CN109170235A - Probiotic microcapsule and the preparation method and application thereof - Google Patents
Probiotic microcapsule and the preparation method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
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- A—HUMAN NECESSITIES
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- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
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- A23K20/163—Sugars; Polysaccharides
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K40/00—Shaping or working-up of animal feeding-stuffs
- A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
- A23K40/35—Making capsules specially adapted for ruminants
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Abstract
The present invention provides a kind of probiotic microcapsule and the preparation method and application thereof.The present invention is directed to the Micro-Encapsulation Technique of ruminant, using the natural controlled-release material with good biocompatibility and high molecular material and mild technique to pH sensitive, carries out microencapsulation preparation to probiotic cell.It is granulated using microcrystalline cellulose blank pellet, as the skinning core particles that pellet is granulated, not only effectively solves the problems, such as that proper sphere degree is low in granulation, granulation uniformity is poor, but also have both the effect of dilution-three mixture of bonding-disintegration.The erosion of rumen microorganism decomposition and abomasum gastric acid can be resistant to respectively using the double-deck coating technique, be a kind of method that promotion microorganism is effectively colonized in milk cow enteron aisle so that thallus as much as possible be made to reach enteron aisle.The method of the present invention is easy to operate, and industrialized microcapsules may be implemented by fluidized bed bilayer coating and prepare, and is coated with probiotic cell for the factorial production, establishes a set of effective technological parameter.
Description
Technical field
The present invention relates to feed additive fields, specifically, being related to probiotic microcapsule and the preparation method and application thereof.
Background technique
Used antibiotic nearly 60 years in feed, significant effect, it is cheap and by favor, but also bring simultaneously
Hidden danger, as in drug resistance problems, livestock products residual and food safety risk, cause animal immune inhibit and suprainfection, resist
The use of raw element has become global problems.European Union just completely forbade from 2006 and uses antibiotic in feed, and feed is prohibited anti-
Afterwards, the main substitute probiotics of antibiotic are applied and are given birth to.It is well known that now widely used one kind Tiny ecosystem viable bacteria
Preparation is mainly developed with a variety of probiotics such as lactic acid bacteria, Bifidobacterium, enterobacteria and bud pole bacterium, main function
It is to supplement the microflora for reducing or lacking in enteron aisle in quantity or type, restores the microecological balance in enteron aisle, in turn
Microorganism is promoted to stick in enteron aisle, be colonized, breeding to achieve the purpose that health care.
These inoculating microbes will be colonized in animal intestinal tract, must just be resistant to the effect of gastric acid, cholate, digestive ferment etc..It is right
For ruminant, it is also necessary to overcome the decomposition of rumen microorganism.Meanwhile as a kind of Tiny ecosystem active bacteria formulation normal
Temperature storage easily loses activity, and reduces the shelf life of product.There are many methods at present to improve microorganism to poor environment
Resistance, wherein the use of microcapsules coating technique is rather extensive, becomes one of new and high technology of focus development.It is obtained after coating high
Concentration, the microencapsulation probiotic products of high activity can go directly enteron aisle fixed point release, and quick occupy-place field planting effectively plays prebiotic work
With.
According to it has been reported that microcapsules are coated with the methods of coating before mostly using emulsion process, extrusion, spray drying process, fermentation.
CN1969889A discloses a kind of method that extrusion prepares enteric-coated multilayer encapsulated probiotic microcapsule, and most literature is more newborn
The advantage and disadvantage of change method and extrusion, it is believed that it is relatively difficult that extrusion forms sphericity when preparing microcapsules, and is unfavorable for outer layer
Coating；In general the partial size for preparing microcapsules using extrusion is bigger (2-3mm), and it is dynamic that excessive particle is suitable only for simple stomach
Object will pass through ruminant the grinding of omasum, be easy microcapsules is made to burst apart and the protective effect that loses coating.
