Antifungal Activity of Pleurotus Ostreatus (Jacq

International Journal of Advancements in Research & Technology, Volume 9, Issue 3, March-2020 ISSN 2278-7763 14

Antifungal activity of Pleurotus ostreatus (Jacq. Ex: Fr.) Kummer from Syria

Rana Butrus1, Ramia Saoud2 1 Master student, Department of biology, Faculty of Science, Al Baath Universit, Syria. 2 Teacher, Department of biology, Faculty of Science, Al Baath University, Syria. Email:[email protected]

ABSTRACT In this study, the antifungal activity of crude Ethanolic and Methanolic extracts of Pleurotus ostreatus which grow on trunk apples in Syria (west Homs), against yeasts as Candida albicans (ATCC 10321), Geotrichum candidum, and pathogenic fungi, Aspergillus brasiliensis (ATCC 16404), Aspergillus flavus, Cladosporium cladosporioides, Fusarium oxysporum, Alternaria citri , using the agare well method for C.albicans and dilution in agar for others fungi. All tested microorganisms affected differently, except C.albicans, and showed the extracts, the maximum inhibition against F.oxysprum (13-14mm) and percentage (15,2%), (16,9%), and C.cladosporioides were the least tested fungi sensitive to both Ethanolic and Methanolic extracts.

Keywords: Ethanolic and Methanolic extracts, oyster mushroom, Pleurotus ostreatus, apple tree trunks, pathogenic fungi.

1 INTRODUCTION In addition to its great environmental importance in the The species of the genus Pleurotus spp. belonging to Fungi recycling of agricultural waste biological conversion of lignine kingdom Basidiomycota phylum, Agaricales order, Pleurotaceae and cellulose wastes, disassociation of industrial dyes, and family, About 70 species were Recorded around the world, And production of enzymes [19],[3]. grow saprophyte at temperatures between (12 - 32 ° C), and Few This present study to investigate antifungal activity of fruiting are parasitic P.eryngii [1],[2],[3],[4]. body of P.ostreatus growing on apple stump against some Their colors vary from white, gray, pink, bluish gray, cream, pathogenic fungi and yeast in west Homs (Syria). brown, and gold [1],[3] P.ostreatus is one of the most important species of oysters, food and medicine, spread in temperate to 2 MATERIALS AND METHODS tropical and subtropical regions, except in the western Pacific 2.1 Collection of the fungal samples: where the polar climate [5], it is called in China by sea eagle (abalone) [6], And in Syria called with apple mushroom, poplar Fresh P.ostreatus samples (oyster mushroom) were collected IJOARTfrom apple tree trunks (strain gray) [picture 1],from western parts mushroom, or fig mushroom depending on where they grow. after Agaricus bisporus, the oyster mushroom is the second of Homs (Syria, during 20 October, to 20 January). Identification most important mushroom in production in the world. [1],[6],[3] and classification of mushroom in Syria was studies by [20] . It grows repeatedly were the high humidity in the late autumn, and in the wet places of the trunk, and the fallen branches, it has a great ability to digest wood, the cap is kidney or oyster shape, with a short stem tapering downward from one side, or not. Often several groups occur simultaneously, and they can be found on the same stumps or long for several years[7],[2],[5],[8]. in Syria it spreads in most of the coastal areas (Baniyas,Lattakia,Tartous) [7], and North (Aleppo, Idlib)[9], the middle (Homs, Hama) [10], during late autumn, on cruel tree stumps, wide and falling leaves Such as figs, berries, oaks, poplar, apples ,In addition to elm trees, willow, beech, birch [5]. Some research was conducted on its geographical distribution and its spread in Syria, and its genetic diversity [7], [9], in addition to studying the biological activity of Ethanolic and Methanolic extracts of the fruiting bodies of the oyster growing wild on figs and berries [11], and the mycelium invitro [12], the fruiting bodies and mycelium has many chemical compounds such as volatile compounds, alkaloids, phenols, tannins, saponins, fatty acids, glycosides, and lactins, give its therapeutic Fig 1: p.ostreatus growth on apples tree trunks and biological properties [13], as an anti-tumors, Immunomodulatory, anti-inflammatory, anti-hypertensive, antimicrobial, antiviral and parasite [14],[15],[16],[17],[18].

