CN103082294A - Application of bacteroides fragilis in preparation of composition for treating diarrhea - Google Patents
Application of bacteroides fragilis in preparation of composition for treating diarrhea Download PDFInfo
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- CN103082294A CN103082294A CN2013100209474A CN201310020947A CN103082294A CN 103082294 A CN103082294 A CN 103082294A CN 2013100209474 A CN2013100209474 A CN 2013100209474A CN 201310020947 A CN201310020947 A CN 201310020947A CN 103082294 A CN103082294 A CN 103082294A
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- bacteroides fragilis
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Abstract
The invention relates to an application of bacteroides fragilis in preparation of a composition for treating diarrhea; the application can be used for the preparation of a vector and also can be used for the preparation of the composition of food or medicine containing such a micro-organism; and experiments show that the bacteroides fragilis has an excellent therapeutical effect to diarrhea.
Description
Technical field
The present invention relates to the purposes of bacteroides fragilis, be specifically related to bacteroides fragilis is treated diarrhoea for the preparation of a kind of carrier with adding in food or medicine, belong to the technical field of microbe application of bacteroides fragilis.
Background technology
Diarrhoea is the modal symptom of large intestine disease, mainly be divided into acute diarrhea and chronic diarrhea, it was 1 ~ 2 week that children with acute diarrheal disease is sent out period, chronic diarrhea is more than 2 months, the common reason that causes diarrhoea has bacterium infection, virus infections, parasite, drug response, intestines problem, catches a cold, diet etc., cause diarrhoea in various degree, sometimes with stomachache, feel sick, the symptom such as vomiting.The main way of present treatment diarrhoea is ORT, correction Electrolyte imbalance, the loose antimicrobial therapy etc. that reaches of oral administration of montmorillonite powder.Within decades recently, probio is as the safety for the treatment of gastrointestinal disease and can utilize form to receive very big concern.
Humans and animals had the bacterial strain of fine effect after absorption by the Bifidobacterium of extensive concern and lactic acid bacteria in probio, their specific bacterial strain can be settled down on intestinal mucosa, the built-in group of enteron aisle and prevent that harmful microorganism from adhering to thereon, by keeping the microorganism formulation of growing of natural microorganisms fauna promotion bion health in intestines, to help to keep the health of humans and animals.File CN1350460A discloses a kind of lactic acid bacteria strains of suffering from diarrhoea of can preventing; File CN1420781A discloses a kind of Bifidobacterium of suffering from diarrhoea of can preventing.
Bacteroides fragilis (Bacteroides fragilis) is as a kind of Gram-negative, shaft-like, blunt round and dense the dying in two ends, pod membrane arranged, without brood cell, motorless obligate anaerobes, minute produces enterotoxin type and non-product enterotoxin type.Bacteroides fragilis mainly is stored in colon as the part of people and animal intestinal normal flora, in addition, but the also field planting growth of respiratory tract, intestines and stomach and urogenital tract mucous membrane.Studying more is that bacteroides fragilis is as a kind of conditioned pathogen, when host's mucous membrane is impaired, can invade submucosa, cause infection, also can flow through blood, cause other organ of health such as enteron aisle, abdominal cavity, liver, lung, brain tissue pyogenic infection and the abscess and cause acute and chronic diarrhea etc. of occurring together.In addition, its generation to colorectal cancer also has facilitation.
But, Zhang Jijie had delivered " separation of a kind of nontoxic bacteroides fragilis is identified " in 1991, reported and separated the large and purer bacterium of output about September nineteen eighty-three from a well-developed infant faeces, through evaluation, basic test and the observation on Clinical Application with series, belong to a strain in the bacteroides fragilis group without toadstool, be the probio of human body, name the bacterial strain into BF839.
The present invention is applied to bacteroides fragilis to prepare in the medicine for the treatment of diarrhoea, and effective, cost is low.
Summary of the invention
The purpose of this invention is to provide the application of a kind of novel microbial bacteroides fragilis in preparation treatment diarrhoea medicine.
In order to address the above problem, the technical solution adopted in the present invention is:
The application of bacteroides fragilis in preparation treatment diarrhoea composition.
Described purposes refers to can be applied to prepare a kind of carrier, also can be for the preparation of the composition of the food that contains this microorganism or medicine.
The content of described bacteroides fragilis in carrier is 10
8~ 10
10Cfi/mL.
Described carrier is the food composition that is selected from milk powder, cheese, curdled milk, yoghourt, milk base fermented product, ice cream, fermented cereal product, milk powder, baby food or pet food.