CN105105144A discloses a kind of method for preparing microcapsules using emulsion process, but the embedding rate of emulsion process itself is lower,
And excessively high shearing force also can generate injury to probiotic cell in emulsion process.CN103283975A discloses a kind of spray
The dry method for preparing probiotic microcapsule of mist, this method is little to microbiological effect resistant to high temperature, but thermally sensitive cream
For sour bacterium, the temperature of spray dryer is more than 40 DEG C or more, and the viable bacteria rate of lactic acid bacteria will decline, in actual operation, temperature
Every 10 DEG C of raising is spent, viable bacteria rate will decline an order of magnitude.CN103275966A is coated with before disclosing a kind of fermentation, passes through coating
Being further cultured for control the microencapsulation technology of growing microorganism and metabolism and obtain good development afterwards, but preparation cost is high.
Summary of the invention
The present invention is insufficient for the above prior art, a kind of probiotic microcapsule is provided, especially for ruminant
Microcapsules and the preparation method and application thereof.
In order to achieve the object of the present invention, in a first aspect, the present invention provides a kind of preparation method of probiotic microcapsule, including
Following steps:
A, the preparation of probiotics bacterium mud mixed liquor: probiotics being activated, is cultivated, and obtains probiotics fermention liquid, centrifugation,
Supernatant is abandoned, thallus is collected, obtains bacterium mud；Appropriate skimmed milk power protection liquid is added into the bacterium mud, obtains bacterium mud mixed liquor；
B, it pelletizes: being added in fluidized bed, make using microcrystalline cellulose blank pellet (MCC) and sodium alginate as skinning core particles
The bacterium mud mixed liquor injects in the fluidized bed according to certain flow rate, under certain condition mixing granulation；
C, primary coating: by particle obtained by a coating solution even application to step B, a coated granule is obtained；
D, secondary coating: secondary coating on particle obtained by secondary coating solution even application to step C, will be completed, after dry
Up to probiotic microcapsule finished product.
Wherein, a coating solution be 1.5-2% sodium alginate aqueous solution or 1.5-2% sodium alginate and
Chitosan aqueous solution (mass ratio of sodium alginate and chitosan is 2-4:1).
The secondary coating solution is the acrylic resin aqueous solution of 10-12%, the ethyl cellulose solution of 1-5% or hydrogenation
Grease (such as C18H36O2)。
In granulation, primary coating and secondary coating process, mixed material temperature is controlled at 25-40 DEG C.The temperature is
Probiotics is suitable for the optimum temperature of breeding, can effectively avoid high temperature and significantly destroys to probiotics viable bacteria rate.
The volume ratio of method above-mentioned, bacterium mud described in step A and skimmed milk power protection liquid is 1:1-3.
Preferably, the skimmed milk power protects the concentration of liquid for 10%-30%, and preferably 20%.
The amount ratio of method above-mentioned, skinning core particles described in step B and bacterium mud mixed liquor is 1g:1-2mL, preferably 1g:
1mL。
Preferably, microcrystalline cellulose blank pellet and the weight ratio of sodium alginate are 11:1-5 in the skinning core particles, excellent
Select 11:1.
Preferably, the condition of step B progress fluidized bed granulation is as follows:
Note: heat engine purpose is that temperature of charge is made to reach 30 DEG C or more；Air quantity and flow velocity are adjusted depending on granulation situation, if
When heat engine or granulation beginning air quantity is excessive, is easy to blow MCC into high (exceeding spray head), and material damage is caused to increase and pelletize uneven
It is even.If the flow velocity of bacterium mud mixing at the beginning is excessive, it is added excessively, moisture can not be taken away in time under small air quantity, make MCC powder
Agglomeration rapidly, influences to pelletize.
Preferably, the sodium alginate aqueous solution that a coating coating solution used is 1.5-2%, secondary coating solution are 10-
12% acrylic resin aqueous solution.
Preferably, it is as follows to carry out the condition that fluidized bed is once coated by step C:
| Sample volume | Intake | It is coated flow velocity | Atomizing pressure | Temperature of charge | Inlet air temperature |
| Heat engine | 20m3/h | ---- | ---- | To 30 DEG C | 70℃ |
| 0-10ml | 20-30m3/h | 6-8r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| 10-20ml | 30-38m3/h | 8-12r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| 20-32ml | 38-45m3/h | 12-18r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
Wherein, the amount ratio of the particle and coating solution is 1.5-2g:1mL, preferably 2g:1mL.
