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, 6â€”8). However, these differed from organisms with â€œstatistical The present study was designed to compare the fecal significanceâ€• 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 â€œgiantâ€• 2 Presenter. Bacteroidesstrains).

NOVEMBER 1975 3407

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)% I0â€•/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 â€œotherâ€•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. However, when such a growth was obtained. This may mean that identification is large number of comparisons are involved, a few so-called not always accurate. It is planned to restudy these strains, â€œstatisticallysignificantâ€•differencesmay occur by chance utilizing various growth additives in hopes of obtaining alone. While the same genera (Streptococcus, Eubacterium, better growth. BijIdobacterium, and Peptostreptococcus) were often noted

3408 CANCER RESEARCH VOL.35

Table 2 Facultative or aerobic Gram-negative bacilli

subjectsNo.Control subjects Polyp

ofspeci-of No. speci mens Con- mens Con vertedboringhar- verted har- mean0Citrobactersp. Rangeâ€• meanÂ° boring Rangea

2.90Enterobacter 1 8 2.76 1 9 2.70Enterobacteraerogenes 0 1 7 2.93Enterobacterliquefacienscloacae I 4 2. 11 2 4-8 0Escherichiacoli I 4 2. 17 8.25Klebsie//apneumoniae 23 4-11 8.10 24 5-lI 2.64Klebsiellasp. 0 1 7 4.54Proteusmirabiis 12 4-10 4.81 8 5-10 2.81Proteus 1 6 2.42 I 8 0Pseudomonasmorganii I 4 2. 11 2.60Total aeruginosa I 4 2.17 3 3-6

8.32a 8.26

Log10 no. of organisms per g (rounded offrange).Table to nearest log in the case of

3Facultative streptococciControl

subjectsNo. subjects Polyp

ofspeci-of No. speci mens Con- mens Con vertedboringhar- verted har- mean0Enterococcus Range0 mean0 boring Range0

groupStreptococcusdurans 2.22Streptococcusfaeca/is 0 1 5 6.36varfaecalisStrepiococcusfaeca/is18 5-11 6.89 19 4-lI

3.55var 3 4-8 3.10 4 4-11 liquefaciensStreptococcusfaecalis 3.45var 5 5-9 3.69 5 4-8 zymogenesStreptococcusfaecium 4.15Other 5 3-10 4.03 6 5-10

Group D strepto cocciStreptococcusbovis 3.61Other 7 4-10 4.04 3 6-Il

streptococci.@treptococcus 0Streptococcusequisi-avium I I I 3.02 3.30mi/isStreptococcuslactis 1 7 2.67 3 5-10

4.24Sireptococcusmitis 8 6-11 4.84 6 7-10 4.26Streptococcus 9 5-11 4.76 10 5-8 3.42Streptococcuspyogenesmutans 5 6â€”10 3.84 4 6-7 0Streptococcussalivarius I 7 2.71 3.88Streptococcus 2 8-9 3.29 5 6-10 3.29StreptococcusGroupGsanguis 0 3 7 0Streptococcus I 6 2.57 IOther Group 0 0 2 6-9 3. I 4.50Total 7 4-Il 4.01 8 6-10

9.09 8.20

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

NOVEMBER 1975 3409

Table 4 Otherfacultative or aerobic organisms

subjectsNo.Control subjects Polyp

ofspeci-of No. speci mens Con- mens Con vertedboringhar- veiled har- mean0Micrococcus Range0 mean0 boring Range0

2.85Staphylococcussp. 2 4 2.3 1 1 9 4.41Staphy/ococcusaureusepidermidis 9 4- 10 4.44 10 4-9 3.26Bacillussp. 2 4-6 2.61 2 8-9 5.92Candida 23 4-10 6.42 23 3-10 2.61Otheryeastsalbicans 0 1 7 4.21Filamentous 9 4-5 3.39 II 3-8 3.35Total fungi I 4 2.00 3 6â€”8

6.94a 7.12

Log10 no. of organisms per g (rounded offrange).Table to nearest log in the case of

5LactobacillusControl

subjectsNo. subjects Polyp

ofspeci-of No. speci mens Con- mens Con vertedboringhar- verted har- mean0Lactobacillusacidophi/us Range0 meanÂ° boring Range0

