CN102191192B - Animal Bifidobacterium and use method thereof - Google Patents
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Abstract
The invention relates to an animal Bifidobacterium and a use method thereof. The animal Bifidobacterium is the Bifidobactrium amimalis Subsp Lactis B174, the CGMCC No. is 4182, the conservation date is September 16, 2010 and the conservation unit is the China General Microbiological Culture Collection Center. The Bifidobactrium amimalis Subsp Lactis B174 can colonize in human body and resist the gastric acid and cholate of human body. Meanwhile, the strain has better inhibitory action on Escherichia coli and can survive for 30 days at the room temperature. The strain and other lactic acid bacteria have better synergic action for growth; the strain has better combination effect with lactic acid bacteria; the product has better flavor, can be used to prepare fermented yoghurt, animal feed additives and antibacterial solution and can be widely used in the fields of other foods and biological products.
Description
Technical field
The present invention relates to a kind of animal bifidobacteria and application method thereof.
Background technology
Bifidus bacillus is a typical probiotics in the human intestinal, is the human physiological bacterium of finding the earliest, and its growth and breeding is present among people's the life, is the probiotic bacterium of group energy field planting in the healthy subjects enteron aisle.Its existence can reduce system's pH value in the human body, and intestinal microflora is changed, and suppress and kill pathogenic bacteria, thus enhances human body immune disease-resistance power, bifidus bacillus can be synthesized multivitamin in enteron aisle.Relax bowel reducing cholesterol and triglyceride, anticancer effect, the anti-ageing effect of waiting for a long time.Therefore confirmed on science that bifidus bacillus quantity and quality are one of major criterions of HUMAN HEALTH in the human body.
The different physiological roles of bifidus bacillus can be used as the source of host's nitrogen.Bifidus bacillus is one type of obligatory anaerobic bacteria, and is high to the environment of oxygen and nutritional requirement, and it is low to produce sour power when being used to make fermented-milk, long between curdled milk, and belongs to opposite sex fermentation and make product special flavour not good enough.When thalline is acidproof, advance less than enteron aisle during the bile tolerance scarce capacity, will lose its effect.
Therefore, bring into play these prebiotic effects, must at first can tolerate human intestines and stomach's digestive process, can resist gastric juice and bile its effect of killing.Report foods such as Berrada under one's belt the time of emptying be 90min, pH value is 3.0 in the stomach, minimumly reaches 1.3, human small intestine's bile fluid concentration is between 0.03%-0.3%.
According to present relevant report, obtain can the cholate of anti-the human body the bifidus bacillus method have three kinds: the one, screening from nature (human excrement and urine), the 2nd, from spontaneous fermentation food, screen, the 3rd, tame having bacterial classification now.But existing bifidus bacillus because differing greatly between the bacterial strain, often can't can at room temperature survive possessing under the function of acidproof, cholate for a long time, also possess simultaneously antibacterial, can with multiple advantages such as milk-acid bacteria is composite.
Summary of the invention
The present invention provides a kind of leavened food that derives from; Acid, cholate, room temperature there is tolerance preferably; Coliform there is stronger inhibition effect, symplastic growth effect is preferably arranged with the milk-acid bacteria of other types, and better with the composite effect of milk-acid bacteria; Product has the animal bifidobacteria of preferable local flavor, and the application method of this animal bifidobacteria.
For realizing above-mentioned purpose, the present invention adopts following technical scheme:
This animal bifidobacteria; It is bifidobacterium animalis subspecies (Bifidobactrium animalis) B174; Preserving number: CGCC NO.4182; Preservation date: on September 16th, 2010, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
Said bifidobacterium animalis subspecies B174 has tolerance to human body hydrochloric acid in gastric juice and cholate.
Said bifidobacterium animalis subspecies B174 has tolerance to room temperature.
Said bifidobacterium animalis subspecies B174 has restraining effect to the intestinal bacteria flora.
