Characterization of the Isolate of Scedosporium Species Complex Population from Soil and water
Our results showed that the species of Scedosporium were recovered from environmental samples from soil and water collected from 18 Sudanese states. The pH of the soil samples ranged between 6.0 and 8.0, with nitrate, concentrations varying from 0–20 mg/100 g (table-1). As shown in figure-2, we found colonies with different morphologies on each PDA, OA, MEA, and SDA agar plate. From each plate, we selected one representative colony of each morphological type of Scedosporium, which yielded 3,059 single colonies: 2,891 colonies from 800 soil samples and only 168 colonies from 100 water samples.
These strains were identified according to their microscopic morphology  and further confirmed by BT2 gene (98-100%sequence similarity) and those sequences for which identification was ≤ 98.% were confirmed by ITS and another region of the β-tubulin gene sequencing [5,33]. Scedosporium apiospermum was the microorganism most commonly recovered with 1662 isolates, followed by Scedosporium. boydii with 1168, Scedosporium aurantiacum with 192 isolates, Scedosporium dehoogii with 22 isolates and Scedosporium angustum with 15 isolates .
Colonies of Scedosporium boydii grow rapidly at 25°C on PDA, as white aerial mycelia and later becomes dark gray or smoky brown. Wooly to cottony from the surface, and it is pale with brownish black colour from the reverse. Microscopy: Hyphae hyaline and septate. Conidia Unicellular, finely smoothened, ovoid with truncate bases formed singly at the ends of the conidiophores  (figure3-1).
The cleistothecia of Scedosporium boydii are dark brown fruiting bodies (completely closed ascocarps) measuring 100 to 200 µm in diameter.. Asci are subglobose to globose measure 8 to 13 µm by 12 to 18 µm and bear 8 ascospores inside. Ascospores are unicellular, ovoid to ellipsoidal, smooth, and pale yellow brown to copper in color measuring 4 to 5 µm by 7 to 8 µm .
Material examined: This fungus was isolated from Urban gardens, agricultural areas, sports parks, recreational parks Urban industrialized area, sandy riverbanks of the White , house plant , surface soil samples from houses , soil from animals manure , surface river water , sewage water , and wastewater treatment plants samples taken from River Nile (250 isolates), El-Obeid (193 isolates), North Darfour (188 isolates), Sennar (135 isolates), El Geteina (125 isolates), Dongla (122 isolates) ,Khartoum (85 isolates), El- Gazira state (70 isolates).
Colonies of Scedosporium apiospermum grew at 25°C on PDA as white gray, and later becoming darker gray or brown in colour from the surface .The colour from the reverse, it is dark brown or gray to black .Microscopy: Hyphae hyaline and septate. conidiogenous cells percurrent, lateral or terminal, subhyaline, smooth-walled, usually cylindrical, producing obovoidal or ellipsoidal, 5-14 x 3-5 µm, smooth-walled conidia(figure 3-2) Scedosporium apiospermum has large cleistothecia 140-480 μm .
Material examined: This fungus was isolated from Urban gardens, agricultural areas, sports parks, recreational parks Urban industrialized area, sandy riverbanks of the White Nile , house plant, surface soil samples from houses , soil from animals manure , surface river water , sewage water , and wastewater treatment plants samples taken from Khartoum ( 350 isolates), North Darfour (235 isolates), El- Gazira (270 isolates), River Nile (225 isolates). El Geteina (210 isolates), Sennar (175 isolates), El-Obeid (122 isolates) and Dongla state (75 isolates)
Scedosporium aurantiacum colonies at 25°C on PDA, expanding, cottony, with a light yellow diffusible pigment; reverse in orange colour. Microscopy:. Conidia produce directly from undifferentiated hyphae abundant, sessile or on short protrusions, brown, usually obovoidal 6-10 x 3-5 µm, thick-walled  (figure 3-3).
Material examined: This fungus was isolated from soil samples from Urban gardens, agricultural areas, sports parks, recreational parks, Urban industrialized area and sandy riverbanks of the White Nile taken from Khartoum (75 isolates),El- Gazira ( 62 isolates), and El Geteina (55 isolates).
Scedosporium dehoogii colonies at 25°C on PDA growing rapidly, cottony to lanose, initially dirty white, becoming pale grey with age; reverse pale, greenish or whitish. Microscopy.:Conidiogenous cells arising from undifferentiated hyphae, cylindrical to somewhat flask-shaped, 6-50 x 1.0-1.5 µm, producing slimy heads of 1-celled, smooth-walled, subhyaline to brown, ovoidal conidia, 6-12 × 4-5 µm  (figure 3-4).
Material examined: This fungus was isolated from soil samples from agricultural areas, house plant , surface soil samples from houses , soil from animals manure taken from AlQadarif state ( 22 isolates).
For the distribution of Scedosporium species complex, between the area studied, we found that The highest density of Scedosporium species was found in urban gardens (30%), agricultural areas(21%), industrial parks (16%), sports parks (12%), home gardens (8%) surface soil samples from houses(5%) , soil from animals manure (4%) and wastewater treatment plant activated sludges (4%) . In contrast, the lowest densities of these fungi were encountered in areas in contact with water (riverbanks) sewage water and surface river water, house plant. No Scedosporium boydii / Scedosporium apiospermum complex colonies were recovered from the evaluated water samples of drinking water and animal drinking water.
For the distribution of Scedosporium complex species over the country, we find the following: while most isolates of Scedosporium apiospermum (78.5%) were obtained from the north and centre of the country. On the other hand, Scedosporium boydii was equally distrusted in the centre, west and north of the country. Scedosporium aurantiacum was recovered in 85.7% of the soil samples from the centre of the country and 14.3% from river bank water. The isolation of Scedosporium dehoogii was restricted to the samples collected from the southern east. where Scedosporium angustum was found in the centre of the country from Khartoum and El- Gazira state and was only isolated from wastewater treatment plant activated sludges.
The phylogenetic relationships of Scedosporium species complex were studied using combined sequences of thee-locus alignment (ITS, CAL, and BT2) on the basis of a dataset consisting of 30 sequences of Scedosporium. apiospermum and related species (figure-4)`. The concatenated alignment consisted of 989 characters (including alignment gaps).The ML analysis was congruent with the BI analysis, both displaying a similar topology. In the combined ITS- BT2 tree, our strains were distributed across four clades, into a well-supported main clade grouping all Scedosporium species (figure-4). 2.814 of the isolates (92%) were placed into the Scedosporium apiospermum species complex clade A , and distributed across several terminal clades: A terminal clade containing the type strains of Scedosporium boydii ,Scedosporium apiospermum and Scedosporium angustum and our strains Clade B containing the type strain of Scedosporium apiospermum and our strains .Clade C included only two type strain of Scedosporium dehoogii and 12 of our strains. Clade D included four type strain of Scedosporium aurantiacum and our strains .