16/11/2025
📌 Understanding the Biochemical Identification Chart for Bacteria
This chart is a quick reference used in microbiology labs to identify Gram-negative bacteria, especially those belonging to the Enterobacteriaceae family (like E. coli, Salmonella, Shigella, etc.).
Each row represents a bacterium, and each column is a biochemical test. The pattern of positive (+) or negative (–) reactions helps identify the organism.
🧪 Key Biochemical Tests in the Chart
1. Urea Test
Checks whether the bacterium can break down urea using the enzyme urease.
(+) pink color → urease is produced (e.g., Proteus, Morganella).
(–) → urease not produced.
2. TSI (Triple Sugar Iron) Agar
This test has two areas:
Slant
Butt
Results show:
A (Acid) → Yellow (sugar fermentation)
K (Alkaline) → Red (no sugar fermentation)
This helps determine which sugars the organism ferments.
3. Gas & H₂S Production
Gas (+) → bubbles or cracks in the medium.
H₂S (+) → black color (e.g., Salmonella, some Proteus species).
4. Motility Test
(+) organism moves away from the stab line (motile).
(–) stays in place (non-motile, like Klebsiella).
5. Indole Test
Checks whether bacteria can break down tryptophan into indole.
(+) → red ring forms (e.g., E. coli).
(–) → no color change.
6. Mannitol Fermentation
(+) bacteria can ferment mannitol sugar.
(–) cannot ferment mannitol.
7. Citrate Test
Tests ability to use citrate as the only carbon source.
(+) medium turns blue (e.g., Klebsiella, Enterobacter).
(–) remains green (e.g., E. coli).
📌 What This Chart Helps With
Microbiologists use these test patterns like a fingerprint to identify bacteria from:
Urine cultures
Stool samples
Blood cultures
Environmental samples
Each species shows a unique combination of positive and negative results.
For example:
E. coli → Indole +, Motile +, Citrate –
Klebsiella → Urease ±, Non-motile, Citrate +
Salmonella → H₂S +, Motile +, Indole –
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