One of said bifidobacterium animalis subspecies B174 and following milk-acid bacteria or more than one are composite, and the symplastic growth effect is arranged;
Lactobacterium acidophilum (Lactobacillus acidophius);
(Lactobacillus delbrueckii) plants in lactobacillus delbruckii Bulgaria;
Lactobacterium casei subspecies (Lactobacillus casei subsp.Casei);
Thermophilus streptococcus (Streptococcus thermophius);
Streptococcus acidi lactici (L.lactis);
Lactococcus lactis subspecies lactis (L.lactis subsp.lactis);
Plant lactobacillus (L.plantarum).
The best compounded combination of said bifidobacterium animalis subspecies B174 and milk-acid bacteria is: bifidobacterium animalis subspecies B174+ thermophilus streptococcus+Lactobacterium acidophilum, or bifidobacterium animalis subspecies B174+ thermophilus streptococcus+lactobicillus bulgaricus.
The application method of this animal bifidobacteria, the application of said bifidobacterium animalis subspecies B174 in preparation fermented yogurt, animal feedstuff additive and antibacterial liquid.
The preparation method of this animal bifidobacteria fermented yogurt; Respectively milk-acid bacteria and the described bifidobacterium animalis subspecies of claim 1 B174 are inoculated in the sterilization degreasing milk medium, 37 ℃ of constant temperature culture go out fermentation to curdled milk; 2～3 generations of activation; Be seeded to after the assembly in the sterilising fresh milk, 42 ℃ of fermentations, after the curdled milk as for preserving in the refrigerator.
The preparation method of this animal bifidobacteria antibacterial liquid; The bifidobacterium animalis subspecies B174 that places low temperature refrigerator is put into 40～50 ℃ of water-baths activation of thawing; Under aseptic condition, be inoculated in the MRS liquid nutrient medium 37 ℃ of constant temperature culture 12～14h, 2～3 generations of activation; The bifidobacterium animalis subspecies B174 fermented liquid that activation is good adopts the method for low-temperature centrifugation, and the preparation fermented supernatant fluid obtains the animal bifidobacteria antibacterial liquid.
The preparation method of this animal bifidobacteria fodder additives; With bifidobacterium animalis subspecies B174 and other probiotic bacterium assembly, the ratio between each probiotic bacterium is: bifidobacterium animalis subspecies B174: Lactobacillus acidophilus: streptococcus acidi lactici=(1～2): (2～3): (1～1.5).
Beneficial effect of the present invention is following:
Physiological condition seed selection one strain from spontaneous fermentation food that the present invention is directed to the human body stomach can tolerate the animal bifidobacteria of human body hydrochloric acid in gastric juice and gallbladder salinity, through being accredited as bifidus bacillus breast subspecies.Animal bifidobacteria of the present invention can field planting in human body, the performance prebiotic effect.This bacterial strain has good inhibitory effect to colibacillus simultaneously, can play and regulate enteric microorganism flora function.In addition, bifidobacterium animalis subspecies B174 of the present invention at room temperature can be survived 30 days, had enlarged the range of application of bifidus bacillus, had prolonged the shelf-lives of bifidus bacillus product, can reduce production costs.The milk-acid bacteria of bifidobacterium animalis subspecies B174 of the present invention and other types has symplastic growth effect preferably, and better with the composite effect of milk-acid bacteria, and product has preferable local flavor.
Bifidobacterium animalis subspecies B174 of the present invention has tolerance to human body stomach environment, and survival is 2 hours under SGF PH=2.0 condition, and viable count is from 2.52 * 10
8Cfu/ml drops to 3.06 * 10
6/ ml is less than two one magnitude.And under the PH=1.3 condition, still have viable bacteria to exist after 2 hours.
Bifidobacterium animalis subspecies B174 of the present invention has tolerance to cholate.Survival is 24 hours under the environment of simulated intestinal fluid gallbladder salinity 0.2%, and viable count shows a rising trend, from 1.5 * 10
7Cfu/ml to 9.4 * 10
7Cfu/ml.
