CN105192250B - A kind of feeding type probiotics and its application - Google Patents
A kind of feeding type probiotics and its application Download PDFInfo
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Abstract
The invention discloses a kind of feeding type probiotics and its applications.Active constituent Cheesecake lactobacillus, candida tropicalis and the bacillus subtilis composition of feeding type probiotics of the present invention, it is big that the feeding type probiotics carry bacterium amount, stable structure, it can promote ciliophoran quick breeding after Rumen acid poisoning, so that Volatile fatty acid contents persistently increase in Rumen liquid, goat is restored to the digestion power of cellulose family feed and has shortened recovering process, Rumen acidosis symptom can be effectively relieved.Environment in Rumen can be quickly rebuild using feeding type probiotics of the invention, restores its physiological function, has great importance for Animal husbandry production.
Description
Technical field
The present invention relates to a kind of feeding type probiotics and its applications.
Background technique
In recent years, with the development of breeding scale industry, concentrated feed proportion is dramatically increased in ruminant, although
Concentrated feed can significantly improve the growth performance of animal, but long-term a large amount of feeding concentrated feeds can cause animal rumens acid poisoning.Tumor
Gastric acid, which is poisoned, can cause the dehydration of stomach and gut epithelium, and serious person can cause the pH value of blood to reduce, and body is dehydrated
Etc. serious consequences.Data shows that acute acid poison is tight every year because the mortality for suffering from subacute acid poisoning cows can often be higher than 45%
Weight even can cause death, and the death rate may be up to 90%.To the main control method of rumen ecology first is that regulation daily ration,
Rationally feeding concentrated feed, to reduce the incidence of rumen ecology, but this method takes a long time, and fits to ruminant
Should be able to power dependence it is larger.Antibiotic inhibits microorganism lactic acid producing to have a certain effect, but many antibiotic such as Monensins etc. from
Subcarrier antibiotic inhibits degradation of the Fibrolytic bacteria to roughage simultaneously；Furthermore due to safety problem, antibiotic is gradually prohibited
With.In recent years, microbe research relevant to acid poisoning is just being paid more and more attention, and only prevention rumen microorganism crosses volume production acid especially
It is lactic acid, could fundamentally prevents the generation of acid poisoning.
When acid poisoning due in cud multiple-microorganism reduce more sensitive to pH and inactivate, in turn result in volatile fat
The generation of acid content is reduced, and volatile fatty acid important role in the growth course of ruminant.Therefore, how
Enough quick-recovery micro organism quantities fast after rumen ecology, improve rumen volatile fatty acid content, rebuild Rumen Internal Environment,
Restore its physiological function to have great importance.
Ruminant tumor gastric infusorian can stablize Rumen Internal Environment, and cud is made to keep suitable fermentation level, promote machine
Body improves breeding performonce fo animals to the digestibility of fiber-like, protide feed, while can reduce nitrite, nitrate
Toxic action.
Summary of the invention
The technical problem to be solved by the present invention is to how treat or assist in the treatment of animal rumens acid poisoning.
In order to solve the above technical problems, present invention firstly provides a kind of feeding type probiotics.
Feeding type probiotics provided by the present invention, active constituent Cheesecake lactobacillus, candida tropicalis and
Bacillus subtilis composition.
In above-mentioned feeding type probiotics, the Lactobacillus casei is Lactobacillus casei (Lactobacillus
Casei) ACCC10640, the candida tropicalis are candida tropicalis (Candida tropicalis) ACCC21161,
The bacillus subtilis is bacillus subtilis (Bacillus subtilis) ACCC02157.
In above-mentioned feeding type probiotics, in the active constituent, the Lactobacillus casei, the candida tropicalis
Colony Forming Unit ratio with the bacillus subtilis can be (3-7): (1-4): (5-11), concretely 6.61:(1-4):
(5-11), (3-7): 1:(5-11), (3-7): (1-4): 10.12 or 6.61:1:10.12；In the feeding type probiotics
Active component content be Lactobacillus casei described in feeding type probiotics described in every gram, the candida tropicalis and institute
The sum of the content of this three plants of bacterial strains of bacillus subtilis is stated, can be (3-9) × 105Cfu/g, concretely 4.56 × 105cfu/g。
In above-mentioned feeding type probiotics, the feeding type probiotics are made of the active constituent and auxiliary material；
The auxiliary material is made of sodium alginate, calcium chloride and chitosan.
