They were incubated at 30°C at 160rpm for 21 days. A control devoid of the bacterial isolate was prepared for each set of experiments.(17) Table-6 Biochemical Reagents Test Medium Reagent Observation 1.Carbohydrate Glucose, maltose, Phenol red Red yellow fermentation test Sucrose, Lactose, color ( Gas Mannitol, Xylose production) 2. Urea utilization Urea broth, Phenol red Pinkish red color test 3. H2S Production 2% Peptone Lead acetate Blackish of paper strip test 4. Gelatin hydrolysis Nutrient gelatin broth Liquefaction at – test 4°C 5. Citrate utilization Simmons Citrate agar Bromothymole Green-Blue test blue Peptone nitrate broth Sulfanylic acid Red color 6. Nitrate reduction test + a-Naphylamine Nutrient Agar Slant Oxidase strip Violet 7. Oxidase test 8. Catalase test Nutrient Agar Slant 3% H2O2 Formation of bubbles 9. M-R test Glucose Phosphate Methyl red Red color 10. V-P test Glucose Phosphate 40% KOH Pink + a- Naphthol 1% Peptone Kovac`s reagent Red ring 11. Iodole production test production 12.TSI slant Triple Sugar iron – – agar Slant 13. Macconkey`s Macconkey’s agar – – Agar plate plate 14. Gram`s Grams iodine, Microscopic – stainining Crystal violet, observation. Ethanol, D/W, Safranin
They were incubated at 30°C at 160rpm for 21 days. A control devoid of the bacterial isolate was prepared for each set of experiments.(17) Table-6 Biochemical Reagents Test Medium Reagent Observation 1.Carbohydrate fermentation test Glucose, maltose, Sucrose, Lactose, Mannitol, Xylose Phenol red Red yellow color ( Gas production) 2. Urea utilization test Urea broth, Phenol red Pinkish red color 3. H2S Production test 2% Peptone Lead acetate strip Blackish of paper 4. Gelatin hydrolysis test Nutrient gelatin broth – Liquefaction at 4°C 5. Citrate utilization test Simmons Citrate agar Bromothymole blue Green-Blue 6. Nitrate reduction test Peptone nitrate broth Sulfanylic acid + a-Naphylamine Red color 7. Oxidase test Nutrient Agar Slant Oxidase strip Violet 8. Catalase test Nutrient Agar Slant 3% H2O2 Formation of bubbles 9. M-R test Glucose Phosphate Methyl red Red color 10. V-P test Glucose Phosphate 40% KOH + a- Naphthol Pink 11. Iodole production test 1% Peptone Kovac`s reagent Red ring production 12.TSI slant Triple Sugar iron agar Slant – – 13. Macconkey`s Agar plate Macconkey’s agar plate – – 14. Gram`s stainining – Grams iodine, Crystal violet, Ethanol, D/W, Safranin Microscopic observation.
Dissertation-on-Petroleum-Biodegradation-In-Natural-Environment.docx