DEPARTMENT OF PHARMACEUTICAL MICROBIOLOGY Onyenwe Nathaniel E.1, Adeniyi-Ake, M.A.2, Akpoyibo EmmamuzJ.1, Okoro, J.C.3 ANTIMYCOTIC EFFECT OF MELALEUCA ALTERNIFOLIA (TEA TREE) PLANT ON CUTANEOUS MYCOSIS BY DEPARTMENT OF PHARMACEUTICAL MICROBIOLOGY  

INTRODUCTION Recently, cutaneous fungi infection poses serious medical issues and more than a hundred thousand fungal species have been considered as natural contaminants (Kacaniova, 2003). Antifungal agents are widely distributed among higher plants (Caceres,et al., 1991), but only a few have been evaluated for their activity against human pathogenic fungi

INTRODUCTION CONTD. Therefore, new prototype antimicrobial agents are needed to address this situation (Sati and Joshi, 2010). Recently, data now shows that a range of yeasts, dermatophytes, and other filamentous fungi are susceptible to TTO (Nenoff, et al, 2014)

Introduction contd. The antifungal activity of Melaleuca alternifolia, and the comprehensive investigations of the susceptibility of fungi to TTO have only recently been completed (Nenoff, et al, 2014). Prior to this, data were somewhat piecemeal and fragmentary. Early data were also largely limited to Candida albicans, which was a commonly chosen model test organism. Thus, the aim to investigate on the effects of the crude soxhlet extract of the tea tree Melaleuca alternifolia on cutaneous mycosis

Material and methods Extraction of the plant The leaves of Melaleuca alternifolia was dried at room temperature (20-250C) for 2weeks and further dried in the oven at 500C for 2 days. Phytochemical screening The plant extracts were assessed for the existence of the phytochemical constituents

Methods continue Preparation of Mcfarland standard Mcfarland standard is used as a reference to adjust the turbidity of fungal suspension so that fungal organisms will be within a given range. Exactly, 0.5 Mcfarland equivalent turbidity was prepared Disc diffusion method Color and percentage yield of crude extract

MATERIALS & METHODS CONTD. Well in Agar diffusion method Morphological / microscopic characterization of fungi isolates

RESULTS Table 1: phytochemical constituents of crude extract Melaleuca alternifolia sample. S/N Phytochemical compound Result 1 Saponin + 2 Terpenes 3 Flavonoids 4 Alkaloids 5 Cardiac glycoside 6 Tannins and phenolic compounds 7 Phenols 8 Anthraquinones -

Table 2. Cutaneous fungi isolates representation of the macroscopic and microscopic identification of the test organism A, B, C and D. Fungi Samples Macroscopic/ microscopic Examination of isolates Identified isolates   A B C D Colonies with previously creamy white surface which turns ink to wine, having reverse side pink to wine colorations on SDA. They Produce conidia Colonies previously white cotton like surface with radial segments, which gradually turn greyish green with white at the edge, having reverse side creamy yellow and radial segmented. They Produce conidia Colonies previously white cotton like surface with radial segments, which gradually turn bluish green with white at the edge, having reverse side creamy yellow and radial segmented. They Produce conidia and filaments Colonies previously white cotton like surface with radial segments, which gradually turn black with whiteat the adge, having reverse side creamy yellow and radial segmented. They Produce conidia and filaments. Rhodotorulasp. ( yeast Aspergillus fumigatus  Aspergillus flavus Aspergillus niger .

RESULT CONTIN.  A (macroscopy of Rhodotorular (yeast)A2

A2( microscopy of Rhodotorular (yeast)

Result continue B (macroscopy of Aspergillus fumigatus) B2( microscopy of Aspergillus)

RESULTS CONT. B2( microscopy of Aspergillus)

RESULTS CONTINUE C (macroscopy of Aspergillus flavus)

RESULTS CONT. C2( microscopy of Aspergillus flavus)

D (macroscopy of Aspergillus

D2( microscopy of Aspergillus niger)

FIG 1.The in vitro antifungal activity of crude extract of Melaleuca alternifolia using agar well diffusion method

FIG 2. The in vitro antifungal activity of ketoconazole using well in agar method

Fig.3.Chart Representation of the Antifungal Activity of Ketoconazole Using Disc Diffusion Method

Fig 4. Chart of the Fig 1. Chart of the antifungal activity of crude extract of Melaleuca alternifolia antifungal activity of crude extract of Melaleuca alternifolia

DISCUSSION AND CONCLUSION The Present study was conducted to analyze the phytochemical, cytotoxic and antifungal potential of leaf extract of Melaleuca alternifolia. Knowledge of the phytochemical constituents of plants is desirable, not only for the discovery of therapeutic agents, but also because such information may be of value in disclosing new sources of such economic materials as tannins, oils, alkaloids, flavonoids, saponins, essential oils precursors for the synthesis of complex chemical substances (Akrout, 2010).

REFERENCES Abad, M. J., Ansuategui, M. and Bermejo, P. (2007). Active antifungal substances from natural sources. (ARKIVOC) 7: 116-145. Abascal K. and Yarnell E. (2002). Herbs and drug resistance: Alternate and complementary therapies. In: Blumenthal, Mark, editor. Potential of botanicals in drug-resistant microbes.

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