CN102959075B - There is the bifidus bacillus of the minimizing effect of corrupt substance in intestines - Google Patents

There is the bifidus bacillus of the minimizing effect of corrupt substance in intestines Download PDF

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CN102959075B
CN102959075B CN201180033393.0A CN201180033393A CN102959075B CN 102959075 B CN102959075 B CN 102959075B CN 201180033393 A CN201180033393 A CN 201180033393A CN 102959075 B CN102959075 B CN 102959075B
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intestines
corrupt substance
bifidobacterium breve
minimizing
fos
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CN102959075A (en
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天藤公子
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Meiji Co Ltd
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Meiji Co Ltd
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Abstract

1st problem of the present invention is that the useful microorganism effectively utilizing security high effectively reduces corrupt substance in intestines.Present inventor is conceived to usual in vivo (in intestines) the dominant bifidus bacillus people and screens, to the minimizing effect of indoles, the strong and bacterial strain confirming the minimizing effect of the indoxyl sulfate in serum is selected, thus completes the present application.

Description

There is the bifidus bacillus of the minimizing effect of corrupt substance in intestines
Technical field
The present application relates to the ameliorative way of the state of intestinal bacterium.More specifically, relate to the useful microorganism that reduces the indoles based compound as corrupt smell reason that intestinal bacterium causes select and this selects the microorganism of gained.
Background technology
Due to irregular, the pressure etc. of the confusion of dietetic life, life, there is constipation repeatedly, diarrhoea, or the people of the symptom such as abdomen rises, aerogenesis is on the increase.These symptoms are considered to affected by intestines environment, and in other words, its reason is that in intestinal flora (group), spoilage organism becomes advantage.Further, intestinal flora under anaerobic produces the corrupt odour components such as indoles or derivatives thereof by tryptophane.Large quantity research (non-patent literature 1) has been carried out to the relation of the generation of this intestinal flora and spoilage product.
In addition, think that indoles is produced by tryptophane by spoilage organism in intestines and by after intestinal absorption, is metabolised to indolol in liver, and then be combined with sulfuric acid, form indoxyl sulfate.Think for the excretion of Healthy People indoxyl sulfate in urine, but dialysis patients etc. is then become to the cause of disease (non-patent literature 2) of the complication such as uremia.
For the removing of the indoles as intestinal toxic material etc., such as, bifidobacterium thermophilum's (PVC Off ィ De バ Network テ イ Le system/サ ー モ Off ィ ラ system) with indoles minimizing ability is used in patent documentation 1.In addition, describe in patent documentation 2 and the extract of Salacia prinoides (Willd.) DC. platymiscium or micro mist minced with milk-acid bacteria and/or bifidus bacillus and is used for improving intestines environment.And then, describe in patent documentation 3 and utilize the composition containing bacterium acidi propionici to promote the growth of the bifidus bacillus in enteron aisle, prevent from releasing foul gas.In addition, there was reported synanthrin, oligose contributes to improving intestinal flora (non-patent literature 3).
Prior art document
Patent documentation
Patent documentation 1: Japanese Unexamined Patent Publication 2006-158216
Patent documentation 2: Japanese Unexamined Patent Publication 2007-031345
Patent documentation 3: Japanese Unexamined Patent Publication 2007-186529
Non-patent literature
Non-patent literature 1: intestinal bacterium learns magazine the 19th volume 169-177 page (2005)
Non-patent literature 2: dialysis can will 24 volume No. 3 312-316 pages (1991)
Non-patent literature 3:TheJournalofNutrition129 (7Suppl): 1438S-1441S (1999).
Summary of the invention
The technical problem that invention will solve
The corrupt substance that in vivo (intestines in) of known person generate can promote that cancer occurs, aging, intravital corrupt substance can be reduced energetically as long as think, the health of people can be contributed to.Therefore first, the problem in the present application is, the useful microorganism effectively utilizing security high reduces corrupt substance in intestines effectively.
When above-mentioned corrupt substance mainly generates in large intestine, as an example of the superior microorganism (bacterial classification) in large intestine, there are milk-acid bacteria, bifidus bacillus (genus bifidobacterium).This bifidus bacillus is used to food as the microorganism playing useful function in vivo since ancient times, has the real result of diet experience.Therefore, the 1st problem of the application is, selects the microorganism (bacterial strain) that effectively can reduce corrupt substance by vivo (in intestines) dominant bifidus bacillus of people.