Above-mentioned feeding type probiotics are prepared as follows: by Lactobacillus casei microbial inoculum, Candida tropicalis
Agent and bacillus subtilis microbial agent carry out being mixed to get the feeding type probiotics；The activity of the Lactobacillus casei microbial inoculum
Ingredient is the Lactobacillus casei；The active constituent of the Candida tropicalis agent is the candida tropicalis；It is described withered
The active constituent of careless gemma bacillus agent is the bacillus subtilis.
In above-mentioned feeding type probiotics, the Lactobacillus casei microbial inoculum, the Candida tropicalis agent and described
The mixed proportion of bacillus subtilis microbial agent meets the Lactobacillus casei in the feeding type probiotics, the torrid zone
The Colony Forming Unit of Candida and bacillus subtilis ratio can be (3-7): (1-4): (5-11), concretely
6.61:(1-4): (5-11), (3-7): 1:(5-11), (3-7): (1-4): 10.12 or 6.61:1:10.12.
In above-mentioned feeding type probiotics, the Lactobacillus casei microbial inoculum, the Candida tropicalis agent and described
The mixing mass ratio of bacillus subtilis microbial agent can be (1-4): (1-3): (1-4), concretely 1:(1-3): (1-4), (1-
4): 1:(1-4), (1-4): (1-3): 1 or 1:1:1.
In above-mentioned feeding type probiotics, the Lactobacillus casei microbial inoculum is prepared by following methods: by Lactobacillus casei-
Sodium alginate liquid, which is added drop-wise in calcium chloride-chitosan liquid, obtains the Lactobacillus casei microbial inoculum, the Lactobacillus casei-sea
Mosanom liquid is the liquid being mixed to get by the bacterium solution of sodium alginate aqueous solution and the Lactobacillus casei；The calcium chloride-
Chitosan liquid be by (40g-100g) calcium chloride (such as 40g, 50g, 60g, 80g or 100g) and (10g-20g) chitosan (such as
Liquid obtained in the acetum that the pH value for 10g) being dissolved in 1L is 4；Seaweed in the Lactobacillus casei-sodium alginate liquid
The proportion of sour sodium and Lactobacillus casei satisfaction (10-30) g sodium alginate: 8.6 × 1010Lactobacillus casei described in cfu, specifically
It can be 10g sodium alginate: 8.6 × 1010Lactobacillus casei described in cfu, 15g sodium alginate: 8.6 × 1010The cream of cheese described in cfu
Bacillus, 20g sodium alginate: 8.6 × 1010Lactobacillus casei described in cfu, 30g sodium alginate: 8.6 × 1010The cream of cheese described in cfu
Bacillus；The Lactobacillus casei-sodium alginate liquid and the calcium chloride-chitosan liquid volume ratio can be (1-2): (1-
3), concretely 1:(1-3), 2:(1-3), (1-2): 1, (1-2): 2, (1-2): 3,1:3,1:2,1:1 or 2:1；The cheese
The content of Lactobacillus casei described in lactobacillus microbial inoculum can be (1.6-6.7) × 105Cfu/g, concretely 5.1 × 105cfu/g。
The Candida tropicalis agent is prepared by following methods: candida tropicalis-sodium alginate liquid is added drop-wise to
The Candida tropicalis agent, the candida tropicalis-sodium alginate liquid are obtained in the calcium chloride-chitosan liquid
It is the liquid being mixed to get by the bacterium solution of the sodium alginate aqueous solution and the candida tropicalis；The false silk ferment in the torrid zone
The proportion of sodium alginate and candida tropicalis satisfaction (10-30) g sodium alginate in mother-sodium alginate liquid: 7.8 ×
1011Candida tropicalis described in cfu, concretely 10g sodium alginate: 7.8 × 1011Candida tropicalis described in cfu, the sea 15g
Mosanom: 7.8 × 1011Candida tropicalis described in cfu, 20g sodium alginate: 7.0 × 107Candida tropicalis described in cfu,
30g sodium alginate: 7.8 × 1011Candida tropicalis described in cfu；The candida tropicalis-sodium alginate liquid and the chlorine
Changing calcium-chitosan liquid volume ratio can be (1-2): (1-3), concretely 1:(1-3), 2:(1-3), (1-2): 1, (1-2):
2, (1-2): 3,1:3,1:2,1:1 or 2:1；The content of candida tropicalis described in the Candida tropicalis agent can be
(4.6-9.5)×104Cfu/g, concretely 7.7 × 104cfu/g。
The bacillus subtilis microbial agent is prepared by following methods: bacillus subtilis-sodium alginate liquid is added drop-wise to
