CN106361778A - Lactobacillus preparation and application thereof in treatment of infantile diarrhea - Google Patents
Lactobacillus preparation and application thereof in treatment of infantile diarrhea Download PDFInfo
- Publication number
- CN106361778A CN106361778A CN201610809305.6A CN201610809305A CN106361778A CN 106361778 A CN106361778 A CN 106361778A CN 201610809305 A CN201610809305 A CN 201610809305A CN 106361778 A CN106361778 A CN 106361778A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- preparation
- lactobacillus preparation
- acidophilus
- bacillus acidophilus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the technical fields of microbiological screening and application, in particular to a novel lactobacillus preparation. The lactobacillus preparation has a good treatment effect for infantile diarrhea, and can shorten the duration of the infantile diarrhea; furthermore, after being treated by using the lactobacillus preparation, a baby tends to have less stool discharge. Lactobacillus acidophilus in the lactobacillus preparation has very strong bacteriostatic ability, has an obvious inhibiting effect for salmonella, staphylococcus aureus and helicobator pylori, and has a highest obvious inhibiting effect especially for escherichia coli; the diameter of an antibacterial circle reaches up to 24mm. The lactobacillus acidophilus can be efficiently propagated in gastric acid and is highly tolerant to bile salt. The lactobacillus acidophilus can strongly remove cholesterol, and the removal rate is as high as 91.4%; the strain also has very high oxidation resistance, and the removal rates of fermentative supernatant liquid of the strain for DPPH and hydroxyl free radical respectively reach up to 92.6% and 94.1%, so that an unexpected technical effect is achieved.
Description
Technical field
The present invention relates to microbe to screen technical field, specifically provide a kind of lactobacillus preparation and its control in infantile diarrhea
Application in treatment.
Background technology
Probiotic bacteria is the quasi-microorganism being beneficial to host health, and nineteen sixty-five is suggested on science first.Intestinal
Be balance be to human nutrition and healthy vital factor, probiotic bacteria has good adjustment effect to human body intestinal canal fungus strain,
This makes the research of probiotic bacteria become a big focus of food research.In recent years, with the further investigation to probiotic bacteria, prebiotic micro-
Biological species also gets more and more, and it is broadly divided into three major types: the first kind is lactic acid bacteria, mainly includes bacillus acidophilus, excrement chain
Coccus and Bifidobacterium lactis etc.；Equations of The Second Kind is bacilluss；3rd class is yeast.
Lactic acid bacteria is avirulence, the common name of gram-positive bacterium that a class can produce lactic acid using carbohydrate,
Including at least 23 genus such as lactobacilluss, Lactococcus and bacillus bifiduss.Be considered as in selection-breeding lactobacilli strain some: (1)
The detached object of thalline should be the human or animal of health, and the bacterial strain that the mankind use is preferably derived from people.(2) it is resistant to the bile of people
Hydrochlorate and sour environment.(3) there is suitable immunoregulation effect, the untoward reaction such as inflammation will not be caused.
Lactic acid bacteria, as the important strain of microbial ecological agent, is being safeguarded humans and animals intestinal health, is being promoted internal Tiny ecosystem
Balance aspect plays an important role, and numerous studies show, lactic acid bacteria can adjust body gastrointestinal tract normal flora, keep micro- life
State balances, and improves food digestion rate and biological value, reduce serum cholesterol, control machine vivotoxin, putrefaction bacteria in suppression intestinal
Growth and breeding and the generation of spoilage product, manufacture nutrient substance, stimulate tissue development, thus to the nutritional statuses of body, physiology
Function, cell infection, drug influence, toxic reaction, immunoreation, tumor generation, aging course and unexpected emergency reaction etc.
Generation acts on.In addition, lactic acid bacteria has been widely used in the industries such as medicine, food and feedstuff as important probiotic bacteria, public
It is considered safe food-grade microorganisms.
For normal adults, microorganism in its intestinal is colonized on intestinal wall in certain population ratio, is in one
Plant in stable colony balance.But the use due to some factors such as antibiotic, radiotherapy, chemotherapy, dietary habit are bad, spiritual pressure
The reasons such as power may result in the change of normal intestinal flora type and quantity, and enteric microorganism ecosystem is destroyed, and is drawn
The pathological state rising is referred to as alteration of intestinal flora.After lactic acid bacteria enters intestinal, growth and breeding can be carried out rapidly, metabolism produces
Lactic acid, carbon dioxide make that enteral ph value is rapid to be reduced it is suppressed that the breeding of the antibacterial of pathogen and harmful health, thus rising
Effect to prevention infection, maintenance intestinal flora balance.
