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Day 1st,

Strains taken:

Two strains of antinomyces were taken from Khumbu region of Nepal designated as
J13e
K82

These samples/strains were taken as provided by RLABB. Pure culture was obtained by streaking on starch-casein agar (SCA) media.


gm/lt
Soluble starch 10.0
Casein 0.3
Potassium nitrate 2.0
Dihydrogen orthophosphate 2.0
Magnesium sulphate 0.05
Calcium carbonate 0.02
Ferrous sulphate 0.1
Agar 20.0
ph 7.2


To avoid fungal contamination cyclo-hexamide was added to the SCA media (50µgm/ml).

Observation:
J13e: growth seen with high cell mass and pigmented
Substrate: size 0.4 to 1 mm, entire margin, pink color, leathery consistency
Ariel: white color, convex elevation

K82: growth was seen with high cell mass and white color
Substrate: yellowish white in color, entire margin, leathery consistency
Ariel: white color, convex elevation


K82 showed antifungal activity: showed zone of inhibition against the fungus growth


Day 2nd,

Mutant development:

Media (SCA) containing different concentration of sodium azide was prepared and culturing of the strains was done by streak plate method.

Observation summary:

· Different concentrations of sodum azide were prepared viz, 1ppm, 2ppm, 3ppm, 4ppm, 5ppm, 6ppm, 7ppm, 8ppm, 9ppm, 10ppm, 20ppm, 30ppm, 40ppm, 50ppm, 60ppm, 70ppm, 80ppm, 90ppm, 100ppm.
· Growth were seen only upto 40ppm for K82
· Growth were seen only upto 50ppm for J13e
· Strain J13e showed pigmentation (pink color)
· Strain K82 showed antifungal activity