But, mainly utilized external system to find minimizing in vivo (intestines in) material of corrupt substance of generating or microorganism in the past.Due to intestinal flora can't be reproduced in vitro at present, thus to whether reflecting situation in actual intestines and carry out experiment to have query.Therefore, the 2nd problem of the present application is, the checking of provide the system of suitable in vitro and in vivo to be used as the reducing material of corrupt substance that in vivo (intestines in) generate or the action effect of microorganism or searching system.
And then the 3rd problem of the present application is, provides, to dialysis patients etc., there is the microorganism reduced as the effect of the concentration of indoxyl sulfate in serum of the complication cause of disease.
For the means of technical solution problem
As the useful microorganism that security is high, present inventor, based on the screening method of the present application, is separated the bacterial strain effectively reducing corrupt substance in intestines, thus completes the present application from milk-acid bacteria and bifidus bacillus.
As the checking system reducing the material of corrupt substance that in vivo (intestines in) generate or the action effect of microorganism, present inventor have studied following method: use damping fluid reproduction thalline to be difficult to the condition of breeding when screening in vitro, evaluate the minimizing effect (minimizing ability) of corrupt substance.And, present inventor establishes external selection method and/or the evaluation method of the microorganism (milk-acid bacteria, bifidus bacillus etc.) reducing the corrupt substance that intestinal bacterium produces, the method is characterized in that, be milk-acid bacteria by reaction soln setting (design), the bifidus bacillus condition of substantially not breeding, and the glucose amount of the energy of minimum needed for minimizing corrupt substance.
In addition, as the checking system reducing the material of corrupt substance that in vivo (intestines in) generate or the action effect of microorganism, present inventor uses mouse when have studied and screening in vivo, measures indoxyl sulfate by serum.And, selection method and/or evaluation method in the body that present inventor establishes the microorganism (milk-acid bacteria, bifidus bacillus etc.) of the generation of the indoxyl sulfate reduced in serum, the method is characterized in that, oligofructose (FOS) and/or oligomeric galactose (GOS) and tryptophane (L-Trp) are added in feed (powder feed) and raise mouse, evaluate the concentration of the indoxyl sulfate in serum.
And then, in order to select the microorganism (bacterial strain) that effectively can reduce corrupt substance, first, present inventor is conceived to usually in vivo (in intestines) dominant bifidus bacillus of people, screening is implemented to total 196 bacterial strain of 32 bacterial strains of bifidobacterium breve (Bifidobacteriumbreve), 92 bacterial strains of bifidus longum bb (Bifidobacteriumlongum), 72 bacterial strains of bifidobacterium adolescentis (Bifidobacteriumadolescenti), evaluate in vitro, pick out 9 strains in advance as the high useful microorganism of security.Then, in order to verify reduce corrupt substance in vivo effect, effect, experimentation on animals is implemented for above-mentioned 9 strains selected, carry out interior evaluating, pick out indoles minimizing effect strong and confirm the bifidobacterium breve OLB6129 of the minimizing effect of the indoxyl sulfate in serum, thus complete the present application.
In addition, in the present application, comprise medicine and food, the bifidus bacillus of the minimizing effect of the indoxyl sulfate wherein in, display serum strong containing the above-mentioned indoles minimizing effect selected is as effective constituent.
This specification sheets comprises the content as recording in the specification sheets of No. 2010-152962, the Japanese patent application on the application's preference basis and/or accompanying drawing.
Invention effect
The bifidobacterium breve (B.breve) of the present application is the bacterial strain deriving from people, in the system that this bacterial strain self is not bred, achieves the excellent effect significantly reducing the indoxyl sulfate in indoles and serum.In addition, high by the security of the B.breve eaten for a long time, bifidus bacillus OLB6129 of the present invention also extensively can be suitable for or application at field of food, medicine field.