Bifidobacterium animalis subspecies B174 of the present invention has tolerance to room temperature, at room temperature survives 30 days, and viable count is from 4.8 * 10
9Cfu/ml only drops to 3 * 10
8Cfu/ml still has a large amount of viable bacterias to exist.
Bifidobacterium animalis subspecies B174 of the present invention can be applicable to prepare fermented yogurt, animal feedstuff additive and antibacterial liquid, also can be widely used in other food and field of biological product.
Embodiment
Embodiment; This animal bifidobacteria; It is bifidobacterium animalis subspecies (Bifidobactrium animalis) B174, preserving number: CGCC NO.4182, preservation date: on September 16th, 2010; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
Separation and the authentication method of bifidobacterium animalis subspecies B174 are following:
1, separation method
Take by weighing in the aseptic buffer salt solution of sample (spontaneous fermentation food) 5g to 45ml (adding an amount of granulated glass sphere) as mother liquor; The abundant mixing of concussion 10min is done gradient dilution with aseptic buffer salt solution, selects three suitable gradients; Draw 0.1ml and in the NPNL-BL substratum, roll the pipe separation; Each dilution gradient is done two repetitions, and the anaerobism pipe is put into 37 ℃ of incubators and cultivated 48～72 hours, selects smooth, protruding garden, neat in edge, white or oyster white bacterium colony microscopy.Doubtful bacterium colony is carried out this separation, purifying by above-mentioned steps again in the BL substratum, up to the complete purifying of bacterium colony.
Morphology is identified: gramstaining, cellular form has or not gemma
Physiology and biochemistry is identified: catalase, and oxydase, fructose-6-phosphoric acid salt phosphoric acid cuprase, glucide produces acid
Molecular biology identification: 16SrDNA sequential analysis
2. experimental result
2.1 physio-biochemical characteristics and morphological specificity
The physio-biochemical characteristics of table 1 B174
2.2 16SrDNA sequencing
Make up the phylogenetic tree of this bacterial strain according to above-mentioned 16SrDNA sequencing result, qualification result B174 is the bifidobacterium animalis subspecies.
3. qualification result
In conjunction with above qualification result, animal bifidobacteria of the present invention is the bifidobacterium animalis subspecies.
Acidproof, bile tolerance, the heatproof of bifidobacterium animalis subspecies B174 are tested as follows:
1. experimental technique
1.1 cholate resistance test
Bifidobacterium animalis subspecies B174 cultivates 15h with 37 ℃ of strictly anaerobics of modified MRS nutrient solution of sterilization.The activation strain cultured solution is inserted with 10% inoculum size in the aseptic MRS liquid culture medium that contains the various biliary salt concn (gallbladder salinity is 0.2%, 0.3%, 0.5%, 1.0%, 1.6%), is contrast with the modified MRS nutrient solution that does not contain cholate simultaneously.Sampling and measuring viable count after 37 ℃ of anaerobism are cultivated 0,2,4,6,24 hour.
2.2 sour tolerance test
Animal bifidobacteria B174 cultivates 15h with 37 ℃ of strictly anaerobics of aseptic modified MRS nutrient solution; The activatory nutrient solution is inserted with 10% inoculum size (PH is respectively 1.3 in the aseptic modified MRS of different PH; 2.0,3.0), the nutrient solution with PH7.0 is contrast simultaneously.37 ℃ of anaerobism are cultivated, respectively at 0min 30min 60min 90min 120min. sampling and measuring viable bacteria.
2.3 room temperature tolerance test
Animal bifidobacteria such as subspecies B174 change the room temperature held after cultivating 15 h with 37 ℃ of aseptic modified MRS nutrient solutions, measure the viable count of a nutrient solution every three days, do contrast with 0 day viable count.
