Documente Academic
Documente Profesional
Documente Cultură
VOL.
8: 259-266 (1995)
The effects of alginate consumption on the faecal microbiota and faecal metabolic activity were studied in eight
healthy male volunteers who consumed a mixed free choice diet (control diet) for 1 wk before and after alginate
consumption, and the diet with additional 10 g alginate once a day for 2 wks. During alginate consumption, the
levels of bifidobacteria increased significantly (P<0.05), while the levels of Enterobacteriaceae and the frequency of
occurrence of lecithinase-negative clostridia showed tendencies to decrease. No detectable change occurred in the
levels or the incidence of other microorganisms throughout the experimental period. Faecal sulphide, phenol,
p-cresol, and indole were significantly decreased (P<0.05) during alginate consumption. After 2 wks of alginate
consumption, faecal concentrations of ammonia and skatole were significantly reduced (P<0.05),whereas the values
of acetic and propionic acids were significantly increased (PcO.05). The water content and weight of the faeces were
slightly increased to some extent during the consumption. The odour of the faeces decreased markedly during
alginate consumption.
KEY WORDS:
INTRODUCTION
Alginates are widely distributed in the cell wall and
cytoplasmic membrane of sea vegetables, l 1 such
as Ecklonia cava, E. maxima, Eisenia bicyclis,
Laminaria, Lessonia, Macrocystis, and Ascophyllum nodosum etc., and extracted from sea
vegetables with sodium carbonate. These are composed of a polymer with p-1, 4-D-mannuronate
and a- 1, 4-L-guluronate.
Alginates are used as emulsifier and stabiliser
in foods, medicine, cosmetics, feeds and various
industries, and suppress intestinal absorption of
heavy metals,31have an antitumour e f f e ~ t ,lower
'~
cholester01'~~~
and lower blood p re~ su re.~ '
Pratt et uI.*~ found that seaweed extract had
antibacterial activity in vitro against various organreported that Staphytocoisms. Burkholder et
ccus aureus is more susceptible to the inhibitory
products of algae than were Escherichia coli or
Candida albicans. Akiyama et
have showed a
significant increase in the growth of bifidobacteria
in vitro by the addition of depolymerised alginate.
*Author to whom correspondence should be addressed.
CCC 0891-060)<195/060259-08
0 1995 by John Wiley & Sons, Ltd
A, TERADA ET AL.
260
Table 1. Mean daily energy and nutrient consumption
Diet ingested
Before
consumption
CN-1"
Alginate consumption
AS- 1
AS-2"
After
consumption
CN-2"
2698 f 175Id
85.6 f 4.3
82.3 f 5.3
749 f 28
11.1 f 1.3
2.0 f 0.3
25.3 f 1.6
27.6 f 1.5
2624 f 154
83.9 f 4.1
81.0 f 5.7
736 f 25
10.9 f 1.1
2.1 zt 0.3
25.1 k 1.3
26.0 f 1.8
2641 f 136
84.3 f 3.9
83.1 f 6.3
738 zt 23
11.23~1.3
2.0 f 0.2
23.8 f 1.5
26.8 f 2.1
~~
Energy (KcaVday)
Protein (g/day)
Fat (glday)
Calcium (mglday)
Salt @day)
Crude fibre (g/day)
Saturated fatty acid (g/day)
Unsaturated fatty acid (g/day)
described p r e v i o u ~ l y .The
~ ~ ' results
~~
are expressed
as the log,, of the number of bacteria per gram wet
weight of faecal material.
Analysis of faecal metabolic products
Faecal ammonia was determined by potentiometer ILO-30 (DKK Co. Ltd, Tokyo) with an
ammonia gas sensing electrode 1716 (DKK Co.
Ltd, Tokyo) using the method of Terada et aZ.34
For the measurement of sulphide, a portion of 1 g
of faeces was suspended in 44 ml of distilled water,
mixed thoroughly with 5 ml of S-DIMAB (NaOH
40 g, L-ascorbic acid 10 g, sodium ethylenediaminetetra acetate 9.3 g, and glycerin 500 ml in
1000 ml of distilled water). The faecal sulphide was
measured using a sulphide electrode 7100 (DKK
Co. Ltd, Tokyo). Faecal concentrations of phenols
and indole were examined by gas chromatography
using the method of Y ~ s h i h a r a Faecal
. ~ ~ analysis
of short chain fatty acids (SCFA) were carried out
by the high-performance liquid chromatographic
method of Hara et al. l 6
Measure of faecal properties
Faecal moisture contents were measured using
approximately 1 g samples, weighed before and
after drying in a vacuum oven at 105C for 2 h.