The bacillus subtilis microbial agent, the bacillus subtilis-sodium alginate liquid are obtained in the calcium chloride-chitosan liquid
It is the liquid being mixed to get by the bacterium solution of the sodium alginate aqueous solution and the bacillus subtilis；The bacillus subtilis
The proportion of sodium alginate and bacillus subtilis satisfaction (10-30) g sodium alginate in bacterium-sodium alginate liquid: 9.6 ×
1010Bacillus subtilis described in cfu, concretely 10g sodium alginate: 9.6 × 1010Bacillus subtilis described in cfu, the sea 15g
Mosanom: 9.6 × 1010Bacillus subtilis described in cfu, 20g sodium alginate: 9.6 × 1010Bacillus subtilis described in cfu,
30g sodium alginate: 9.6 × 1010Bacillus subtilis described in cfu；The bacillus subtilis-sodium alginate liquid and the chlorine
Changing calcium-chitosan liquid volume ratio can be (1-2): (1-3), concretely 1:(1-3), 2:(1-3), (1-2): 1, (1-2):
2, (1-2): 3,1:3,1:2,1:1 or 2:1；The content of bacillus subtilis described in the bacillus subtilis microbial agent is
(2.7-8.6)×105Cfu/g, concretely 7.8 × 105cfu/g。
Above, the content of Lactobacillus casei described in the bacterium solution of the Lactobacillus casei is 8.6 × 109cfu/mL；Institute
The content for stating candida tropicalis described in the bacterium solution of candida tropicalis is 7.8 × 1010cfu/mL；The bacillus subtilis
Bacterium solution described in bacillus subtilis content be 9.6 × 109cfu/mL。
Above, the concentration of sodium alginate can be 10g/L-30g/L in the sodium alginate aqueous solution, concretely 10g/
L, 15g/L, 20g/L or 30g/L.
Above, the concentration of calcium chloride can be 40g/L-100g/L in the calcium chloride-chitosan liquid, concretely
40g/L, 50g/L, 60g/L, 80g/L or 100g/L；The concentration of chitosan can be 10g/L- in the calcium chloride-chitosan liquid
20g/L, concretely 10g/L.
In order to solve the above technical problems, the present invention also provides the product for treating or assisting in the treatment of animal rumens acid poisoning,
Its active constituent is the feeding type probiotics.
The said goods have the function of following 4 kinds:
1) increase the content of acetic acid in rumen fluid；
2) increase the content of propionic acid in rumen fluid；
3) increase the content of butyric acid in rumen fluid；
4) increase ciliophoran quantity in rumen fluid.
The present invention also provides the feeding type probiotics answering in the product that preparation has following 4 kinds of functions
With:
1) increase the content of acetic acid in rumen fluid；
2) increase the content of propionic acid in rumen fluid；
3) increase the content of butyric acid in rumen fluid；
4) increase ciliophoran quantity in rumen fluid.
It is demonstrated experimentally that feeding type probiotics load bacterium amount of the invention is big, cheese in every gram of feeding type probiotics
The sum of content of this three plants of bacterial strains of lactobacillus, candida tropicalis and bacillus subtilis is 4.56 × 105Cfu/g, structure are steady
It is fixed, Rumen acidosis symptom can be effectively relieved, in the 2nd day experimental group Rumen of feeding type probiotics application
Ciliophoran quantity has been significantly higher than control group (t=10.613, P < 0.05) in liquid, illustrates that feeding type probiotics can promote
Ciliophoran quick breeding after into rumen ecology, has restored goat to the digestion power of cellulose family feed and has shortened extensive
Multiple process.To the measurement result of Volatile fatty acid contents in Rumen liquid show in experimental group Rumen liquid acetic acid to
Give being significantly higher than for the 4th day control group (t=9.281, P < 0.05) for feeding type probiotics, propionic acid (t=5.574, P <
0.05) control has been significantly higher than it the 2nd day for giving feeding type probiotics with the content of butyric acid (t=5.568, P < 0.05)
Group, and persistently raised trend is showed during testing.Mountain can be quickly rebuild using feeding type probiotics of the invention
Sheep Rumen Internal Environment restores its physiological function, has great importance for Animal husbandry production.