Lactic acid bacteria has special physiological action to human body, is requisite normal physiological flora in body, expands strain
Source, the existing excellent fermentation character of selection-breeding, there are high, excellent, the safe lactic acid bacteria culturerss of the viable count of special health care again；Should
With molecular biotechnology developing food products level lactobacilli strain and engineering bacterial strain, huge economic benefit, society will be brought for the mankind
Can benefit and ecological benefits.
Content of the invention
It is an object of the invention to provide a kind of lactobacillus preparation, described lactobacillus preparation is from a kind of acid from natural fermentation
The acidophilic lactobacilluss (lactobacillus acidophilus) filtering out in vegetable juice, not only stomach juice-resistant and bile saltss, tool
There is very strong non-oxidizability, and notable to colibacillary inhibitory action, have a extensive future.
One aspect of the present invention provides a kind of lactobacillus preparation, comprises bacillus acidophilus (lactobacillus
acidophilus).
The preferred bacillus acidophilus kdb-03 (lactobacillus acidophilus kdb-03) of described bacillus acidophilus,
It is preserved within 29th the China typical culture collection center of Wuhan, China Wuhan University in August in 2016, deposit number is
cctcc no:m2016429.
Described bacillus acidophilus are lactobacillus acidophilus freeze-drying powder.
The preparation method of the lyophilized powder of described bacillus acidophilus, concrete steps include:
1) liquid fermentation medium, sterilizing are prepared；
2) culture medium is preheated to 50-55 DEG C, carries out homogenizing under 20-25mpa pressure condition, TRANSIENT HIGH TEMPERATURE sterilizes；
3) press 5% inoculating lactobacillus acidophilus kdb-03 of amount of fermentation, after having inoculated, continue stirring 15min；
4) control 37-40 DEG C of fermentation temperature, ferment 10 hours；
5) fermentation liquid is centrifuged, vacuum lyophilization to water content is less than 3%, obtains lactobacillus acidophilus freeze-drying powder.
In described liquid fermentation medium each component and its mass volume ratio be respectively as follows: milk powder 10%, white sugar 0.5%,
Glucose 0.5%；
Described lactobacillus preparation also includes starch, Hydroxypropyl Cellulose, dead plaster, maltodextrin, oligofructose, local flavor
One of agent or the combination of two or more.
Described lactobacillus preparation is capsule, tablet or powder.
Application in treatment infantile diarrhea for the described lactobacillus preparation.
Beneficial effect
The lactobacillus preparation that the present invention provides has good therapeutic effect to infantile diarrhea, can significantly shorten infantile diarrhea
Duration, and baby tends to there is relatively low stool output after treatment.Acidophilus breast in described lactobacillus preparation
Bacillus bacteriostasis are very strong, and it all has obvious inhibitory action to Salmonella, staphylococcus aureuses and helicobacter pylori,
Especially strong to colibacillary inhibitory action, antibacterial circle diameter reaches 24mm.Described bacillus acidophilus can be had in gastric acid
Effect breeding, and to bile saltss, there is strong tolerance.Described bacillus acidophilus kdb-03 energy strength Cholesterol removal, clearance is up to
91.4%；This bacterial strain also has very strong oxidation resistance, the clearance rate to dpph and hydroxyl radical free radical for its fermented supernatant fluid
It is up to 92.6% and 94.1% respectively, achieve unexpected technique effect.
Specific embodiment
Embodiment 1 bacillus acidophilus' kdb-03 separation screening and identification:
1st, screening sample
The sauerkraut juice collecting natural fermentation from Shenyang Su Jiatun rural area is no dirty as separating sample, described sauerkraut juice outward appearance
Dye, good taste, fermentation time is more than 1 year, standby after filtration.
2nd, lactic acid bacteria primary dcreening operation
Take the sauerkraut juice 5ml after filtration to add 45ml sterilized water, mix, coat the mrs flat board adding 0.5% Calcium Carbonate
On, 37 DEG C of culture 48h, separation and Culture has the bacterial strain of transparent circle, obtains 42 strains of lactic acid bacteria.
3rd, antibacterial type lactic acid bacteria screening
1) lactobacillus suspension preparation: 32 strains of lactic acid bacteria that primary dcreening operation is obtained are inoculated in 100ml mrs fluid medium respectively,
37 DEG C of quiescent culture 48h；
2) pathogenic bacterium bacterium solution preparation: escherichia coli, Salmonella, staphylococcus aureuses and (four kinds of helicobacter pylori
Pathogenic bacterium are given by Shandong University), strain is inoculated in nutrient broth, 37 DEG C of shaking table cultures are overnight；
3) bacteriostatic experiment double-layer plate, Odontothrips loti: often (50 DEG C about) plus 100 μ of 5ml bactericidal nurishing agar culture medium
L pathogenic bacterium bacterium solution (bacterium amount 106The order of magnitude), mix hypsokinesis and make double-layer plate down to nutrient agar panel, in culture after solidification
Place Oxford cup on base, add the cultured lactic acid bacterial liquid of 200 μ l in Oxford cup, after bacterium solution diffusion, put into 37 DEG C of cultures
Cultivate 20h in case, observe antibacterial circle diameter.