The difference in weight was regarded as the water
content. The pH values were determined by inserting a flat glass electrode (DKK Co. Ltd, Tokyo)
directly into faeces.
26 1
Organism
Before
consumption
Day 0
Total bacteria
Bifidobacteria
10.95 f 0.17"
9.97 f0.26
Bacteroidaceae
10.66 f 0.21
(100)
9.98 f 0.3 1
(100)
9.34 f 0.66
(100)
8.87 f 0.37
(63)
8.73 f 0.61
(38)
6.10 f 0.20
(38)
Eubacteria
Peptococcaceae
Megasphaerae
Curved rods
Veillonellae
Clostridia
Lecithinase-positive
Lactobacilli
Enterobacteriaceae
Pseudomonads
Streptococci
Staphylococci
Bacilli
Yeasts
Moulds
~
After
consumption
Day 28
Day 14
Day 21
10.96 f 0.10
10.27 f0.12*
( 100)
10.69 f 0.12
(100)
9.94 f 0.31
( 100)
9.69 f 0.29
( 100)
9.23 f 0.35
(75)
8.94 f 0.70
(63)
5.63 f 1.89
(38)
10.86 f 0.10
10.22 f 0.21*
(100)
10.62 f 0.10
(100)
9.62 f 0.41
( 100)
9.49 f 0.57
(100)
9.13 f 0.37
(75)
8.80 f 0.50
6.40 f 0.78
(38)
11.03 f 0.13
9.98 f0.18
( 100)
10.85 f 0.14
(100)
9.82 f 0.33
( 100)
9.53 f 0.25
(100)
9.02 f 0.69
(75)
8.94 iz 0.58
(50)
5.90 f 1.16
(38)
3.55 f 0.90
(75)
9.06 f0-55
(38)
7.39 f 1.28
(88)
7.93 f 1.00
( 100)
3.43 f 0.62
(38)
8.11 f 1.54
(100)
2.95 f 0.79
4.18 f 1.17
(75)
9.13 f 0-49
5.39 f 1.83
(88)
9.30 f 0.33
4.81 f 1.21
( 100)
8.98 f 0.62
(88)
6.11 f 1.76
( 100)
8.37 f 0.84
( 100)
3.29 f 0.68
(50)
7.94 f 1.47
( 100)
3.36 f 0.77
(63)
3.49 f 0.92
(38)
2.94 f 1.13
(50)
2.40
(13)
Lecithinase-negative
~~~~~~~
During consumption
(50)
2.79 f 0.70
(38)
2.98 f 0.72
(50)
(0)
<
(50)
(50)
6.17 f 1.29
(88)
7.78 f 0.92
(100)
2.95 f 0-23
(50)
7.89 f 1.00
(100)
3.14 f 0.98
(75)
3.40 f 1.06
(63)
3.03 f 0.52
(38)
(0)
(100)
7.06 f 1.94
( 100)
8.72 f 1.15
(100)
3.02 f 0-40
(63)
8.28 f 0.84
( 100)
4.23 f 1.19
(75)
3.34 f 1.51
(63)
3.06 f 0.44
(50)
2.30
(13)
RESULTS
Faecal mkrobiota analysis
The effects of alginate on the faecal microbiota
analysis of eight volunteers are shown in Table 2.
The numbers of bifidobacteria were significantly
increased (P<0.05) during days seven and 14 of
262
A . TERADA ET AL.
10 -
9
Day 0
Day 14
Day 21
Before
During consumption
consumption
Day 28
After
consumption
days seven (P<0.05) and 14 (P<O.Ol) of consumption. Total faecal SCFA, acetic acid and propionic
acid significantly increased (W0.05) on day 14
of consumption (Figure 4). The odour of faeces
was noted to be less offensive during alginate
consumption.