Accompanying drawing explanation
[Fig. 1] gives the impact of the concentration on the indoxyl sulfate in serum that bifidobacterium breve (viable bacteria) causes to mouse
[Fig. 2] gives the impact of the concentration on the indoxyl sulfate in serum that bifidus longum bb (viable bacteria) causes to mouse
[Fig. 3] gives B.breveOLB6129(viable bacteria to Mouse oral) in, B.breveOLB6129(viable bacteria) the impact of the concentration on the indoxyl sulfate in serum that causes of concentration
[Fig. 4] detect in TOS propionic acid nutrient agar just in the colony number of B.breveOLB6129 in (Fecal) and cecal content (cecal)
[Fig. 5] gives B.breveOLB6129(viable bacteria to Mouse oral) in, with or without the impact of the concentration on the indoxyl sulfate in serum added FOS and cause
[Fig. 6] detect in TOS propionic acid nutrient agar just in B.breveOLB6129 in (Fecal) colony number with or without the relation of adding FOS.
Embodiment
1. first
The corrupt substance that in vivo (intestines in) of known person generate can promote that cancer occurs, aging, intravital corrupt substance can be reduced energetically as long as think, the health of people can be contributed to.As the main corrupt substance generated in large intestine, phenol, p-cresol, indoles, skatole, ammonia etc. can be enumerated.Among these corrupt substances, particularly indoles, skatole etc. are odor pollutants, still likely bring dysgenic composition to health in addition, thus expect to reduce indoles, skatole.
As an example of the superior microorganism (bacterial classification) in large intestine, there are lactobacillus, bifidus bacillus (genus bifidobacterium).Bifidus bacillus is used to food as the microorganism playing useful function in vivo since ancient times.In addition, bifidus bacillus is considered to have whole intestines effect, is thus also used to medicines for relieving intestinal disorders etc., also contributes to the improvement of intestinal flora, thinks by improving intestines environment, to suppression diarrhoea, prevent pachylosis, improve pollinosis, the Taoism of immunizing power etc. is also effective.
2. for verifying, finding the appraisement system of the action effect of the microorganism reducing corrupt substance in intestines
2-1. reduces in-vitro screening or the prescreen of the microorganism of corrupt substance in intestines
In order to use microorganism, reduce corrupt substance in intestines (generate in intestines and produce the material of odor pollutant etc.) under intestines environment, importantly this specific microorganism is preferentially to exist in intestines.That is, as the character that this microorganism should possess, the patience for the condition adapting to intestines environment is required.
Therefore, it is desirable to be suitable for screening method, the evaluation method of the state (practical situation) meeting the microorganism (thalline) existed in intestines environment.Therefore, when screening reduces material or the microorganism of corrupt substance in intestines in vitro, the condition that first present inventor uses damping fluid to be difficult to breed to reproduce microorganism, have studied the evaluation method of the minimizing effect (minimizing ability) of corrupt substance in intestines.By adopting above-mentioned condition, do not breed even if microorganism can be selected, that also can remove corrupt substance, excellent under the condition closer to intestines environment microorganism.Such as, to set condition that object microorganism do not breed substantially and to reduce the such condition of the energy of the minimum needed for corrupt substance, the composition of reaction soln, concentration can be decided.
More specifically, for the use level of the medium component as energy source, such as, make the common culture condition of the concentration ratio object microorganism of glucose, lactose, fructose etc. low, and as removing the degree of corrupt substance in intestines, applicable 0.3 ~ 0.7(w/v) %.
Should illustrate, in screening (evaluation is selected) method (in vitro tests), the result that the concentration exemplified with the reaction soln of the carbon source to glucose and so on is studied, even but other composition, also can determine composition, the concentration of reaction soln equally.
2-2. reduces the interior screening of body of the microorganism of corrupt substance in intestines
In the past, failed to implement experimentation on animals by the minimizing effect fully confirming the indoxyl sulfate in serum in body.The minimizing effect of the indoxyl sulfate in serum, although expect to play in intestines, even if but microorganism (thalline) is given in vivo, before also thinking in arrival intestines its major part likely dead or in arrival intestines after its likely can not stable maintenance (maintenance).Therefore, in the present application, microorganism is being given to after in vivo, when expecting to contribute to the raising of its viability, stability etc. and try oligofructose (FOS) to be added in feed, the minimizing effect of the indoxyl sulfate in serum can confirmed.