The condition that step D carries out the secondary coating of fluidized bed is as follows:
| Sample volume | Intake | It is coated flow velocity | Atomizing pressure | Temperature of charge | Inlet air temperature |
| 0-5ml | 55m3/h | 15r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| 5-16ml | 55m3/h | 16r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| Dry 10min | 50m3/h | ---- | ---- | 31-40℃ | Closing temperature |
Wherein, the amount ratio of a coated granule and coating solution is 2-4g:1mL, preferably 4g:1mL.Secondary coating knot
Shu Hou closes fluidized bed, utilizes the dry 10min of the residual temperature of instrument.
Probiotics of the present invention includes lactic acid bacteria and its adheres to separately, as Lactobacillus casei, lactobacillus plantarum, streptococcus fecalis,
Bacillus licheniformis, bacillus subtilis, bifidobacterium bifidum, lactoenterococcus, lactobacillus acidophilus, lactoenterococcus, lactic acid
Lactobacillus, Pediococcus acidilactici, lactobacillus bulgaricus etc..
Second aspect, the present invention provide the probiotic microcapsule prepared according to the method described above.
The third aspect, the present invention provide following any application of the probiotic microcapsule:
1) application in feed preparation；
2) application in probiotics, especially Ruminat s Microecological Agent preparation；
3) restore the application in enteron aisle in microecological balance.
By above-mentioned technical proposal, the present invention at least have following advantages and the utility model has the advantages that
(1) the present invention provides a kind of Micro-Encapsulation Techniques for ruminant, using compatible with good biological
Property natural controlled-release material and high molecular material and mild technique to pH sensitive, micro-capsule is carried out to probiotic cell
Change preparation.
(2) the method for the present invention is easy to operate, and industrialized microcapsules may be implemented by fluidized bed bilayer coating and prepare,
It is coated with probiotic cell for the factorial production, establishes a set of effective technological parameter.
(3) heat labile problem in lactic acid bacteria pelletization is successfully solved using low temperature cold circulator；Using porous
The stability of granulation can be improved as carrier matrix in microcrystalline cellulose cellulose crystal, and has preferable mobility, the water suction of appropriateness
Property, higher mechanical strength and sphericity.
(4) it is directed to the distinctive physiological structure of ruminant, is coated with using the packaging material to pH sensitive, after coating
Microcapsules not only can be to avoid the decomposition of rumen microorganism, but also can prevent the destruction of abomasum gastric acid, promotes microorganism
It is discharged in milk cow intestinal-specific, promotes Microorganism colonization in enteron aisle.
(5) present invention carries out cold temperature to fluidized bed and handles, develop a kind of temperature using the lactic acid bacteria of thermal sensitivity as core-clad material
And technique, it is applicable not only to lactic acid bacteria, and equally effective to the coating of other high temperature resistant probiotics.Using microcrystalline cellulose sky
White capsule core is granulated, and as the skinning core particles that pellet is granulated, not only effectively solves that proper sphere degree (roundness) is low in granulation, particle is uniform
The problem of difference is spent, and has both the effect of dilution-three mixture of bonding-disintegration.It can be resistant to respectively using the double-deck coating technique
Rumen microorganism is decomposed and the erosion of abomasum gastric acid, is a kind of promotion microorganism to make thallus arrival enteron aisle as much as possible
In the method that milk cow enteron aisle is effectively colonized.Using animal vivo test, (in vivo studies is the gold mark for assessing probiotics coating effect
It is quasi-), actual response lactic acid bacteria plays the physiology course of prebiotic effect experience.
(6) functional using probiotic microcapsule uniform particle sizes, encystation made of fluidized bed coating technique of the present invention,
Operating procedure is simple, is conducive to large-scale production, can be widely applied to food, pharmacy, chemical industry and feed preparation field.