Faecal weight, water content and p H value
Stool weights and water content showed a slight
tendency to increase during alginate consumption compared with those before consumption
(Table 3). No great difference occurred in faecal
pH values throughout the experimental period.
DISCUSSION
The indigenous faecal microbiota and its metabolic activities are thought to be important in
human
In particular, the increase of
bifidobacteria in the intestinal tract is thought to
exert a beneficial effect for the host by production
of short chain fatty acids, lowering the pH in the
bowel, decreasing putrefactive products, inhibiting
the growth of potential pathogens and immunopotentiation.
The intestinal bacteria (such as
bifidobacteria, ruminococci, clostridia and some
bacteroides) are known to play an important role
on the utilisation of dietary
such as plant
cell ~ a l l s ~ (cellulose,
, ~ ~ . ~ hemicellulose,
~
xylan,
'5320,27
263
d
v
'z
0
300
W
250
Em 200
v)
Y-
Bc
150
100
-0.
3
3 :0
.-
LI
u
C
P 50
W
0
Day14
Day21
During consumption
Day 0
Before
consumption
Day 28
0 !,
"
After
consumption
........I.
!!:::::-.!
264
A. TERADA ET AL.
...
Butyric acid
Propionic acid
Acetic acid
Day0
Day 14
Day21
Day28
Before
During consumption
After
consumption
consumption
Succinic acid
c]Lactic acid
..
Figure 4. Effect of alginate consumption on faecal short chain fatty acids (SCFA). Data are expressed
as mean of eight adults. Significant difference from the values before consumption; *P<0.05
Table 3. Effect of alginate consumption on human faecal weight, pH and water content
During consumption
Product
Before
consumption
Day 0
Day 14
Day 21
After
consumption
Day 28
108.4 z t 24.9
6.6 f 0.4
72.9 f 3.1
135.8 k 39.2
6.5 f 0.5
75.5 & 5.5
127.3 f 31.7
6.6 zt 0.6
76.1 f 4.3
103.6 zk !0.3
6.5 & 0.4
74.1 + 4.3
concentrations of Enterobacteriaceae and the hemicellulose.30Faecal sulphide formed by deamilower detection rates of lecithinase-negative nation of cysteine and methionine in the intestine
clostridia during alginate consumption is similar decreased significantly during alginate consumpto that seen during lactosucrose16 and ~ h i t o s a n ~tion
~ similarly to that seen during lactosucrose6
consumption, respectively.
consumption. Phenolic and indolic compounds
The principal bacterial end-products in the produced by deamination of tyrosine, phenylhuman intestine are ammonia, sulphide, amines, alanine and tryptophan are detoxified by glucurophenols, the SCFA, fermentation gases and nide or sulphate conjugation in the liver and
energy, which the organisms utilise for rowth and excreted in urine. The decrease of faecal phenol,
the maintenance of cellular function! The end- p-cresol and indole in this study were similar
products of the metabolites produced during the to those seen with o l i g o s a ~ c h a r i d e s and
~~~~
catabolism of proteins are potentially harmful to ~ h i t o s a nconsumption.
~~
the host. Ammonia formed by deamination of
Cummings and MacFarlane8 reported that little
amino-acids, peptides and protein is absorbed change in faecal SCFA occurred with variation in
from the gut and detoxified by urea formation in diet, but that the total SCFA increased in parallel
the liver. In this study, the reduction of faecal with faecal weight. Adding corn bran to a low fibre
ammonia have resulted from either the sup- diet increased total SCFA in proportion of faecal
pression of ammonia production in the intestine, output. In the present study, the concentration
or from enhanced ammonia absorption from of total SCFA, acetic and propionic acids were
the large intestine as seen in humans fed corn increased during alginate consumption as has been
265
11.
12.
13.
REFERENCES
1. Abe S, Kaneda T. (1973). Studies on the effect of
marine products on cholesterol metabolism in
rats-IX. Effect of betaines on plasma and liver
cholesterol levels. Bulletin of the Japanese Society
of Scientific Fisheries 39, 391-393.
2. Akiyama H, Enda T, Nakakita R, Murata K,
Yonemoto Y, Okayama K. (1992). Effect of depolymerized alginates on the growth of bifidobacteria. Bioscience Biotechnology Biochemistry 56,
355-356.
