1. 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

2. 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

3. 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)

4. 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

5. 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

6. 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

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

8. 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 -

9. 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 .

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

11. A2( microscopy of Rhodotorular (yeast)

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

13. RESULTS CONT. B2( microscopy of Aspergillus)

14. RESULTS CONTINUE
C (macroscopy of Aspergillus flavus)

15. RESULTS CONT.
C2( microscopy of Aspergillus flavus)

16. D (macroscopy of Aspergillus

17. D2( microscopy of Aspergillus niger)

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

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

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

21. 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

23. 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).

24. REFERENCES
Abad, M. J., Ansuategui, M. and Bermejo, P. (2007). Active antifungal substances from natural sources. (ARKIVOC) 7:
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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