In addition, in experimentation on animals, usual used mouse is the healthy individuals in children's age, and thus intravital corrupt substance can be suppressed to minute quantity, and most cases can lower than detectability.Therefore, in the present application, in the experimentation on animals using mouse, tryptophane (L-Trp) in vivo as benzazole precursor is added in feed (powder feed), during to attempt the baseline improving the indoles generated in vivo, for the baseline of indoles, confirm significance rising.That is, become the concentration can stablized as the indoxyl sulfate in the serum of the index (mark) of corrupt substance and evaluate definitely.Further, for specific bacterial strain, the minimizing effect can evaluating, confirm corrupt substance with the form of the significant difference compared with control group is become.
More specifically, oligofructose (FOS) and/or oligomeric galactose (GOS) and tryptophane (L-Trp) can be added in feed (powder feed) with the concentration specified and feed common mouse (experimentation on animals with), evaluate the concentration of the indoxyl sulfate in serum.Now, as the concentration of the total amount of oligofructose (FOS) and/or oligomeric galactose (GOS), be set as 0.5 ~ 20(w/v) %, be preferably set to 0.5 ~ 15(w/v) %, and be more preferably set as 0.5 ~ 10(w/v) %.Should illustrate, concrete experimental result is omitted, but is set as 0.5(w/v as the concentration of oligofructose (FOS)) % time, as the viable count in ight soil, the viability having confirmed B.breveOLB6129 significantly improves.In addition, as the concentration of tryptophane, be set as 1 ~ 4(w/v) %, be preferably set to 1.5 ~ 3.5(w/v) %, and be more preferably set as 2 ~ 3(w/v) %.Thus, can select in vivo and/or evaluate or confirm microorganism (milk-acid bacteria, bifidus bacillus etc.), particularly bifidus bacillus (microorganism of genus bifidobacterium) that the generation of the indoxyl sulfate made in serum reduces.
3. make the microorganism that in intestines, corrupt substance reduces
As making the microorganism that in intestines, corrupt substance reduces, in order to select the microorganism (bacterial strain) that effectively can reduce corrupt substance, present inventor is that object is studied with usual in vivo (in intestines) dominant milk-acid bacteria people, bifidus bacillus, particularly bifidus bacillus.Further, as bifidus bacillus, total 196 bacterial strain for 32 bacterial strains of B.breve, 92 bacterial strains of B.longum, 72 bacterial strains of B.adolescentis implements screening, evaluates in vitro.As its result, pick out total 9 bacterial strain of 6 bacterial strains of B.breve and 3 bacterial strains of B.longum.
And then, in order to verify reduce corrupt substance in vivo effect, effect, to above-mentioned select 9 bacterial strains implement experimentation on animalies, evaluate in vivo.As its result, pick out the bacterial strain playing the minimizing effect of the indoles in intestines.
Should illustrate, screen the bifidobacterium breve OLB6129(B.breveOLB6129 that obtains) be preserved in Independent Administrative Leged Industrial Technology Complex Inst on May 26th, 2010 and specially permit biological sustenance center (contact address: east, つ く ば city, postcode 305-8566 Hitachinaka County, Japan 1-1-1 つ く ば center central the 6th), this microorganism carries out preservation with deposit number FERM BP-11257.
4. the food containing the microorganism of corrupt substance in minimizing intestines or medicine
4-1. contains the food of the microorganism reducing corrupt substance in intestines
In the minimizing intestines of the present application, the microorganism (milk-acid bacteria, bifidus bacillus etc. are preferably B.breveOLB6129) of corrupt substance can be added in food.Namely, as the food of the present application, any food being added with the microorganism reducing corrupt substance in intestines can be enumerated, preferably can enumerate the cultured milk prod or leben that are added with the microorganism reducing corrupt substance in intestines, preferably can enumerate further to be added with and reduce in intestines the microorganism (B.breveOLB6129) of corrupt substance and the cultured milk prod of oligofructose (FOS) and/or oligomeric galactose (GOS) or leben.Should illustrate, known GOS can carry out utilizing (http://www.nyusankin.or.jp/scientific/hosono_3.html) identically with FOS.In addition, for the addition of oligofructose (FOS) and/or oligomeric galactose (GOS), with the densitometer of total amount, can 0.5 ~ 20(w/v be set as) %, be preferably set to 0.5 ~ 15(w/v) %, and be more preferably set as 0.5 ~ 10(w/v) %.