3. experimental result
3.1 bile tolerance experimental result
B174 sees table 2 to the tolerance of various biliary salt concn, gallbladder salinity smaller or equal to 0.3% o'clock to its unrestraint effect, viable count constantly increases in the 6h, growth has certain tolerance in the gallbladder salinity of 0.5%-1.6%.
Table 2 B174 is to the tolerance of cholate
| Gallbladder salinity | 0h (cfu/ml) | 2h | 4h | 6h | 24h |
| 0 | 1.54 * 10 7 | 7.84 * 10 8 | 1.25 * 10 9 | 4.37 * 10 9 | 1.05 * 10 9 |
| 0.2% | 1.54 * 10 7 | 1.52 * 10 8 | 2.17 * 10 8 | 2.2 * 10 8 | 1.84 * 10 8 |
| 0.3% | 1.54 * 10 7 | 1.4 * 10 8 | 1.7 * 10 8 | 1.25 * 10 8 | 9.4 * 10 7 |
| 0.5% | 1.54 * 10 7 | 3.05 * 10 7 | 2.95 * 10 7 | 2.08 * 10 7 | 8.4 * 10 6 |
| 1.0% | 1.54 * 10 7 | 1.87 * 10 6 | 3.5 * 10 6 | 1.85 * 10 6 | 9.8 * 10 5 |
| 1.6% | 1.54 * 10 7 | 1.25 * 10 6 | 3.35 * 10 6 | 1.49 * 10 6 | 6.3 * 10 5 |
3.2 acidproof experimental result
B174 viable count in different acid nutrient solutions changes sees table 3, under the PH2.0 condition, cultivates and still has a large amount of viable bacterias existence in 2 hours, explains that this bacterial strain has very strong tolerance to acid.
Table 3 B174 is to the tolerance of acid
| 0min(cfu/ml) | 30min | 60min | 90min | 120min | |
| PH7.0 | 2.52×10 8 | 3.31×10 8 | 3.5×10 8 | 3.6×10 8 | 3.8×10 8 |
| PH 3.0 | 2.52×10 8 | 2.48×10 8 | 1.71×10 8 | 1.73×10 8 | 1.76×10 8 |
| PH 2.0 | 2.52×10 8 | 1.28×10 8 | 3.06×10 | 1.25×10 7 | 3.06×10 6 |
| PH 1.3 | 2.52×10 8 | 4.9×10 6 | 2.2×10 5 | 6.0×10 3 | 1.0×10 3 |
3.3 heatproof experimental result
General bifidobacterium preparations preservation period at room temperature is about 7 days, and B174 still had more viable bacteria to exist in 30 days in the room temperature placement, had prolonged the shelf-lives of this bacterium greatly, and under the room temperature preservation condition, the variation of viable count and the relation of fate are seen table 4.
Table 4 B174 is to the tolerance of room temperature
| Store fate (d) | Viable count (CFU/ML) |
| 0 | 4.8 * 10 9 |
| 6 | 1.83 * 10 9 |
| 12 | 1.2 * 10 9 |
| 18 | 1.0 * 10 9 |
| 22 | 1.3 * 10 9 |
| 27 | 5.3 * 10 9 |
| 30 | 3.0 * 10 9 |
This experiment separates the bifidobacterium animalis subspecies B174 that obtains all has good tolerance to acid, cholate and room temperature.PH value is between 2-3 in people's the stomach, eats minimumly behind the acidic food to reach 1.5.B174 was survived 2 hours in the sour environment of PH=2, and viable count is from 2.52x10
8Cfu/ml drops to 3.06x10
6Cfu/ml, two one magnitude have only descended; And under the acidic conditions of PH=3, the order of magnitude of viable count does not change after two hours.And food (especially fluid) is shorter relatively through the time of stomach, and general 1～2h just gets into small intestine.So the acid resistance of bifidus bacillus breast subspecies B174 can guarantee that it has a considerable amount of viable bacterias to arrive enteron aisle through stomach.
