THE EFFECT OF A} STRICT VEGETARIAN DIET ON THE FAECAL FLORA AND FAECAL STEROID CONCENTRATION Vivienne C. Ants, J. 8, Crowrier, B. $, Daasar, M.J. HiLt aNb FR. Ets Department of Bacteriology, St Mary's Hospital Medical School, London, ‘and Kingston Group Hospital, Kingston-upon-Thames, Surrey CanceR of the colon is primarily a disease of developed countries; the incidence is highest in Europe and North America and low among the indigenous natives of Asia, Africa and ‘South America (Doll, Payne and Waterhouse, 1966). This variation in incidence, supported asit is by other observations (e-., Wynder ef al., 1969), suggests that diet may be among the ‘most important factors involved.” The effect of diet might be mediated by the action of in- testinal bacteria if, for example, some strains have the capacity to produce carcinogens from dietary components or from intestinal secretions (Aries er al, 19698). ‘AS part of our study of this hypothesis we have investigated the faecal flora and faecal steroids in a group of strict vegetarians who do not eat animal products of any kind, and ‘compared the findings with those in English people living on a normal mixed diet. Though thereas no gross difference in the faecal bacterial florain the two groups, there were interest ing differences in the concentration and composition of the feecal acid steroids and in the ability of the gut bacteria to degrade bile acids MATERIALS AND METHODS Facces were collected according to the method of Aries e al. (19696); bacterial specimens ‘were cultivated by the methods described by Drasar (1967) and Drasar and Crowther (1970); faecal steroid extractions and analyses were performed by the technique of Hill and Aries (unpublished), and the activity of bacteria in degrading bile salts was iovestigated by the ‘method of Aries etal. (19692). The results are presented in tables I and Il. Discussion “The one striking diference betven the faecal bacterial flora inthe two groups was the ‘courreace of anaerobic sarcinas in faeces from the strict veeetarians (table; these organ- isms have not previously been reported as being present in faess from healthy adults though ‘We have isolated them in large numbers from fasces ofa high proportion of people living on substantially vegetarian diets in Uganda and South Indi, Apart from this fnding, there ‘was no highly significant diferece in the numbers ofthe major groups of bacteria in faeces from the two groups of people “Although te feel flora ofthe strict vegetarians was similar to that of people ona mixed diet, the strains of bacteria isolated from them were less active in degrading bile salts and acids in vir Only 20 percent. ofthe cultures of Bacteroides spp. isolated from faeces of Strict vegetarians bad the enzyme 7dshydronplase (Aries and Hl, 1970) compared with 44 per cent. of those soated from people on a mixed diet. The dehydroxplase enzyme pro- duces mono and un-substituted bile acids from the di and trisubstituted cholanic acids secreted inthe bile, Only 30 percent, by weight of the faecal bile acids ofthe strct vegetarians fre mono- or unsubstituted compared with 49 percent. ofthe faccal bile acids of people ‘on a mixed dit (table ID. Further, the total concentration of acid steroids is much loier in the faces of strict vegetarians than inthe faeces of people on a mixed diet Tt has been shown that he faecal bile acid concentration depends on the amount of deiary fat Hil, ‘Recetved 17 Ape, 1970; accepted 15 July 1970, 2 rartevon, 103 G70) *y. Be ae ae ane EFFECT OF DIET ON FAECAL FLORA AND FAECAL STEROIDS 55 a pres) and that sret vegetacans consume a smaller amount of fat than do people on ined det (Elis and Mumford, 1967) ‘Tass 1 Vile count of bactrim feces fro 40 adults ling on mixed diets and 18 adits living om src vegetarian det Mean lpie orem count (stan dis Gao) argv weet fess a aaa P for difference. Organison |__| between A and B (Tames er] 25am | smn i IL wD — aaa 1 Bacteroides ors06 | 92. >oost Bitabaceia 