Original Article

診療と新薬 2018; 55: 115-123

The Functional Food Containing Lactobacillus andDietary Fiber for Amelioration of Bowel Movementin Healthy Japanese:

診療と新薬 WebA Randomized, Double-blind, Placebo-controlled Study

Masatomo NAJIMA1) ／ Akinobu MIYATA2) ／ Kimitoshi YAMAGUCHI3)● Abstract
Objectives: The objective of this study is to examine how the ingestion of the functional food containing
Enterococcus faecalis (EC- BabyM®) and indigestible dextrin (dietary ﬁber), Dietary ﬁber Nuusankin 5,000
oku-ko ( DFN ) contributes to improve bowel movement.
Methods: A randomized, placebo-controlled, double-blind study was conducted to elucidate an eﬀect of food.
In this study we measured frequency of bowel movement, days of defecation, volume of stool, fecal properties,
and intestinal ﬂora as the primary outcome. We also evaluated blood biochemical parameters as the secondary
outcome, and adverse events were collected by means of a written questionnaire during the study.
Results: From all of 63 applicants, 8 were eliminated due to not meeting inclusion criteria. 50 subjects wererandomly assigned to an intervention group and made a start with ingestion. 22 subjects discontinued due topersonal reasons, and the remaining 28 subjects completed the study. 1 was eliminated because of inadequacyof the records of bowel movement, thus data obtained with 27 subjects (DFN: 15, Placebo: 12) was used forthe analysis of eﬃcacy. After 12-weeks of ingestion, the intergroup analysis showed signiﬁcant diﬀerences inFrequency of bowel movement , Days of defecation , and Bifidobacterium share in the intestinal floracompared to the Placebo. No adverse events were observed after the ingestion of the test product.
Conclusion: We found out that the ingestion of the functional food containing Enterococcus faecalis (ECBabyM®) and indigestible dextrin (dietary fiber) for 12 weeks improved bowel movement. In addition, no
safety-related matter occurred during the 12-week test period.
Key Words: indigestible dextrin, dietary fiber, lactobacillus, Enterococcus faecalis , EC-BabyM®,
constipation, bowel movement

1. INTRODUCTION
Improving the intestinal environment is considered to bea key element in our healthcare. A number of studies
i n d i c a t e t h a t t h e i n t e s t i n a l f l o r a i s i n vo l ve d i npathogenesis of gastrointestinal disorders such as
inflammatory bowel disease, metabolic diseases such as
obesity and diabetes mellitus, as well as neuronal
disorders including multiple sclerosis. On the other hand,according to the Comprehensive Survey of Living
Conditions conducted by Ministry of Health, Labor andWelfare in 2016, constipation is prevalent in 16.4％ ofmales and females of 65 years and over 1). Needless to say,
it is important to change the lifestyle habits, increase theamount of exercise, or reduce stress, in order to improvebowel movement. In addition, it is also effective toactively ingest dietary ﬁber (recent Japanese are said tolack this) or the nutrition which stimulates the intestinal
1) JACTA (Japan Clinical Trial Association)2) Nihonbashi M s Clinic3) M. Foods Corporation

functions. Moreover to supply non-viable microbial
ingredients as well as live one in gastrointestinal
disorders has been illustrated in many studies 2)-5).
In this study, we conducted a randomized, placebocontrolled, double-blind study to verify the efficacy andthe safety of the functional food containing Enterococcus
faecalis (EC- BabyM®) and indigestible dextrin (dietaryﬁber) for the amelioration of bowel movement in healthy
Japanese.

2. METHODS
2.1.Trial design
A randomized, placebo-controlled, double-blind study was
conducted with the aid of a fund from M. Foods
Corporation (Tokyo) at Japan Clinical Trial Association( JACTA, Tokyo). 12 days pre-observation period was
carried out, followed by a 12 weeks intervention (fromJune 17 th to September 10 th, 2017). This study was
conducted in accordance with the ethical principles of thedeclaration of Helsinki and the Ethical Guidelines for
Medical and Health Research Involving Human Subjects.
The study protocol was approved by the Institutional36 （116）

診療と新薬・第 55 巻

第 2 号（2018 年 2 月）

Table 1 Ingredients（per 12 g / daily intake）
Item

DFN

PlaceboEnterococcus faecalis（EC-BabyM®）
indigestible dextrin（dietary ﬁber）
whole milk powder
Isomaltooligosaccharide
fructosemaltodextrin

