Abstract
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.
Highlights
Cryptococcus neoformans are the most widespread opportunistic fungi in the world affecting humans and most animals [1]
When the yeast of C. neoformans was grown on the Staib agar at 37°C, the results showed that the yeast had grown in spherical colonies, smooth mucus, and brown structure
The browned C. neoformans were colored brown due to phenoloxidase, which oxidizes the phenolic compounds found in the medium, depositing the melanin pigment on the wall of the yeast, giving colonies of brown, and distinct colonies from C. albicans Fig. 2
Summary
Cryptococcus neoformans are the most widespread opportunistic fungi in the world affecting humans and most animals [1]. The genus Cryptococcus belongs to yeast-like, an opportunistic yeast that affects immunocompromised people, which causes high morbidity and mortality, and affects people who are immunocompetent [2]. C. neoformans cause cryptococcosis, a globally dangerous disease [8]. This disease is more severe in people with immunosuppressive weakness, especially those with T-cell (CD4) cellmediated immunity and people with human immunodeficiency virus [9]. Yeast grows on the Sabouraud dextrose agar (SDA) in colonies of creamy, smooth, and mucous form. When the yeast grows on Staib agar, it is formed in spherical, brown, and mucous shaped colonies. We used in this study plant (at this 1st time) which contains phenolic compounds in prepared differential medium for this yeast
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