5.81Lactobacillus 5 6-11 4.15 9 5-11 0Lactobacillus brevis 1 9 2.91 3.27Lactobacilluscaseivarcaseibuchneri 0 3 5- 10 0Lactobacil/us 2 6-10 3.19 3.27rhamnosusLactobadillusfermentumcasei var 0 2 8â€”10

5.22Lactobacillus 7 5-10 4.27 9 6-1 1 2.40La@tobadillusleichmannii 0 I 5 0Lactobacillusplantarumminutus 3 8- 10 3.70 4.19Lactobadillus 5 7-10 4.16 6 5-10 2.73var. salivarius 2 7- I I 3.26 1 8 salivariusLactobacillus 3.59Total sp. I 6 2.46 3 6-10

6.90 7.15

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

3410 CANCER RESEARCH VOL.35

Table 6 Anaerobic cocci

subjectsNo.Control subjectsPolyp

ofspeciofNo. speci mensCon mensCon vertedboringRange0mean0boringRangeÂ°mean0Acidaminococcusfermenhar vertedhar

104.43tansAcidaminococcus 94- I I5.0168-

13.02MegasphaeraelsdeniiI102.92192.87Peptococcussp. I011

0182.72cusPeptococcusconste/lazus0210-113.47Peptococcusasaccharo/yti

125.26Peptococcusprevotii87-115.1849-114.08Peptococcusmagnus510- 114.5 I810-

I4.22Peptococcusvariabiis29-113.43210-113.43Peptococcussp.saccharolyticus057- 1

I0192.89Peptococcussp.301Il3.00Peptococcussp.5I102.950Peptococcussp.

10182.800Peptococcus 11102.990Peptococcussp.sp. I 120192.90Peptostreptococcus I92.830biusPeptostreptococcusinteranaero

123-lI5.91II8-Il6.05mediusPeptostreptococcusmicros47-lI3.9454-103.93Peptostreptococcusparvulus46-103.88210-lI3.46Peptostreptococcuspro

45-113.96II4-125.81duct usPeptostreptococcussp. I310-113.840Peptostreptococcus 20182.82Peptostreptococcussp. 13.04Peptostreptococcussp.sp. I I011 1301102.92Peptostreptococcussp.26I102.920Peptostreptococcussp.27I92.910Ruminococcusalbus152.19192.91Ruminococcus

bromiiI52.240RuminococcusfiavefaciensII I0Sarcina I3.0 ventriculi0172.65Sarcinasp. I035-83.31Veil/onellaparvula057- IUnknown 114. I Gram-negative1102.940cocciTotal9.949.67

a Log,0 no. of organisms per g (rounded off to nearest log in the case of range).

NOVEMBER 1975 3411

Table 7 Bacteroides

subjectsNo.ofNo.ofspeciControl subjectsPolyp

speci mensCon mensCon vertedboringRange0mean0boringRange0mean0har vertedhar

Bacteroidesamylophilus 1 9 2.87 2 8-10 3.31 Bacteroidescapi//osus 6 9-Il 4.69 3 9-lI 3.78 Bacteroidesclostridiiformis I 10 2.92 2 7-9 3.19 55. clostridiiformis Bacteroides coagulans 0 I I 1 3.01 Bacteroidesfragilisss.dis- I7 10-11 8.10 15 7â€”11 7.31 tasonis Bacteroidesfragi/isss.fra- 15 8-11 7.34 17 10-12 8.11 giis Bacteroidesfragilisss.ova- 8 8-11 5.29 12 7-12 6.46 tus Bacteroidesfragilisss.zhe- 22 6-12 9.26 21 6-12 9.17 taiotaomicron Bacteroidesfragilisss.vul- 17 9-12 8.04 17 10-11 8.19 gatus Bacteroidesfragi/isss. II 9-12 6.16 15 9-lI 7.56 other Bacteroideshypermegas I 9 2.83 I 9 2.91 Bacteroides hypermegas 2 10- 11 3.47 0 type I Bacteroidesoralis 0 2 10â€”lI 3.50 Bacteroides pneumosintes 5 10- 11 4.43 6 8- 10 4.47 Bacteroidesputredinis 5 10-11 4.50 0 Bacteroidesruminico/ass. 3 10-11 3.85 1 10 2.95 brevis Bacteroidessp.5 I 10 2.94 0 Bacteroidessp. 10 1 5 2.25 0

Total 10.81 10.99

a Log,, no. of organisms per g (rounded off to nearest log in the case of range).