33303 38 3008 pict gers | ag ae Aeobesiepocosi| 70208 & San Pacrooee ss Etc goete | aE Soa Giese asun9 a S005 Sainte aSu15 6 50058 Yeats 19313 30 30083 Flapentous fungi | e07 iy Sone Sanne Noitoma | #4 : Data trom Ares tol, 09630, SVL bile fom Sento test andthe x est Vales shied from the rok test andthe get Segal elated rom ons of 18 specinens ‘ane Tt The fccol acd and neural sterols of strlet vegetarians and of people ‘ona normal mixed det cre (ox per by weight cee) 280 sero Tota sri, 7 zl won|" | mE stot, - tit, | dtu unsab, | Sptecatt Sites | Semaod | tT | Ssed | stated | 2¥Bs 4 | Sted | Fasos) Mined a2 | 7 | wal ¢ | a | w | s | ox Sins it | $B] Mt) ak | S| mw) 2 | So veetaan ‘Annumber of bile acids have been shown o be caeinogeni (Druckrey, Richter and Vier~ shale, 941s, Lazassagne, Buu-Ho\and Zajdels, 961, 1960, The ileacid, deoxyeholcacid, ean be chemically converted into the potent carclnogen, 20-methylcolanthrene (addow, 1958), and its posible thet bacteria can perform the same conversion. Such eactions would te mere likely fo ocer in te intestine of people on a ined det since their gut bacteria are row active aguinat ble acid, andthe reaction could be more important in these people Since te substrate concentration i thei intestine Is higher. ks 5ARIES, CROWTHER, DRASAR, HILL AND ELLIS SUMMARY As part of a study of the aetiology of cancer of the large bowel the faecal bacterial and fungal flora and the faecal steroids of English people living on a strict vegetarian diet have been compared with those of English people living on a mixed diet. There was no gross, difference in the number or the species of organisms present in the two groups, but fewer Of the species of Bacteroidesin the faeces of strict vegetarians could dehydroxslate di and ti substituted cholanic acids, and the faeces of strict vegetarians contained relatively smaller ‘quantities of mono- and un-substituted acid steroids, and lower concentrations of total bile acids, than those of persons on a mixed diet. ‘We would like to thank the strict vegetarians for their helpful co-operation in this study and to acknowledge the excellent technical assistance of Miss Deirdre Blann and Miss Rosalind Steward. This work was supported by the British Empire Cancer Campaign for Research. REFERENCES ‘Anuzs, V. C., CrowrHer, J. S., Deasaz, B.S, aNo Hint, M, J. 1969a.. Degradation of bile salts by human intestinal bacteria. "Gur, 10, 575. Anits, V. C., Crowrurr, J. S,, DRASAR, B. 8, HILL, M. J., AND Wittians, R. E. O, 19598, Bacteria and the aetiology of cancer of tite large bowel. Gur, 10, 33 ‘Ans, V. C,, ano Hunt, M. J. 1970. Degradation of steroids by intestinal bacteria, 11, Enzymes catalysing’ the oxidoreduction of the 32-, Ta- and I2a-hydroxyl groups in cholic acid, and the dehydroxylation of the Te-hydroxyl group. Biochim. Biophys. deta, 202, 535. Dott, R, Payne, P., ano WATERHOUSE, J. 1966. Cancer incidence in five continents: technical report, Ber Drasar, B.S. 1967.” Cultivation of anaerobic intestinal bacteria, J. Path. Bact, 94, 417. Drasa, B.S., aNb Crownuen, J.$. 1970. Cultivation of intestinal bacteria. Soe. Applied ‘Bact. Techn. Ser. no. 5: Isolation of anaerobic organisms (in press). Davcxsey, H., Ricatex, R., an Viexruater, R. 1941, Zur endogenen Entstehung krebser~ regenden Stoffe beim Menschen. Klin, Wsckr., 20, 781. Euus, F. R,, axD Muntrorn, P. 1967, The nutritional status of vegans and vegetarians, Proc. Nutr. Soe. 26, 205. Happow, A, 1958. Chemical carcinogens and their modes of action. Br. Med. Bull, 14,79. Lacassacns, A., Buu-Hot, N. P., aN ZalbEL, F. 1961, Carcinogenic activity of apochoic acid. Nature, Lond, 190, 1007. LacassaGns, A., Buu-Hot, N. P., AND ZasosLa, F, 1966, Carcinogenic activity in situ of further steroid compounds. ‘Nature, Lond, 209, 1026. Wyner, E. L., Kasttany, T,, Iskixawa, S., Dono, HL, AND TaxANo, A. 1969. Environ ‘mental factors of cancer of the colon and rectum, Il. Japanese epidemiological data, Cancer, Philad., 23, 1210.