505 billion（number）

−

7g
○
○
○
−

−
−
−
−
○

Review Board of Pharmaceutical Law Wisdoms (Tokyo).
Written informed consent was obtained from all Subjects.
This trial was registered at UMIN Clinical Trial Registry(Trial ID: UMIN 000027526). The allocation of the test
product to the subjects was carried out by the person in
charge of allocation. The allocation list was sealed andstrictly controlled in a safe deposit box of JACTA until
the end of the study.
2.2. Subject
Healthy subjects participated in the present study. All of
the subjects in this study were public volunteers who had
enrolled in the monitor bank of TRIBELATECORPORATION (Tokyo), recruited from April throughMay, 2017.
2.2.1. Inclusion criteria(1) Japanese aged between 20 and 60 years;(2) Japanese within normal level of defecation or with atendency toward constipation.
2.2.2. Exclusion criteria(1) Subjects with chronic constipation;(2) Subjects with food allergies;(3) Subjects who are pregnant or lactating;(4) Subjects who consume medicinal products which may
inﬂuence the outcome of the study;(5) Subjects who consume food which may inﬂuence theoutcome of the study;(6) Subjects who are judged as unsuitable for the study by
the principle investigator.
2.3. Randomization
From all 63 applicants, 8 were declined and 5 were
eliminated. The inclusion/ exclusion criteria was judged
by the principle investigator. All subjects weresequentially allocated to Group A (n ＝ 25, M; 12, F; 13)
and Group B (n ＝ 25, M; 12, F; 13) using a random
number table. In the process of subject assignment,
background factors such as gender, age, and frequency ofbowel movement were taken into consideration to avoid
biased distribution. Subjects in Group A ingested placeboand subjects in Group B ingested the test sample, DFN
for 12 weeks.
2.4. Description of test foods and blinding
The test product was Dietary fiber Nyuusankin 5,000
oku-ko ( DFN ) containing Enterococcus faecalis (ECBabyM®) and indigestible dextrin (dietary fiber),
prepared by M. Foods Corporation. The amount of daily

intake was 3 spoonfuls of powdery DFN (weighing 12 g).
The placebo ( Placebo ) does not include indigestible
dextrin (dietary fiber) and Enterococcus faecalis (ECBabyM®). Table 1 shows the ingredients of the sample.
Both powders were indistinguishable in color, ﬂavor, or
taste. All involved were blinded.
2.5. Experimental procedures
2.5.1. Experimental protocol
Subjects consumed 3 spoonfuls of powder directly, or
drinking by dissolving it in water after every dinner for
12 weeks. Subjects were instructed as follows: to takethe assigned concoction; to maintain their usual lifestyles
and habits; to avoid taking other supplements; tomaintain their usual lifestyles and habits; to avoid
excessive amounts of food, drink, or alcohol; to avoid
excessive intake of lactobacillus food (such as yogurt,
lactobacillus beverage, and Natto), oligosaccharide, or
dietary ﬁber which may inﬂuence fecal ﬂora; to maintaina daily record of one s bowel movement (frequency andvolume of stool) and fecal properties (scent, sensation,color, and texture); and to send the diary to the study
coordinator.
2.5.2. Outcome
The objective of this study is to elucidate the change forthe improvement of bowel movement by taking the foodcontaining Enterococcus faecalis (EC-BabyM®) andindigestible dextrin (dietary fiber). To evaluate thisobjective, frequency of bowel movement, days ofdefecation, volume of stool, fecal properties, andintestinal ﬂora were measured as the primary outcome.
As the secondary outcome, blood biochemical parameters
were recorded to evaluate the safety of DFN.
Furthermore, adverse events were collected by means ofa written questionnaire during the study. According to
the schedule shown in Figure 1, we measured
parameters on efficacy and safety. These assessments
were conducted upon pre- and post- intervention.
2.5.2.1. Bowel movement
Dairy reporting of Frequency of bowel movement ,Days of defecation , and Volume of stool were
calculated. Frequency of bowel movement is theamount of times one defecated during 1 week. Days ofdefecation is the number of days in which defecation
occurred during 1 week. Regarding Volume of stool ,subjects compared volume to a 5 x 2.5 cm cylindrical診療と新薬・第 55 巻