Table 8 Fusobacterium

subjectsNo.Control subjectsPolyp

ofspeciofNo. speci mensCon mensCon vertedboringRangeÂ°mean0boringRange0mean0Fusobacteriumhar vertedhar

mortiferum2103.39172.68Fusobacterium necrogenesI82.800Fusobacterium necrophorumI82.81152.38Fusobacterium prausnitzii28-93.26192.83Fusobacterium russiiI82.7226â€”103.14Fusobacterium 501102.98Fusobacteriumsp. 8182.74I72.58Total4.374.00sp.

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

3412 CANCER RESEARCH VOL.35

Table 9 Eubacterium

Control subjects Polyp subjects

No. of No.of speci speci mens Con mens Con har- verted har- verted boring Range0 mean0 boring Range0 meanÂ°

Eubacterium aerofaciens 7 8-Il 4.86 II 9-11 6.17 Eubacterium combesii 8 8-lI 5.21 2 10â€”lI 3.50 Eubacterium contortum 5 10-Il 4.45 3 10â€”11 3.82 Eubacterium cy/indroides 7 8-lI 4.84 5 7-10 4.08 Eubacterium /entum 20 8-11 8.61 17 6â€”lI 7.73 Eubacterium limosum I 10 2.96 2 9 3.34 Eubacterium moniliforme I 8 2.81 0 Eubacterium multiforme I 9 2.87 0 Eubacterium nitritogenes I 7 2.71 I 4 1.99 Eubacterium rectale 3 9-10 3.66 II 7â€”Il 5.84 Eubacterium tenue 6 6-lI 4.43 I II 3.02 Eubacterium ventriosum 4 8-lI 3.95 2 9-10 3.37 Eubacterium sp. 0 I II 3.06 Eubacterium sp. I 2 8-10 3.35 4 8-Il 4.03 Eubacterium sp. 6 2 7-9 3.19 2 8 3.20 Eubacterium sp. 7 0 2 9-10 3.34 Eubacterium sp. 8 4 8-10 3.94 3 6-11 3.63 Eubacterium sp. 11 I 9 2.91 0 Eubacterium sp. 14 2 9-12 3.45 I 8 2.72 Eubacterium sp. 17 2 9â€”10 3.37 2 7-11 3.35 Eubacterium sp. 20 0 I 8 2.72 Eubacterium sp. 21 I 10 2.95 0 Eubacterium sp. 24 2 6-11 3.20 2 II 3.51 Eubacterium sp. 28 I 11 3.00 0 Eubacterium sp. 30 2 6â€”lI 3.22 0 Eubacterium sp. 32 I 8 2.79 0 Eubacterium sp. 33 I 10 2.97 I 8 2.72 Eubacterium sp. 36 0 I 10 2.97 Eubacterium sp. 37 I 10 2.98 0 Eubacterium sp.41 3 5-Il 3.63 3 7-Il 3.66 Eubacterium sp. 43 0 10 2.98 Eubacterium sp. 44 0 2 10-Il 3.48 Eubacterium sp. 46 0 3 7â€”Il 3.57 Eubacterium sp.47 0 10 2.95 Eubacterium sp. 48 0 2 9-Il 3.41 Eubacterium sp.49 0 II 3.02 Eubacterium sp. 50 0 II 3.06 Eubacterium sp. 51 I 6 2.49 7 2.62 Eubacterium sp. 52 0 6 2.55 Eubacterium sp. 53 0 6 2.55 Eubacterium sp. 54 I 9 2.91 0 Eubacterium sp. 55 0 2 8-9 3.28 Eubacterium sp. 56 0 8 2.79

Total 10.46 10.38

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

NOVEMBER 1975 3413

Table 10 Bijidobacterium

subjectsNo.ofNo.ofspeciControl subjectsPolyp

speci mensCon mensCon vertedboringRange0meanÂ°boringRange0meanÂ°Bifidobacteriumhar vertedhar

14.84centis ado/es 77â€”1 14.7878â€”1 ABifidobacterium 38-103.6148-103.85centisado/es BBifidobacteriumadoles 29-113.40108-115.61centis CBifidobacterium 103.73centis ado/es 393.6539- DBifidobacterium 13.79BifidobacteriumbijIdumII I3.06310-1 103.38I92.84Bijidobacteriumbreve29- eriksonii0I82.82Bifidobacterium 13.49055. infants210- 1 infantisBifidobacterium 103.3755. infantis49- 114.0529- liberorumBifIdobacterium I3.41otherBifidobacteriuminfantis,66- I I4.4929-1