Described food or composite medicine contain the bacteroides fragilis bacterial strain that can treat diarrhoea at least.
Above-mentioned composition is selected from milk powder, cheese, curdled milk, yoghourt, milk base fermented product, ice cream, fermented cereal product, milk powder, baby food or pet food, tablet, liquid bacterial suspension, dried oral supplement, wet oral supplement, main is raised or the tube feed that wets.
Above-mentioned composition is tablet, liquid bacterial suspension, dried oral supplement, wet oral supplement, main is raised or the form of the tube feed that wets.
The invention has the beneficial effects as follows: experimental results show that bacteroides fragilis has good effect for treatment diarrhoea.
The specific embodiment:
Below by embodiment, the present invention is described in further details, these embodiment only are used for illustrating the present invention, do not limit the scope of the invention.
Embodiment 1
(1) by separating bacteroides fragilis in infant faeces
Take 0.5 gram stool by " normal flora inspection technique ", be placed in the test tube that 4.5ml physiological saline first is housed, become 10 with this serial dilution
-1~ 10
-6, dripped on the Bd culture medium flat plate to low rare long-pending degree by the rare long-pending degree of height, be placed in the anaerobic gas generation box and add the anaerobic gas generation bag and carry out anaerobism and cultivate.Put 37 ℃, can opening inspection after 2 ~ 7 days.Menu one bacterium colony carries out pure culture, dyeing microscopic examination, cultural character, Physiology and biochemistry detection.
(2) bacterium that increases of bacteroides fragilis is cultivated and bacterium solution preparation
The single colony inoculation of choosing after purifying increases bacterium, the centrifugation of gained bacterium liquid, rotating speed 3000r/min in the tryptone meat soup of improvement, centrifugal 15min goes the supernatant precipitation to wash with physiological saline, the sediment normal saline dilution, do the bacterium number with the Maxwell opacity tube and measure, be diluted to 10
8Cfi/ml, 10
9Cfi/ml, 10
10Cfi/ml.
Embodiment 2
The bacteroides fragilis virulence experiment
(1) chmice acute virulence experiment
Get body weight 22 ± 2 60 of gram C57BR/C mouse (male and female half and half) and be divided into high dose group, low dose group and control group, each organizes 20.Every of high dose group gavage every day 10
12Cfi/ml bacterium liquid 0.5ml, low dose group gavage every day 10
9Cfi/ml bacterium liquid 0.5ml, control group gavage every day 0.5ml physiological saline.Observed 14 days.
Experimental result: the mouse normal growth, have no dead, LD50 does not measure.
(2) rat virulence experiment
Get 30 of 150 ± 20 gram SD rats (male and female half and half).Be divided into 3 groups: high low dose group and control group.Every of high dose group gavage every day 10
12Cfi/ml bacterium liquid 2ml, low dose group gavage every day 10
9Cfi/ml bacterium liquid 2ml, control group gavage every day 2ml physiological saline.Continuous Observation 14 days phlebotomizes after 14 days and surveys blood picture, liver kidney merit.Get stomach and intestine liver spleen tissue and do the pathological section electron microscopic observation.
Experimental result: be showed no obvious abnormalities.
(3) rhesus macaque virulence experiment: get 10 of rhesus macaquies (male and female half and half).Be divided into 2 groups: high low dose group and control group.Every every day oral 10 of high dose group
12Cfi/ml bacterium liquid 100ml, low dose group every day oral 10
9Cfi/ml bacterium liquid 100ml.Continuous Observation 60 days.Phlebotomize after 14 days, 30 days, 45 days, 60 days and survey blood picture, liver kidney merit, thought-read electrograph.Get stomach and intestine liver spleen tissue and do the pathological section electron microscopic observation.
Experimental result: be showed no obvious abnormalities.
Embodiment 3
Bacteroides fragilis is for the therapeutic action of diarrhoea
(1) ampicillin sodium (adding the physiological sodium chloride solution preparation) 0.2g gavage is used in animal fasting feedwater before modeling subsequently, and every 0.5ml Bid causes diarrhea of mouse, draining sample or mushy stool.
Diarrhea mice is used respectively 10
8Cfi/ml, 10
9Cfi/ml, 10
10The bacterium liquid gavage of cfi/ml, each 0.5ml Bid, the blank group is given isopyknic physiological saline gavage, every group each 20 of Bid().
* compare P＜0.05 with the blank group.
The fresh stool of (2) getting 10 mouse before modeling does the faecal microbiota analysis, and after treatment, the 5th day each group lived and to be killed 10 mouse and get cecal content and do the flora analysis, the results are shown in following table:
Analysis of intestinal microflora Lg g-1, n=10
* compare P＜0.05 with the blank group.