<Pre-observation period><Ingestion of test foods>

12 days

12 weeks〔Time line〕〔Visit〕▲
1)
Visit 1

〔Blood drawing〕

37 （117）▲
2)
Visit 2▲Visit 3△△
1 week

1 week〔Defecation〕〔Fecal sample〕

第 2 号（2018 年 2 月）

Within 3 days
after the period

Within the last
3 days of the period
13 weeks〔Diary〕

1) Informed consent and screening
2) Selection and/ or allocation

Figure 1 Schedule for the study

Table 2 Questionnaire about fecal properties
Item

Choices and score

Unit

Valuation method

Scent of stool

1; Very strong, 2; Strong, 3; A little strong, 4; Odorless

Score/
1 defecation

Average score at
1 defecation for a week

Color of stool

1; Yellow, 2; Ocher, 3; Light brown, 4; brown,
5; Dark brown, 6; Blackish brown

Score/
1 defecation

Average score at
1 defecation for a week

Texture of stool

1; Watery, no solid pieces, 2; Mushy, 3; Soft blobs withclear-cut edges, 4: Like a banana, smooth and soft,
5; Lumpy or cracks in the surface, 6; Separate hard
lumps, like a nut

Score/
1 defecation

Average score at
1 defecation for a week

object and recorded every time of defecation, then totaled
the amount of 1 week.
2.5.2.2. Fecal properties
As for fecal properties, subjects rated 3 items; Scent ofstool , Color of stool , and Texture of stool every timeof defecation on an ordinal scale, which are illustrated inTable 2. The subjects assessment of Color of stool
used a color chart. In the way of Texture of stool , weused Bristol stool form scale 6) and modiﬁed to Japanese.
Then the average score was calculated during a week.
2.5.2.3. Intestinal ﬂora
Subjects collected their fecal by the brush-type of feces
sampling kit containing preservative solution twice justbefore starting to intake (at the end of pre-observation
period) and just after 12 weeks of intake period. We
sampled feces excreted within 72 hours just before andafter the test period, for the reason that it is consideredthat it takes 36-72 hours for ingested food to be excreted
as feces 7), and subjects with constipation defecate once

every few or three days8) (Figure 1).
A fecal analysis was carried out in the Techno Suruga
Laboratory Co., Ltd, by means of Terminal restriction
fragement length polymorphism (the Nagashima methodof T-RLFP) 9) 10), and the ratio to the total number ofbacteria (occupancy) of Biﬁdobacterium , Lactobacillales,
Bacteroides , Prevotella , Clostridium cluster IV,
Clostridium subcluster XIVa, Clostridium cluster IX,
Clostridium cluster XI, Clostridiu m cluster XVIII,
Clostridium cluster XVIII, and others were measured.
2.5.2.4. Biochemical blood test
At visit 2 (0-week) and visit 3 (12-week), the subjects had
their blood taken. 12 hours prior to the visit, the subjects
were not allowed to eat or drink (except water). The
subjects took a 10 minute rest before proceeding withblood drawing to check blood biochemical parameters.
2.6. Data analysis
Per protocol set (PPS) was adopted in the study and nosample size design was used. All statistics were38 （118）

診療と新薬・第 55 巻

第 2 号（2018 年 2 月）

Assessed for eligibility (n ＝ 63)〔M:28, F:35〕

Enrollment

Excluded (n ＝ 13)〔M:4, F:9〕
¨Declined to participate (n ＝ 8)〔M:3, F:5〕
¨Did not meet inclusion criteria (n ＝ 5)¨¨Doctor̀s judgement (obesity, constipation)(n ＝ 5)〔M:1, F:4〕

Randomized (n ＝ 50)〔M:24, F:26〕

Group AGroup B

Allocation
Allocated to Placebo (n ＝ 25)〔M:12,F:13〕¨Received allocated intervention (n ＝ 25)¨Did not receive allocated intervention (n ＝ 0)

Allocated to DFN (n ＝ 25)〔M:12,F:13〕¨Received allocated intervention (n ＝ 25)¨Did not receive allocated intervention (n ＝ 0)

Follow - Up
Lost to follow - up (n ＝ 0)
Discontinued intervention (n ＝ 12)¨Personal reason,¨¨business reason (n=8)〔M:6, F:2〕¨¨Feeling too much forthe demanding tetsing expectation (n=4)
〔M:2, F:2〕

Lost to follow - up (n ＝ 0)
Discontinued intervention (n ＝ 10)¨Personal reason,¨¨business reason (n=4)〔M:2, F:2〕¨¨Feeling too much forthe demanding tetsing expectation (n=6)
〔M:3, F:3〕

AnalysisAnalysed (n ＝ 12)〔M:4, F:8〕
¨Excluded from analysis (n ＝ 0)〕¨¨Insuﬃcient diary (bowel movement) (n ＝ 1)
〔M:0, F:1〕