104.29longumTotal8.317.36longum ss.610- I I4.8059-

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

in this list of significant organismsfrom this study and from tistically significant increase in Eubacterium). Finally, Hill our earlier comparison of Japanese Americans on a Japa (7) notes that steroid nuclear-dehydrogenating clostridia nese diet versus those on a western diet (5), the species (the Closiridium paraputrificum group) are found in larger involved were always different. Data analysis is continuing, numbers in stools of British and Americans than is true in seeking to find significant relationships between various the case of people from low bowel cancer incidence organisms or groups of organisms among the 4 popula countries. We have looked specifically for all of the above tions studied. To date, no really striking relationships have relationships in the 4 populations we havestudied(Japanese been uncovered. This raises some questions concerning the diet, Western diet, polyp subjects, and controls) and have British studies in which fecal specimens from various high failed to observe them. Differences may be related to the and low-risk groups were said to show significant differ population differences involved, as well as to differences ences in fecal flora. Thus, Aries et a!. (1) noted signifi in the detail in which the bacteriology was studied and in cantly more Bacteroides and Bifidobacterium in fecal the number of studies performed. specimens from English subjects than in specimens from Additional studies are certainly needed to investigate Ugandans. A subsequent paper from this. group (8) in further the fascinating interrelationships between diet, intes cluded a very large number of specimensfrom high bowel tinal bacteria, and bowel cancer. In the meantime, these cancer-risk subjects (British and American) and low-risk studies provide valuable data on the makeup of the normal subjects (Ugandans, Indians, and Japanese); the high-risk fecal flora and on methods for studying it. population yielded many more Bacteroides, whereas the low-risk group had many more aerobic streptococci and Acknowledgments enterobacteria. The enterococci included a much higher percentage of Streptococcus faeca!is in the English subjects We would like to acknowledge the excellent technical assistance of Walker Carter, Joe Trammell, Stan Cooper, Larry Jensen, and Paul than in the Indians. A more recent paper (10) indicates Sugihara. Computing assistance was obtained from the Health Sciences that Western diet country subjects appeared to have a Computing Facility, UCLA, sponsored by NIH Special Research Re larger proportion of Gram-negative organisms (Bacteroides sources Grant RR-3. We wish to thank Dr. Robert Mah of the UCLA and Fusobacterium), and subjects from low-risk countries School of Public Health for his assistance in the studies relating to methane had a larger proportion of Gram-positive organisms (a sta bacteria.

3414 CANCER RESEARCH VOL.35

Table I I Clostridium

Control subjects Polyp subjects

No.of No.of speci speci mens Con mens Con har- verted har- verted boring Range0 meanÂ° boring Range0 mean0

