Biochemical Test of Clostridium botulinum

(Type A and proteolytic strains of types B and F)

Biochemical Test of Clostridium botulinum

Basic Characteristics Properties (Clostridium botulinum)
Ammonia Production Positive (+ve)
20% Bile Negative (-ve)
Capsule Negative (-ve)
Catalase Negative (-ve)
Flagella Peritrichous
Gelatin Hydrolysis Positive (+ve)
Gram Staining Gram-positive
H2S Positive (+ve)
Hemolysis Positive (+ve). β-hemolytic
Indole Negative (-ve)
Milk Digestion Mostly Positive (+ve)
Meat Digestion Mostly Positive (+ve)
Motility Mostly Positive (+ve)
6.5% NaCl Negative (-ve)
Oxidase Negative (-ve)
Shape Straight to slightly curved rods, 0.6–1.4 × 3.0–20.2 μm.
Spore Positive (+ve)

Fermentation of

Arabinose Negative (-ve)
Cellobiose Negative (-ve)
Fructose Negative (-ve)
Galactose Negative (-ve)
Glucose Positive (+ve)
Glycogen Negative (-ve)
Inulin Negative (-ve)
Lactose Negative (-ve)
Maltose Negative (-ve)
Mannitol Negative (-ve)
Mannose Negative (-ve)
Melibiose Negative (-ve)
Raffinose Negative (-ve)
Rhamnose Negative (-ve)
Ribose Negative (-ve)
Salicin Negative (-ve)
Sorbitol Negative (-ve)
Starch Negative (-ve)
Sucrose Negative (-ve)
Trehalose Negative (-ve)
Xylose Negative (-ve)

Amino Acid Utilization

Arginine Positive (+ve)
Glycine Positive (+ve)
Phenylalanine Positive (+ve)
Proline Positive (+ve)
Serine Positive (+ve)
Tyrosine Positive (+ve)
Tryptophan Positive (+ve)

Enzymatic Reactions

Casein Hydrolysis Positive (+ve)
Esculin Hydrolysis Positive (+ve)
Lecithinase Negative (-ve)
Lipase Positive (+ve)
Superoxide dismutase Positive (+ve)
Tryptophan Deaminase Negative (-ve)


  1. Bergey, D H, William B. Whitman, Vos P. De, George M. Garrity, and D Jones. Bergey’s Manual of Systematic Bacteriology: Vol. 3. New York: Springer, 2009. Print.

About Author

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Sagar Aryal

Sagar Aryal is a microbiologist and a scientific blogger. He attended St. Xavier’s College, Maitighar, Kathmandu, Nepal, to complete his Master of Science in Microbiology. He worked as a Lecturer at St. Xavier’s College, Maitighar, Kathmandu, Nepal, from Feb 2015 to June 2019. After teaching microbiology for more than four years, he joined the Central Department of Microbiology, Tribhuvan University, to pursue his Ph.D. in collaboration with Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Saarbrucken, Germany. He is interested in research on actinobacteria, myxobacteria, and natural products. He has published more than 15 research articles and book chapters in international journals and well-renowned publishers.

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