Analysed (n ＝ 15)〔M:7, F:8〕
¨Excluded from analysis (n ＝ 0)

Figure 2 Flow diagram of subject disposition

Table 3 Subject demographics
Item

Unit

DFN

Placebo

Subjects
Male: Female *Age *Frequency of bowel movement *numbers
numbers
years
times/ a week15
7:8
41.7 ± 11.9
10.5 ± 3.8

12
4:8
49.3 ± 5.5
7.4 ± 4.6

mean ± SD* No signiﬁcant diﬀerence

expressed as mean ± standard deviation (SD). For thefrequency of bowel movement, days of defecation,
volume of stool, fecal properties, intestinal flora, andblood biochemical parameters, Student s t-test was used
for intergroup comparisons, and paired t-test was used
for intragroup analysis. A chi-square test and Student s

t-test were used to compare subject backgrounds
between groups. Multiplicity according to the occasions
was not adjusted. Any subjects with missing values were
eliminated from the analysis. Statistical analyses were
performed using Statcel 4 (Yanai, 2015) and Excel Tokei
2015 (SSRI). The results were considered signiﬁcant at a

診療と新薬・第 55 巻

Table 4Item
Frequency of bowel
movement

第 2 号（2018 年 2 月）

39 （119）

Results of bowel movement

Unit

Time point

DFN (n ＝ 15)

Placebo (n ＝ 12)

0-week10.5 ± 3.8

7.4 ± 4.6

12-week

11.9 ± 4.3

8.3 ± 4.8 ＃number/ 1 wk

Days of defecation

day/ 1 wk

0-week12-week

6.4 ± 0.9
6.4 ± 0.8

5.3 ± 1.5 ＃4.9 ± 1.7 ＃＃Volume of stool

piece/ 1 wk

0-week12-week

27.5 ± 11.8
38.0 ± 10.4 ＊＊24.6 ± 19.3
30.9 ± 14.1

Values are expressed as the mean ± SD.
＊＊＃

p ＜ 0.01 against 0-week.

p ＜ 0.05, p ＜ 0.01 between-group diﬀerence in measured value.

Table 5 Results of fecal properties
Item

Unit

Time point

DFN (n ＝ 15)

Placebo (n ＝ 12)

Scent of stoolscore/
1 defecation

0-week12-week

2.7 ± 0.4
2.9 ± 0.6

2.9 ± 0.6
2.8 ± 0.6

Color of stoolscore/
1 defecation

0-week12-week

4.1 ± 0.4
4.0 ± 0.4

4.1 ± 0.5
3.9 ± 0.6

Texture of stoolscore/
1 defecation

0-week12-week

3.4 ± 0.5
3.7 ± 0.4

3.5 ± 0.6
4.1 ± 0.7 ＊Values are expressed as the mean ± SD.
＊

p ＜ 0.05 against 0-week.

＜ 5％ level in the two-sided test.

3. RESULTS
3.1. Participant demographics
The 50 subjects were randomly assigned to anintervention group and made a start with ingestion. 22
subjects were withdrawn due to personal reason (businessreasons:12, or feeling too much for the demanding testing
expectation:10), and the remaining 28 subjects completed
the study. Due to inadequacy of the records of bowel
movement, 1 (Placebo) was eliminated, thus dataobtained with 27 subjects (DFN: 15 ＜M; 7, F; 8＞ ,Placebo: 12 ＜M; 4, F; 8＞) was used for the analysis ofefficacy (Figure 2). The subject s age range was 23-58
years old (mean age 45.1±10.1 y.o.). There was no
significant difference in gender, age, and frequency ofbowel movement between groups (Table 3). The test
sample intake rates were 100％ and 100％ , respectively.
3.2. Bowel movement
Table 4 depicts the results of bowel movement. As forFrequency of bowel movement , a signiﬁcant diﬀerence
was yielded in the intergroup analysis at week 12,
without intragroup analysis. In Regard to Days ofdefecation , a significant difference was observed at thebaseline. At week 12, DFN maintained almost the sameconsistency. In contrast Placebo decreased, which
realized the between-group diﬀerence in measured value.

