Comparative gas chromatography-mass spectrometry study of the composition of microbial chemical markers in feces

Figures & data

Table I. List of fatty acids, aldehydes, and sterols detected in the feces indicating the most likely microorganisms in whose cells they usually were found.

No.	Abbreviation^*	Chemical name	Most likely microorganisms
1	C10	Decanoic	Streptococcus
2	C12:0	Dodecanoic	Non-specific
3	iC13	iso-Ridecanoic	Bacillus subtilis
4	al3	ansteiso-Tridecanoic	Bacillus cereus
5	il4	iso-Tetradecanoic	Peptostreptococcus anaerobius, Streptomyces, Bacillus, Bacteroides
6	14:1ω07	7,8-Tetradecenoic	Haemophilus parahaemolyticus, Arcobacter
7	14:1ω05	9,10-Tetradecenoic	Clostridium
8	14:1ω03	11,12-Tetradecenoic	Nocardia
9	10Mel4	10-Methyl-tetradecanoic	Actinomycetes
10	15:1 ω06	9,10-Pentadecanoic	Clostridium
11	il5	iso-Pentadecanoic	Propionibacterium
12	al5	anteiso-Pentadecanoic	Staphylococcus, Bacillus
13	1 OMe15	10-Methyl-pentadecanoic	Actinomycetales
14	il6:l	iso-Hexadecenoic	Pseudonocardia
15	16:1ω09	7,8-Hexadecenoic	Clostridium ramosum
16	16:1ω05	11,12-Hexadecenoic	Ruminococcus
17	il6:0	iso-Hexadecanoic	Streptomyces, Corynebacterium betae, Curtobacterium, Cellulomonas
18	16:0	Hexadecanoic	Non-specific
19	1 OMe16	10-Methyl-hexadecanoic	Rhodococcus
20	i17:1 I	iso-Heptadecenoic I	Campylobacter mucosalis
21	17:1	Heptadecenoic	Candida albicans, Mycobacterium
22	il7:0	iso-Heptadecanoic	Bacillus, Prevotella, Propionibacterium
23	al7:0	aHJmo-Heptadecanoic	Corynebacterium CDC group
24	17 eye	cycfo-Heptadecanoic	Enterobacteriaceae
25	17:0	Heptadecanoic	Non-specific
26	1 OMe17	10-Methyl-heptadecanoic	Actinomadura
27	18:1ω07	11,12-Octadecenoic	Lactobacillus
28	il8	iso-Octadecanoic	Clostridium difficile, Bacillus subtilis,
29	1 OMe18	10-Methyl-octadecanoic	Mycobacterium, Nocardia, Corynebacterium bovis, C. xerosis group, C. urealyticum, Actinomycetes
30	19 eye	cycfo-Nonadecanoic	Enterococcus,
31	20:1	Eicosenoic	Propionibacterium jensenii, Actinomyces
32	il9	iso-Nonadecanoic	Bacillus subtilis, Bacteroides hypermegas
33	al9	aHJezso-Heptadecanoic Hydroxy-acids	Staphylococcus
34	hlO	Hydroxy-decanoic^†	Pseudomonas, Comamonas
35	hill	Hydroxy-iso-undecanoic	Stenotrophomonas maltophilia
36	hl2	Hydroxy-lauric	Acinetobacter, Moraxella
37	hil3	Hydroxy-iso-tridecanoic	Stenotrophomonas maltophilia
38	hl3	Hydroxy-tridecanoic	Selenomonas
39	hl4	Hydroxy-myristic	Fusobacterium, Haemophilus
40	hi15	Hydroxy-iso-pentadecanoic	Prevotella, Flavobacterium, Capnocytophaga
41	hl6	Hydroxy-palmitic	Bacteroides, Wolinella
42	hi17	Hydroxy-iso-heptadecanoic	Bacteroides fragilis
43	hl7	Hydroxy-heptadecanoic	Bacteroides ruminicola
44	10hl8:l	10-Hydroxy-octadecenoic	Clostridium perfringens
45	hl8	Hydroxy-stearic	Helicobacter pylori
46	10hl8	10-Hydroxy-stearic	Clostridium perfringens
47	hal5	Hydroxy-aHJmo-pentadecanoic	Bacteroides
48	hal7	Hydroxy-aHJmo-heptadecanoic	Bacteroides
49	2hl2	2-Hydroxy-lauric	Pseudomonas aeruginosa, Alcaligenes
50	2hl4	2-Hydroxy-myristic	Sphingomonas
51	2hil5	2-Hydroxy-iso-pentadecanoic	Flavobacterium, Flexibacter
52	2hl6	2-Hydroxy-palmitic	Pseudomonas cepacea, Flexibacter
53	2hi17 Fatty aldehydes	2-Hydroxy-iso-heptadecanoic	Flexibacter
54	14a	Tetradecanoic	Eubacterium lentum, Bifidobacterium
55	il5a	iso-Pentadecanoic	Propionibacterium acnes
56	al5a	aHJezso-Pentadecanoic	Eubacterium, Peptostreptococcus
57	15a	Pentadecanoic
58	16:1a	Hexadecenoic	Eubacterium moniliforme
59	il6a	iso-Hexadecanoic	Eubacterium lentum
60	16a	Hexadecanoic	Non-specific
61	17a	Heptadecanoic	Propionibacterium freudenreichii
62	i17a	iso-Heptadecanoic	Propionibacterium
63	a17a	anteiso-Heptadecanoic	Eubacterium lentum
64	18:1ffl9a	9,10-Octadecenoic	Bifidobacterium
65	18:1ωo7a	11,12-Octadecenoic	Eubacterium, Clostridium ramosum
66	19cyc a Sterols	cyclo-Nonadecanoic	Enterococcus
67	Coprostanol	Dehydro-cholesterol	Eubacterium
68	Cholestendiol		Herpes virus
69	Campesterol		Microscopic fungi
70	β5-Sitosterol		Microscopic fungi
71	Cholestadienon		Cytomegalovirus
72	Ergosterol		Microscopic fungi

