Technique	CLOSTRIDIAL TOXIN ,(/,)/,(
Principle	Remove a few drops of the culture supernate in the egg meat medium after 72 to 96 hours of incubation and administer orally to a mouse with a syringe. Observe the animal over a period of a few hours to a few days. Flaccid paralysis and death are the usual symptoms. Inject 0.5 ml of 10% lactic acid intra- muscularly into the shaved thigh of a guinea pig to produce necrosis. In about 5 to 10 minutes the same volume of supernate from the egg meat culture should be injected at the same site. The animal is examined over a period ofa week. If the strain is toxigenic, the skin at the injection site will be tight and the subcutaneous tissue broken down and crepitant. An incision in the area releases a thin red fluid and a foul- smelling gas. On microscopic examination the fluid is found to contain large numbers of bacterial cells. Inject 0.5 ml of 10% lactic acid intra- muscularly into the shaved thigh of a guinea pig to produce necrosis. In about 5 to 10 minutes the same volume of supernate from the egg meat culture should be injected at the same site. The animal is examined over a period of a week. If the strain is toxigenic, the skin at the injection site will be tight and the subcutaneous tissue broken down and crepitant. An incision in the area releases a thin red fluid and a foul- smelling gas. On microscopic examination the fluid is found to contain large numbers of bacterial cells. Inject a white mouse in the thigh muscle with a 0.1 to 0.2 ml of 10% lactic acid to produce necrosis of the tissue. In 5 to 10 minutes inject the same volume of supernate from the egg meat culture at the same site. Observe the animal for 2 to 4 days. Symptoms of toxigenicity are rigidity of the tail and the limb followed by death.
Cautions
Method
Results
Positive control
Negative control
Reagents
Reference