In Volume of stool , DFN increased significantly after
12-weeks of ingestion, whereas the Placebo showed nochange.
3.3. Fecal properties
Table 5 depicts the results of the state of stool. All items
showed no significant difference, whereas Texture ofstool indicated a tendency between two groups. As for
Texture of stool , DFN as well as Placebo were
evaluated between type 3 (Soft blobs with clear-cut
edges) and type 4 (Like a banana, smooth and soft) atbaseline. At week 12, Placebo showed a significant
hardness.
3.4. Intestinal ﬂora
Table 6 shows changes of compositions of the intestinal
flora. Bifidobacterium share of DFN was significantly
increased after 12-weeks of ingestion, so that signiﬁcantdiﬀerence was yielded in the intergroup analysis. As for
Lactobacillales , Placebo showed decreased tendency,
meanwhile DFN stayed about the same as pre-intake.
Other parameters did not show any signiﬁcant diﬀerence.
3.5. Biochemical blood test
Though an abnormal value was detected on CK (CPK) of
pre-ingestion of DFN, the principle investigator judged itas the range of physiological variation, because of
returning to its normal state after 12-weeks of ingestion.
Moreover, DFN showed a significant difference inGlucose, Neutral fat (TG), Sodium, Chloride, and40 （120）

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第 2 号（2018 年 2 月）

Table 6 Results of intestinal ﬂora
Item

DFN (n ＝ 15)

Placebo (n ＝ 12)

0-week

8.4 ± 9.0

9.4 ± 6.7

12‑week⊿ 0 ‑12 w

10.0 ± 9.7
1.6 ± 3.4

6.1 ± 5.0 ＊− 3.3 ± 5.6 ＃Lactobacillales

0-week12‑week⊿ 0 ‑12 w

2.2 ± 2.9
2.3 ± 2.3
0.1 ± 3.4

3.0 ± 2.8
1.4 ± 1.9
− 1.6 ± 3.2

Bacteroides

0-week12‑week⊿ 0 ‑12 w

43.5 ± 15.5
45.4 ± 17.1
1.9 ± 8.9

40.6 ± 11.3
42.9 ± 16.8
2.3 ± 17.9

Prevotella

0-week12‑week⊿ 0 ‑12 w

6.5 ± 14.8
5.5 ± 12.6
− 0.9 ± 4.7

8.8 ± 15.7
9.5 ± 15.2
0.6 ± 3.5

Clostridium cluster IV

0-week12‑week⊿ 0 ‑12 w

6.5 ± 3.9
6.3 ± 6.3
− 0.2 ± 3.3

5.5 ± 3.6
4.6 ± 3.3
− 0.9 ± 4.7

Clostridium subcluster XIVa

0-week12‑week⊿ 0 ‑12 w

15.9 ± 7.3
15.0 ± 6.7− 0.9 ± 3.9

16.4 ± 8.4
19.0 ± 14.8
2.6 ± 14.4

Clostridium cluster IX

0-week12‑week⊿ 0 ‑12 w

8.6 ± 8.5
8.1 ± 7.8
− 0.5 ± 7.3

6.8 ± 6.4
8.1 ± 9.1
1.3 ± 6.3

Clostridium cluster XI

0-week12‑week⊿ 0 ‑12 w

0.5 ± 1.1
0.3 ± 0.6
− 0.3 ± 0.9

0.4 ± 0.7
0.2 ± 0.5
− 0.2 ± 0.7

Clostridium cluster XVIII

0-week12‑week⊿ 0 ‑12 w

1.2 ± 0.7
1.0 ± 0.6
− 0.3 ± 0.6

1.6 ± 1.7
1.7 ± 1.7
0.1 ± 2.1

0-week12‑week

6.8 ± 2.4
6.3 ± 3.1

⊿ 0 ‑12 w− 0.5 ± 2.5

7.4 ± 4.2
6.6 ± 3.8
− 0.8 ± 3.3

Biﬁdobacterium

OthersTime point

Unit; ％, Values are expressed as the mean ± SD.
＊

p ＜ 0.05 against 0-week.

＃p ＜ 0.05 between-group diﬀerence in change from 0-week.

Calcium. In either case, since the diﬀerence was a minor
one, the principle investigator judged it clinically safe.
There was no significant difference in change between
two groups (data not shown).
3.6. Safety
No adverse events associated with the test product wereobserved in the course of the reporting.