The fresh stool of getting 10 mouse before modeling does the faecal microbiota analysis, and after treatment, the 5th day each group lived and to be killed 10 mouse and get cecal content and do the flora analysis, the results are shown in following table:
Analysis of intestinal microflora Lg g-1, n=10
* compare P＜0.05 with the blank group.
This studies show that, the mouse of bacteroides fragilis gavage group is than the sx↓ of the diarrhea of mouse of physiological saline gavage, and the diarrhoea duration shortens, and illustrates that bacteroides fragilis has certain therapeutic action to diarrhoea, during at 106cfi/ml-1010cfi/ml, result for the treatment of is best when bacteroides fragilis dosage.The demonstration of mouse intestinal flora analysis result, bacteroides fragilis can promote Bifidobacterium, Bacillus acidi lactici, enterococcal growth, equally diarrhoea is had prevention and therapeutic action.
Embodiment 4
The protective effect of bacteroides fragilis to the RV virus infected cell
Test is with viral: attenuated strain hRV Wa, pRV OSU virus;
1.IEC-6 cell is with 1*10
5/ cm
2Concentration plant 25 cm
2Blake bottle after form good cell monolayer at 48 h, the cell number of every bottle is about 3.5 ~ 4*10
6
2. be divided into two schemes:
Scheme one: process cell with Bd before the RV infection cell:
(1) Bd (1*108 cfu/mL) evenly is suspended in the maintain base of 2.5 mL, inoculating cell individual layer and at 5% CO2 is cultivated 24 h for 37 ℃;
(2) use PBS rinsing cell monolayer 3 times, wash the bacterium that is not attached on cell off;
(3) cell monolayer of then virus inoculation of 20 MOI (multiplicity of infection) being processed to probio at 37 ℃ of viral 30 min of activation with 5 μ g/mL pancreatin and at 37 ℃ of 5% CO2 cultivation 1 h;
(4) virus liquid that discards inoculation is also used PBS rinsing cell monolayer 1 time;
(5) add 2.5 mL to contain the maintain base of 0.5 μ g/mL pancreatin, at 37 ℃ of 5% CO2 effect 24 h;
Scheme two: after the RV infection cell, Bd processes cell:
(1) activate viral 30 min with 5 Lg/mL pancreatin at 37 ℃.Then the cell monolayer of the virus inoculation of 20 MOI being processed to probio and at 37 ℃ of 5% CO2 cultivation 1 h;
(2) virus liquid that discards inoculation is also used PBS rinsing cell monolayer 1 time;
(3) Bd (1*108 cfu/mL) evenly is suspended in the maintain base of 2.5 mL, inoculating cell individual layer and at 5% CO2 is cultivated 24 h, is contained 0.5 μ g/mL pancreatin in bacterium liquid for 37 ℃.
Control group does not add Bd, and all the other processing are identical with experimental group.
The collecting cell culture supernatant is also removed at centrifugal 10 min of 3000 r/min, bacterium and the cell fragment that comes off, with the supernatant packing and be stored in-80 ℃ until use to analyze, check the integrality of cell monolayer after effect finishes and determine the activity of cell with the trypan blue counting method.
The cytoactive that experimental result demonstration bacteroides fragilis was processed is than the obvious height of control group.Bacteroides fragilis has the activity of stronger anti-rotavirus.During at 106cfi/ml-108cfi/ml, the most obvious to the protective effect of cell when the quantity of bacteroides fragilis.
Claims (7)
1. the application of bacteroides fragilis in preparation control diarrhoea composition is characterized in that: the application of bacteroides fragilis in preparation treatment diarrhoea composition.
2. the application of bacteroides fragilis according to claim 1 in preparation control diarrhoea composition is characterized in that described purposes refers to can be applied to prepare a kind of carrier, also can be for the preparation of the composition of the food that contains this microorganism or medicine.
3. the application of bacteroides fragilis according to claim 1 in preparation treatment diarrhoea composition is characterized in that the content of described bacteroides fragilis in carrier is 10
8~ 10
10Cfi/mL.
4. the application of bacteroides fragilis in preparation control diarrhoea composition according to claim 2, it is characterized in that: described carrier is the food composition that is selected from milk powder, cheese, curdled milk, yoghourt, milk base fermented product, ice cream, fermented cereal product, milk powder, baby food or pet food.
5. contain bacteroides fragilis food or composite medicine, it is characterized in that: described food or composite medicine contain the bacteroides fragilis bacterial strain that can treat diarrhoea at least.
