Histamine and 5-hydroxytryptamine were determined in the intestinal tract of germfree and conventional rats and mice. Comparable histamine data were collected in Clostridium perfringens mono-associated rats, while 5-hydroxytryptamine determinations were extended to include the chicken. In rats and mice harbouring an intestinal microflora, bacterial formation of histamine occurs mainly in the caecum. Compared to values found in germ-free animals, histamine levels in the wall of the small intestine of the conventional animal tend to be higher, though in the rat the data are not consistent. Mono-association (harbouring of one microbial species) of germ-free rats with a histamine-producing strain of Clostridium perfringens resulted in high histamine concentrations in the caecal contents, but failed to increase the histamine levels in the wall of the small intestine. 5-Hydroxytryptamine levels in the intestinal wall in the presence of an intestinal flora were generally lower than those in germ-free animals. Modification of the flora by dietary administration of penicillin to mice partly abolished its depressing effect.Comparison of -the intestinal tract of the germ-free animal with that of its conventional counterpart shows a lower relative weight, partially caused by a lower tissue hydration (Gordon & Wostmann, 1960). The absence of a microbial flora strikingly reduces the various reticulo-endothelial elements, notably the plasma cells and the lymphocytes (Gordon & Bruckner-Kardoss, 1961). Apparently the presence of the flora imparts elements of a mild (physiological) inflammation to the gut as an essential part of the mechanism with which the host animal adapts itself to its environment. Results obtained by Gilbert (1959) and by Des Prez, Fallon & Hook (1960) seem to indicate that, depending on the species, histamine and/or 5-hydroxytryptamine are released upon contact with bacterial endotoxins. Histamine formation is accelerated under the influence of Escherichia coli endotoxin (Hinshaw, Jordan & Vick, 1961), evidently through adaptive stimulation of histidine decarboxylase (Schayer, 1962). As both amines are known to induce vasodilatation, increase permeability and produce oedema, they could be important factors in regulating the " adaptive " inflammation. Both histamine and 5-hydroxytryptamine are known to be relatively abundant in the intestinal mucosa of many species, the mucosa being the site of the most intimate contact between the microbial flora and the host.In this paper we present histamine and 5-hydroxytryptamine values in the intestinal tract of animals reared with and without a "normal" microbial flora. Rats and mice were killed by cardiac exsanguination under pentobarbitone anaesthesia. Chickens were exsanguinated after electro-shock. Germ-free animals were taken out of the germ-free environment and sacrificed within 1 hr. Tissues were collected in ice-cold acid saline (0.1 N hydrochloric acid) and stored at -25°C. Homogenization was in acid saline. Aliquots of the total homogenates were used in the...

Germfree gerbils were associated with a murine-derived hexaflora which produced only minor changes in the primary bile acid pattern of rats. These hexaflora-associated gerbils had relatively small ceca (4% of body weight) and reproduced well. Although serum cholesterol levels of both conventional and hexaflora-associated gerbils increased in response to dietary cholesterol, the hexaflora-associated gerbil showed a greater elevation in serum cholesterol than the conventional gerbil when maintained on a diet containing 0.1% cholesterol. This increase in serum cholesterol manifested itself almost totally in the very low density lipoprotein and low density lipoprotein fractions. The fecal bile acids of the hexaflora-associated gerbil were largely deconjugated, but very little further modification of either cholic or chenodeoxycholic acid had taken place. The data suggest that in the absence of elements of the intestinal microflora that can express a bile acid-modifying potential, and particularly a 7-alpha-dehydroxylating capacity, catabolism of cholesterol to bile acids is reduced, and cholesterol accumulates in the very low density and low density serum lipoprotein fractions.

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Feeding of lactose in amounts comparable to the adult human intake in developed countries (6% of diet, and in later studies 10%) had no major effect on cholesterol and bile acid metabolism of germ-free and conventional rats. However, when lactose-containing casein-starch diets were sterilized by autoclaving, changes in intestinal and/or fecal bile acids were found. Both germ-free and conventional rats demonstrated some increase in intestinal beta-muricholic acid concentrations ascribable to the mere presence of lactose in the diet. Autoclaving of the diet produced additional changes, especially in the fecal bile acid pattern of conventional rats. Here the ratio between the beta-muricholic-derived secondary bile acids hyodeoxycholic and omega-muricholic acids changed from the usual 5:3 to approximately 1:10, with omega-muricholic acid becoming the major fecal bile acid. These changes point to a notable effect of lactose-derived products, formed during steam-sterilization, on the microbial modification of intestinal bile acids in the lower gut. Similar changes have been observed after oral administration of aureomycin and other, unrelated antibiotics that inhibit growth of gram positive organisms.

In recent years, the use of milk products and the concomitant intake of lactose have been tentatively linked to the etiology of cardiovascular disease. An effect of lactose on the microbial modification of acid and neutral sterols has been suggested. In the present study lactose intake, ranging up to 30% of total diet increased beta-muricholic (beta-MC) but not cholic acid concentrations in conventional (CV) rat small intestine to the extent that at the 20% and 30% intake level, the intestinal cholic: beta-MC ratio approached that in germ-free (GF) rats. Total intestinal bile acid (BA) content increased by approximately 1/3, but remained at less than half the value found in GF rats. At lactose intake levels within a range corresponding to the consumption of dairy products often recommended for adult man (5% to 10%) only moderate changes in intestinal, and little change in fecal BA were found during and after the 3 months experimental period. Intestinal beta-MC was increased in the presence and in the absence of an intestinal microflora. Experiments with GF rats fed 10% lactose or 10% maltose indicated that this increase is evoked similarly by both carbohydrates. The slight increase in serum cholesterol levels seen with disaccharide feeding, which became evident only in the GF rats, was again not specific for lactose. No influence was found of lactose feeding on liver cholesterol values. Comparison of CV rats fed nonsterile and radiation-sterilized lactose-containing diets suggested that this mode of sterilization has only a minor influence on the resulting data. When GF experiments are to be incorporated, sterilazation of diet by irradiation with 3.5 to 4.0 X 10(6) Rad is preferable to autoclaving. The present data indicate that no major effect specifically related to a normal dietary intake of lactose on cholesterol and BA metabolism of the adult rat could be demonstrated for the duration of these experiments.