PREPARATION DIFFERENTIAL CULTURE MEDIUM FOR CRYPTOCOCCUS NEOFORMANS FROM AQUEOUS EXTRACT OF LEAVES AND FLOWERS OF CHRYSANTHEMUM CINERARIAEFOLIUM
Objective: A new medium was prepared to isolate and diagnose the yeast Cryptococcus neoformans from flower and leaves aqueous extract of chrysanthemum.
Methods: Prepared differential culture medium for C. neoformans from aqueous extract of leaves and flowers of Chrysanthemum cinerariaefolium and chemical, spectral tests of the extracts were tested, in addition of gas chromatography (GC)–mass was used to diagnose phenolic compounds in both leaves and flowers.
Results: Showed that the yeast was grow with typical colonies on the new medium compared with other media which using in diagnosed of this yeast such as Staib agar and Sabourauds dextrose agar and unlike the yeast Candida albicans (as a negative control), which appeared in cream to white on this medium. Furthermore, the colonies are dark brown in color on flower chrysanthemum medium and light brown color on leaves chrysanthemum medium. In addition, the results of the chemical and spectral tests of the extracts confirmed that the plant contains many active compounds such as alkaloids, turbines, tannins, and phenols. The analysis of the extracts of phenolic compounds using GC–mass led to the diagnosis of five compounds in the leaf extract and nine compounds in the flower extract of this plant.
Conclusions: The media was prepared is differential medium that use to diagnosis of Cryptococcus such as Staib agar. Moreover, low economic cost, which consists of leaves and flowers of a plant available, abundance and the method of preparation is very simple.
2. Chen J, Varma A, Diaz MR, Litvintseva AP, Wollenberg KK, Kwon- Chung KJ. Cryptococcus neoformans strains and infection in apparently immunocompetent patients, China. Emerg Infect Dis 2008;14:755-62.
3. Fonseca A, Boekhout T, Fell JW. Cryptococcus Vuillemin. In: Kurtzman CP, Fell JW, Boekhout T, editors. The Yeasts, a Taxonomic study. 5th ed. Amsterdam, Netherlands: Elsevier; 2011. p. 1661-738.
4. Baddley JW, Dismukes WE. Cryptococcosis. In: Dismukes WE, Pappas PG, Sobel JD, ediotrs. Clinical Mycology. Ch. 12. Oxford: Oxford University Press; 2003. p. 519.
5. Kwon-Chung KJ, Varma A. Do major species concepts support one, two or more species within Cryptococcus neoformans? FEMS Yeast Res 2006;6:574-87.
6. Vu K, Eigenheer RA, Phinney BS, Gelli A. Cryptococcus neoformans promotes its transmigration into the central nervous system by inducing molecular and cellular changes in brain endothelial cells. Infect Immun 2013;81:3139-47.
7. Koneman EW, Allen SD, Jawa WM, Sachreckeber PC. Color Atlas and Textbook of Diagnositic Microbiology. 4th ed. Philadelphia, PA, United States: J.B. Lippincott; 1992. p. 838-55.
8. Barnett JA. A history of research on yeasts 14: Medical yeasts part 2, Cryptococcus neoformans. Yeast 2010;27:875-904.
9. Jarvis JN, Casazza JP, Stone HH, Meintjes G, Lawn SD, Levitz SM, et al. The phenotype of the Cryptococcus-specific CD4+ memory T-cell response is associated with disease severity and outcome in HIV-associated cryptococcal meningitis. J Infect Dis 2013;207:1817-28.
10. Leal AL, Faganello J, Fuentefria AM, Boldo JT, Bassanesi MC, Vainstein MH. Epidemiological profile of cryptococcal meningitis patients in Rio Grande do Sul, Brazil. Mycopathologia 2008;166:71-5.
11. Staib F, Seibold M, Antweiler E, Fröhlich B, Weber S, Blisse A. The brown colour effect (BCE) of Cryptococcus neoformans in the diagnosis, control and epidemiology of C. neoformans infections in AIDS patients. Zentralbl Bakteriol Mikrobiol Hyg A 1987;266:167-77.
12. Kalia R, Kaur J, Kaur A. Antitumor activity of aqueous extract of different cultivars of C. morifolium R using potato disc tumor assay. J Pharm Sci Res 2016;18:1262-5.
13. Harborn J. Phytochemical Method. 2nd ed. New York: Chapman and Hall; 1984. p. 278.
14. Tiwari P, Kumar B, Kaur G, Kaur H. Phytochemical screening and extraction. A review. Int Pharm Sci 2011;1:98-106.
15. Hetal S, Disha P, Mahendra V. Evaluation of conventional and serological methods for rapid diagnosis of cryptococcal meningitis in HIV seropositive patients at tertiary care hospital. Natl J Community Med 2011;3:354-7.
16. Sharma R, Das A, Duggal N, Hans C. Cryptococcal meningitis in an immunocompetent patient. Sch J Med 2014;6:414-6.
17. Lungran P, Devi AV, Singh WS, Damroulien S, Mate H, Golmei A. Cryptococcosis: Its prevalence and clinical presentation among HIV positive and negative patients in Rims, Manipur. IOSR J Dent Med Sci 2014;13:38-41.
18. Dulaimi HA, Shafiq SA. Use of the fruits of some plants to prepare media cultures to isolate and diagnose the yeast of Cryptococcus neoformans. J Mustansiriya Sci 2011;1:251-62.
19. Minhas N, Patil S, Sharma PC. Novel preparation of culture media for cultivation of Cryptococcus neoformans with extracts of different plant products. Int J Pharm Sci Rev Res 2013;24:70-5.
20. Katiyar R, Deorukhkar SC, Saini S. Comparison of different media for the pigment production of C. neoformans. J Clin Diagn Res 2011;5:1187-9.
21. Ajah HA. New culture medium, fava bean (Vicia faba)agar for cultivation and identification of C. neoformans. Int J Adv Res 2015;3:964-71.
22. Ughari JH, Elmahmood AM, Mazara S. Studies on the antibacterial activity of root extracts of Carica papaya L. Afr J Microbiol Res 2007;7:37-41.
23. Kareem RA. Effect of Organic Solvents and Secondary Compounds of Dodi Leaves and Flowers in Some Aspects of Culex Mosquito Thesis. Faculty of Science University of Qadisiyah; 2016.
24. Silverstein R, Webster F, Kiemle D. Spectrometric Identification of Organic Compounds. 7th ed. USA: John Wiley and Sons; 2005.
25. Mohammad NS, Srinivasulu A, Chittibabu B, Rao VU. Isolation and purification of antibacterial principle from Avicennia Marina L in methanol. Int J Pharm Pharma Sci 2015;7:38-41.
26. Han AR, Kim HY, So Y, Nam B, Lee IS, Nam JW, et al. Quantification of antioxidant phenolic compounds in a new Chrysanthemum cultivar by high-performance liquid chromatography with diode array detection and electrospray ionization mass spectrometry. Int J Anal Chem 2017;2017:1254721.
27. Sheikh RE, Amin AS, Atwa MA, Gouda AA, Abdullah AA. Determination of phenolic components and antioxidant activity of some Egyptian tea samples. Int J Pharm Pharm Sci 2015;7:198-202.
This work is licensed under a Creative Commons Attribution 4.0 International License.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.