4. DISCUSSION
We conducted a randomized, placebo-controlled, doubleblind study for examining the effect of food containing
Enterococcus faecalis (EC- BabyM®) and indigestible
dextrin (dietary fiber) on bowel movement. As theprimary outcome, the intergroup analysis showedsignificant differences in Frequency of bowel
movement , Days of defecation , and Biﬁdobacterium
share in the intestinal flora between the groups, after
12-weeks of ingestion. As the secondary outcome, it
proved that no abnormal change was triggered by theingestion of the test product.
Main Findings
This study examined the efficacy of ingesting foodcontaining Enterococcus faecalis (EC- BabyM®), aspecies of lactobacillus, and indigestible dextrin, a type ofwater soluble dietary ﬁber. After 12-weeks of ingestion,
DFN showed a significant difference in Frequency ofbowel movement , Days of defecation , andBiﬁdobacterium share in the intestinal ﬂora compared診療と新薬・第 55 巻

to Placebo.
The Japanese society of Gastroenterology developed anevidence-based clinical practice guideline for irritablebowel syndrome 11), which followed Roma Ⅲ 12) (currently
updated Rome Ⅳ 13) ). This guideline covers chronic
abdominal pain, abdominal discomfort, abnormal bowel
movement. In this guideline, diet therapy and behavioral
modification play important roles as step Ⅰ therapy.
These considerations should be highly referred to in theintestinal environment improvement study of Japanesewithin normal level of defecation or with a tendency
toward constipation.
The Food and Agricultural Organization of the United
Nations and the World Health Organization definedprobiotics as living microorganisms, which whenadministered in adequate amounts confer health beneﬁts
on the host 14) . Nobel laureate Elie Metchnikoff
linked the longevity of Bulgarians with consumption of
fermented milk products containing viable Lactobacilli,
and introduced the concept of probiotics to the scientiﬁccommunity. Since then, probiotics had been widely
consumed, mostly as functional foods or dietary
supplement. Recently, increasing evidences have showedthat not only viable Lactobacilli but also non-viable
Lactobacilli is suﬃcient to modify biological responses15).
Dietary nutrients may be converted into metabolites byintestinal microbes that serve as biologically active
molecules aﬀecting regulatory functions in the host. They
could improve the intestinal environment, namely,
manipulation of intestinal microbial communities,
suppression of pathogens, immunomodulation, stimulationof epithelial cell proliferation and differentiation andfortiﬁcation of the intestinal barrier 16) 17).
Enterococcus faecalis is a spherical bacteria that is partof the normal ﬂora of humans, and is commonly found in
the vaginal and intestinal regions of about 40 to 80％ of
individuals. EC-BabyM® is one of the non-viable
Enterococcus faecalis isolated from newborn Japanesebabies. The orally administered non-viable Enterococcus
faecalis cell wall preparations stimulate the gut immune
system18). Moreover EC-BabyM® is a kind of strain, forwhich the trade mark is registered, and recorded asEnterococcus faecalis EC-BabyM in NITE Patent
Microorganisms Depositary (NPMD), National Instituteof Technology and Evaluation (NITE).
In the contrast, the indigestible dextrin is a watersoluble dietary fiber taken out and prepared from theindigestible components in the resulting mush. Dextrin isconsidered a prebiotic, compounds that feed probiotics orgood bacteria19). It has been shown to aid lactobacilli, a
powerful beneficial bacteria responsible for supporting
digestion20). As the good bacteria continues to proliferate
throughout the digestive system, a balanced ratio of goodto bad bacteria is established.
Prebiotics are indigestible dietary ﬁbers that probiotic
bacteria use as a source of fuel to ﬂourish. The discovery

第 2 号（2018 年 2 月）

41 （121）of prebiotics and supporting theories is relatively recent
and the term of prebiotics was defined first in 1995 21).
Since then, numerous researches have shownthat adequate prebiotic intake has many potential
beneﬁts. Microecology of the gastrointestinal tract is thephysiologic basis for the effect of dietary fiber. The
ecology consists in the intestines, the foods that are fed
into the tract, and the flora living within. Within thisecology, normal ﬂora and ferment dietary ﬁber produce ashort chain fatty acids, which is very effective for thetreatment of gastrointestinal disorders 22).
In the present study it was observed that the share of
Biﬁdobacterium in the stool was improved, moreover theingestion of the DFN increased the stool frequency andthe number of days with bowel movement. Thisobservation suggests that Enterococcus faecalis (ECBabyM®) can make feeds for intestinal bacteria to reachthe intestine, followed by indigestible dextrin (dietaryﬁber) in DFN.
Secondary Findings
In this study we examined the safety of the test productby blood test. Although both groups showed significant
diﬀerences in the levels of glucose, natural fat, sodium,
chloride, and potassium after 12 weeks, the differenceswere minor and the principle investigator judged them as
the range of physiological variation (or clinically safe).
During the test period 22 subjects discontinued the test.
The reasons for discontinuance were personal
circumstances such a business trip. A subject waswithdrawn from analysis due to inadequacy of the recordsof bowel movement, and had nothing to do with theingestion of the test product.
Therefore, based upon the blood test and the diaries ofthe subjects, we observed no harmful influence against
biochemical and/or physiological matters of the subjects,
and this result indicated the safety of the ingestion of thetest product for 12 weeks of test period.
General Information
Roma Ⅲ 12) defined Functional Constipation Diagnostic
criteria as follows [Criteria fulﬁlled for the last 6 months
with symptom onset at least 6 months prior to diagnosis]; 1. Must include two or more of the following: a.
Straining during at least 25％ of defecations, b. Lumpy or
hard stools in at least 25％ of defecations, c. Sensation ofincomplete evacuation for at least 25％ of defecations, d.
Sensation of anorectal obstruction/blockage for at least
25％ of defecations, e. Manual maneuvers to facilitate atleast 25％ of defecations (e.g., digital evacuation, supportof the pelvic ﬂoor), f. Fewer than three defecations per
week; 2. Loose stools are rarely present without the useof laxatives; 3. Insufficient criteria for irritable bowel
syndrome. On the other hand, according to The JapaneseSociety of Internal Medicine, constipation is a casewhere the number of times of defecation is signiﬁcantly
reduced and/or exhibits diﬃculty in defecation . Decreasein stool frequency is defined as frequency of stool less