Human small intestine's gallbladder salinity is between 0.03-0.3%, and gallbladder salinity did not have restraining effect less than 0.3% o'clock to B174 in this experiment, and viable count constantly increases in the 6h.B174 has very strong tolerance to cholate, can arrive large intestine through small intestine.
The product room temperature quality guaranteed period of generally containing bifidus bacillus is about a week, and this experiment B174 deposited 30 days in room temperature, still had very high viable count, can increase the product shelf-lives, reduced cost.
Bifidobacterium animalis subspecies B174 is following to the biological antagonist property experiment of intestinal bacteria, milk-acid bacteria:
1. materials and methods
1.1 bacterial classification
Animal bifidobacteria B174, intestinal bacteria, Lactobacterium acidophilum, lactobacterium casei, plant lactobacillus, streptococcus acidi lactici, lactobacillus delbruckii Bulgaria plants, JCM 1192T, bifidobacterium adolescentis, bifidumbacterium bifidum all come from this laboratory.
1.2 substratum
MRS liquid and solid medium, tomato juice solid and liquid nutrient medium, nutrient broth solid and liquid nutrient medium
1.3 actication of culture
Bifidus bacillus activation: will place the bifidus bacillus of low temperature refrigerator to be put into 40～50 ℃ of water-baths and thaw, and under aseptic condition, draw 0.1ml and be inoculated in the MRS liquid nutrient medium, 37 ℃ of constant temperature culture 12～14h, 2～3 generations of activation.
The activation of milk-acid bacteria: same bifidus bacillus, lactobacillus spp are adopted the activation of MRS liquid nutrient medium, and streptococcus acidi lactici adopts the activation of tomato juice substratum.
Colibacillary activation: will place the intestinal bacteria in the refrigerator to be transferred to the liquid nutritional broth culture, 37 ℃ of constant temperature culture 12 h, two generations of activation according to 2% inoculum size.
1.4 coating nutrient agar medium plate
The intestinal bacteria that activation is good are applied on the nutrient agar plate with coating method uniformly, and the viable bacteria amount when requiring coating is 10
6Cfu/ml.
1.5 tilt-pour process is cultivated milk-acid bacteria
Lactobacillus spp that activation is good and streptococcus acidi lactici respectively with tilt-pour process be layered on uniformly on the MRS flat board with the tomato juice solid plate on, the viable bacteria amount when requiring coating is 10
6Cfu/ml.
1.6 the preparation of bifidus bacillus fermented liquid
The bifidus bacillus fermented liquid that activation is good adopts the method for low-temperature centrifugation, the preparation fermented supernatant fluid, and centrifugal condition is: 4 ℃, 7500rpm, 5min.
1.7 the bifidus bacillus fermented liquid fungistatic effect is observed
Oxford agar diffusion method: with nutrient agar plate, MRS solid plate and the tomato juice solid plate seasoning 30min of the indicator of completing; Under aseptic condition, Oxford cup (internal diameter is 6mm) evenly is positioned on the flat board again, presses down a little, make itself and plate culture medium contact surface tight; Draw the 0.2ml fermented supernatant fluid then in cup; 37 ℃ of constant temperature culture 24h, the size of observing inhibition zone, and measure antibacterial circle diameter.Each sample do three parallel, results averaged.Control group adds MRS liquid nutrient medium 0.2ml in the cup of Oxford, culture condition is identical.
1.8 experimental result
The result shows that most of bifidus bacillus all has restraining effect to coliform, and the inhibition zone transparency is high, and antibacterial circle diameter is bigger.Milk-acid bacteria also shows antibacterial phenomenon, but the part inhibition zone is transparent inadequately, and of the present invention particularly evident to the animal bifidobacteria fungistatic effect, and concrete outcome is seen table 5.