(7) use microcrystalline cellulose as skinning core particles in pelletization, it is therefore an objective in lactic acid bacteria coating, Ke Yi
Outside one approximate circle thallus, equably coating solution is sprayed up.And conventional method is mostly by means of mold, so that bacterium
Mud is squeezed into almost spherical, but this method is larger to the shearing force of thallus, and viable bacteria rate and embedding rate is made to be affected.This hair
It is bright used by microcrystalline cellulose blank pellet, belong to hollow fiber carrier, it is spherical in shape, can save extruding production model this
Process；During coating, about 80% lactic acid bacteria thallus is all adsorbed among microcrystalline cellulose blank pellet, is efficiently solved
The problem that proper sphere degree (roundness) is low in granulation, granulation uniformity is poor.
Detailed description of the invention
Fig. 1 is to pelletize (A) in the embodiment of the present invention 1, be once coated (B), secondary coating (C) and microcapsules finished product (D)
Pictorial diagram.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment
Used in the conventional means that are well known to those skilled in the art of technological means, raw materials used is commercial goods.
The outer diameter of microcrystalline cellulose blank pellet particle used in the following embodiment is 850-1000 μm, high purchased from Hangzhou
At biological nutrition Technology Co., Ltd..
The preparation of 1 lactobacillus plantarum of embodiment (single bacterium) microcapsules
1) activation and proliferation of strain:
Lactobacillus plantarum JCM-1149 (is derived from into the Japanese bacterium of Microbiological Lab of agricultural college of China Agricultural University preservation
Strain) it is inoculated in progress activation culture three generations in MRS fluid nutrient medium, it is centrifuged using tube centrifuge (10000r/min or more)
10-15min collects bacterium mud.
2) bacterium mud mixed liquor is prepared:
Take bacterium mud 15ml that 20% skimmed milk power aqueous solution 45ml (bacterium mud: 20% skimmed milk power solution=1:3) is added, sufficiently
It mixes to get protectant bacterium mud mixed liquor is added.
3) it pelletizes:
It is mixed well using microcrystalline cellulose blank pellet and sodium alginate as matrix (60g microcrystalline cellulose and the sea 5.4g
Mosanom mixes well), it is added bacterium mud mixed liquor 60ml (bacterium mud mixed liquor: microcrystalline cellulose ≈ 1:1), fluidized bed granulation, item
Part is as follows:
Note: heat engine purpose is that temperature of charge is made to reach 30 DEG C or more；Per stage finely tunes intake and flow velocity with situation；Sample introduction
When measuring big, flow velocity can be reduced.
4) preparation of coating solution
The preparation of coating solution: the sodium alginate aqueous solution containing 1.5% is prepared；
The preparation of secondary coating solution: the acrylic resin aqueous solution containing 12% is prepared.
5) fluidized bed coating
Primary coating: being coated using top spray, and a coating solution dosage is particle: coating solution=2:1；After first step granulation
Particle 64g, coating solution 32ml used.Coating conditions are as follows:
| Sample volume | Intake | It is coated flow velocity | Atomizing pressure | Temperature of charge | Inlet air temperature |
| Heat engine | 20m3/h | ---- | ---- | To 30 DEG C | 70℃ |
| 0-10ml | 20-30m3/h | 6-8r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| 10-20ml | 30-38m3/h | 8-12r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| 20-32ml | 38-45m3/h | 12-18r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
Secondary coating: secondary coating solution dosage is a coated granule: secondary coating solution=4g:1mL.Coating conditions are such as
Under:
| Sample volume | Intake | It is coated flow velocity | Atomizing pressure | Temperature of charge | Inlet air temperature |
| 0-5ml | 55m3/h | 15r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| 5-16ml | 55m3/h | 16r/min | 1.2kg/cm3 | 31±1℃ | 75℃ |
| Dry 10min | 50m3/h | ---- | ---- | 31-40℃ | Closing temperature |
The pictorial diagram of granulation, once coating, secondary coating and microcapsules finished product is shown in Fig. 1.
After secondary coating, fluidized bed is closed, utilizes the dry 10min of the residual temperature of instrument.
Coating front and back viable count compares and embedding rate is shown in Table 1.
Table 1
As shown in Table 1, through the present invention wrap processed lactobacillus plantarum after coating viable count still up to 109, and embed
Rate is up to 80% or so.