In addition, in minimizing intestines containing the present application, the food of the microorganism (milk-acid bacteria, bifidus bacillus etc., be preferably B.breveOLB6129) of corrupt substance can being suitable for for the health-care food (functional foodstuff) prevented or improve dialysis complication as renal dialysis patients.
4-2.
In minimizing intestines containing the present application, the medicine of the microorganism (milk-acid bacteria, bifidus bacillus etc., be preferably B.breveOLB6129) of corrupt substance can be suitable for as the preventive of the dialysis complication of the minimizing agent of corrupt substance in intestines or renal dialysis patients or activator.And then the medicine of the present application also can be used as the activator that abdomen rises and is suitable for.
The medicine of the present application can be made as the suitable formulation such as powder, granule, tablet, capsule, solution.Oligofructose (FOS) and/or oligomeric galactose (GOS) can be contained in the medicine of the present application.Should illustrate, with the densitometer of total amount, the content of oligofructose (FOS) and/or oligomeric galactose (GOS) can be set as 0.5 ~ 20(w/v) %, be preferably set to 0.5 ~ 15(w/v) %, and be more preferably set as 0.5 ~ 10(w/v) %.In formulation, vehicle, tackiness agent, disintegrating agent, lubricant etc. can be added in order to formulation and conventional auxiliary.As vehicle, such as, there are starch, lactose, white sugar, methylcellulose gum, carboxymethyl cellulose, sodiun alginate, secondary calcium phosphate, synthetic aluminium silicate, Microcrystalline Cellulose, Polyvinylpyrolidone (PVP) (PVP), hydroxypropylated starch (HPS) etc.In addition, as tackiness agent, such as, there is the aqueous solution or their hydrous ethanol solution etc. of starch, Microcrystalline Cellulose, Xylo-Mucine, Polyvinylpyrolidone (PVP) (PVP), gum arabic powder, gelatin, glucose, white sugar etc.Further, as disintegrating agent, such as, starch, carboxymethyl cellulose, calcium carboxymethylcellulose, Microcrystalline Cellulose, hydroxypropylated starch, calcium phosphate etc. are had.And then, as lubricant, such as, there are carnauba wax, light anhydrous silicic acid, synthetic aluminium silicate, natural aluminium silicate, synthetic magnesium silicate, winterized stearin, hydrogenated vegetable oil derivative (SterotexHM), sesame oil, beeswax, titanium oxide, Aluminium Hydroxide stearic acid, calcium stearate, Magnesium Stearate, talcum, secondary calcium phosphate and sodium lauryl sulphate etc.
The medicine of the present application it is desirable to oral giving, for bifidus bacillus, because security is determined, thus arbitrary amount can be given, such as, its administered dose can be set to every 1 day and give bifidus bacillus 1,000,000,000 ~ 5,000 hundred million, is preferably 10,000,000,000 ~ 5,000 hundred million.
Embodiment
［ reference example 1 ］ has the condition setting of the selection method " in vitro tests " of the microorganism of the minimizing effect of corrupt substance
First, for the composition of reaction soln, substantially do not breed, play the minimizing effect of the indoles index of corrupt substance (in the intestines) with bifidus bacillus and the mode can evaluating this effect clearly decides the composition of reaction soln.
In the reaction soln used in this experiment, such as, glucose is set as 1.0(w/v) % time, bifidus bacillus can excessive proliferation, fully can not reproduce the environment in intestines.Namely, because the microorganisms such as bifidus bacillus (thalline) substantially do not breed in intestines, thus in order to evaluate the minimizing effect of the corrupt substance that bifidus bacillus etc. realizes under the state close to intestines environment, the condition that bifidus bacillus does not breed substantially should be set, to reproduce above-mentioned environment.Therefore, glucose is set as such as 0.7(w/v) below %, be preferably set to 0.6(w/v) below %, and be more preferably set as 0.55(w/v) below %.
In addition, for the reaction soln used in this experiment, bifidus bacillus should be set and substantially do not breed and condition that the minimum energy needed for the minimizing of corrupt substance is provided, thus think that it is inappropriate for originally being coordinated as substrate by medium component.