42 （122）

診療と新薬・第 55 巻

than 3 times per week, and diﬃculty in defecating means
straining during defecation , feeling of incomplete
evacuation , cooped-up feeling of anal (uncomfortablefeeling in perineum) . Many constipation patients mainlycomplain about difficulty in defecating rather thanfrequency of stool 23). Accordingly, the thought that bowel
movement is an essential part of QOL enhancement canbe supported. While Japanese tend to suffer from
irregular life in modern stress-sick society, it can bedifficult to change one s lifestyle. On the other hand,
habits such as taking supplements do not require them tochange their lifestyle substantially, and therefore they
are a much easier way for them to continue for a long
period. Moreover, DFN is a powder which might beingested soluble in drinks or foods, so we are able to take
DFN divided several times without any damage of thetaste. DFN is expected to be a convenient healthsupplement, and is easy to ingest by people with noconstipation depending on the effect of supporting goodbacteria.
Limitations
In this study we examined the effect of the foodcontaining Enterococcus faecalis (EC- BabyM®) andindigestible dextrin (dietary fiber) by evaluating
frequency of bowel movement, days of defecation,
volume of stool, fecal properties, and intestinal ﬂora. The
screening was performed to choose subjects with normal
level of defecation or with a tendency toward
constipation. Meanwhile the principle investigator
excluded the subjects with chronic constipation by aninterview, because subjects were not able to be diagnosed
with constipation judging only by the number ofdefecations. It is thought that many subjects with normal
level of defecation were included by reason that the
number of defecation for a week (at baseline) of all
subjects (n 27) was 9.1±4.3 times. Moreover, it isassumed that at the baseline many subjects were normalin fecal properties such as scent of stool, color of stool,and texture of stool. Due to these conditions, the eﬀect of
DFN observed in this study should be applied not only for
subjects with a constipation tendency but also subjects
with normal level of defecation. In addition, regardingdays of defecation, a significant difference between two
groups was found at the baseline. Because of this, theeﬀect of DFN for days of defecation would have limited.

5. CONCLUSION
In conclusion, we found out that the ingestion of Dietaryﬁber Nyuusankin 5,000 oku-ko containing Enterococcus
faecalis (EC- BabyM®) and indigestible dextrin (dietaryfiber) for 12 weeks improved bowel movement. In
addition, no safety- related matter occurred during thetest period.

第 2 号（2018 年 2 月）
CONFLICT OF INTEREST
All parts of this study were funded by M. Foods Corporation. Kimitoshi
Yamaguchi is the CEO. All authors state that the study was conducted in
the absence of any other relationships that could be interpreted as a
conﬂict of interest.

