An effective endotoxin was isolated from two Bact. tularense strains. The cultures were poured onto blood-cystin-agar in 'petri' plates, strained, the culture soaked in sterile, distilled water, and this emulsion was autolysed for 45 days at 37C and shaken twice daily, after which all bacteria appeared dead. The emulsion was centrifuged after sterility test, and the resulting clear autolysate was dried in a vacuum over concentrated sulfuric acid. This dried endotoxin was dissolved in sterile distilled water, 50 milligrams to every cc, and this bouillon colored solution after a second centrifuging was applied to experimental testing.
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Talât Vasfi Öz (1940) studied this question.