Effect of Some Plant Extracts on Candida albicans

Transcription

1 Effect of Some Plant Extracts on Candida albicans Nabil S.B. Al-Habib Nihad A.M. Al-Rashedi Biology Department, College of Science, Al-Muthanna University Abstract The aim of this study is to estimate the efficacy of seven plant extracts on inhibition of Candida albicans growth in vitro, and the comparison with standard antibiotic Nystatin. This study shows in details that plant extracts with (80%) ethanol had higher biological activity than hot aqueous extracts against C. albicans. The ethanolic extract of Juglans nigra fruit in studied concentrations, 10, 0 and 30% showed high average of diameter inhibition zone 13, 1, 17 and 18 mm, respectively. During the comparison with antifungal activity of nystatin found that average of diameter inhibition of this extract is approximately near to standard antibiotic activity. Ethanolic extracts were most potent of minimal inhibitory concentration (0.781 mg/ml) of Callendula officinalis, Echinophora platyloba and Juglans nigra. The studied plants could provide some activity against C. albicans. These extracts may be potential for use in antifungal treatment. Introduction Plants have been used extensively throughout history to treat medical problems. Numerous studied have been carried out to extract various natural products for screening antibacterial and antifungal activity, but attention has been also focusing on the comparison of these products with standard antibiotic nystatin (1, ). The objective of this study is examining the antifungal effect of seven plant extracts on Candida albicans growth; this is necessary at curing several diseases of C. albicans.. Materials and Methods Plant Collection and extraction: The seven plants were collected from Al-Jami'a clinic in Samawha city for alternative drugs and herbal medicine. The studied parts of

2 plants were separated, dried up and crushed into powder using pestle and mortar, after that they are extracted with hot water, and ethanol 80%. Filtration of extracts by Millipore filtration (0. µ), then extracts were evaporated to dryness and weighed. They are kept in dark and tight closed containers (3). Isolation of Candida albicans: The isolates of Candida albicans were collected from the previous study (). The identification of Candida albicans by culture on sabouraud's agar+chloramphencol, germ tube formation test, api Candida test (biomerieux, France), and confirmation of species by PCR kit IC 00/730 (Sacace, Italy). Assay of Antifungal activity: (Ι) paper disc diffusion method: The aqueous and ethanolic extracts of the plants were evaluated by the paper disc diffusion method on Mueller Hinton agar (). Cultures were adjusted to 0. McFarland turbidity standards and inoculated into agar plates. Sterile filter paper discs (diameter 6 mm) soaked in a known concentration of extract (,, 10, 0 and 0 mg/ml per disc in each case) were applied over each of the culture plates previously seeded with the 0. McFarland turbidity standard cultures of the test Candida albicans. The cultures were then incubated at 37 ºC for 8 hr. Paper discs soaked in a known concentrations (1.,, and 0 mg/ml per disc) of nystatin in distilled water, as standard antibiotic, were used for comparison. Antifungal activity was determined by measurement of zone of inhibition around each paper disc. (ΙI) Broth dilution method: It's determine the minimal inhibitory concentration (MIC) mg/ml. MIC of the plant extracts were determine by making sequence of decreasing concentrations, then broth inoculated with (fungi inocula were adjusted to 0. McFarland turbidity standard) Candida albicans after 8 hr. of incubation. The end point of each plant extract had sequence of dilution is taken as the greatest dilution show no visible growth. Final concentration from 6. to mg/ml was used for each plant extract (6). Result and Discussion The results indicated that the plant extracts of all the species studied showed antifungal activities in different levels towards Candida albicans. These results are consistent with previous reports on related plants regarding yeast (7). The sensitivity of Candida albicans to plant extracts was not unexpected as; in general, Candida

3 albicans is more sensitive than other species. Such sensitivity or resistance could be due to the different compositions between yeast species (8). This study shows that ethanolic extracts of plants extracts had highest biological activity than aqueous extracts against C. albicans. These results had some differences with previous studies (9, 10). All aqueous extracts of plants with concentration, and 10% (w/v) did not showed any biological activity against C. albicans, while inhibited zone for leaf extract of Aloe ferox and fruit extract of Juglans nigra concentration 0% were 1 mm. In concentration 30% of extracts of Aloe ferox leaves, Citrus unshia root bark and Juglans nigra fruit were mm average of diameter inhibited zone, while in the same concentration of extract of Callendula officinalis leaves was 1 mm and the rest plant extracts did not show any effect. The ethanolic extract of Juglans nigra fruit in studied concentrations, 10, 0 and 30% showed the highest which were 13, 1, 17 and 18 mm, respectively (table 1).

