MA, Hildebrand KS (2003). Ben Khalifa A., Guerradi M., Tirichine A., Quantitative Determination of Phenolic Belguedj M Et Labgaa L. (2003). Diterpenes in Rosemary Extracts. 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