Detailed description of the invention
Fig. 1 is the map of the HPLC of acetic acid, propionic acid and butyric acid.Wherein, A is the hybrid standard liquid of acetic acid, propionic acid, butyric acid
Chromatogram；B is the chromatogram of Rumen liquid；1 represents acetic acid, 2 represents propionic acid, 3 represents butyric acid.
Specific embodiment
The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining
The bright present invention, the range being not intended to be limiting of the invention.
Experimental method in following embodiments is unless otherwise specified conventional method.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Sodium alginate in following embodiments, chitosan and calcium chloride are the production of Sinopharm Chemical Reagent Co., Ltd.
Product；Acetic acid (ρ=1.04g/L), propionic acid (ρ=0.993g/L) and butyric acid (ρ=0.959g/L) are chromatographically pure, are the U.S.
The product of J.T.Baker company；Potassium dihydrogen phosphate is chromatographically pure, is the product of U.S. ACS En Ke chemical company；Phosphoric acid is chromatography
It is pure, it is the product of U.S. Fisher company.
1100 series liquid chromatograph instrument system of Agilent in following embodiments is that the product of agilent company (is furnished with
G1379A type automatic deaerating machine, G1311A type quaternary pump, G1313A type autosampler, G1316 type column oven, G1312A type can
Become wavelength UV detector and chromatographic work station)；BSA224S assay balance is the production of Sai Duolisi scientific instrument Co., Ltd
Product；Genpure UV-WC laboratory ultrapure water system is the product of Thermo company；EYEL4 MG-2200 nitrogen evaporator is Tokyo
The product of RIKAKIKAL company；XH-D turbula shaker is the product of Wuxi Wo Xin Instrument Ltd..
Lactobacillus casei (Lactobacillus casei) ACCC10640 (hereinafter referred cheese cream in following embodiments
Bacillus), China Committee for Culture Collection of Microorganisms agricultural microorganism center was concealed in front of the applying date of the application, also known as
Chinese agriculture Microbiological Culture Collection administrative center (Agricultural Culture Collection of China, referred to as
ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, Chinese Academy of Agricultural Sciences's agricultural resource and agricultural regionalization research
Institute, postcode 100081), collection day is on 01 01st, 2008, and from collecting, the public can be from Chinese Microbiological Culture Collection
Administration committee agricultural microorganism center obtains the bacterial strain.
(the hereinafter referred torrid zone candida tropicalis (Candida tropicalis) ACCC21161 in following embodiments
Candida), China Committee for Culture Collection of Microorganisms agricultural microorganism center was concealed in front of the applying date of the application,
Also known as Chinese agriculture Microbiological Culture Collection administrative center (Agricultural Culture Collection of China,
Abbreviation ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, Chinese Academy of Agricultural Sciences's agricultural resource are ground with agricultural regionalization
Study carefully institute, postcode 100081), collection day is on 01 01st, 2008, and from collecting, the public can protect from Chinese microorganism strain
Hiding administration committee's agricultural microorganism center obtains the bacterial strain.
Bacillus subtilis (Bacillus subtilis) ACCC02157 (hereinafter referred withered grass bud in following embodiments
Spore bacillus), China Committee for Culture Collection of Microorganisms agricultural microorganism center was concealed in front of the applying date of the application, again
Claim Chinese agriculture Microbiological Culture Collection administrative center (Agricultural Culture Collection of China, letter
Claim ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, Chinese Academy of Agricultural Sciences's agricultural resource and agricultural regionalization research
Institute, postcode 100081), collection day is on 07 20th, 2007, and from collecting, the public can be from Chinese Microbiological Culture Collection
Administration committee agricultural microorganism center obtains the bacterial strain.
The preparation of embodiment 1, feeding type probiotics
Feeding type probiotics provided in this embodiment, are made of active constituent and auxiliary material；Active constituent Cheesecake cream
Bacillus, candida tropicalis and bacillus subtilis composition；Auxiliary material is made of sodium alginate, calcium chloride and chitosan.