The preparation of 2 compound lactobacillus of embodiment (compound bacteria) microcapsules
1) activation and proliferation of strain:
Compound lactobacillus is mainly made of lactobacillus, galactococcus and bacillus etc. (by agricultural college of China Agricultural University
Microbiological Lab provides), it is inoculated in progress activation culture three generations in MRS fluid nutrient medium, using tube centrifuge
(10000r/min or more) is centrifuged 10-15min, collects bacterium mud.
2) bacterium mud mixed liquor is prepared:
Take 1 kilogram of addition 3 kilograms of 20% skimmed milk power aqueous solution (bacterium mud: 20% skimmed milk power of the above-mentioned bacterium mud processed
Solution=1:3), it mixes well to get protectant bacterium mud mixed liquor is added.
3) it pelletizes:
It is mixed well using microcrystalline cellulose blank pellet and sodium alginate as matrix and is pelletized, 3.6 kilograms of microcrystalline cellulose
0.36 kilogram of sodium alginate (microcrystalline cellulose: sodium alginate=10:1) is added in element, adds 4 kilograms of (bacterium of bacterium mud mixed liquor
Mud mixed liquor: microcrystalline cellulose ≈ 1:1).It finally pelletizes to obtain about 8 kilograms of wet granular material.
Fluidized bed granulation technological parameter is as follows:
| Sample volume | Intake | Flow velocity | Atomizing pressure | Temperature of charge | Inlet air temperature |
| Heat engine | 12m3/h | ---- | ---- | To 30 DEG C | 65℃ |
| 0-20ml | 15-24m3/h | 5-8r/min | 1.2kg/cm3 | 29±1℃ | 65℃ |
| 20-40ml | 25-40m3/h | 8-13r/min | 1.2kg/cm3 | 29±1℃ | 65℃ |
| 40-60ml | 40-50m3/h | 13-10r/min | 1.2kg/cm3 | 29±1℃ | 65℃ |
| Dry 10min | 45m3/h | ---- | ---- | 31-40℃ | Closing temperature |
Note: heat engine is that temperature of charge is allowed to reach 30 DEG C or more；Per stage finely tunes intake and flow velocity with situation.After granulation
About 7 kilograms of wet granular.
4) preparation of coating solution
The preparation of coating solution: the sodium alginate and chitosan aqueous solution of 1.5-2% are prepared, wherein sodium alginate and shell
The mass ratio of glycan is 4:1.
The preparation of secondary coating solution: 3% ethyl cellulose solution is prepared.
5) fluidized bed coating
Primary coating: being coated using top spray, and coating solution is 1.5% sodium alginate soln, according to particle: 1.5% alginic acid
Sodium=4:3 dosage is coated, and one time coating parameter is as follows:
| Sample volume | Intake | It is coated flow velocity | Atomizing pressure | Temperature of charge | Inlet air temperature |
| Dry 10min | 50m3/h | ---- | 1.2kg/cm3 | 26℃ | 55℃ |
| 0-20ml | 50m3/h | 10r/min | 1.2kg/cm3 | 27℃ | 55℃ |
| 20-40ml | 50m3/h | 12-13.5r/min | 1.2kg/cm3 | 27±1℃ | 55℃ |
| 40-60ml | 55m3/h | 15r/min | 1.2kg/cm3 | 27±1℃ | 55℃ |
| 60-105ml | 55m3/h | 18r/min | 1.2kg/cm3 | 27±1℃ | 55℃ |
| 105-150ml | 60m3/h | 18r/min | 1.2kg/cm3 | 27±1℃ | 55℃ |
| Dry 5min | 60m3/h | ---- | ---- | 29-34℃ | Closing temperature |
Particle is before coating first in fluidized bed drying 10min, entire coating process about 1~1.5h of time-consuming.In commodity production
In can be selected and large-scale fluidized bed be coated with according to the demand of package amount.