Experiment shows, the animal bifidobacteria under the present invention and most of milk-acid bacteria can both symplastic growths, and this has enlarged the range of application of bifidus bacillus, and concrete outcome is seen table 6.
Table 5. bifidobacterium animalis subspecies B174, milk-acid bacteria and colibacillary antagonistic effect
The compatibility of table 6. animal bifidobacteria and milk-acid bacteria (antagonistic effect)
Preparation method between bifidobacterium animalis subspecies B174 and the milk-acid bacteria is following:
1. material
Bacterial classification: make up between bifidobacterium animalis subspecies B174 and the milk-acid bacteria, be re-dubbed fermented yogurt.
Material: the fresh milk of three tyndallizations, sterilization skimmed milk substratum.
2. experimental technique
2.1 preparation method
Respectively with bifidus bacillus and lactobacillus inoculum in the sterilization degreasing milk medium, 37 ℃ of constant temperature culture go out fermentation to curdled milk; In 2～3 generations of activation, the ratio assembly according to different is seeded in the sterilising fresh milk; 42 ℃ of fermentations go out fermentation after the curdled milk, preserve as for 4 ℃ of refrigerators; Carry out sensory evaluation and flavour substances behind the 12h and detect, mainly comprise acetaldehyde and dimethyl diketone Determination on content.
2.2 the measuring method of acetaldehyde
2.2.1 reagent: (1) 1% Starch Indicator (2) 1.2% sodium sulfite solution (3) 0.1N hydrochloric acid (4) 1M carbonic acid hydracid (5) 0.1N iodine liquid (6) 0.01N iodine liquid.
2.2.2 method
100m sour milk sample is placed matrass, add 50ml water and distill, get the 100ml distillate in the 350ml iodine flask, add 1.2% sodium sulfite solution 15ml; 0.1N hydrochloric acid 7ml, shake up 1h in the dark place, add 1% Starch Indicator 2ml, fixed with 0.1N iodine drop; During neighbour's terminal point, use 0.01N iodine reference liquid titration to light blue violet instead and occur, add 1M sodium hydrogen carbonate solution 20ml; Crack bottle stopper sways 0.5min, till occurring once more to lavender with 0.01N iodine standard drop.Make blank as stated above with 100ml fresh milk sample simultaneously.
Acetaldehyde (ppm)
X=[(V
1-V
2)N×0.022]/100
V
1: iodine reference liquid consumption
V
2: blank iodine reference liquid consumption
N: the equivalent concentration of iodine reference liquid
0.022: the quality of every milliliter of equivalent acetaldehyde (g)
100: style amount (ml).
2.3 the measuring method of dimethyl diketone
The mensuration of typical curve
Prepare the dimethyl diketone solution of 20 mg/L, as standard, the preparation mass concentration is 1.0,2.0,4.0,8.0,12.0,16.0, the solution of 20.0mg/L.
Respectively get the solution that two tube comparison tubess add the above-mentioned mass concentration of 10 ml respectively, to wherein 1 add 0.5 ml, 1% o-phenylenediamine solution, fully shake up, another does not add, and places dark place 20-30 minute, adds 2ml and 2.5ml 4mol/L HCL solution mixing again.And then the 335nm place, make reference with the solution that does not add O-Phenylene Diamine.
With ultraviolet spectrometry degree instrumentation light absorption value.
2.3.1 the measuring method of di-acetyl in the sour milk
Operation steps:
⑴ distillation: in 25 ml graduated cylinders, put 2ml left and right sides zero(ppm) water, place the distiller condenser lower end mouth of pipe.In the 250ml triangular flask, add 100ml sour milk sample, add 50ml water again, put into pre-heated boiling water rapidly, distill, take off graduated cylinder during near 25ml, complement to 250ml with zero(ppm) water up to distillate.