Result shows, to escherichia coli, husky 8 Salmonellas, staphylococcus aureuses and pylorus in the lactic acid bacteria that primary dcreening operation obtains
The strain altogether of the bacterial strain all more than 15mm for the antibacterial circle diameter of pylori.Further anti-biotic resistance screening is carried out to it.Through two
Secondary delete choosing, final applicant screens from above-mentioned 8 plants of bacterium and obtains the lactic acid bacteria the strongest to colibacillary growth inhibition effect, life
Entitled kdb-03.
The inhibitory action to pathogenic bacterium for the table 1 lactic acid bacteria kdb-03
| Pathogenic bacterium | Escherichia coli | Salmonella | Staphylococcus aureuses | Helicobacter pylori |
| Antibacterial circle diameter | 24mm | 20mm | 21mm | 21mm |
As can be seen from the above table, the lactic acid bacteria kdb-03 that the present invention filters out to Salmonella, staphylococcus aureuses and
Helicobacter pylori all has obvious inhibitory action, and especially strong to colibacillary inhibitory action, antibacterial circle diameter reaches
24mm.
3rd, identification of strains
Extract the genome dna of above-mentioned lactic acid bacteria kdb-03 using test kit, expand 16s rrna sequence using pcr technology
Row.Sequencing result is carried out in genbank nucleic acid database blast and compare discovery, the 16s rrna sequence of lactic acid bacteria kdb-03
Row are higher than 99% with the sequence similarity of bacillus acidophilus (lactobacillus acidophilus).It is thus identified that it is thermophilic
Lactobacillus lactiss (lactobacillus acidophilus), are named as bacillus acidophilus kdb-03 (lactobacillus
Acidophilus kdb-03), and it is preserved within 29th the Chinese Typical Representative culture guarantor of Wuhan, China Wuhan University in August in 2016
Tibetan center, deposit number is cctcc no:m2016429.
Embodiment 2 bacillus acidophilus kdb-03 is acidproof, bile tolerance experiment
1st, stomach juice-resistant method for testing performance:
Take the bacillus acidophilus kdb-03 bacteria suspension 40ml after activation to add in 50ml centrifuge tube, be centrifuged (5000g, 5min)
Collects thalline, with adding 40ml simulated gastric fluid (to take dilute hydrochloric acid 16.4ml to add water to shake in pbs buffer solution twice backward centrifuge tube
Even be diluted to 1000ml, with concentrated hydrochloric acid or 10% naoh adjustment ph be 2.0, in 121 DEG C sterilize 30min, in sterilizing room with 1g/
100ml ratio adds pepsin), 3h are cultivated in 37 DEG C of water-baths, shake up once every 30min, live in 0h, 2h and 3h are separately sampled
Bacterium counts, and with 0h sample for comparison, calculates the survival rate of 2h and 3h sample thalline, survival rate=(viable count/0h during 2h or 3h
When viable count) × 100%.
2nd, the method for testing performance of resistance to bile saltss:
Take the bacillus acidophilus kdb-03 bacteria suspension 40ml after activation to add in 50ml centrifuge tube, be centrifuged (5000g, 5min)
Collects thalline, with adding 40ml simulated intestinal fluid (compound method: take in pbs buffer solution twice backward centrifuge tube
kh2po46.8g, plus distilled water 500ml dissolving, adjust ph value to 6.8 with the naoh solution of 0.4% (w/v), add water to 1000ml,
0.3g fowl cholate is added, after fully dissolving, in 115 DEG C of 15min that sterilize in every 100ml solution.), 2h is cultivated in 37 DEG C of water-baths, every
30min shakes up once, makees count plate in 0h, 2h and 3h are separately sampled, with 0h sample for comparison, calculates 2h and 3h sample thalline
Survival rate, survival rate=(viable count during viable count/0h during 2h or 3h) × 100%.
Meanwhile, carried out using bacillus acidophilus (lactobacillus acidophilus) cgmcc1.1854 as comparison
Above-mentioned stomach juice-resistant and resistance to bile saltss performance detection.Concrete outcome is shown in Table 2.