Secondary coating: secondary coating, secondary coating solution dosage are carried out after primary coating are as follows: a coated granule:
12% acrylic resin aqueous solution=4g:1mL is coated, and coating conditions are as follows:
| Sample volume | Intake | Flow velocity | Atomizing pressure | Temperature of charge | Inlet air temperature |
| 0-5ml | 60m3/h | 18r/min | 1.2kg/cm3 | 28-30℃ | 55℃ |
| 5-20ml | 60-70m3/h | 18r/min | 1.2kg/cm3 | 28-30℃ | 55℃ |
| Dry 10min | 60-70m3/h | ---- | ---- | 28-30℃ | Closing temperature |
After secondary coating, fluidized bed is closed, utilizes the dry 10min of the residual temperature of instrument.
Coating front and back viable count compares and embedding rate is shown in Table 2.
Table 2
As shown in Table 2, through the present invention wrap processed compound lactobacillus after coating viable count still up to 109, and embed
Rate is up to 80% or so.
It is worth noting that this method could be applicable to other unmentioned probiotics in the present invention.
The performance test of 3 microcapsule product of embodiment
Existing report is all made of in-vitro simulated gastrointestinal tract environment, come detect coating after thallus whether can be resistant to gastric acid and
Cholate environment, final Targeting delivery.And this research is using the method verified in vivo, it is true to assess lactic acid bacteria in performance prebiotic effect
When bioprocess experienced, be embodied as follows:
Test in In Shunyi District of Beijing breeding stock fine breed cow Technology Park choose 4 body conditions it is good, three similar in weight
Tire He Sitan dry milk ox installs permanence lymphoma stomach fistulization pipe and duodenum fistula, for adopting for cud and duodenum content
Collection.After postoperative care January, animal body restores to normal condition to be tested.
Test was divided into for three phases, and each issue of 7 days formal phase, interval 20 days, 4 Fistu- lated cows fed 2 (early 9:00 and evenings daily
21:00) fully mixed diet.Using own control experimental design, the first phase is blank control group, does not add any probiotics, the
Two, the 1h (10:00) after morning raises opened rumen fistula plug and fed and be not coated with lactobacillus plantarum and implementation respectively three phases (test group)
(living bacteria count is respectively 8 × 10 to the coating lactobacillus plantarum bacterium powder of example 19Cfu/g and 3.8 × 109cfu/g).Test group is continuous
It feeds 7 days, daily throwing bacterium amount is that (head d), then every ox feeds viable bacteria amount daily and respectively reaches 6.40 × 10 80g11Cfu and
3.04×1011cfu.Each issue of the last continuous sampling in 3 days of test, daily sampling time point are to raise preceding 8:00 morning to start to adopt for the first time
Sample then carries out dynamic acquisition rumen fluid, Duodenal digesta and rectum excrement sample for 24 hours at interval of 3h, using flow cytometer
Detect the quantity (table 3, table 4) that wherein viable bacteria ratio and quantitative PCR detection lactic acid bacteria reach enteron aisle.
3 rumen fluid of table and duodenal juice viable bacteria ratio (%)
Note: the different letters of colleague's shoulder mark indicate significant difference (P < 0.05)；It is identical or without letter indicate difference it is not significant (P >
0.05)。
As shown in Table 3, it in rumen fluid, compared with the control group, is coated in 11:00,2:00,5:00 and whole day average value
Lactobacillus plantarum JCM-1149 viable bacteria ratio significantly increases (P < 0.05)；In duodenal juice, 8:00,11:00,20:00 and
In whole day average value, coating lactobacillus plantarum JCM-1149 viable bacteria ratio significantly increases (P < 0.05).
Table 4 quantifies cud and enteron aisle lactobacillus plantarum JCM-1149 content (ng)
Note: the different letters of colleague's shoulder mark indicate significant difference (P < 0.01)；Identical expression difference is not significant (P > 0.01).
As shown in Table 4, in rumen fluid, duodenal juice and rectum excrement, coating lactobacillus plantarum JCM-1149 is fixed
Amount PCR value is significantly higher than control group and is not coated with lactobacillus plantarum JCM-1149 (P < 0.01).In cud, 3 groups of Q-PCR's
Value illustrates just to put into coated lactobacillus plantarum JCM-1149 also not release completely, explanation in cud all in an order of magnitude
Coating is played the role of protecting lactic acid bacteria in cud, avoids the degradation of rumen microorganism；And in duodenum, coated plant
Object lactobacillus JCM-1149 resists the erosion of abomasum gastric acid, can discharge in enteron aisle, so quantitative PCR value increases；In rectum
In, coated lactobacillus plantarum JCM-1149 discharges substantially, and the value of Q-PCR significantly increases, respectively than in cud and duodenum
High 1 and 2 order of magnitude of the value of Q-PCR, illustrates that coated lactobacillus plantarum JCM-1149 is largely discharged in enteron aisle.
