Research Article

Evaluation of novel culture media prepared from plant substrates for isolation and identification of Cryptococcus Neoformans Species Complex

Ellabib M*, Krema ZA, Mokthar ES, El Magrahi HS, Eshwika A and Cogliati M

Published: 14 August, 2018 | Volume 1 - Issue 1 | Pages: 009-013

Background: Melanin production due to phenoloxidase activity is a distinctive property of Cryptococcus neoformans and Cryptococcus gattii species complex yeasts. Therefore, an agar medium containing a precursor of melanin pigment is potentially useful to identify and differentiate cryptococcal colonies from other yeasts.

Background: This study aimed to evaluate and compare the ability of Cryptococcus neoformans species complex isolates to produce brown-pigmented colonies when grown on media prepared from various plant leaves or seeds extracts.

Material and Methods: Forty-six C. neoformans species complex isolates which were obtained from various environmental and clinical samples were inoculated on different media containing coriander, cumin, soybean, lupine, flax, pumpkin, basil, peppermint, and marjoram, were observed for the rate of growth and pigment production during a five-day period.

Results: All isolates were pigmented on all media within 24-48 hours, and brown or dark brown colonies were observed in less than five days, while C. albicans grew but did not produce any pigment.

Conclusion: The differential media tested in the present study are simple and inexpensive, and represent alternative valid tools for presumptive identification of C. neoformans species complex from clinical and environmental samples

Read Full Article HTML DOI: 10.29328/journal.ijcmbt.1001002 Cite this Article Read Full Article PDF


Cryptococcus neoformans; Melanin production; Differential medium; Seeds, Leaves


  1. Rajasingham R, Smith RM, Park BJ, Jarvis JN, Govender NP, et al. Global burden of disease of HIV-associated cryptococcal meningitis: an updated analysis. Lancet Infect Dis. 2017; 17: 873-881. Ref.: https://tinyurl.com/y9kkgk8e
  2. Heitman J, Kozel TR, Kwon-Chung KJ, Perfect JR, Casadevall A. Cryptococcus: From Human Pathogen to Model Yeast. ASM Press, Washington, DC, USA. 2011. Ref.: https://tinyurl.com/yd53oxge
  3. De Hoog GS, Guarro J, Gene J, Figueras MJ. Atlas of clinical fungi. ASM Press, Washington, DC, USA. 2000.
  4. Lacaz CS, Porto E, Martins JEC. Micologia media: fungos, actinomicetos ealgas de interesse medico. 8th ed. Sao Paulo: Sarvier. 1991.
  5. Chaskes S, Tyndall RL. Pigment production by Cryptococcus neoformans from para and ortho-diphenols: effect of the nitrogen source. J Clin Microbiol. 1975; 1: 509-514. Ref.: https://tinyurl.com/y8us93ja
  6. Pal M, Mehrotra BS. Studies on the efficacy of sunflower seed agar for the isolation and identification of Cryptococcus neoformans. Arogya J Health Sci 1982; 8: 74-79.
  7. Strachan AA, Yu RJ, Blank F. Pigment production of Cryptococcus neoformans grown with extracts of Guizotia abyssinica. Appl Microbiol. 1971; 22: 478-489.
  8. Hopfer RL, Blank F. Caffeic acid-containing medium for identification of Cryptococcus neoformans. J Clin Microbiol. 1976; 2: 115-120. Ref.: https://tinyurl.com/ycwcbdn4
  9. Nandhakumar B, Kumar G, Prabhu CP, Menon T. Mustard seed agar,a new medium for the differentiation of Cryptococccus neoformans. J. Clin. Microbiol. 2006; 44: 674. Ref.: https://tinyurl.com/y78ymenx
  10. Nandhakumar B, Menon T, Kumar G. A new henna-based medium for the differentiation of Cryptococcus neoformans. J Med Microbiol. 2007; 56: 568. Ref.: https://tinyurl.com/yba8zu4q
  11. Tendolkar U, Tainwala S, Jog S, Mathur M. Use of a new medium - tobacco agar, for pigment production of Cryptococcus neoformans. Indian J Med Microbiol. 2003; 21: 277-279. Ref.: https://tinyurl.com/ybtory4b
  12. Hamzia Ali Ajah. Two new media apple leaves agar and eggplant leaves agar for identification of Cryptococcus neoformans. J Biology, Agriculture and Healthcare. 2014; 4. Ref.: https://tinyurl.com/y7al3pva
  13. Ellabib MS, Krema ZA, Allafi AA, Cogliati M. First report of two cases of cryptococcosis in Tripoli, Libya, infected with Cryptococcus neoformans isolates present in the urban area. J Mycol Med. 2017; 27: 421-424. Ref.: https://tinyurl.com/ycwn9gb4
  14. Ellabib MS, Aboshkiwa MA, Husien WM, D'Amicis R, Cogliati M. Isolation, identification and molecular typing of Cryptococcus neoformans from pigeon droppings and other environmental sources in Tripoli, Libya. Mycopathologia. 2016; 181: 603-608. Ref.: https://tinyurl.com/ycbvydzl
  15. Springer DJ, Mohan R, Heitman J. Plants promote mating and dispersal of the human pathogenic fungus Cryptococcus. PLoS One. 2017; 12: e0171695. Ref.: https://tinyurl.com/y8sr4ctv
  16. Rajeshwari CU, Andallu B. Reverse phase HPLC for the detection of flavonoids in the ethanolic extract of Coriandrum sativum L seeds. International Journal of Basic and Applied Sciences. 2012; 1: 21-26.
  17. Claudia-CT, Neli-Kinga O, Laurian V, Cristina M. Comparative Studies on Polyphenolic Composition, Antioxidant and Diuretic Effects of Nigella sativa L. (Black Cumin) and Nigella damascena L. (Lady-in-a-Mist) Seeds. Molecules. 2015; 20: 9560-9574. Ref.: https://tinyurl.com/y88sgcq9
  18. Montserrat D, Teresa H, Sergio R, Grzegorz L, Isabel E, et al. Bioactive Phenolic Compounds of Soybean (Glycine max cv. Merit): Modifications by Different Microbiological Fermentations. Pol J Food Nutr Sci. 2012; 62: 241-250. Ref.: https://tinyurl.com/ya8azlua
  19. Aleksander S, Jaroslaw C, Piotr K, Krzysztof D, Eleonora LS, et al . Antioxidant activity and phenolic content in three lupin species. Journal of Food Composition and Analysis. 2012; 25: 190-197. Ref.: https://tinyurl.com/y7fvumzg


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