^*In 17:1, 17 is the number of carbon atoms, the figure after the colon denotes the number of double bonds; h, hydroxy-acid; a,I, indicates methyl-branching; cyc, cyclopropanoic acid. For example, 2hi15 means 2-hydroxy-iso-pentadecanoic acid.

^†3-Hydroxy-acids, if position of hydroxyl is not indicated.

Table IV. Comparison of data for analysis of fecal microbiota by genetic, cultural and biochemical, and mass-spectrometric studies.

	Composition of adult fecal microbiota, cells/g wet weight
Parameter	Mass spectrometry (this paper)	Genetic study, Harmsen et al. (51)	Cultural study
			Schaechter et al. (52)	Wittmann (dry weight) (53)
Total number	0.3-4 ×10^11	2.1 ×10^11	10^10-10^12	2 ×10^11
Proportion of anaerobes, %	70-90	Up to 100	Up to 100	33–100
Eubacterium	0.3-3 ×10^10	1.4 ×10^10	10^9-10^12	3 ×10^10
Bacteroides	0.2-2.4 ×10^10	3.6 ×10^10	10^10-10^12	10^11
Clostridium	10^9-10^11	2.9 ×10^10	10^5-10^11	3 ×10^10
Bifidobacterium	10^8-10^10	6 ×109	10^8-10^12	2 ×10^8

Table II. Results of examination of the microbial composition (cell/g ×10⁶) of feces from healthy persons aged 12-60 years, neonatal meconium, and transitional stool of newborn and IBS patients before and after treatment.