On the other hand, for the reaction soln used in this experiment, such as, glucose is set as 0.1(w/v) % time, the activity (activity) of bifidus bacillus reduces, and is difficult to the minimizing effect of performance indoles.Like this, although the minimizing effect etc. of each bacterial classification to indoles of bifidus bacillus cashes out difference originally, this difference becomes and is difficult to embody, and is difficult to evaluate its effect clearly.Therefore, glucose is set as, such as, 0.3(w/v) more than %, preferred 0.4(w/v) more than %, more preferably 0.45(w/v) more than %.In addition, other composition is also determined in the same manner to the composition of reaction soln.
As the composition of reaction soln, can according to each 20 μ g of phenol p-cresol indoles skatole, Bis-Tris0.1M, magnesium sulfate (MgSO 4) 0.75mM, glucose 0.5(w/v) % uses, pH is about 6.5.
［ embodiment 1 ］ has selecting " in vitro tests " of the microorganism of the minimizing effect of corrupt substance
For total 199 bacterial strain of 34 bacterial strains of B.breve, 93 bacterial strains of B.longum, 72 bacterial strains of B.adolescentis, when carrying out quiescent culture, in 2 bacterial strains of B.breve, 1 bacterial strain of B.longum, it is extremely slow to grow.Therefore, using above-mentioned 3 bacterial strains as experimental subjects, total 196 bacterial strain finally for 32 bacterial strains of B.breve, 92 bacterial strains of B.longum, 72 bacterial strains of B.adolescentis has carried out screening (evaluation is selected).
Above-mentioned bacterial strains (viable bacteria) each 1mg is suspended in the reaction soln (pH:6.5, the phenol p-cresol indoles skatole: each 20 μ g, Bis-Tris:0.1M, magnesium sulfate (MgSO that determine in reference example 1 4): 0.75mM, glucose: 0.5(w/v) %) in 2ml.Further, by this suspension liquid (reaction soln) after 37 DEG C of standing 24h react, centrifugation, removing bifidus bacillus (thalline), is adjusted to 7 by the pH of its supernatant.
Then, this supernatant diethyl ether is divided 2 extractions, after this extracting solution is complemented to 5ml, by the concentration of Gas chromatography (GC) to this measured in solution (detection) indoles, phenol, p-cresol, skatole.
Should illustrate, the analysis condition of Gas chromatography (GC) is as described below.Namely, be equipped with the detector of flame ionization detector: GC-2014 (Shimadzu), post (size): TC-1 (GLSciencesInc.), I.D.=0.25mm, L=30m, dF=0.25mm, column temperature: 60-170 DEG C (raising with 5 DEG C/min), 170-250 DEG C (raising with 20 DEG C/min), carrier gas: nitrogen, flow velocity: 71.3ml/min, injector temperature: 240 DEG C, detector temperature: 250 DEG C, volume injected: 1 μ l.
The results are shown in table 1(B.breve 32 bacterial strains), table 2(B.longum 92 bacterial strains), table 3(B.adolescentis 72 bacterial strains).
Should illustrate, the bacterial strain that arrangement numbering starts with " JCM " is preserved in following preservation mechanism.
Preservation mechanism name: Physical Chemistry Inst バ イ オ リ ソ ー ス セ Application タ ー microbial material exploitation room (contact address: He Guang city, postcode 351-0198 Saitama County, Japan Wide swamp 2-1, telephone number 048-467-9560).
[table 1]
[table 2]
[table 3]
In the above results, select the bacterial strain that the minimizing concentration of the indoles relative to microorganism (thalline) 1g is more than 13mg/g-cell, what result met is 6 bacterial strains of B.breve, total 9 bacterial strain with 3 bacterial strains of B.longum.Should illustrate, the minimizing concentration of indoles is that more than 13mg/g-cell refers to that the decrement of indoles is more than 65%.
［ embodiment 2 ］ has selecting " in vivo test (experimentation on animals) " of the microorganism of the minimizing effect of corrupt substance
9 bacterial strains selected in above-described embodiment 1 are undertaken screening (evaluation is selected) by experimentation on animals (in vivo test).
