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Botulinal neurotoxin in and around colonies of Clostridium botudinum types A, B, and E and of toxigenic Clostridium butyricum was detected by an enzyme-linked immunoassay procedure whereby the toxin was transferred from the agar medium to a nitrocellulose support and the immobilized toxin was probed with type-specific antibodies. The method identified the toxin types of the colonies grown from a mixed inoculum of C. botulinum serotpes. The specificity of the antitoxins for tpe A and B toxins was improved by adsorption of the antitoxins with the antigens of heterologous type cultures.

Bacillus coagulans was described at the Iowa Agricultural Experiment Station (Hammer (1915)) as the cause of an outbreak of coagulation in evaporated milk packed by an Iowa condensery. Cordes (1928) found B. coagulans responsible for an outbreak of "flat-sours" in evaporated milk. The cans from which the organism was secured had been subjected to 114.40C. (238°F.) for twenty minutes in a batch sterilizer. However, milk cultures of the organism were killed when exposed in an autoclave to a temperature of 111.70 to 112.80C. (2330 to 2350F.) for fifteen minutes, the milk itself reaching 112.20C. (234°F.), being between 1000 and 112.20C. (2120 and 234°F.) for six minutes, and between 107.2°and 112.20C. (2250 and 2340F.) for four minutes. B. coagulans was recently found to be the cause of an outbreak of coagulation in evaporated milk packed by a condensery in a neighboring state. A number of the cultures isolated were studied in order to (1) check and enlarge the description of the organism, and (2) secure additional information on the changes it produces in evaporated milk. GENERAL CHARACTERS OF OUTBREAKS OF COAGULATION IN EVAPORATED MILK DUE TO B. COAGULANS In 1915 an outbreak of coagulation in evaporated milk occurred in an Iowa condensery and extended over a period of several months, with the May and June milk showing the highest percentage of spoilage. B. coagulans was isolated from the spoiled milk. At the time coagulation was first encountered the heat Journal Paper Number B1 of the Iowa Agricultural Experiment Station.

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In studies on the metatrophic bacteria, action on carbohydrates and proteins has been given much more attention than action on fats. This has been due, in part, to the inability of many of the common bacteria to attack fats, but the relative inconvenience of I The work reported was supported by a grant from the Rockefeller Fluid Research Fund.