F.oxysporum was the most sensitive, with inhibition diameter 2.2 MICROORGANISMS AND GROWTH CONDITIONS: (13-14.4 mm) at (1875- 1250 μg/ml) respectively table 1, fig(4,A C. albicans (ATCC 10321) and A. brasiliensis (ATCC 16404) and B) and percentage (15.2, 16.9 %) fig (2 and 3) respectively. were Test microorganisms included the yeast obtained from labs G.candidum showed clear sensitivity to extracts in low company Medico medical– Homs–Syria. concentrations (62.5 and 312 μg/ml), Table 1, fig(4,C and D) Cultures of these were growed on Sabouraud Dextrose Agar at Compared to other fungi. temperature (28°C) and (37°C) respectively. Similary in our study, [25] report that the ethanolic extract of And F.oxysporum isolated from Seeds of Cicer sp,)Department P.ostreatus exhibited different level of antifungal activity against of biology-Al-Baath University-Homs), and Aspergillus flavus, A.fumigatous, Pencillium chrysogenum, C.cladosporioides ,G.candidum were obtained from Sporotricum carnis, and Humicola grisea. microbiology lab (Department of Plant, Tishreen University, Lattakia). TABLE 1 : Average diameters inhibition of pathogen tested fungi of these culture were growed on Potato Dextrose Agar at P.ostreatus in mm temperature (28°C). 2.3 MICROORGANISMS AND GROWTH CONDITIONS: C. albicans (ATCC 10321) and A. brasiliensis (ATCC 16404) were Test microorganisms included the yeast obtained from labs company Medico medical– Homs–Syria. Cultures of these were growed on Sabouraud Dextrose Agar at temperature (28°C) and (37°C) respectively. And F.oxysporum isolated from Seeds of Cicer sp,)Department of biology-Al-Baath University-Homs), and C.cladosporioides ,G.candidum were obtained from microbiology lab (Department of Plant, Tishreen University, Lattakia). these culture were growed on Potato Dextrose Agar at temperature (28°C). 2.4 SAMPLES PREPARATION: Fruiting bodies were cut into bits and dried in dehydrator at (40°C) for three days, Then it's were homogenized in a house hold blender until they turned to powder. 2.5 PREPARATION OF CURD EXTRACTS: (50 g) powdered mushroomsIJOART were extracted separately in (200 ml) methanol and ethanol, in flasks.The flasks were covered with aluminum foil and left for (72 hours) at room temperature. sample were extracted twice, then filtered through whatman No.1, the solvent evaporated and concentrated extracts at Figure 2: The percentage of inhibition of Methanol extracts of P.ostreatus (40°C).The concentration was stored at (4°C) until study, this against tested pathogenic fungi. concentrations methods modified from [21]. 2.6 ANTIFUNGAL ACTIVITY: The concentraction dissolved in DMSO (2 ml), to obtain the final .(concentration (62.5،312،625،937،1250،1875 μg/ml The antifungal activity of extracts were determined using agar well diffusion for C.albicans [22] and agar dilution methods for others pathogenic fungi [23]. Inhibition diameters were measured in millimeters (diameters of control – diameters of treatment with extracts), and the percentage inhibition was calculated according to [24].

Each experiment was performed at least three replications. Figure 3: The percentage of inhibition of Ethanol extracts of P.ostreatus against tested pathogenic fungi. 3 RESULTS AND DISCUSSION All tested pathogenic fungi were inhibited by Ethanolic and Jahhar [12] showed ability the pure extract of lamella Methanolic extracts differently, except C.albicans, and was P.ostreatus, inhabition F.oxysporum (17 mm) [18] indicated to diameters inhibition between (3.4-14.4 mm), table 1. P.ostreatus (gray) against Trichoderma harzianum (5-10 mm), Ethanol extract was more effective than Methanol towards and Pyhium sp (3 mm), Verticillum sp (1-3 mm). tested fungi except A.brasiliensis. C.cladosporioides was least affected by the methanolic extract, compared to the tested fungi, in inhibition diameter (3.7 mm), at

(1875 μg/ml), while the ethanol extract has the biggest effect in [2]- Huffman,D.M., Tiffany, L. H., Knaphus, G., and Healy, R. A., 2008- low concentration (937 μg/ml). Mushrooms and others fungi of the midcontinental united stated, chin, I. Lowapress, 361pp. And it is known that C.cladosporioides contains fungal [3]- Owaid,M.N.,Al-Saeedi,S.S.,Abed,I.A. 2015- Mineral elements of white, antibiotics used to inhibit the growth of many plant pathogenic grey,yellow and pink oyster mushrooms (higer Basidiomycetes), Gida, 40(6):319- fungi especially apples scabes [27]. 326pp. While C.albicans was resistant to action of ethanolic and [4]- Yonis، Fang-Sheng Wu, Hussien H. El Shikh .2015-antimicrobial activity of extracts of the oyster culinary medicinal mushroom Pleurotus ostreatus and methanolic extracts table 1,probably as aresult the type of solvent identification of a new antimicrobial compunds.int J Med mushroom.pub med. used, and the part of fungus. 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