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Bacillus. Bacillus sp. GP rod Spore former Examples B. anthracis B. cereus B. subtilis B. megatarium Exoenzymes Lipase Amylase Casease Gelatinase. Spores. Agar Characteristics. Starch Hydrolysis. Enzymes Amylase Maltase Substrate Starch Reaction Clear zone.
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Bacillus sp. • GP rod • Spore former • Examples • B. anthracis • B. cereus • B. subtilis • B. megatarium • Exoenzymes • Lipase • Amylase • Casease • Gelatinase
Starch Hydrolysis • Enzymes • Amylase • Maltase • Substrate • Starch • Reaction • Clear zone
Protein Hydrolysis • Enzymes • Proteases • Caseases • Substance • Casein • Reaction • Clear zone
Lipid Hydrolysis • Enzymes • Lipase • Substrate • Tributyrin • Reaction • Clear zone
Collagen Hydrolysis • Enzyme • 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 • 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 • Starch (amylase, maltase) • Lipid (lipase) • Proteins (Casease, protease) • Collagen (Gelatinase) • Reaction • (+) = clearing • (-) = no clearing
BEA and Citrate Results + + Bile Esculin Citrate
Hydrolysis Results Summary casein lipid Gelatin Starch