		Colonization level, cells/g × 10^6, mean value
No.	Microorganism	Meconium (n = 2)	Newborn (n = 2)	Adults (n = 7)	Elderly (n = 1)	IBS before treatment (n = 5)	IBS after treatment (n = 3)
1	Eubacterium lentum	144	83	8213	2792	3223	978
2	Eubacterium	0	0	13 743	11 579	14 251	4693
3	Propionibacterium	18	0	13 442	7632	7875	4377
4	Ruminicoccus	315	368	506	30	1280	93
5	Peptostreptococcus	5519	539	2514	2991	6636	6384
6	Clostridium histolyticum	6	45	112	6	78	5
7	Clostridium propionicum	0	828	4055	384	829	198
8	Clostridium ramosum	638	884	3281	2096	1635	706
9	Clostridium difficile	1333	622	1046	820	390	377
10	Clostridium perfringens	6	34432	45470	9401	55209	11452
11	Bacteroides hypermegas	0	34	151	99	364	101
12	Porphyromonas	0	0	55	82	95	38
13	Prevotella	74	0	3137	8149	3452	2430
14	Bacteroides fmgilis	0	0	1451	3223	2763	1575
15	Bacteroides ruminicola	52	0	2208	2130	6764	2822
16	Fusobacterium	0	601	385	428	907	280
17	Bifidobacterium	78	122	6589	1769	2441	2131
18	Lactobacillus	1054	2102	26 354	4545	19 070	3549
19	Helicobacter pylori	33	251	9968	368	2427	505
20	Pseudomonas aeruginosa	0	0	21	42	5	4
21	Acinetobacter	0	27	182	539	318	132
22	Stenotrophomonas	0	8	14	0	0	0
23	Alcaligenes	142	638	112	159	116	71
24	Flavobacterium	3	16	19	25	29	22
25	Enterobacteriaceae spp.	0	365	359	0	138	19
26	Campylobacter	125	14	44	0	9	30
27	Bacillus cereus	94	27	445	181	247	220
28	Bacillus megaterium	6	66	623	0	431	774
29	Enterococcus	10	1643	855	921	292	154
30	Staphylococcus	556	226	531	1991	271	175
31	Streptococcus (oral)	160	1169	4503	0	421	443
32	Streptococcus mutans	455	44	818	0	542	601
33	Enterococcus faecalis	0	0	1573	3587	1927	0
34	Streptococcus intermedius	0	685	2659	0	1371	2322
35	Coryneform CDC group	0	306	521	0	221	172
36	Nocardia spp.	54	18	146	70	172	86
37	Actinomycetes 10Mel5	69	42	48	34	78	61
38	Pseudonocardia	133	41	43	477	128	153
39	Streptomyces	828	164	2378	518	1420	900
40	Rhodococcus	414	283	395	74	140	166
41	MycobacteriumlCandida	41	331	771	65	140	32
42	Actinomadura	9	0	22	0	7	11
43	Nocardia asteroides	235	126	868	0	159	56
44	Actinomycetes 10Mel4	155	472	167	54	425	155
45	Microscopic fungi 1^a	160	5350	2462	1173	783	500
46	Microscopic fungi 2^b	290	6240	5118	8	1958	1526
47	Cytomegalovirus	16	0	59	0	29	19
48	Herpes virus	204	68	20	13	54	209
	Total	13435	59292	168490	68484	141569	51770

Gas chromatography-mass spectrometry of microbial markers was used. Accuracy of measurements is 20% reproducible.

^aMicroscopic fungi producing campesterol.

^bMicroscopic fungi producing sitosterol.

Table III. Grouping of microorganisms (cell/g × 10⁶) of feces from healthy persons aged 12-60 years, neonatal meconium, and transitional stool of newborn, and IBS patients before and after treatment.

	Colonization level, cells/g ×10^6, mean
Microorganism group	Meconium (n = 2)	Newborn (n = 2)	Adults (n = 7)	Elderly (n = 1)	IBS before treatment (n = 5)	IBS after treatment (n = 3)
Total	13435	59292	168490	68484	141569	51770
Total anaerobes	9273	40918	142701	58544	129721	42734
Anaerobes, %	69	65	79	85	90	84
Aerobic gram-negative bacteria	271	1068	752	765	616	278
Cocci and bacilli	1281	3861	12 007	6680	5502	4690
Actinobacteria	1938	1783	5361	1292	2890	1793
Fungi	450	11 591	7579	1181	2741	2026
Viruses	220	68	80	13	83	228
Bacteroides	126	34	7002	13683	13439	6965
Clostridium	1982	36810	53964	12707	58140	12738
Proportion, %
Aerobic gram-negative	1.9	4.4	0.5	1.1	0.6	0.5
Cocci and bacilli	9.3	9.8	7.2	9.8	4.7	7.3
Actinobacteria	14.3	7.8	3.5	1.9	2.4	3.2
Fungi	3.5	12.3	9.4	1.7	2.6	4.9
Viruses	1.6	0.3	0.1	0.02	0.1	0.5
Enterobacteria	0.0	2.5	0.2	0.0	0.1	0.1
Bifidobacteria	0.5	0.2	4.6	2.6	2.4	3.9
Lactobacilli	8.0	17.1	13.6	6.6	10.7	5.9
Clostridium, %	14	39	25	18.6	29	25

Harmsen HJM, Raangs GC, He T, Degener JE, Welling GW. Extensive set of 16S rRNA probes for detection of bacteria in human feces. Appl Environ Microbiol 2002;68:2982–90.

 PubMed Web of Science ®Google Scholar

Schaechter M, Medoff G, Eisenstein BJ, Mechanisms of microbial disease. Baltimore: Williams & Wilkins. 1993.

 Google Scholar

Wittmann DH. Intra-abdominal infections. New York: Marcel Dekker, Inc. 1991.

 Google Scholar