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Lab 17 Msc. Shaimaa obead Clostridium Division: Firmicutes Class: Clostridia Order: Clostridiales Family: Clostridiaceae Genus: Clostridium Species: C. tetani C. botulinum C. perfringens Clostridia are spore-forming, Gram-positive, anaerobes. They are known to produce a variety of toxins, some of which are fatal. There are three species of clostridia that cause widely recognized and often-deadly diseases. Clostridium tetani is the etiological agent of tetanus, Clostridium botulinum is the etiological agent of botulism, and Clostridium perfringens is one of the etiological agents of gas gangrene. C. tetani: C. tetani the causative organism of tetanus. Its produces terminal spores, giving it the appearance of drumsticks or tennis rackets. This species is motile and produces a single antigenic type of exotoxin. Spore wide spread in soil and animal feces. Tetanus Toxin, tetanospasmin, causes tetanus: unopposed muscle spasm, contraction due to interference of toxin with neurotransmitters. Tetanus (sometimes called lockjaw) is caused by the toxin (poison) of a bacterium. Tetanus: A disease caused by chemicals which are produced by a bacterium (Clostridium tetani) and are toxic to 1 the nerves. The infection usually occurs when the bacteria enter the body through a deep wound - these bacteria are anaerobic and hence don't need oxygen to survive C. botulinum: C. botulinum produces oval subterminal spores and is motile. This organism producing a toxin in food/wound that causes botulism. C. perfringens: C. perfringens produces large rectangular spores and is nonmotile.formerly called C. welchii, causes a wide range of symptoms, from food poisoning to gas gangrene. Also responsible for enterotoxemia (also known as "pulpy kidney disease") in sheep and goats and other animals Gas gangrene Diagnosis: Members of the genus Clostridium can be differentiated from other bacteria by laboratory techniques including enzymatic digestion on eggyolk agar plates. Diagnosis of C. perfringens: Egg yolk agar: opacity appears around the colony because C. perfringens produce lecithinase. Litmus milk agar: after incubation the stormy fermentation will appear because fermentation of glucose and digestion of casein. Naglers reaction: plate contain egg yolk agar with human serum divided in 2 parts. In one part adding 3 dope of antiserum and leave to dry then culturing each part with bacteria after incubation at 37 under anaerobic condition the part without antiserum show opacity around the colony while the part with antiserum will not. 2 Culture: Anaerobically at 37C On Robertson's cooked meat medium → blackening of meat will observed with the production of H2S and NH3 Difference characteristic between Clostridium spp.: Clostridium sp. Haemolysis Motility Terminal spore C. tetani + + + C. perfringens + _ _ C. botulinum + + _ Biochemical test: Clostridium sp. Egg yolk agar Digestion of fermentation casein G L S M lecithinase lipase C. tetani _ _ _ _ _ _ _ C. perfringens + _ + + _ _ + C. botulinum _ + + + + + + G=glucose L=lactose 3 S=sucrose M=maltose