Clostridium acetobutylicum 0 8 2.76 Clostridium aminovakricum 4 6-10 3.78 8 4-10 4.54 Clostridium barkeri 2 10-11 3.45 0 Clostridium bifermentans 9 7-11 5.49 14 4â€”12 6.52 Clostridium cadavers I 7 2.70 0 Closiridium chauvoei 2 10-lI 3.45 0 Clostridiumfallax 4 6-10 3.72 10 2.95 Clostridiumft/sineum I 8 2.78 Clostridium ghoni I 10 2.99 0 Clostridium glycolicum 3 5â€”6 3.03 8 2.76 Clostridium haemolyticum I II 3.02 0 Clostridium indolis 0 10 2.94 C/ostridium innocuum 4 8-9 3.86 7 6-10 4.55 C/ostridium mu/mum I 6 2.42 0 Clostridium irregulars I 7 2.65 Clostridium limosum 2 4-10 3.10 8 2.74 Clostridium malenominatum I 5 2.40 5-9 3.45 Clostridium mangenoti 4 4-10 3.43 II 3.02 Clostridium oroticum 2 6-10 3.23 2 4-8 2.91 Clostridium paraperfringens I 5 2.40 4 4-9 3.56 Clostridium paraputrificum 2 4-9 2.96 3 4-8 3.08 Clostridium pasteurianum I 10 2.96 0 Clostridium perenne 2 8-10 3.25 0 Clostridium perfringens II 4â€”Il 4.93 9 4-Il 4.41 Clostridium p/agarum 0 9 2.91 Clostridium propionicum I 9 2.91 0 Clostridiumpseudotetanicum 2 4-11 3.11 2 4-6 2.72 Clostridium putrefaciens I 10 2.97 2 10â€”Il 3.45 Clostridium ramosum 14 8-lI 6.78 II 6-Il 5.80 Clostridium rectum I 5 2.40 0 Clostridium sartagoformum 0 9 2.88 Clostridium septicum I 9 2.91 0 Clostridium sordellii 0 2 9-Il 3.43 C/ostridium sporosphaeroides 4 5-9 3.43 2 7-10 3.22 Clostridium subterminale I 7 2.71 0 Clostridiumsp. 0 4 6-9 3.61 Clostridium sp. I 2 9-lI 3.40 7 2.66 Clostridium sp. 2 1 7 2.65 0 Clostridium sp. 5 0 8 2.74 Clostridium sp. 7 3 4â€”S 2.81 0 Clostridium sp. 8 0 4-Il 3.26 Clostridiumsp.18 0 6 2.54 Clostridium sp. 25 I 7 2.70 0 Clostridium sp. 27 I 7 2.62 0 C/ostridium sp. 28 1 6 2.44 0 Clostridium sp. 32 I 8 2.75 0 Clostridium sp. 48 0 9 2.91 Clostridium sp. 51 I 6 2.44 2 7-10 3.22 Clostridium sp. 56 0 10 2.96 Clostridium sp. 58 0 8 2.80 Clostridium sp. 59 1 9 2.84 10 2.92 Clostridium sp. 60 I 9 2.83 0 Clostridium sp. 62 I 7 2.70 0 Clostridium sp. 63 1 10 2.97 8 2.81 Clostridiumsp.64 0 10 2.95

Total 10.38 9.99

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

NOVEMBER 1975 3415

Table 12 Miscellaneous anaerobes

subjectsNo.ofNo.ofspeciControl subjectsPolyp

speci mensCon mensCon vertedboringRangeÂ°meanaboringRange0mean0Actinomyceshar vertedbar

I01102.94Arachniapropionica01113.01Butyrivibriofibriso/vens0182.72Propionibacteriumsp.

avidum01102.96

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

Table I 3 Clostridium paraputrificum group Analysis of variance reveals no significant differences between polyp subjects and controls, whether results are analyzed by individual species or as a group.

subjectsNo.ConControl subjectsPolyp

No.Con vertedboringRangeÂ°mean0boringRange0mean0Closiridiumparaputrijlcum24-92.9634-83.08Clostridiuminnocuum48-93.8676-104.55Clostridiumhar vertedbar

butyricum00Clostridium tertium00Clostridium indolis01102.94Clostridium sartagoformum0192.88

a Log10 no. of organisms per g (rounded off to nearest log in the case of range).

Table 14 Significant differences in poh'p fecal flora study

Controlsub

subjects0pbStreptococcus jects0Polyp

mitis4.764.260.024Bifidobacterium C3.405.610.009Eubacteriumadolescents 13Eubacteriumrectale3.665.840.0 tenue4.433.020.049Eubacterium combesii5.213.500.037Bacteroidesputredinis4.5000.025Peptostreptococcusproductus3.965.810.031Veillonellaparvula

@ 1I Peptococcus saccharolvticus0 04. 4.22 1025

a Log10 no. of organisms per g (converted mean). b Based on contingency table analysis (Fisher's exact probability statistic).

3416 CANCER RESEARCH VOL.35

Table IS Most prevalent species Only organismsharboredby8 or moreindividualsineachgroupwereconsidered.Altogether,theseorganismsaccountfor22%of thetotal bacterial count in the control group and 27% ofthe total in the polyp group.