⑵ reaction: mix distillate, draw 10ml respectively and add in two tube comparison tubess, first pipe is put into 0.5ml 1% o-phenylenediamine solution, and thorough mixing places dark place 20min, adds 4N HCL solution 2ml again; Second pipe adds 4N HCL solution 2.5ml
Mixing.
⑶ measure:, use the 2cm cuvette to inhale second pipe and be blank in the 335nm place with ultraviolet spectrophotometer, measure the absorption value of first pipe (being sample).This mensuration should be accomplished in 20min.
⑷ calculate: di-acetyl (PPM)=E * 1.2-0.01
The E-absorption value
1.2-reduction factor
0.01 – IC
Reagent: 1. 1% O-Phenylene Diamine: accurately take by weighing O-Phenylene Diamine 0.25 height, be dissolved among the 4N HCL, place in the 25ml volumetric flask, complement to scale, be stored in the brown bottle with 4N HCL, interim with preparation.
2. 4N HCL: get the 34ml concentrated hydrochloric acid, be diluted with water to 100ml.
3. experimental result
Judge test through antagonistic effect and composite; From 39 groups of compounded combination, choose two groups, sense organ state, the combination of better flavor; Be respectively animal bifidobacteria+thermophilus streptococcus+Lactobacterium acidophilum; Animal bifidobacteria+thermophilus streptococcus+lactobicillus bulgaricus, and measure the two acetaldehyde and dimethyl diketone content, can be applicable in the fermented yogurt to prove this bifidus bacillus.Concrete outcome is seen table 7
Table 7 acetaldehyde and dimethyl diketone assay result
| Strain combinations | Acetaldehyde (ppm) | Dimethyl diketone content (ppm) |
| Animal bifidobacteria+thermophilus streptococcus+Lactobacterium acidophilum | 22.45 | 0.69 |
| Animal bifidobacteria+thermophilus streptococcus+lactobicillus bulgaricus | 24.29 | 0.76 |
The best sour milk of local flavor is given the credit to acetaldehyde at 23～41ppm, and when the ratio of acetaldehyde and dimethyl diketone surpassed 2.8, the sour milk local flavor was best.More than two groups record that flavour substances content meets these two conditions and the sensory evaluation result is consistent.
The method that bifidobacterium animalis subspecies B174 is used to prepare fodder additives is following:
Bifidobacterium animalis subspecies B174 prebiotic effect of the present invention is obvious, can with other probiotic bacterium assembly, be used for animal-feed.Test shows that composite probiotics preparations can effectively be avoided the colibacillary poison of attacking, and improves colibacillary resistibility.Ratio between each probiotic bacterium is: bifidobacterium animalis subspecies B174: Lactobacillus acidophilus: streptococcus acidi lactici=1.5:2.5:1.
Claims (10)
1. animal bifidobacteria; It is characterized in that: it is bifidobacterium animalis subspecies (Bifidobacterium animalis) B174; Preserving number: CGMCC No.4182; Preservation date: on September 16th, 2010, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center.
2. animal bifidobacteria according to claim 1 is characterized in that: said bifidobacterium animalis subspecies B174 has tolerance to human body hydrochloric acid in gastric juice and cholate.
3. animal bifidobacteria according to claim 1 is characterized in that: said bifidobacterium animalis subspecies B174 has tolerance to room temperature.
4. animal bifidobacteria according to claim 1 is characterized in that: said bifidobacterium animalis subspecies B174 has restraining effect to the intestinal bacteria flora.
5. animal bifidobacteria according to claim 1 is characterized in that: one of said bifidobacterium animalis subspecies B174 and following milk-acid bacteria or more than one are composite, and the symplastic growth effect is arranged;
Lactobacterium acidophilum (Lactobacillus acidophius);
(Lactobacillus delbrueckii) plants in lactobacillus delbruckii Bulgaria;
Thermophilus streptococcus (Streptococcus thermophius);
Streptococcus acidi lactici (L.lactis);
Lactococcus lactis subspecies lactis (L.lactis subsp.lactis).
