Metabolic Reactions Enzymology Catabolism Phototrophy Anabolism Microbial Metabolism.
BACTERIAL · PDF fileBacterial Metabolism Metabolism –Sum up all the chemical processes...
Transcript of BACTERIAL · PDF fileBacterial Metabolism Metabolism –Sum up all the chemical processes...
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BACTERIAL METABOLISM
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Bacterial Metabolism
Metabolism
– Sum up all the chemical processes that occur within a cell
1. Anabolism: Synthesis of more complex compounds and use of energy
2. Catabolism: Break down a substrate and capture energy
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All microbial metabolisms can be arranged according to three principles:
1. obtaining carbon for synthesis
– Autotrophic: carbon is obtained from CO2
– Heterophic: Corbon is obtained from organic compounds
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All microbial metabolisms can be arranged according to three principles:
• 2. Obtaining reducing equivalents for enegry conservation and synthesis
– Lithotrophic: reducing equivalents are obtained from inorganic compounds
– Organotrophic: reducing equivalents are obtained from inorganic compounds
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All microbial metabolisms can be arranged according to three principles:
• 3. Source of the energy
– Chemotrophic: energy is obtained from external sources
– Phototrophic: energy is obtained from light by photosynthesis
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Biochemical tests can be used for the identification of the bacteria
1. Oxidase Test
2. Catalase Test
3. Fermentation tests: Glucose, lactose, sucrose,…….
4. Enzyme tests: urease, tryptophan deaminase, DNase,….
5. Utilization tests: Citrate, malonate, asetate
6. Decorboxylation tests: Lysine, Arginine, Ornitine
7. Hydrolysis tests: Gelatin, esculin
8. Others: H2S production, motility, growth in KCN,….
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Catalase test
Streptococci
Staphylococci
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Oxidase test
Positive Negative
P. aeruginosa E. coli
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Lactose fermentation
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On the front bench
Oxidase test Catalase test
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.
Citrate medium
Pseudomonas
E. coli
Klebsiella
Urease medium
E. coli Proteus
Oxidative Fermentative medium
On every bench
Triple Sugar Iron Agar
Pseudomonas
E. coli
Proteus
Pseudomonas
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Inoculation to Fermentation medium
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Inoculation to Urease medium and Citrate medium
Urea agar Citrate agar
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Inoculation to the Triple Sugar Iron Agar (TSIA)
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Fermentation medium
Urea agar Citrate agar
1
2
TSIA
Don’t tighten the cap of the tube
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BACTERIAL METABOLISM II
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Oxidation-Fermentation Test (OF test)
Carbohydrate (Glucose)
Bromthymol blue (pH indicator)
Mineral oil
(anaerobic environment)
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Mineral oil
(anaerobic environment)
Glucose fermentation
Acid production
pH of the medium
pH indicator
green yellow
Gas production-Durham tube
Glucose fermentation (-) Glucose fermentation (+)
Glucose fermentation
Acid production
pH of the medium doesn’t change.
pH indicator
green
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Fermentative
Oxidative
Non-fermentative
Oxidative
Non-fermentative
Non-oxidative
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Urease Test
Urea agar:
Urea
Phenol red (pH indicator)
Urea agar
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Urea
Ammonia + CO2
pH
Urease
pH indicator
yellow pink
Urease (+) Urease (-)
Urea
Urease
The pH of the medium doesn’t change.
pH indicator
yellow
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Citrate utilization
Citrate medium
Citrate medium:
Citrate (Carbon source)
Ammonium salts (Nitrogen source)
Bromthymol blue (pH indicator)
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Citrate utilization
Ammonium salts
Ammonia (pH )
pH indicator
green blue
Citrate(+) Citrate (-)
Citrate utilization
Ammonium salts
No ammonia production
pH doesn’t change.
pH indicator
green
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Triple Sugar Iron Agar (TSI)
Glucose %1
Lactose %10 Acid and gas production
Sucrose %10
Phenol red (pH indicator)
Sodium thiosulfate (-S source)
Ferrous sulfate (H2S indicator)
Peptone (N source-Oxydation) Triple Sugar Iron Agar
(TSI)
Acid-gas and H2S production
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Only glucose fermentation
Glucose (%1) fermentation
Acid production
The pH of the medium
pH indicator
Red yellow
Peptids amines
pH of the slant
pH indicator
red pink
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Glucose and lactose and / or sucrose fermentation
Gas production
Glucose and lactose and/or sucrose fermentation
Acid production
The pH of the medium
pH indicator
red yellow
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Nonfermentative bacteria
Carbohydrate fermentation (-)
pH of the medium doesn’t change.
The colour of the medium remains same
Peptids amines O2
pH
pH indicator
Red pink
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H2S production
Thiosulfate (sulfur source)
H2S
H2S + ferrous sulfate (H2S indicator)
Ferric sulfide
(black precipitate)
Enzymatic activation
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Automated System depends on biochemical and metabolic reactions