Isolation and enumeration of actinomycetes were performed on different media.
A measured area of sponge tissue (1-cm2 area) was excised using a sterile scalpel from the internal mesophyl area.
The tissue was then homogenized in sterile ASW using a tissue homogenizer.
The homogenate was serially diluted and all dilutions were placed on the media.
The isolation was performed on the selective media such as marine sponge agar (MSA)
(raffinose: 10 g; L-histidine: 1 g; ferrous sulphate: 0.01 g; dipotassium hydrogen phosphate: 1 g;
calcium carbonate: 0.5 g; agar: 15 g; sodium chloride: 20 g; aqueous host sponge extract: 100 ml;
double distilled
water: 900 ml; pH 7.8; autoclaved at 15 lbs for 15 min) and
standard media such as Emerson agar (EA) and modified
marine agar (MMA).