In a continuing inquiry into the problems of intra-specitic natural resistance to infection, this laboratory, since 1940, has focused on mouse salmonellosis as an infection model, with nutritional manipulations of the host as the main interest of study. During the course of these investigations it was found that a nutritionally arranged difference in survivorship had its operational base in what was named "the double strain phenomenon" (1). Briefly, this involved the infection of the mice by the combined use of two strains of Salmonella typhimurium, one avirulent and the other virulent. On this information an assay was devised to detect survivorship differences with maximal sensitivity, the so called double strain inoculation test. Using this method of assay a program was begun to identify chemically the nutritional factor or factors promoting increased survivorship.In appfication it has been found that the double strain inoculation test has not been unburdensome in its statistical demands on the number of animals needed and in the amount of nutritional material which had to be prepared during the course of the various chemical fractionations. For practical reasons, therefore, an attempt was made to improve the method of assay. This improved assay, as will be shown, is based on the measurement of the population sizes of the virulent S. typhimurium in the spleens of mouse hosts in the presence of the avirulent form. This was achieved by the use of a distinguishing genetic marker. It is the purpose of the present paper to set forth this improved and more practicable form of the double strain inoculation method of assay.
Materials and MethodsAnimals.--The mice used were from a strain of W-Swiss mice, the same as employed in previous investigations (1). The program of outbreeding, previously described (2), has been continued and the stock was free of Sa/mond/a.