Bacillus

1
Bacillus
2
Bacillus sp.

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

3
Spores
4
Agar Characteristics
5
Starch Hydrolysis

• Enzymes
• Amylase
• Maltase
• Substrate
• Starch
• Reaction
• Clear zone

6
Protein Hydrolysis

• Enzymes
• Proteases
• Caseases
• Substance
• Casein
• Reaction
• Clear zone

7
Lipid Hydrolysis

• Enzymes
• Lipase
• Substrate
• Tributyrin
• Reaction
• Clear zone

8
Bacillus Tri-Plate
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Collagen Hydrolysis

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

10
MSA

• Selective
• Differential
• Substrate
• Mannitol
• Salt (7.5)
• pH phenol red
• Reaction
• Growth
• Fermentation

11
Nitrate Reduction

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

12
Citrate

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

13
Bile Esculin Agar (BEA)

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

14
Results

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

15
Starch Hydrolysis
16
Lipid Hydrolysis
17
Protein Hydrolysis
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Gelatin (Collagen) Hydrolysis
19
BEA and Citrate Results


Bile Esculin
Citrate
20
(No Transcript)
21
Hydrolysis Results Summary
casein
lipid
Gelatin
Starch