6. composition according to claim 5 is characterized in that: described composition is selected from milk powder, cheese, curdled milk, yoghourt, milk base fermented product, ice cream, fermented cereal product, milk powder, baby food or pet food, tablet, liquid bacterial suspension, does oral supplement, wet oral supplement, main is raised or the tube feed that wets.
7. composition according to claim 5 is characterized in that: described composition is tablet, liquid bacterial suspension, do oral supplement, wet oral supplement, main is raised or the form of the tube feed that wets.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN2013100209474A CN103082294A (en) | 2013-01-21 | 2013-01-21 | Application of bacteroides fragilis in preparation of composition for treating diarrhea |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103156887A (en) * | 2013-03-18 | 2013-06-19 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for promoting bifidobacterium and lactobacillus to grow |
| WO2017020784A1 (en) * | 2015-07-31 | 2017-02-09 | 广州知易生物科技有限公司 | Bacteroides fragilis and application thereof |
| WO2018014832A1 (en) * | 2016-07-18 | 2018-01-25 | 广州普维君健药业有限公司 | Applications of bacteroides fragilis in preparing composition for preventing vibrio parahaemolyticus infection |
| CN109481473A (en) * | 2017-09-11 | 2019-03-19 | 广州知易生物科技有限公司 | Application of the bacteroides fragilis extract in the drug or food of preparation prevention and treatment irritable bowel syndrome |
| CN114558036A (en) * | 2022-01-25 | 2022-05-31 | 广州知易生物科技有限公司 | Application of bacteroides fragilis in improvement and treatment of diarrhea |
| WO2023061154A1 (en) * | 2021-10-12 | 2023-04-20 | 广州知易生物科技有限公司 | Use of bacteroides fragilis in prevention and treatment of cancer-related diarrhea |
Citations (2)
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| CN1029857C (en) * | 1990-05-03 | 1995-09-27 | 张季阶 | Bacterial strain and its utilization |
| CN1420781A (en) * | 1999-08-05 | 2003-05-28 | 雀巢制品公司 | Bifidobacteria capable of preventing diarrhea |
-
2013
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1029857C (en) * | 1990-05-03 | 1995-09-27 | 张季阶 | Bacterial strain and its utilization |
| CN1420781A (en) * | 1999-08-05 | 2003-05-28 | 雀巢制品公司 | Bifidobacteria capable of preventing diarrhea |
Non-Patent Citations (2)
| Title |
|---|
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103156887A (en) * | 2013-03-18 | 2013-06-19 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for promoting bifidobacterium and lactobacillus to grow |
| CN103156887B (en) * | 2013-03-18 | 2015-06-24 | 广州知易生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for promoting bifidobacterium and lactobacillus to grow |
| WO2017020784A1 (en) * | 2015-07-31 | 2017-02-09 | 广州知易生物科技有限公司 | Bacteroides fragilis and application thereof |
| CN106399141A (en) * | 2015-07-31 | 2017-02-15 | 广州知易生物科技有限公司 | Bacteroides fragilis and applications thereof |
| CN106399141B (en) * | 2015-07-31 | 2019-07-05 | 广州知易生物科技有限公司 | A kind of bacteroides fragilis and its application |
| WO2018014832A1 (en) * | 2016-07-18 | 2018-01-25 | 广州普维君健药业有限公司 | Applications of bacteroides fragilis in preparing composition for preventing vibrio parahaemolyticus infection |
| CN109481473A (en) * | 2017-09-11 | 2019-03-19 | 广州知易生物科技有限公司 | Application of the bacteroides fragilis extract in the drug or food of preparation prevention and treatment irritable bowel syndrome |
| CN113730443A (en) * | 2017-09-11 | 2021-12-03 | 广州知易生物科技有限公司 | Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome |
| CN113730443B (en) * | 2017-09-11 | 2022-10-04 | 广州知易生物科技有限公司 | Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome |
| CN109481473B (en) * | 2017-09-11 | 2023-03-21 | 广州知易生物科技有限公司 | Application of bacteroides fragilis extract in preparation of medicine for preventing and treating irritable bowel syndrome |
| WO2023061154A1 (en) * | 2021-10-12 | 2023-04-20 | 广州知易生物科技有限公司 | Use of bacteroides fragilis in prevention and treatment of cancer-related diarrhea |
| CN114558036A (en) * | 2022-01-25 | 2022-05-31 | 广州知易生物科技有限公司 | Application of bacteroides fragilis in improvement and treatment of diarrhea |
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Application publication date: 20130508 |