3. Aoe S, Ohta F, Ayano Y. (1988). Effect of watersoluble dietary fiber on intestinal microflora in rats.
Nippon Eiyo Shokuryo Gakkaishi 41, 203-21 1 (in
Japanese).
4. Bayliss CE, Houston AP. (1984). Characterization
of plant polysaccharide- and mucin-fermenting
anaerobic bacteria from human feces. Applied and
Environmental Microbiology 48, 626632.
5. Benno Y, Endo K, Miyoshi H, Okuda T, Koishi H,
Mitsuoka T. (1989). Effect of rice fiber on human
fecal microflora. Microbiology and Immunology 33,
43540.
6. Burkholder PR, Burkholder LM, Almodovar LR.
(1960). Antibiotic activity of some marine algae of
Puerto Rico. Botanic Marina 2, 149-156.
7. Cummings JH, Hill MJ, Jenkins DJA, Pearson JR,
Wiggins HS. (1976). Changes in fecal composition
and colonic function due to cereal fiber. American
Journal of Clinical Nutrition 29, 1468-1473.
8. Cummings JH, MacFarlane GT. (1991). The control and consequences of bacterial fermentation in
the human colon. Journal of Applied Bacteriology
70,443459.
9. Drasar BS, Jenkins DJA, Cummings JH. (1976).
The influence of a diet rich in wheat fibre on
the human faecal flora. Journal of Medical
Microbiology 9, 42343 1.
10. Endo K, Kumemura M, Nakamura K, Fujisawa T,
Suzuki K, Benno Y, Mitsuoka T. (1991). Effect of
high cholesterol diet and polydextrose supplementation on the microflora, bacterial enzyme activity,
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
266
24. Moore WEC, Cat0 EP, Holdeman LV. (1969).
Anaerobic bacteria of the gastrointestinal flora and
their occurrence in clinical infections. Journal of
Infectious Disease 119, 641-649.
25. Pratt R, Mautner H, Gardner GMG, Hsien S,
Dufrenoy J. (1951). Antibiotic activity of seaweed extracts. Journal of American Pharmacology
Association 40, 575.
26. Roberton AM, Stanley RA. (1982). In vitro utilization of mucin by Bacteroides fragilis. Applied and
Environmental Microbiology 43, 325-330.
27. Rowland IR, Wise A. (1985). The effect of diet on
mammalian gut flora and its metabolic activities.
CRC Critical Reviews in Toxicology 16, 31-103.
28. Salyers AA. (1979). Energy sources of major intestinal fermentative anaerobes. American Journal of
Clinical Nutrition 32, 158-1 63.
29. Salyers AA, Leedle JAZ. (1983). Carbohydrate
utilization in human colon. In: Hentges DJ (ed)
Human Intestinal Microflora in Health and Disease.
London: Academic Press, pp. 129-146.
30. Sugawara M, Suzuki K, Endo K, Kumemura M,
Takeuchi M, Mitsuoka T. (1990). Effect of the
dietary supplementation of corn hemicellulose on
faecal flora and bacterial enzyme activities in
human adults. Agricultural Biological Chemistry
54, 1683-1688.
31. Sutton A. (1967). Reduction of strontium absorption in man by the addition of alginate to the diet.
Nature 216, 1005-1007.
32. Tanaka R, Takayama H, Morotomi M,
Kuroshima T, Ueyama S, Matsumoto K, Kuroda
A, Mutai M. (1983). Effects of administration
of TOS and Bijidobacterium breve 4006 on the
human faecal flora. Bijidobacteria and Microflora 2,
17-24.
33. Terada A, Hara H, Kataoka M, Mitsuoka T.
(1992). Effect of lactulose on the composition
and metabolic activity of the human faecal
flora. Microbial Ecology in Health and Disease 5,
43-50.
34. Terada A, Hara H, Hemmi A, Kataoka M,
Mitsuoka T. (1993). A simple and rapid method
for the determination of excretal ammonia using
ammonia gas sensing electrode. Animal Science and
Technology 64,719-722.
35. Terada A, Hara H, Ikegame K, Sasaki M,
Mitsuoka T. (1994). Recommended method of
A. TERADA ET AL.
36.
37.
38.
39.
40.
41.
42.
43.
44.