Table 2 bacillus acidophilus' kdb-03 stomach juice-resistant, resistance to bile saltss performance test results
Data from table 2 can be seen that compared with bacillus acidophilus cgmcc:1.1854, the acidophilus that the present invention screens
Lactobacilluss kdb-03 has very strong stomach juice-resistant, resistance to bile saltss ability, and especially its survival rate during 3h in gastric acid is up to
237.52%, not only significantly larger than control strain, and the survival rate in 2h apparently higher than bacillus acidophilus kdb-03, this says
Bright described bacillus acidophilus kdb-03 is not only able to effectively survive in gastric acid, and also achieves obvious propagation, achieves
Unexpected effect.
The removal effect to cholesterol for the embodiment 3 bacillus acidophilus kdb-03
This experiment is with reference to the method for (1998) such as brashears, and slightly improves.
It is inoculated in mrs culture medium, 37 DEG C of overnight incubation, as seed liquor after bacillus acidophilus' kdb-03 bacterial strain is activated.
Prepare the mrs culture medium containing 5% (v/v) egg yolk liquid, take wherein 2ml culture medium, 5000r/min is centrifuged 7min, takes
1ml supernatant is placed in centrifuge tube, adds 9ml dehydrated alcohol, oscillation treatment 5min, 10000r/min is centrifuged 10min, takes 2ml
Supernatant adds 2ml p-fe-s reagent, and ice bath mixes reaction 30min, surveys od550, and in calculating culture medium, cholesterol initially contains
Amount.
Then, the seed liquor of bacillus acidophilus kdb-03 is seeded in above-mentioned mrs culture medium, cultivates under the conditions of 37 DEG C
After 24h, measure the final content of cholesterol in culture medium again, calculate the clearance to cholesterol for the bacillus acidophilus kdb-03.
Simultaneously using bacillus acidophilus (lactobacillus acidophilus) cgmcc:1.1854 as control strain, using above-mentioned
Same operation, calculates the clearance to cholesterol for this bacterium.
Removal rate of cholesterol=(initial content-final content)/initial content × 100%
Result shows: the bacillus acidophilus kdb-03 that the present invention screens is high to the clearance of cholesterol in mrs culture medium
Reach 91.4%；And be equally that the control strain of bacillus acidophilus is only 19.8% to the clearance of cholesterol.Thus this is described
Bright screen bacillus acidophilus kdb-03 can strength Cholesterol removal, achieve unexpected technique effect.
Embodiment 3 bacillus acidophilus' kdb-03 antioxidation in vitro is tested
Bacillus acidophilus kdb-03 is carried out passing on after activation, mrs liquid culture is inoculated in 5% (mass ratio) inoculum concentration
Base, 37 DEG C of quiescent culture 18h, 3 000r/min, it is centrifuged 15min, collect fermentation supernatant and thalline respectively.
Prepare cell-free extract: with phosphate buffer solution (pbs) washing thalline 3 times for 7.4 for the ph value, be resuspended in
In phosphate buffer solution, adjustment bacterium number to 109cfu/ml；Then ultrasound wave ice bath crushes thalline, and 10000r/min is centrifuged
10min, supernatant is cell-free extract.
3.1 dpph remove experiment
Dpph is a kind of very stable free radical centered on nitrogen, if tested material can remove it then it represents that tested material
Antioxidant effect is notable.
Dpph solution；Respectively take 1ml fermented supernatant fluid and cell-free extract, being separately added into 1ml concentration is 0.2mmol/l
Dpph, mix, under room temperature standing 30min after, 517nm at survey absorbance change.
The clearance rate (%) of dpph=[1- (a1-a2)/a3] × 100
In formula: a1 represents the original absorbance of the dpph solution of the product of sample-adding；
A2 represents absorbance when measuring wavelength for the sample itself；
A3 represents the absorbance of dpph solution after sample-adding.
Result shows: the fermented supernatant fluid of bacillus acidophilus kdb-03 and cell-free extract all can strongly remove
Dpph, its clearance rate is up to 92.6% and 81.2% respectively, thus illustrating the bacillus acidophilus kdb-03's that the present invention provides
Fermented liquid supernatant liquid and cell-free extract are respectively provided with stronger oxidation resistance, and the removing of this strain fermentation supernatant effect
Fruit will be apparently higher than cell-free extract, and the metabolite of this explanation bacillus acidophilus' kdb-03 growth period is clear to free radical
Except playing prior effect, oxidation resistance is higher.
3.2 hydroxyl radical free radicals remove experiment
In ros free radical, hydroxyl radical free radical is the most activated, exists in metal ion (as copper ion or iron ion)
Under the conditions of, superoxide anion and peroxidating Hydrogen Energy generate hydroxyl radical free radical.Hydroxyl radical free radical is a kind of very strong freedom of oxidisability
Base, can cause on biological cellular macromolecule to damage and affect the normal function of cell.Therefore, the Scavenging activity to hydroxyl radical free radical
It is a leading indicator of antioxygenic property.