After buying Balb/c mouse, make it freely to absorb feed (powder CRF-1/FOS(and the pure pharmaceutical worker's industry (strain) of light): 10(w/v) %, L-Trp: 2.5(w/v) %) 1 week, tame.After terminating between this domestication, the bacterial strain (viable bacteria) of regulation is suspended in physiological saline, gives 2 weeks according to the frequency of 1 day 1 time continuously at morning.Now, the capacity that gives of physiological saline, the capacity that gives of each bacterial strain, the concentration that gives of each bacterial strain are respectively 10 10cfu/ml, 0.2ml/ pcs/day, 2x10 9cfu/ pcs/day.Carry out adopting whole blood at the end of this gives period (2 weeks), measure the concentration of the indoxyl sulfate in serum.
Should illustrate, the bacterial strain (viable bacteria) giving mouse selects (screening) in testing in vitro.Particularly, pick 6 bacterial strains (MEP22023004, OLB6129, MEP22023007, MEP22023012, MEP22023015, MEP22023016) of B.breve, with 3 bacterial strains (MEP22023039, MEP22023061, MEP22023078) of B.longum.Further, the physiological saline of the capacity such as use as a control group, compares with the minimizing effect of group to corrupt substance that give of regulation bacterial strain (viable bacteria).The results are shown in Fig. 1 (6 bacterial strains of B.breve), Fig. 2 (3 bacterial strains of B.longum).
In the above results, by 6 bacterial strains (Fig. 1) of B.breve, give mouse with 3 bacterial strains (Fig. 2) of B.longum with viable bacteria, the concentration of the indoxyl sulfate in the serum of mouse is measured at the end of this gives period, result gives in group at B.breveOLB6129, confirms the remarkable minimizing compared with control group of its concentration.That is, for B.breveOLB6129(FERM BP-11257) give group, compared with control group, in the concentration of the indoxyl sulfate in serum, confirmed significant minimizing.By this result, imply that B.breveOLB6129 can reduce corrupt substance in vivo.
［ embodiment 3 ］ is on the impact of the concentration of the B.breveOLB6129 that live body gives on the minimizing effect of corrupt substance
［ experimental procedure ］
After Balb/c mouse (SLC, ♀, 6 week age) is tamed one week, as shown in table 4, being divided into 4 groups based on body weight, to each group, the B.breveOLB6129(viable bacteria by physiological saline and physiological saline) suspension liquid orally gives 2 weeks at morning continuously.Domestication chien shih is all organized and is freely absorbed feed (powder CRF-1/FOS(and the pure pharmaceutical worker's industry (strain) of light): 10(w/v) %, L-Trp: 2.5(w/v) %).
[table 4]
(1) B.breveOLB6129 give 2 weeks of period after, take a blood sample by Wei Jing Veins, the concentration of indoxyl sulfate in mensuration serum.After serum ultrapure water (Milli-Q) is diluted to 100 times, filter, according to existing report (Takayamaetal., AmJKidneyDis.2003Mar with SYRINGELESSFILTERDEVICE0.2 μm (Whatman); 41(3Suppl1): S142-5), measured by high performance liquid chromatography (HPLC).
(2) B.breveOLB6129 give period at the end of gather just, gather cecal content at when dissected, measure separately contained B.breveOLB6129(administration bacterium) bacterium number.
［ experimental result ］
B.breveOLB6129 give period at the end of, confirm the concentration of indoxyl sulfate in serum, for the minimizing effect of the concentration of indoxyl sulfate, 2x10 8cfu/ pcs/day give group (III) and 2x10 9cfu/ pcs/day give in group (IV), with physiological saline give compared with group (contrast) (I), there is significant difference, but 2x10 7cfu/ pcs/day give group (II) cfu/ only compared with the giving of physiological saline group (contrast) (I), not there is significant difference (Fig. 3).
In addition, just in the B.breveOLB6129(administration bacterium in cecal content) detect (Fig. 4) in the mode of consumption interdependence.
［ embodiment 4 ］ gives the presence or absence of FOS to the impact of the minimizing effect of corrupt substance on live body
［ experimental procedure ］
By Balb/c mouse (SLC, ♀, 6 week age) tame one week after, as shown in table 5,4 groups are divided into based on body weight, for each group, the B.breveOLB6129(viable bacteria by the physiological saline and physiological saline that do not add FOS) the B.breveOLB6129(viable bacteria of suspension liquid, the physiological saline that with the addition of FOS and physiological saline) suspension liquid orally give 2 weeks at morning continuously.That all group freely absorbs feed (powder CRF-1/L-tryptophane: 2.5(w/v) % between domestication).Should illustrate, for B.breveOLB6129(viable bacteria) concentration, be set as in embodiment 3 about corrupt substance minimizing effect display necessity minimum consumption namely, 2x10 8cfu/ pcs/day.
