Confirmation was obtained for the occurrence of an interaction between high and low molecular weight fractions of milk during its pasteurization, resulting in the formation of germinant(s) for Bacillus cereus. Various milk fractions were added to supernatants or dialysates of raw skim-milk, pasteurized and assayed for germinant. Micellar casein obtained by high speed centrifugation of raw milk stimulated germinant formation, but whole casein obtained by acid precipitation was less stimulatory and individual casein fractions showed no stimulation. A membrane rich ‘fluff’ layer obtained by high speed centrifugation of raw milk was highly stimulatory and suggested the possibility that the somatic cell count of milk may be related to germinant production.

Using milk from endotoxin infused and infected quarters of individual cows, a correlation between cell count and germinant formation was clearly demonstrated. Adjustment of the cell counts of individual milk samples was generally accompanied by corresponding changes in germinant formation, though removal of cells by filtration did not have this effect, possibly since germinant precursor had leached from the cells and remained in the milk. Pasteurization was necessary, not simply to express germinant from cells but to effect an interaction of germinant precursors. It is not clear whether the cell associated germinant precursor is derived from the cells themselves or from a related component. Addition of blood serum proteins was without stimulatory effect.

Since low cell count milks sometimes supported appreciable germination after pasteurization, but showed still higher levels when the cell count was raised, germinant may be formed via 2 pasteurization dependent processes, one between milk constituents alone and the other involving both milk constituents and somatic cells.