Esr method measures the ability removing hydroxyl radical free radical: respectively by the fermented supernatant fluid of 50 μ l bacillus acidophilus kdb-03
Respectively it is added to after the dmpo that 50 μ l concentration are 0.3mol/l with cell-free extract, the system of reaction is transferred to the quartz of sealing
In capillary tube, then plus 50 μ l concentration be 10mol/l h2o2 start reaction.After reaction 2.5min, by er 200d src
Esr spectrometer analysis.Compare the phosphate buffered solution (ph 7.4) for 0.05mol/l.Sample to the scavenging action of oh with
Clearance rate represents.
Clearance rate=(h0—h)/h0× 100%
In formula: h and h0Represent sample respectively and compare spectroscopic signal intensity, the of the relative intensity spectroscopic signal of signal
Two peak values are representing.
Result shows: the removing to hydroxyl radical free radical of the fermented supernatant fluid of bacillus acidophilus kdb-03 and cell-free extract
Rate is up to 94.1% and 88.3% respectively, thus proving that bacillus acidophilus kdb-03 has very strong oxidation resistance further,
And the elimination effect of this strain fermentation supernatant will be apparently higher than cell-free extract.
Antioxidation zoopery in embodiment 4 body
4.1 experimental design
Internal anti-oxidation function experiment adopts high lipid oxidation Stress model.Laboratory animal is using healthy Kunming mouse 40
Only, male and female half and half, body weight 23 scholar 2g, provided by jiangsu wuxi Hui Shan south of the River laboratory animal field.Mice is raised at room temperature, freely
Search for food drinking-water, after the adaptability through 3d is raised, be randomly divided into 4 groups, every group 10, feed normal feedstuff (formula: 89.5% is common
Feedstuff, 10% Adeps Sus domestica, 0.5% cholesterol), matched group daily gavage 0.2ml normal saline, low middle high three dosage of its excess-three component
Gavage variable concentrations bacillus acidophilus kdb-03, as shown in table 3, daily 0.2ml/, continuous 4 weeks.Last is to fasting after sample
24h, gathers rapidly blood, separating red corpuscle and serum, detection erythrocyte superoxide dismutase (sod) activity, whole blood trough Guang
Sweet peptide peroxidase (gsh-px) activity, blood plasma catalase (cat) activity, Plasma MDA (mda) content, all numbers
According to all carrying out statistical procedures with statistical software spss.Test kit is purchased from Nanjing and builds up bio-engineering corporation.
The zoopery that table 3 anti-oxidation function is evaluated
4.2 interpretation of result
4.2.1 superoxide dismutase (sod)
Superoxide dismutase (sod), is to have the biological internal superoxide ion of single-minded removing, can balance the oxygen of body certainly
By base.It can remove excessive ultra-oxygen anion free radical, reduces the disease being produced by ultra-oxygen anion free radical.In the present invention
The impact to mouse red blood cell superoxide dismutase (sod) activity for the gavage bacillus acidophilus kdb-03 is shown in Table 4.
Table 4 bacillus acidophilus kdb-03 is to mouse red blood cell sod effect of vigor
| Group | Sod activity (u/mghb) |
| Matched group | 214.93±1.56 |
| Low dosage | 245.33±1.91 |
| Middle dosage | 268.79±2.16* |
| High dose | 298.38±1.36* |
* p ＜ 0.05
From table 4, it can be seen that with respect to matched group, the erythrocyte sod enzyme activity of middle dosage and high dose mice carries respectively
High by 25.1% and 38.8%, thus illustrating that the bacillus acidophilus kdb-03 of the present invention can significantly improve sod enzyme activity, have relatively
Strong oxidation resistance.
4.2.2 glutathion peroxidase (gsh-px)
Glutathion peroxidase (gsh-px) is that a kind of important catalyzing hydrogen peroxide being widely present in body divides
The enzyme of solution, is catalyzed the reduction reaction to hydrogen peroxide for the reduced glutathion, plays the effect of protection membrane structure and function.
The impact to mouse red blood cell glutathione peroxidase activity for the gavage bacillus acidophilus kdb-03 is shown in Table 5.
Table 5 bacillus acidophilus kdb-03 affects on mouse red blood cell glutathione peroxidase activity
| Group | Gsh-px (enzyme activity unit) |
| Matched group | 131.45±0.67 |
| Low dosage | 151.34±2.21 |
| Middle dosage | 174.03±3.82* |
| High dose | 189.77±2.51* |
* p ＜ 0.05
As can be seen from Table 5, middle and high dosage group mouse red blood cell glutathione peroxidase activity is up to respectively
174.03 and 189.77, improve 32.4% and 44.4% than matched group, thus confirming the bacillus acidophilus that the present invention provides
Kdb-03 can significantly improve the vigor of mouse red blood cell glutathion peroxidase.