REFERENCES
1) Ministry of Health,Labour and Welfare. Ministerial Notiﬁcation No.
430 of the Ministry of Health, Labour and Welfare. 2012 Jul.
http://www.mhlw.go.jp/toukei/saikin/hw/k-tyosa/k-tyosa16/index.
html (referred 2017-12-6)
2) Williams EA, Stimpson J, et al. Clinical trial: a multistrain probiotic
preparation significantly reduces symptoms of irritable bowel
syndrome in a double-blind placebo-controlled study. Aliment
Pharmacol Ther. 2009; 29: 97-103.
3) Enck P, Zimmermann K, et al. A mixture of Escherichia coli (DSM
17252) and Enterococcus faecalis (DSM 16440) for treatment of theirritable bowel syndrome--a randomized controlled trial with primarycare physicians.Neurogastroenterol Motil. 2008; 20:1103-9.
4) OʼMahony L, McCarthy J,et al. Lactobacillus and biﬁdobacterium in
irritable bowel syndrome: symptom responses and relationship tocytokine proﬁles. Gastroenterology. 2005; 128: 541-551.
5) Adams CA. The probiotic paradox: live and dead cells are biologicalresponse modiﬁers. Nutr Res Rev. 2010; 23: 37-46.
6) Lewis SJ, Heaton KW. Stool form scale as a useful guide to intestinal
transit time. Scand J Gastroenterol. 1997; 32: 920-4.
7) Nishino H. Dietary ﬁber and disease (colon diseases) (in Japanese).
Dietary ﬁber. The Japan Dietetic Association. 1995. Tokyo, DAI-ICHISHUPPAN Co.Ltd. 314-327.
8) Matsumoto M, Imai T, et al. Effect of Yogurt with Bifidobacterium
lactis LKM 512 in Improving Fecal Microflora and Defecation of
Healthy Volunteers. Journal of Intestinal Microbiology. 2001; 14: 97102.
9) Nagashima K, Hisada T, et al. Application of new primer-enzyme
combinations to terminal restriction fragment length polymorphism
profiling of bacterial populations in human faces. Appl Environ
Microbiol. 2003; 69: 1251-63.
10) Nagashima K, Mochizuki J, et al. Phylogenetic analysis of 16S
ribosomal RNA gene sequences from human fecal microbiota andimproved utility of terminal restriction fragment length polymorphism
proﬁling. Biosci Microﬂora. 2006; 25: 99-107.
11) Fukudo S, Kaneko H, et al. Evidence-based clinical practice guidelines
for irritable bowel syndrome. J Gastroenterol. 2015; 50: 11-30
12) Drossman DA. The functional gastrointestinal disorders and the Rome
Ⅲ process. Gastroeneterology. 2006; 130: 1377-90.
13) Schmulson MJ, Drossman DA. What Is New in Rome Ⅳ . J
Neurogastroenterol Motil. 2017; 23: 151-163.
14) Food and Agriculture Organizationof the United Nations, World
Health Organization. Guidelines for the Evaluation of Probiotics inFood . Report of a joint FAO/WHO working group on drafting
guidelines for the Evaluation of Probiotics in Food. London Ontario,
Canada. 2002: April 30 and May 1.
15) Mehling H, Busjahn A. Non-viable Lactobacillus reuteri DSMZ 17648(Pylopass™) as a new approach to Helicobacter pylori control inhumans. Nutrients. 2013; 5: 3062-73.
16) Thomas CM, Versalovic J. Probiotics-host communication: Modulationof signaling pathways in the intestine. Gut Microbes. 2010; 1: 148163.
17) Peera Hemarajata, James Versalovic. Effects of probiotics on gut
microbiota: mechanisms of intestinal immunomodulation andneuromodulation. Therap Adv Gastroenterol. 2013; 6: 39-51.
18) Sakai Y, Tsukahara T, et al. Cell preparation of Enterococcus faecalis
strain EC-12 prevents vancomycin- resistant enterococci colonizationin the cecum of newly hatched chicks. Poult Sci. 2006; 85: 273-277.
19) Lefranc-Millot C, Guérin-Deremaux L, et al. Impact of a resistantdextrin on intestinal ecology: how altering the digestive ecosystemwith NUTRIOSE®, a soluble ﬁbre with prebiotic properties, may be
beneﬁcial for health. J Int Med Res. 2012; 40:211-224.

診療と新薬・第 55 巻
20) Slizewska K. The citric acid-modiﬁed, enzyme-resistant dextrin frompotato starch as a potential prebiotic. Acta Biochim Pol. 2013; 60:
671-675.
21) Gibson GR, Roberfroid MB. Dietary modulation of the human colonicmicrobiota: introducing the concept of prebiotics. J Nutr. 1995; 125:
1401-12.

第 2 号（2018 年 2 月）

43 （123）

22) Park J, Floch MH. Prebiotics, probiotics, and dietary fiber ingastrointestinal disease. Gastroenterol Clin North Am. 2007; 36: 4763.
23) Nakajima A. Diagnosis and Treatment for Chronic Constipation ; A
Session 2. The Journal of the Japanese Society of Internal Medicine.
2016; 105: 429-433.