Different concentration gradients of the substrates i.e. cheese whey, cow urine, rain water and tube well water ranges from 10-1 to 10-6 with ZARROUK’S medium were analyzed for Spirulina growth at pH 9.5 ± 2, temperature 30°C ± 2 and photo-period of 10-12 hours. Spirulina was successfully cultivated by using different substrates and maximum chlorophyll and protein content was founded. The results indicate the potentiality of all the four substrates to provide nutrients to culture medium that reduces its valuable cost and make it a cheaper and economic medium to cultivate Spirulina. (Mahavir Joshi,2013)