4.2.3 catalase
Catalase generally existing in body, catalyzing hydrogen peroxide is decomposed into water and oxygen.Hydrogen peroxide is body
The harmful by-products of metabolism, are catalytically converted into the chemicals of other low toxicities through catalase, prevent body from sustaining damage.Fill
Stomach bacillus acidophilus kdb-03 is shown in Table 6 to the impact of mice catalase activity.
Table 6 gavage bacillus acidophilus kdb-03 affects on mice catalase activity
| Group | Cat activity (u/ml) |
| Matched group | 1.65±0.82 |
| Low dosage | 1.88±2.43 |
| Middle dosage | 2.74±2.89* |
| High dose | 2.94±3.15* |
* p ＜ 0.05
As can be seen from Table 6, compared with matched group, the hydrogen peroxide enzyme activity of the treatment group mice of middle and high dosage carries respectively
High by 63.9% and 75.3%, effect is significant, thus illustrate that bacillus acidophilus kdb-03 can be effectively improved the antioxidation of mice
Ability.
4.2.4 malonaldehyde
Malonaldehyde horizontal reverse has answered body inner lipid peroxidating, indirectly reflects the degree of cell injury.Gavage acidophilus
The impact to mouse red blood cell malonaldehyde level for the lactobacilluss kdb-03 is shown in Table 7.
Table 7 gavage bacillus acidophilus kdb-03 affects on mouse red blood cell malonaldehyde level
| Group | Mda content (nmol/ml) |
| Matched group | 3.44±1.87 |
| Low dosage | 3.03±1.89 |
| Middle dosage | 2.67±1.33 |
| High dose | 1.59±1.37* |
* p ＜ 0.05
As can be seen from Table 7, compared with matched group, the treatment group mda content of three dosage all substantially reduces, explanation
The bacillus acidophilus kdb-03 that the present invention provides can mitigate the damage to cell for the mice interior free yl, wherein high dose acidophilus
Lactobacilluss kdb-03 makes the content of malonaldehyde in mouse red blood cell reduce 53.8%, effect highly significant.
Embodiment 5 contains the preparation method of the lactobacillus preparation of bacillus acidophilus kdb-03
Add suitable quantity of water in preparing tank, milk powder, white sugar, glucose are added in preparing tank, stirring and dissolving, finally fixed
Hold, the mass volume ratio (g/ml) of wherein each raw material is respectively as follows: milk powder 3%-10%, white sugar 0.5%-8%, glucose
0.5%-45%.By raw material preheating to 50-55 DEG C, carry out homogenizing under 20-25mpa pressure condition.Using TRANSIENT HIGH TEMPERATURE sterilization
130-135 DEG C of sterilization 4-6s of machine, controls material outlet temperature to be 30-45 DEG C.Open fermentation tank stirring, by the 5-10% of amount of fermentation
Inoculating lactobacillus acidophilus kdb-03.Continue stirring 15min after having inoculated, after fully mixing, close stirring, control fermentation temperature 37-
40℃；Centrifugation, vacuum lyophilization to water content is less than 3%, that is, bacillus acidophilus kdb-03 lyophilized powder is obtained.
Take bacillus acidophilus kdb-03 lyophilized powder 400g, plus 200g amylum pregelatinisatum and 100g low-substituted hydroxypropyl cellulose,
Mix, plus 70% ethanol solution is appropriate, soft material processed, 20 mesh sieve series granules.By wet granular plus 50g dead plaster with 100g is low takes
Mix for Hydroxypropyl Cellulose, be dried, plus appropriate hydrated magnesium silicate micropowder mixes, tabletting, obtains final product lactic acid bacteria tablet, every contains
Bacillus acidophilus kdb-03 about 0.2g, viable count is more than 3,000,000,000/g.
In addition to tablet, can also be by lactobacillus acidophilus freeze-drying powder and maltodextrin and oligomeric xylose mixed in equal amounts, system
Become capsule, or lactobacillus acidophilus freeze-drying powder is crushed powdered, and add appropriate milk powder, oligosaccharide, maltodextrin, local flavor
Powdery product is made in agent etc., for daily consumption or as health product.
The therapeutic effect to infantile diarrhea for embodiment 6 lactobacillus preparation
Diarrhoeal diseasess are the multifactor diseases causing of cause of disease more than a group, are increased with times of defecation and change into feature with stool
One group of clinical syndrome, seriously can cause dehydration and electrolyte disturbance, this disease is often with abdominal distention, stomachache and acute gastrointestinal
The symptoms such as inflammation, to mankind's health risk greatly, are worldwide public health problem.Because sickness rate is high, children's health is endangered
Greatly, it is to cause one of major reason of infantile malnutrition, dysplasia and death, in many developing countries abdomen
Rushing down is first dead reason of children.