Feeding type probiotics provided in this embodiment are prepared as follows: by Lactobacillus casei microbial inoculum, the torrid zone
Candida microbial inoculum and bacillus subtilis microbial agent carry out being mixed to get feeding type probiotics；The work of Lactobacillus casei microbial inoculum
Property ingredient be Lactobacillus casei (Lactobacillus casei) ACCC10640；The active constituent of Candida tropicalis agent is
Candida tropicalis (Candida tropicalis) ACCC21161；The active constituent of bacillus subtilis microbial agent is withered grass bud
Spore bacillus (Bacillus subtilis) ACCC02157.
Lactobacillus casei microbial inoculum is prepared by following methods: Lactobacillus casei-sodium alginate liquid is added drop-wise to calcium chloride-shell
Lactobacillus casei microbial inoculum is obtained in glycan liquid, Lactobacillus casei-sodium alginate liquid is by sodium alginate aqueous solution and cheese
What the bacterium solution of lactobacillus was mixed to get.
Candida tropicalis agent is prepared by following methods: candida tropicalis-sodium alginate liquid is added drop-wise to chlorination
Candida tropicalis agent is obtained in calcium-chitosan liquid, candida tropicalis-sodium alginate liquid is water-soluble by sodium alginate
What the bacterium solution of liquid and candida tropicalis was mixed to get.
Bacillus subtilis microbial agent is prepared by following methods: bacillus subtilis-sodium alginate liquid is added drop-wise to chlorination
Bacillus subtilis microbial agent is obtained in calcium-chitosan liquid, bacillus subtilis-sodium alginate liquid is water-soluble by sodium alginate
What the bacterium solution of liquid and bacillus subtilis was mixed to get.
Specific experiment operation is as follows:
Wherein, the liquid spawn of the Lactobacillus casei in following experiments is the preparation method is as follows: with fermentation medium A in 35-
The culture (all substances in culture vessel) that 37 DEG C of culture 46h-54h are obtained.Specifically the preparation method is as follows: by activated
Lactobacillus casei is inoculated in the 500mL triangular flask of the fermentation medium A equipped with 100mL, 35-37 DEG C, shaking table 160rpm (rotation
Radius 20mm) culture 48 hours, all substances (culture) in triangular flask are the liquid spawn of Lactobacillus casei.The cheese
Lactobacillus casei content is 8.6 × 10 in the liquid spawn of lactobacillus9cfu/mL。
The liquid spawn of candida tropicalis in following experiments is the preparation method is as follows: with fermentation medium A at 28-30 DEG C
The culture (all substances in culture vessel) that culture 32h-40h is obtained.Specifically the preparation method is as follows: by the activated torrid zone
Candida is inoculated in the 500mL triangular flask of the fermentation medium A equipped with 100mL, 28-30 DEG C, 180rpm (radius of turn
20mm) shaken cultivation 36 hours, all substances (culture) in triangular flask are the liquid spawn of candida tropicalis.The heat
Candida tropicalis content is 7.8 × 10 in liquid spawn with Candida10cfu/mL。
The liquid spawn of bacillus subtilis in following experiments is the preparation method is as follows: with fermentation medium A at 30-32 DEG C
The culture (all substances in culture vessel) that culture 18h-24h is obtained.Specifically the preparation method is as follows: by activated withered grass
Bacillus is inoculated in the 500mL triangular flask of the fermentation medium A equipped with 100mL, 30-32 DEG C, 180rpm (radius of turn
20mm) shaken cultivation 20 hours, all substances (culture) in triangular flask are bacillus subtilis bacterium solution.The withered grass gemma
Bacillus subtilis bacterial content is 9.6 × 10 in bacillus bacterium solution9cfu/mL。
Every liter of fermentation medium A is prepared as follows: 10g glucose, 10g peptone, 1.5g KH2PO4、0.80g
MnSO4、1.00g MgSO4, 1.0g NaCl, be settled to l000mL, pH value 6.5 with distilled water；It sterilizes 20 points under the conditions of 121 DEG C
Clock.
Sodium alginate aqueous solution in following experiments is matched the preparation method is as follows: by 15g sodium alginate addition deionized water
It is set to the sodium alginate aqueous solution that concentration is 15g/L.