By table 3 and 4 test result of table it is found that the lactobacillus plantarum JCM-1149 after coating can pass through cud, true
Stomach eventually arrives at enteron aisle, to promote external source lactic acid bacteria in milk cow intestinal colonisation.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be modified or is improved, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. the preparation method of probiotic microcapsule, which comprises the following steps:
A, the preparation of probiotics bacterium mud mixed liquor: probiotics being activated, is cultivated, and obtains probiotics fermention liquid, centrifugation, in abandoning
Clearly, thallus is collected, bacterium mud is obtained；Appropriate skimmed milk power protection liquid is added into the bacterium mud, obtains bacterium mud mixed liquor；
B, it pelletizes: being added in fluidized bed using microcrystalline cellulose blank pellet and sodium alginate as skinning core particles, make the bacterium mud
Mixed liquor injects in the fluidized bed according to certain flow rate, under certain condition mixing granulation；
C, primary coating: by particle obtained by a coating solution even application to step B, a coated granule is obtained；
D, secondary coating: will complete secondary coating on particle obtained by secondary coating solution even application to step C, it is dry after to obtain the final product
Probiotic microcapsule finished product；
Wherein, the sodium alginate and shell of a coating solution is 1.5-2% sodium alginate aqueous solution or 1.5-2% are poly-
Sugar aqueous solution；Wherein the mass ratio of sodium alginate and chitosan is 2-4 in the sodium alginate of 1.5-2% and chitosan aqueous solution:
1；
The secondary coating solution is the ethyl cellulose solution or hydrogenated oil and fat of the acrylic resin aqueous solution of 10-12%, 1-5%
Rouge；
In granulation, primary coating and secondary coating process, mixed material temperature is controlled at 25-40 DEG C.
2. the method according to claim 1, wherein the body of bacterium mud described in step A and skimmed milk power protection liquid
Product is than being 1:1-3；Wherein, the concentration of the skimmed milk power protection liquid is 10%-30%.
3. the method according to claim 1, wherein the use of skinning core particles and bacterium mud mixed liquor described in step B
Amount is than being 1g:1-2mL；Wherein, microcrystalline cellulose blank pellet and the weight ratio of sodium alginate are 11:1- in the skinning core particles
5。
4. the method according to claim 1, wherein the condition that step B carries out fluidized bed granulation is as follows:
5. the method according to claim 1, wherein the alginic acid that a coating coating solution used is 1.5-2%
Sodium water solution, secondary coating solution are the acrylic resin aqueous solution of 10-12%.
6. according to the method described in claim 5, it is characterized in that, step C to carry out the condition that is once coated of fluidized bed as follows:
Wherein, the amount ratio of the particle and coating solution is 1.5-2g:1mL.
7. according to the method described in claim 5, it is characterized in that, the condition of the step D progress secondary coating of fluidized bed is as follows:
Wherein, the amount ratio of a coated granule and coating solution is 2-4g:1mL.
8. method according to claim 1-7, which is characterized in that the probiotics includes lactic acid bacteria.
9. the probiotic microcapsule of any one of -8 the method preparations according to claim 1.
10. following any application of probiotic microcapsule described in claim 9:
1) application in feed preparation；
2) application in probiotics, especially Ruminat s Microecological Agent preparation.
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| CN110074256A (en) * | 2019-06-12 | 2019-08-02 | 瞿瀚鹏 | Improve the feed and preparation method thereof of ruminant milk fat rate |
| CN111436612A (en) * | 2020-04-22 | 2020-07-24 | 绵阳市润土农业科技开发有限公司 | In-vivo enzyme, preparation method thereof and application of in-vivo enzyme in inhibiting novel viruses |
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