In line with voluntary principle, the boy baby that 60 are suffered from the eutrophy of 3-12 month of moderate dehydration acute diarrhea is carried out
Random, double blind, for evaluating the lactic acid bacteria tablet containing bacillus acidophilus kdb-03 provided by the present invention in treatment
Effect in infantile diarrhea.
Matched group: baby 30, takes not lactobacteria-containing Low lactose milk baby preparation；
Treatment group: baby 30, in addition to taking identical with matched group, equivalent Low lactose milk baby preparation, three times a day,
A piece of lactic acid bacteria tablet is provided every time.
Record the daily following data of each baby: defecation volume, stool and denseness and accurate body weight value.Abdomen
The persistent period rushed down (does not have in 24 hours afterwards until discharging last liquid or semiliquid feces after being defined as being admitted to hospital more again
Occur abnormal feces) hourage that passed through.
Statistical result showed, compared with matched group, the infantile diarrhea duration for the treatment of group significantly shorten (treatment group and
Matched group is respectively 1.63 days and 2.45 days, p < 0.05), this just becomes apparent from after the treatment of first day.And, treatment group
Baby through treatment after tend to that there is relatively low stool output (in terms of the grams of every kg body weight per day).Thus illustrating,
The lactic acid bacteria tablet containing bacillus acidophilus kdb-03 that the present invention provides has significant therapeutic effect to infantile diarrhea, can be clinically
Popularization and application.
Claims (8)
1. a kind of lactobacillus preparation is it is characterised in that described lactobacillus preparation comprises bacillus acidophilus (lactobacillus
acidophilus).
2. lactobacillus preparation as claimed in claim 1 is it is characterised in that described bacillus acidophilus' deposit number is cctcc
no:m2016429.
3. lactobacillus preparation as claimed in claim 1 or 2 is it is characterised in that described lactobacillus preparation is bacillus acidophilus
Lyophilized powder.
4. lactobacillus preparation as claimed in claim 1 or 2 is it is characterised in that described lactobacillus preparation also includes starch, hydroxyl
One of third cellulose, dead plaster, maltodextrin, oligofructose, flavoring agent or the combination of two or more.
5. lactobacillus preparation as claimed in claim 1 or 2 is it is characterised in that described lactobacillus preparation is capsule, tablet
Or powder.
6. application in treatment infantile diarrhea for the arbitrary described lactobacillus preparation of claim 1-5.
7. a kind of preparation method of lactobacillus preparation is it is characterised in that described preparation method comprises the steps:
1) liquid fermentation medium, sterilizing are prepared；
2) culture medium is preheated to 50-55 DEG C, carries out homogenizing under 20-25mpa pressure condition, TRANSIENT HIGH TEMPERATURE sterilizes；
3) bacillus acidophilus as described in 5% inoculation claim 1 or 2 of amount of fermentation, continue stirring 15min after having inoculated；
4) control 37-40 DEG C of fermentation temperature, ferment 10 hours；
5) fermentation liquid is centrifuged, vacuum lyophilization to water content is less than 3%, obtains final product lactobacillus preparation.
