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Fecal Bacteriology of Colonic Polyp Patients and Control Patients' [CANCER RESEARCH35,3407-3417,November1975] Fecal Bacteriology of Colonic Polyp Patients and Control Patients' Sydney M . Finegold,2 Dennis J . Flora, Howard R . Attebery, and Vera L. Sutter Medical Service, Wadsworth Hospital Center, Veterans Administration and Department ofMedicine, University ofCalifornia at Los Angeles School of Medicine, Los Angeles, California 9tXl24 Summary and villous adenomata are rare in populations with a low incidence of colon cancer and that they are found with the Feces from 25 subjects with colonic polyps (multiple highest incidence in areas where colon cancer has the adenomatous, large single, or single with atypia) and from highest prevalence. Berge et a!. (3) note a close association 25 matched control subjects were studied by detailed between polyps and carcinoma, both tending to occur in the quantitative aerobic and anaerobic techniques, using a large same distribution. Fifty-nine % of polypoid tumors more battery of culture media and several atmospheric condi than 10 mm in diameter were carcinomas, as were 16.9% of tions. Over 55% of organisms detected on microscopic count those 5 to 10 mm in size. were recovered anaerobically. In several cases, there were There has been much speculation on interrelationships significantly different numbers of organisms of specific between diet, intestinal bacteria, intestinal polyps, and types recovered from the two different populations studied. carcinoma (I, 2, 6—8). However, these differed from organisms with “statistical The present study was designed to compare the fecal significance― noted in a previous study from this laboratory bacterial flora of 25 patients with colonic polyps (chiefly involving two different diet groups (Japanese Americans on multiple adenomatous polyps) with that of 25 subjects either a Japanese or a Western diet). Specific differences in matched as to age, sex, and race but with demonstrated (by bacteriology between groups with different risks of bowel fiberoptic cobonoscopy) absence of polyps. cancer, noted in earlier British studies, were not noted in our present or previous study. Materials and Methods Introduction Subjects. With a few exceptions, polyp patients had multiple adenomatous polyps. Excluded were hyperplastic It has been estimated that in 1973 in the United States, polyps, inflammatory polyps, carcinomatous polyps, and 92,500 new cases of colon and rectal cancer would occur and patients with congenital polyposis. All polyp patients and that large bowel cancer would account for 47,400 deaths, controls had not had any antimicrobial drugs for at least 2 being second only to lung cancer in this regard (2). weeks and had not had a prolonged course of antimicrobial The association of colon polyps and carcinoma of the drugs (2 months or longer) within 1 year of the time colon is a disputed one. However, Buntain et a!. (4), after a sampled. No laxatives or enemas were used for at least I detailed review, conclude that the evidence indicates some week prior to stool collection. Excluded were all subjects as yet undefined relationship between adenomatous polyps who had had prior gastrointestinal tract surgery other than and carcinomatous change. They also indicate that a appendectomy or removal of polyps. All subjects volun significant number of such polyps, depending on size and teered to participate in the study. age, will eventually undergo malignant change. Accord Bacteriology. All specimens were collected at the hospital ingly, they recommend removal ofpolyps, particularly those or a@tthe residences of the subjects. Initial processing and 1 cm in diameter or larger. A comparison of a high-risk transport of the specimens and subsequent weighing and colon cancer population in New Orleans with a low-risk dilution of specimens were carried out as indicated in an population in Cali, Colombia, revealed that the former had earlier paper from this laboratory (5). Culture techniques a higher prevalence of adenomatous polyps at all ages in and media utilized were also as previously described (5) with both sexes (6). This difference is accentuated when the the following exceptions. (a) For aerobic incubation, nitro comparison is limited to multiple adenomas. Also, in New gen-deficient agar, molybdate agar, and tryptose-phosphate Orleans,the adenomasarelarger(averagesize,8 x 4 mm) broth-agar with rabbit serum were eliminated. (b) For and show more atypia (villous pattern). A much higher anaerobic incubation, deleted were blood agar plate, blood percentage of the adenomas in the population of New agar plate (heated dilutions), Bryant Medium 10, Rumen Orleans is concentrated in the sigmoid and rectum. fluid-glucosecellobioseagar, neomycin-Nagleregg yolk MacGregor (9) also pointsout that adenomatouspolyps agar, rifampin-vancomycin blood agar, mitis-salivarius agar, and anaerobic spirochete medium (Virginia Poly technicInstitute).