Bacillus

Bacillus sp. GP rod Spore former Examples Exoenzymes B. anthracis B. cereus B. subtilis B. megatarium Exoenzymes Lipase Amylase Casease Gelatinase

Spores

Agar Characteristics

Starch Hydrolysis Enzymes Substrate Reaction Amylase Maltase Starch Clear zone

Protein Hydrolysis Enzymes Substance Reaction Proteases Caseases Casein Reaction Clear zone

Lipid Hydrolysis Enzymes Substrate Reaction Lipase Tributyrin Clear zone

Bacillus Tri-Plate

Collagen Hydrolysis Enzyme Use loop (vs. needle) Substrate Reaction Gelatinase Use loop (vs. needle) Inoculate several times Stab deep Substrate Gelatin Reaction Clear Zone on Agar Liquefy Gelatin in tube

MSA Selective Differential Substrate pH = phenol red Reaction Mannitol Salt (7.5%) pH = phenol red Reaction Growth Fermentation

Nitrate Reduction Inoculate with loop Incubate Additional solutions Sulfanilic acid (A) Naphthylamine (B) (+) = Red Nitrate to Nitrite No color, add Zinc (+) = not red (-) = red

Citrate Enzyme Citrase Substrate Citrate pH Bromthymol blue Reaction Keto acids Oxaloacetic Pyruvate Acetic acid CO2 CO2 + NaH2CO3 Prussian blue = +

Bile Esculin Agar (BEA) Selective Bile Salts Differential Esculin (CH2O) Fe-NH3-citrate Reaction Growth in bile Hydrolyze esculin Esculetin-Iron Rxn Black precipitate

Results Exoenzymes Reaction Starch (amylase, maltase) Lipid (lipase) Proteins (Casease, protease) Collagen (Gelatinase) Reaction (+) = clearing (-) = no clearing

Starch Hydrolysis

Lipid Hydrolysis

Protein Hydrolysis

Gelatin (Collagen) Hydrolysis

BEA and Citrate Results + + Bile Esculin Citrate

Hydrolysis Results Summary casein lipid Gelatin Starch