8. preparation method as claimed in claim 7 is it is characterised in that each component and its matter in described liquid fermentation medium
Amount volume ratio is respectively as follows: milk powder 10%, white sugar 0.5%, glucose 0.5%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610809305.6A CN106361778B (en) | 2016-09-08 | 2016-09-08 | A kind of lactobacillus preparation and its application in baby diarrhea treatment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610809305.6A CN106361778B (en) | 2016-09-08 | 2016-09-08 | A kind of lactobacillus preparation and its application in baby diarrhea treatment |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106361778A true CN106361778A (en) | 2017-02-01 |
| CN106361778B CN106361778B (en) | 2019-10-22 |
Family
ID=57900456
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610809305.6A Active CN106361778B (en) | 2016-09-08 | 2016-09-08 | A kind of lactobacillus preparation and its application in baby diarrhea treatment |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106361778B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628660A (en) * | 2018-06-22 | 2019-12-31 | 景岳生物科技(中国)有限公司 | Lactobacillus composition for inhibiting gastritis caused by gastric helicobacter pylori and application thereof |
| CN116083300A (en) * | 2022-12-08 | 2023-05-09 | 玫斯江苏宠物食品科技有限公司 | Probiotic composite preparation for preventing and treating canine diarrhea and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101693882A (en) * | 2009-10-29 | 2010-04-14 | 北京博益众科技有限公司 | Lactobacillus acidophilus freeze-drying composite protectant and application method thereof |
| CN105831534A (en) * | 2016-03-28 | 2016-08-10 | 济南康多宝生物技术有限公司 | Compound lactic acid bacteria beverage and a preparation method thereof |
-
2016
- 2016-09-08 CN CN201610809305.6A patent/CN106361778B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101693882A (en) * | 2009-10-29 | 2010-04-14 | 北京博益众科技有限公司 | Lactobacillus acidophilus freeze-drying composite protectant and application method thereof |
| CN105831534A (en) * | 2016-03-28 | 2016-08-10 | 济南康多宝生物技术有限公司 | Compound lactic acid bacteria beverage and a preparation method thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628660A (en) * | 2018-06-22 | 2019-12-31 | 景岳生物科技(中国)有限公司 | Lactobacillus composition for inhibiting gastritis caused by gastric helicobacter pylori and application thereof |
| CN110628660B (en) * | 2018-06-22 | 2021-06-22 | 景岳生物科技(中国)有限公司 | Lactobacillus composition for inhibiting gastritis caused by gastric helicobacter pylori and application thereof |
| CN116083300A (en) * | 2022-12-08 | 2023-05-09 | 玫斯江苏宠物食品科技有限公司 | Probiotic composite preparation for preventing and treating canine diarrhea and application thereof |
| CN116083300B (en) * | 2022-12-08 | 2023-10-17 | 玫斯江苏宠物食品科技有限公司 | Probiotic composite preparation for preventing and treating canine diarrhea and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106361778B (en) | 2019-10-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106399162B (en) | A kind of novel Lactobacillus casei and its application | |
| CN106350468B (en) | A kind of acidophilic lactobacillus | |
| CN106434433B (en) | A kind of novel fermentation lactobacillus and its application in sour milk beverage | |
| CN1297653C (en) | Composition comprising a lactobacillus pentosus strain and uses thereof | |
| CN100552016C (en) | The bifidus bacillus that is used for the treatment of inflammatory diseases | |
| CN105779343B (en) | One plant of lactobacillus plantarum for effectively removing cholesterol and free radical and its application | |
| CN103491969A (en) | Synbiotic compositions for restoration and reconstitution of gut microbiota | |
| CN101144065A (en) | Oxidation resistant Lactobacillus casei capable of resisting hydrogen peroxide and eliminating free radical, and use thereof | |
| CN101560523B (en) | Nutrient composition based on mixed fermented culture of five probiotics | |
| CN105831534B (en) | A kind of compound lactobacillus beverage and preparation method thereof | |
| CN110903995B (en) | Probiotic edible composition and food with digestion promoting effect | |
| CN103445048A (en) | Soybean oligosaccharide product containing probiotics and preparation method thereof | |
| CN105420150A (en) | Lactobacillus acidophilus and application thereof | |
| CN103289942B (en) | Antioxidative animal bifidobacterium and applications thereof | |
| CN106381278B (en) | A kind of lactobacillus preparation and its application in gastric ulcer treatment | |
| CN106417594B (en) | A kind of compound probiotic beverage and its application | |
| CN105802876B (en) | A kind of composite probiotics ferment clover tender shoots powder preparation and its preparation method and application | |
| CN104431352B (en) | A kind of preparation method and application of growth promotion symphysis unit bacterium solution | |
| CN106361778B (en) | A kind of lactobacillus preparation and its application in baby diarrhea treatment | |
| CN105559087A (en) | Probiotic product containing sialic acid and preparation method of probiotic product | |
| CN105567590A (en) | Enzymatic hydrolysis preparation technology of bifidobacteria and lactic acid bacteria mixed fermented bacteria and application thereof | |
| CN105505815A (en) | Lactobacillus mucosae with anti-aging function | |
| CN114921383B (en) | Probiotic preparation with cholesterol removal function and preparation method thereof | |
| CN105725215B (en) | A kind of lactic acid bacteria oral liquor and preparation method thereof | |
| CN114984065A (en) | Probiotic composition for improving immunity and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20200410 Address after: Mulberry road Licheng District 250131 in Shandong city of Ji'nan province No. 11 Co-patentee after: Inner Mongolia Yikang Health Development Co., Ltd Patentee after: JINAN KANGDUOBAO BIOTECHNOLOGY Co.,Ltd. Co-patentee after: Biotechnology Research Center, Shandong Academy of Agricultural Sciences Address before: Mulberry road Licheng District 250131 in Shandong city of Ji'nan province No. 11 Co-patentee before: Biotechnology Research Center, Shandong Academy of Agricultural Sciences Patentee before: JINAN KANGDUOBAO BIOTECHNOLOGY Co.,Ltd. |