Fibrosis and lymphocytosis are common in aflatoxin-induced hepatoma in rainbow trout. It is suggested that these reactions may control the growth of tumors and in some cases destroy them. Nearly all of the hepatomas we have observed are fibrotic to some extent. The extent of the fibrosis seems to be age dependent and in advanced cases fibrosis appears to have disorganized the tumors t o a degree which we believe is suppressive. The lymphocytosis shows a negative correlation with the size and age of the hepatoma, being found almost exclusively in the very "early" (small) hepatomas and preneoplastic nodules. It is suggested that the invasion of the tumors by lymphocytes is a host defense mechanism.Infiltration of hepatomas in the rainbow trout by ductal tissue components (fibrosis) and by lymphocytes has been described by a number of workers (Haddow and Blake, '33; Cudkowicz and Scolari, '55; Nigrelli and Jakowska, '61; Wood and Larson, '61; Rucker et al., '61; Hueper and Payne, '61; Ghittino and Ceretto, '62; Ashley and Halver, '63; Scarpelli et al., '63; Ashley, '69; Wales, '70; Ashley, '70). Although these workers have mentioned the presence of either or both tissues in trout hepatomas, they have not discussed their importance in controlling the growth or causing the regression of tumors. Hellstrom and Hellstrom ('69) have reviewed the subject of lymphocytic antagonism to tumor cells. The body of literature concerned with host immune reaction in neoplasia is already large and growing rapidly, but apparently there have been no published reports of this phenomenon in trout hepatoma.
MATERIALS AND METHODSThe brood stock of Mt. Shasta "strain" rainbow trout (Salmo gairdneri) maintained in our laboratory has been found to be more sensitive to aflatoxin-induced hepatoma than any other animal tested. of carcinogens and cocarcinogens have been tested under a wide variety of experimental conditions and in conjunction with many dietary modifications (Sinnhuber et al., '68). Large numbers of hepatomas can be produced by feeding any nutritionally balanced trout diet to which has been added aflatoxin B1 at levels within the range of 4 to 20 parts per billion (ppb). The carcinogen is added to the diet as soon as possible after feeding starts. Hepatomas may reach macroscopic size in four months although this is temperature-dependent and in water averaging 12" C large numbers of hepatomas are rarely obtained in less than six months. Livers containing hepatomas are routinely fixed in Bouin's fluid, embedded in paraffin and stained with hematoxylin and eosin. Masson's triple stain is occasionally used to emphasize the connective tissue in fibrotic tumors.In the course of this work, a very large number of hepatomas has been examined. Certain carcinogenic diets produce as many as 100 tumors in a single liver, ranging from microscopic size to more than 8 cm in diameter. Unfortunately the observations of fibrosis and lymphocytosis are difficult to treat statistically. Only a rough esti-
OBSERVATIONSNearly all of the neopla...