APPENDIX I. I )! i. 0.3 g Magnesium sulfate (Hydrated) g Calcium carbonate g Ferrous sulfate g Agar-agar - 20.

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1 APPENDIX I 1. Starch casein agar medium Medium was used for isolation, morphological studies, maintaining the cultures and for antitumor antibiotic production from the soil and river sediments isolates. Starch (soluble) Potassium nitrate Sodium chloride Di potassium hydrogen Ortho phosphate I )! i o o o 9 to M M m cro, cro oo 0 0 Casein (vitamin free) 0.3 g Magnesium sulfate (Hydrated) 0.03 g Calcium carbonate 0.02 g Ferrous sulfate 0.01 g Agaragar 20.0 g 1000 ml ph (prior to sterilization) 7 ± Oatmealagar medium (ISP 3) For selective isolation of actinomycetes Oatmeal 60g Agar 12.5g 100ml 3. Potassium tellurite agar medium For selective isolation of actinomycetes 4. Half strength nutrient agar medium For selective isolation of actinomycetes Peptic digest of animal tissue 2.5 g 0.75g Beef extract 0.75g Sodium chloride 2.5 g Agar 10.0 g 1000 ml ph 7.4 ± 0.2

2 5. Production medium I For antibiotic production 05.Og Dextrose 10.5g Starch 20. Og Casein hydrolysate 5.0g Calcium carbonate 4.0g 1000 ml ph 7.2 Production medium H For antibiotic production Soya bean meal 05.5g Com streap liquor 05.5g Soluble starch 10. Og Dextrose 05.5g Calcium carbonate 07. Og 1000 ml ph Malt extract agar (ISP 2) Medium was used for the test of sodium chloride tolerance; to determine cell wall composition only broth was used. Malt extract Dextrose Agar ph. j o o M C w o k o era era ora cro Peptoneiron agar (ISP 6) Medium was used for the test of Melanin production and H2S production. Peptone Proteose peptone Ferric Ammonium Citrate K2HPO4 Na2S203. 5H20 O O cn t o o b o 1 P1 00 CTQ, aa era 0 era era

3 1.0 g Agar 20.0 g 1000 ml ph 7 ± Tyrosine agar (ISP 7) Medium used for the test of tyrosinase reaction. LTyrosine 05 g Glycerol 15.0 g LAsparagine 1.0 g k2hpo4 0.5 g MgS04.7H g NaCl 05 g FeS04.7H g ISP trace salts solution 1.0 ml Agar 20.0 g 1000 ml ph 7.2 to 7.4 ISP trace salts solution FeS04.7H20 MnCl2.4H20 ZnS04.7H20 'QD'OD'OD^ 6 rh r ( i l *55? d o o h 10. Inorganic salts starch agar medium (ISP4) Medium was used to study cultural characteristics. Soluble starch K2HPO4 (anhydrous base) MgS04?7H20 NaCl (NH4)2S04 CaCOs Trace salts solution* Bacto Agar (Difco) The unadjusted ph should be *Trace salts solution FeS04?7H20 MnC12?4H20 ZnS04?7H20 os acrqgrqcrqtocrq 0 1 ggtq (to 2a g o p p Q a l~ > J a m 0 *

4 11. Glycerol asparagine agar (ISP5) Medium was used to study cultural characteristics LAsparagine, l.oog Dipotassium phosphate, l.oog Trace salt solution (ml), l.oog Agar, 20.00g 1000ml ph TryptoneYeast Extract broth (ISP 1) Medium was used for the test of Melanin production Tryptone 5.0 g 3.0 g 1000 ml ph 7 ± Gelatin Agar medium Medium was used for the test of Gelatin hydrolysis. Peptone Beef extract Gelatin Agar ph N2 4^ CO U1 i+ 3 P o o b o o (W cro, oro, to a Milk casein agar medium Medium was used for the test of casein hydrolysis Diy milk, instant nonfat 50.0g Pancreatic digest of casein 5.0g 2.5g Glucose iog Agar 12.5g

5 15. StarchPeptone agar medium Medium was used for the test of Starch hydrolysis. Protease peptone 15.0 g Dextrose 2.0 g Starch 10.0 g Sodium chloride 5.0 g Disodium acid phosphate 3.0 g Gelatin 20.0 g Agar 10.0 g 1000 ml ph 7.3 ± Organic Nitrate Broth (Waksman medium No. 17) Medium was used for Nitrate reduction reaction Peptone 5.0 g Meat extract 3.0 g KNOs 1.0 g 1000 ml ph 7 ± Glycerol nutrient broth The medium was used for the chemical tolerance for the extent of growth. Medium used to cultivate the type cultures and resistant cultures. Peptic digest of animal tissue Beef extract Sodium chloride ph i i i i i i.*vj > j cn h' v j cjt ^ o ui bi o i + o CTQ 0Q CTOi cro

6 18. Basal medium for Nitrogen source utilization DGlucose MgS04.7H20 NaCl FeS04.7H20 K2HPO4 Agar ph Nitrogen source b o b o ^ cn 01 o r( *' O «g.cm cm cmcmcm cm # 1+ < P to 19. Nutrient agar (Hi media labs, Mumbai) Medium was used for agar diffusion assay and cross streak method. Peptic digest of animal tissue 5.0 g 1.5 g Beef extract 1.5 g Sodium chloride 5.0 g Agar 20.0 g 1000 ml ph Bennett s Agar 1 g Beef extract 1 g NZ Amine, Type A* 2 g Glucose 10 g 1000ml Agar 20 g ph Czapek s medium Sucrose Nano3 K2 Hpo4 MgS04. 7H20 Feso4. 7H20 Tap water Ph Medium used to study antibiotic production.