4 Table(1): Antifungal activity of plant extracts against Candida albicans Average of diameter inhibition zone (mm) Plant extracts Concentration of aqueous extracts w/v (%) Concentration of ethanolic extracts w/v (%) 0 0 Aloe ferox Callendula officinalis Citrus unshiu (root bark) 7 9 Echinophora platyloba Juglans nigra (fruit) Lindera benzoin (stem bark) 8 Rhus glabra (root bark) 1 8 = 0.0 mm During comparison with antifungal activity of nystatin found the inhibited diameter of this extract was approximately nearst to standard antibiotic activity (table )

5 Table (): Antifungal activity of standard antibiotic nystatin against Candid albicans Nystatin concentration (%) Average of diameter inhibition zone (mm) In ( table 3), MIC assay indicated that aqueous extracts for all seven plant extracts were not noted inhibited on Candida albicans growth, while ethanolic extracts were most potent of MIC which was of Callendula officinalis, Echinophora platyloba and Juglans nigra,then the MIC fell in 1. of Aloe ferox, Citrus unshiu, Lindera benzoin and Rhus glabra. The ethanolic extracts demonstrated the highest activity against Candida albicans, while aqueous extracts of plant showed the lowest activity. Higher activity against Candida albicans by organic solvent such as ethanol may be an indication that the phytoconstituents are more soluble in ethanol than in the water.

6 Table (3): Minimal inhibitory concentration (MIC) of plant extracts against Candida albicans Plant extract MIC (mg/ml) Aqueous extracts Ethanolic extracts Aloe ferox Callendula officinalis Citrus unshiu (root bark) Echinophora platyloba Juglans nigra (fruit) Lindera benzoin (stem bark) Rhus glabra (root bark) = no effect The studied plants could provide some activity against C. albicans, although it appears that C. albicans was not as sensitive to the aqueous extracts compared with the ethanolic extracts as determined by disc sensitivity and MIC assays. It is not surprising that there are differences in the antifungal activities of the different extracts of tested plants. This could be due to the phytochemical differences between them. The reasons for this could be that components as aromatic or saturated organic compounds from plants active against yeast. They are mostly obtained through initial ethanol extraction (11). Further researches are necessary to identify the active compounds and evaluate their potential for use as antifungal drugs.

7 References 1- Sakagami, Y.; Murata, H. and Nakanishi, T. (001) Inhibitory Effect of Plant Extracts on Production of Verotoxin by Enterohemorrhagic Escherichia coli 17: H7. J. Health Sci.; 7: Velickovic, D.T.; Randjelovi, N.V. and Ristic, M. (0)Chemical constituents and antimicrobial activity of the ethanol extracts obtained from the flower, leaf and stem of Salvia officinalis L. J. Serb. Chem. Soc.; 68: Kambizi, L.; Sultana, N. and Afolayan, A.J. (00). Bioactive compounds isolated from Aloe ferox: A plant traditionally used for the treatment of sexually transmitted infections in the Eastern Cape, South Africa. J. Pharm. Biol.; (8): Al-Rashedi, N.A.M.; A. Al-Hamadani, A.H. and Al-oebadyi, M.A. (009) Evaluation of Candida albicans Diagnosis by using PCR. Proceeding to the Second Conference for Institute of Genetic Engineering and Biotechnology, Baghdad University. - National Committee for Clinical Laboratory Standards (NCCLS) (1997) Performance standards for antimicrobial disk susceptibility tests-sixth Edition: approved standard M-A6. NCCLS. J. Pharmaceut. Res. 1: National Committee for Clinical Laboratory Standards (NCCLS) (000) Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved StandardÑFifth Edition. NCCLS document M7-A. NCCLS: Wayne, PA, USA. 7- Kabir, O.A.; Olukayode, O.; Chidi, E.O.; Christopher. C.I.; and Kehinde, E.F. (00). Screening of crude extracts of six medicinal plants used in South- West Nigerian unorthodox medicine for antimethicillin resistant staphylococcus aureus activity. J. BMC Compl. Alt. Med. (6): Ikram, M. and Inamual, H. (1980)Screening of Medical Plants For Antimicrobial Activity. Part 1.Fitoterapia 1: Shahidi Bonjar, G.H.; Aghighi, S. and Kamiri Nik, A. (00) Antimicrobial and Antifungal Survey in Plants used in Endogenous Herbal-Medicine of South East Regions of Iran. J. Bio. Sci.; (3): 0-1

8 10- Avijgan, M.; Saadat, M. and Nilforoushzadeh, M.A.(00) Effect of extract of Echinophora Platyloba on some common dermatophyts. Proceeding of the Second Symposium of Medicinal Plants; Tehran, Iran. 11- Cowan, M.M.(1999) Plant products as antimicrobial agent. J. Clin. Microbiol. Rev.;1: 6-8 تأثير بعض المستخلصات النباتية على فطر داء المبيضات ب م صبري انحب ب اد انراشدي قسى عه و انح اة, كه ت انعه و, جايعت ان ث ى ان دد يد دا اندةاهدت د حفدد ر كعدامة هدبعت يسخيه داث باح دت عهدى حثبد ل د در دام ان ب داث خداة انجسى, يفاة خ ا بان اد انح ي انف اه ان سخاح 8 ضحج اندةاهت ا ان سخيه داث اال ثا ن دت نه باحداث ن دا عان ت با ه ج ت اكبر ي ان سخيه اث ان ائ ت عهى ر دام ان ب اث 8 ان سخيهص انكح ن نث اة انج ز انخراك س ان دة هت %,, اع ج حثب ل عدان د حدص انحساهد ت 08 07,0,00 يه خدر

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