(c) LBS mediumwassupplementedwith I Presented at the Conference on Nutrition in the Causation of Cancer, May 19 to 22, 1975, Key Biscayne, Fla. Supported in part by USPHS tomatojuice (as in Bifidobacteriummedium),and Bader Contract NIH NCI-E-72-3209. oides medium (Nissui) was added (specifically for “giant― 2 Presenter. Bacteroidesstrains). NOVEMBER 1975 3407 Downloaded from cancerres.aacrjournals.org on September 30, 2021. © 1975 American Association for Cancer Research. S. M. Finegold et al. All specimens were coded so that the bacteriologists did Table I not know the source until the entire bacteriological study General data, polyp study and tabulation were completed. There were 25 subjects in each group. There were no statistically Statistical Analysis. Data were analyzed statistically (by significant differences between polyp and control subjects in any of the following parameters. the UCLA Health Sciences Computing Facility) using a 1-way analysis of variance, t test, x2 analysis, Boolean Control factor analysis, and discriminant analysis. Data were an subjectsS.D.Age subjectsS.D.Polyp alyzed not only in terms of converted mean counts, but also in terms of absence or presence of organisms and high (yr)56.4010.7558.8410.60Specimen (g)72.326.6468.9247.58%wt versus low ( l06/g) counts. Various combinations of solids26.309.6226.238.72pH6.820.416.620.70Microscopic organisms were also studied for possibly significant associa tions, and data from this study were compared with that count6.422.956.804.38(x from an earlier study of Japanese versus Western diet wt)Totall0'1/gdry count3.792.683.712.32(xanaerobic subjects (5). wt)% I0―/g dry 56.1231.2357.8027.31obes,recoveryof anaer Results tomicroscopiccompared count% to17.5616.8022.0723.77anaerobesEOSa organisms Four of the polyp patients had solitary polyps, 15 had 2 (zeroes cx polyps, 3 had 3 polyps, I had 6, and 2 had multiple polyps (8 cluded)Aerobiccount(x in I patient; an unspecified number in the other). In the case l0'/g8.2918.796.1215.32dry of the solitary polyps, 2 were 2.5 cm in diameter, I was 1.2 wt)Ratio cm in diameter, and the 4th was 4 x 3 x 3 mm, but showed to3,587.687,600.475,690.3113,706.73aerobesNo.of anaerobes focal atypia. One subject with 2 polyps also showed with78methaneof subjects superficial atypia. In 4 patients with multiple polyps, villous oncultureNo.bacteria patterns were observed. In 1 case, biopsy was not obtained. The locations of the polyps were as follows: cecum, 6; with10methaneof subjects inbreath detected ascending colon, 7; hepatic flexure, I; transverse colon, 10; (<0.05%)No. splenic flexure, 7; descending c@lon, 9; and sigmoid, 25. I7drogenof subjects with hy There were 21 males and 4 females in both the polyp and inbreath detected control groups. There were 2 blacks in each group; the ( <0.03%) balance of the subjects were Caucasian except for 1 Japanese and 1 American Indian in the control group. Age a EOS, extremely oxygen-sensitive anaerobes. matches were quite close, usually within 3 years for each Differences in counts of Clostridium paraputrijIcum and pair (polyp and control). However, in the case of the blacks, related organisms between the 2 groups of subjects are it was not possible to obtain close matches (52 and 79 years shown in Table 13; they are not significantly different and 49 and 63 years). Three of the control patients had statistically. diverticulosis; the others had normal colons. The species or groups showing statistically significant Certain general characteristics of the 2 groups are noted differencesbetweenpolyp and control subjectsare noted in in Table 1. Included are subject age; data on the specimen Table 14. weight, moisture content, and pH; and general bacteriologi The most prevalent species in the polyp and control cal data. populations are noted in Table 15. Details on the specific bacteria recovered with ranges and converted mean counts are given in Tables 2 through 12. These counts represent a conversion from the means Discussion obtained after using a log transformation on the data values. The log transformation was used because of extremely We have carried out a thorough study of the entire fecal skewed distributions of some organisms, reflecting rela flora of 25 subjects with colonic polyps (or documentation tively small samples. All counts are based on weight of dry of recent polyps) and 25 matched controls. A variety of stool. The numbered species cannot be fit into any of the selective, differential, and nonselective media, as well as established species; these were grouped into apparently various special substrates and atmospheric conditions, were similar groups, with each distinct group being assigned a used. Recovery of over 55% of the microscopic count is separate number. In the case of the facultative streptococci quite good. When extremely oxygen-sensitive anaerobes (Table 3), the “other―categoryalso includes unclassifiable were present, they accounted for about 20% ofthe anaerobic strains, but these were lumped together. flora. In the case of some of the anaerobic bacteria (up to Certain differences between the polyp and control pa one-third of the anaerobic isolates in some specimens), poor tients were significant statistically.
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