Germ-free (GF) status signi'cantly "protected" male C3H mice from the early (before 40 weeks) development of liver-cell tumours in response to 7,12-dimethyE benz (a)Pollard and his colleagues (Pollard, 1963; Pollard and Kajima, 1967;Pollard et al., 1964) showed that tumours could be induced by relatively high doses of the carcinogenic polycyclic aromatic hydrocarbons, 3,4-benzopyrene, 3-methylcholanthrene and 7,12-dimethylbenz(a)-anthracene in germ-free rats and mice. They observed no difference between germ-free and conventionally-maintained rats or mice in susceptibility to the induction of cancer by these chemicals, and reported that the neoplasms induced in germ-free animals were indistinguishable from those induced in conventionally-maintained animals. Other work has established that leukaemia may be induced in germ-free mice by Friend virus (Mirand and Grace, 1963), and in germ-free rats either by Gross passage A virus or by irradiation (Pollard and Kajima, 1966).Despite the reports of the apparently normal response of germ-free animals to various carcinogenic stimuli, we thought it would be worth while to compare the response of germ-free and nongerm-free C3H mice to much lower doses of chemical carcinogens, since, in this way, differences in response which are obscured at high doses might be revealed.Earlier, we and others had found that doses of 7,12-dimethylbenz(a)anthracene (DMBA) of the order of 10-40 pg given to mice shortly after birth