6. animal bifidobacteria according to claim 5; It is characterized in that: the best compounded combination of said bifidobacterium animalis subspecies B174 and milk-acid bacteria is: bifidobacterium animalis subspecies B174+ thermophilus streptococcus+Lactobacterium acidophilum, or bifidobacterium animalis subspecies B174+ thermophilus streptococcus+lactobicillus bulgaricus.
7. the application method like said any one animal bifidobacteria of claim 1～6 is characterized in that: the application of said bifidobacterium animalis subspecies B174 in preparation fermented yogurt, animal feedstuff additive and intestinal bacteria antibacterial liquid.
8. the preparation method of an animal bifidobacteria fermented yogurt; It is characterized in that: respectively milk-acid bacteria and the described bifidobacterium animalis subspecies of claim 1 B174 are inoculated in the sterilization degreasing milk medium, 37 ℃ of constant temperature culture go out fermentation to curdled milk; 2～3 generations of activation; Be seeded to after the assembly in the sterilising fresh milk, 42 ℃ of fermentations, after the curdled milk as for preserving in the refrigerator.
9. the preparation method of an animal bifidobacteria antibacterial liquid; It is characterized in that: will place the bifidobacterium animalis subspecies B174 of low temperature refrigerator to be put into 40～50 ℃ of water-baths activation of thawing; Under aseptic condition, be inoculated in the MRS liquid nutrient medium 37 ℃ of constant temperature culture 12～14h, 2～3 generations of activation; The bifidobacterium animalis subspecies B174 fermented liquid that activation is good adopts the method for low-temperature centrifugation, and the preparation fermented supernatant fluid obtains animal bifidobacteria intestinal bacteria antibacterial liquid.
10. the preparation method of an animal bifidobacteria fodder additives; It is characterized in that: with bifidobacterium animalis subspecies B174 and other probiotic bacterium assembly, the ratio between each probiotic bacterium is: bifidobacterium animalis subspecies B174: Lactobacillus acidophilus: streptococcus acidi lactici=(1～2): (2～3): (1～1.5).
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| CN103275893A (en) * | 2013-05-06 | 2013-09-04 | 哈尔滨美华生物技术股份有限公司 | Complex microbial inoculants of bifidobacterium longum and bifidobacterium animalis and preparation method of complex microbial inoculants |
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| CN116747245B (en) * | 2023-06-14 | 2024-06-21 | 辽宁越秀辉山营养科技有限公司 | Application of bifidobacterium animalis subspecies lactis BX-245 in bacteriostasis and/or production of functional active substances |
| CN116836887B (en) * | 2023-08-09 | 2024-01-02 | 安徽善和生物科技有限公司 | High-activity high-stability starter and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101260377A (en) * | 2008-03-14 | 2008-09-10 | 内蒙古蒙牛乳业（集团）股份有限公司 | Animal bifidobacteria and use thereof |
| CN101262779A (en) * | 2005-06-14 | 2008-09-10 | 爱尔伯股份公司 | Probiotic health or fitness promoting human or animal foodstuff, feed, and/or drinking water additive and use thereof |
| CN101715303A (en) * | 2007-04-27 | 2010-05-26 | 格伦多拉无添加果汁公司 | Fruit drink with probio |
-
2010
- 2010-12-21 CN CN201010598683A patent/CN102191192B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101262779A (en) * | 2005-06-14 | 2008-09-10 | 爱尔伯股份公司 | Probiotic health or fitness promoting human or animal foodstuff, feed, and/or drinking water additive and use thereof |
| CN101715303A (en) * | 2007-04-27 | 2010-05-26 | 格伦多拉无添加果汁公司 | Fruit drink with probio |
| CN101260377A (en) * | 2008-03-14 | 2008-09-10 | 内蒙古蒙牛乳业（集团）股份有限公司 | Animal bifidobacteria and use thereof |
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