My thesis is on the isolation of biosurfactant producing halophilic streptomyces from saline soil.
Use aged sea water for making the medium or you can use artificial salt. But before deciding the salt concentration, analyze the salt content of the soil based on which you can use 50% or 75% seawater. Use ISP medium or Actinomycete Isolation Agar (AIA). It is suggested that cycloheximide (50 µg/ml) andnystatin (50 µg/ml) be added to prevent growth of fungi.
try air drying of soil to reduce bacterial contaminants / dry heat - 120 oC for 2hr - isolation of hardy spore bearers - the rare actinomycetes including rare Streptomybes
I have isolated streptomyces from normal soil but if I were to try and isolate them from saline soil (~0.2%) I would first try to just do it the regular way without adding any extra salt or add ( ~0.2% NaCl) to everything you do.
First dry out the dirt. Leave it in some incubator >50 C for a week then add enough water to mix the dirt and filter through a cotton plug using a syringe.
Dilute this mixture 100 fold and plate the filtrate onto your plates (ISP 1 2 3 4 and TSB etc.) The plates should contain nalidixic acid and cyclohexamide. Grow at 28 C for a bout a week.
This is a general approach. I would be surprised if nothing was on those plates.
thanks,actually I want to isolate biosurfactant producing halophilic streptomyces,when I was searching about this subject I find out salinity has a positive effect on biosurfactant production,Do you know whats the mechanism ?
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