[table 5]
(1) FOS give 2 weeks of period after, take a blood sample by Wei Jing Veins, the concentration of indoxyl sulfate in mensuration serum.After serum ultrapure water (Milli-Q) is diluted to 100 times, filter, according to existing report (Takayamaetal., AmJKidneyDis.2003Mar with SYRINGELESSFILTERDEVICE0.2 μm (Whatman); 41(3Suppl1): S142-5), measured by high performance liquid chromatography (HPLC).
(2) FOS give period at the end of, gather B.breveOLB6129 give group just, measure by intravital B.breveOLB6129(administration bacterium) bacterium number.
［ experimental result ］
FOS give period at the end of, confirm the concentration of indoxyl sulfate in serum, only B.breveOLB6129 give in group, the minimizing effect of the concentration of indoxyl sulfate is little (Fig. 5).In addition, for the minimizing effect of the concentration of indoxyl sulfate, give group for only FOS, with physiological saline give compared with group (I), there is no significant difference (Fig. 5).On the other hand, for the minimizing effect of the concentration of indoxyl sulfate, give in group at B.breveOLB6129 and FOS, with physiological saline give compared with group (I) etc., there is significant difference (Fig. 5).Should illustrate, for 2 groups (OLB6129+) giving to organize that give group and B.breveOLB6129 and FOS of only B.breveOLB6129, measure the B.breveOLB6129(administration bacterium just) bacterium number, result FOS give observe viable count in group and significantly increase (Fig. 6).
[deposit number]
FERM　BP-11257
The whole distribution publications quoted in this specification sheets, patent and patent application are all introduced in this specification sheets as a reference.

Claims (17)

1. bifidobacterium breve ( bifidobacteriumbreve) OLB6129, its deposit number is FERM BP-11257, and it makes corrupt substance in intestines reduce.
2. the minimizing agent of corrupt substance in intestines, its with bifidobacterium breve according to claim 1 for effective constituent.
3. deposit number is the purposes of bifidobacterium breve OLB6129 in the minimizing agent manufacturing corrupt substance in intestines of FERM BP-11257.
4. deposit number is the purposes of bifidobacterium breve OLB6129 in the preventive or activator of the dialysis complication of manufacture renal dialysis patients of FERM BP-11257.
5. deposit number be FERM BP-11257 bifidobacterium breve OLB6129 manufacture renal dialysis patients for prevent or improve dialysis complication health-care food in purposes.
6. deposit number be FERM BP-11257 bifidobacterium breve OLB6129 manufacture renal dialysis patients for prevent or improve dialysis complication functional foodstuff in purposes.
7. the minimizing agent of corrupt substance in intestines according to claim 2, wherein, further containing oligofructose (FOS) and/or oligomeric galactose (GOS).
8. the minimizing agent of the indoxyl sulfate in serum, its with bifidobacterium breve according to claim 1 for effective constituent.
9. deposit number is the purposes of bifidobacterium breve OLB6129 in the minimizing agent manufacturing the indoxyl sulfate in serum of FERM BP-11257.
10. the minimizing agent of the indoxyl sulfate in serum according to claim 8, wherein, further containing oligofructose (FOS) and/or oligomeric galactose (GOS).
11. food with the minimizing effect of corrupt substance in intestines, its with bifidobacterium breve according to claim 1 for effective constituent.
12. food with the minimizing effect of indoxyl sulfate in serum, its with bifidobacterium breve according to claim 1 for effective constituent.
13. cultured milk prods with bifidobacterium breve according to claim 1.
14. lebens with bifidobacterium breve according to claim 1.
Food according to any one of 15. claims 11 ~ 12, wherein, further containing oligofructose (FOS) and/or oligomeric galactose (GOS).
16. cultured milk prods according to claim 13, wherein, further containing oligofructose (FOS) and/or oligomeric galactose (GOS).
17. lebens according to claim 14, wherein, further containing oligofructose (FOS) and/or oligomeric galactose (GOS).
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