subjectsCon Control subjectsPolyp

Con vertedvertedmeanmeancount0Organismcount0Organism

8.61 Eubacterium lentum 9. 17 Bacteroides fragilis ss. thetaiotaomicron 8.10 Escherichia co/i 8.25 Escherichia coli 8.10 Bacteroidesfragiis ss. distasonis 8.19 Bacteroides fragiis ss. vulgatus 8.04 Bacteroides fragiis ss. vulgatus 8.11 Bacteroidesfragilis ss.fragilis 7.34 Bacteroides fragilis ss.fragiis 7.73 Eubacterium lentum 6.89 Streptococcus faecalis var. faecalis 7.56 Bacteroidesfragiisss.other 6.78 Clostridium ramosum 7.31 Bacteroides fragiis ss. distasonis 6.42 Bacillus sp. 6.53 Clostridium bifermentans 6.16 Bacteroidesfragilis ss. other 6.46 Bacteroides fragiis ss. ovatus 5.91 Peptostreptococcus intermedius 6.36 Streptococcus faecalis var. faecalis 5.49 Clostridium bifermentans 6. 17 Eubacterium aerofaciens 5.29 Bacteroidesfragilis ss.ovatus 6.05 Peptostreptococcusintermedius 5.21 Eubacteriumcombesii 5.93 Bacillus sp. @.19 Peptococcus prevotii 5.84 Eubacterium rectale 5.01 Acidaminococcusftrmentans 5.8 1 Peptostreptococcus productus 4.93 Clostridium perfringens 5.80 Clostridium ramosum 4.84 Streptococcus /actis 5.61 Bacillus adolescentSC 4.81 Kiebsiella mitis 5.36 Peptococcusmagnus 4.76 Streptococcus mitis 5.22 Lactobacilus fermentum 4.44 Staphylococcus epidermidis 5.18 LactobaÃ§illus acidophilus 3.39 Yeasts (other) 4.54 Clostridium aminovalericum 4.54 Klebsiella sp. 4.41 Clostridium perfringens 4.41 Staphylococcus epidermidis 4.26 Staphylococcus mitis 4.21 Yeasts (other)

a Log10 no. of organisms per g feces (dry weight).

References Human Fecal Flora: Comparison of Japanese and American Diets. Am. J. Clin. Nutr., 27: 1456-1469, 1974. 6. Haenszel, W., and Correa, P. Cancer ofthe Large Intestine: Epidemio I. Aries, V. C., Crowther, J. S., Drasar, B. S., Hill, M. J., and Williams, logic Findings. Diseases Colon Rectum, 16: 371 -377, 1973. R. E.0. BacteriaandtheAetiologyof Large-BowelCancer.Gut,10: 7. Hill, M. J. Steroid Nuclear Dehydrogenation and Colon Cancer. Am. 334â€”335,1969. J. Clin. Nutr., 27: 1475-1480, 1974. 2. Berg, J. W., Howell, M. A., and Silverman, S. J. Dietary Hypotheses 8. Hill, M. J., Drasar, B. S., Aries, V., Crowther, J. S., Hawksworth, G.. and Diet-related Researchin the Etiology of Colon Cancer. Health and Williams, R. E. 0. Bacteria and Aetiology of Cancer of Large Serv.Rept.,88:915-924,1973. Bowel. Lancet, 1: 95-100, 1971. 3. Berge, T., Ekelund, G., Mellner, C., PihI, B., and Wenckert, A. 9. MacGregor, I. 1. Carcinoma of the Colon and Stomach. A Review Carcinoma of the Colon and Rectum in a Defined Population. Acta with Comment on Epidemiologic Associations. J. Am. Med. Assoc., Chir. Scand. Suppl., 438: 1-86, 1973. 227:911-915,1974. 4. Buntain, W. L., ReMine, W. H., and Farrow, G. M. Premalignancy of 10. Peach, S., Fernandez, F., Johnson, K., and Drasar, B. S. The Polypsof the Colon. Surg. Gynecol.Obstet., /34: 499-508, 1972. Non-Sporing Anaerobic Bacteria in Human Faeces. J. Med. Microbi 5. Finegold, S. M., Attebery, H. R., and Sutter, V. L. Effect of Diet on ol.,7:213-221,1974.

NOVEMBER 1975 3417

Sydney M. Finegold, Dennis J. Flora, Howard R. Attebery, et al.

Cancer Res 1975;35:3407-3417.

Updated version Access the most recent version of this article at: http://cancerres.aacrjournals.org/content/35/11_Part_2/3407

E-mail alerts Sign up to receive free email-alerts related to this article or journal.

Reprints and To order reprints of this article or to subscribe to the journal, contact the AACR Publications Subscriptions Department at [email protected].

Permissions To request permission to re-use all or part of this article, use this link http://cancerres.aacrjournals.org/content/35/11_Part_2/3407. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC) Rightslink site.