Calcium chloride-chitosan liquid in following experiments to the acetum that pH value is 4 the preparation method is as follows: be added appropriate
Then calcium chloride dissolution weighs a certain amount of chitosan dissolution and obtains calcium chloride-chitosan liquid.Calcium chloride-chitosan liquid
Calcium chloride concentration in body is 40g/L or 60g/L；The concentration of chitosan is 10g/L.
One, the preparation method of Lactobacillus casei microbial inoculum (Lactobacillus casei microballoon)
The liquid spawn of the Lactobacillus casei of 10mL is added into the sodium alginate aqueous solution that 1.0L concentration is 15g/L, obtains
To Lactobacillus casei-sodium alginate liquid, 4 DEG C of standings remove bubble, alginic acid in Lactobacillus casei-sodium alginate liquid for 24 hours
The proportion of sodium and Lactobacillus casei meets 15g sodium alginate: 8.6 × 1010Cfu Lactobacillus casei.With syringe by cheese cream bar
Bacterium-sodium alginate liquid is added drop-wise to calcium chloride-chitosan liquid, and (concentration of calcium chloride is 60g/L, and the concentration of chitosan is 10g/
L in), volume ratio meets 1:1, and with 600rmin-1Speed stirring 10min makes chitosan and sodium alginate that glue nuclear reaction occur
Microballoon is formed, i.e. acquisition Lactobacillus casei microbial inoculum (Lactobacillus casei microballoon).Lactobacillus casei contains in Lactobacillus casei microbial inoculum
Amount is 5.1 × 105cfu/g。
Two, the preparation method of Candida tropicalis agent (candida tropicalis microballoon)
The liquid spawn of the candida tropicalis of 10mL is added into the sodium alginate aqueous solution that 1.0L concentration is 15g/L,
Obtain candida tropicalis-sodium alginate liquid, 4 DEG C stand and remove bubble for 24 hours, in candida tropicalis-sodium alginate liquid
The proportion of sodium alginate and candida tropicalis meets 15g sodium alginate: 7.8 × 1011Cfu candida tropicalis.Use syringe
Candida tropicalis-sodium alginate liquid is added drop-wise to calcium chloride-chitosan liquid, and (concentration of calcium chloride is 40g/L, chitosan
Concentration be 10g/L) in, volume ratio meets 1:1, and with 600rmin-1Speed stirring 10min makes chitosan and sodium alginate
Glue nucleus forming reaction microreactor ball occurs, i.e. acquisition Candida tropicalis agent (candida tropicalis microballoon).Candida tropicalis
The content of candida tropicalis is 7.7 × 10 in agent4cfu/g。
Three, the preparation method of bacillus subtilis microbial agent (bacillus subtilis microballoon)
The liquid spawn of the bacillus subtilis of 10mL is added into the sodium alginate aqueous solution that 1.0L concentration is 15g/L,
Obtain bacillus subtilis-sodium alginate liquid, 4 DEG C stand and remove bubble for 24 hours, in bacillus subtilis-sodium alginate liquid
The proportion of sodium alginate and bacillus subtilis meets 15g/L sodium alginate: 9.6 × 1010Cfu/L bacillus subtilis.With note
Emitter by bacillus subtilis-sodium alginate liquid be added drop-wise to calcium chloride-chitosan liquid (concentration of calcium chloride be 60g/L, shell
The concentration of glycan is 10g/L) in, volume ratio meets 1:1, and with 600rmin-1Speed stirring 10min makes chitosan and seaweed
Glue nucleus forming reaction microreactor ball occurs for sour sodium, i.e. acquisition bacillus subtilis microbial agent (bacillus subtilis microballoon).Bacillus subtilis
The content of bacillus subtilis is 7.8 × 10 in bacteria agent5cfu/g。
Lactobacillus casei microbial inoculum, Candida tropicalis agent and bacillus subtilis microbial agent are carried out according to following mass ratio
1:1:1 is mixed, feeding type probiotics are obtained.Lactobacillus casei in the feeding type probiotics, candida tropicalis and
The Colony Forming Unit ratio of bacillus subtilis is 6.61:1:10.12, and active component content is 4.56 × 105cfu/g；Wherein,
Lactobacillus casei 1.70 × 105Cfu/g, candida tropicalis 2.57 × 104Cfu/g, bacillus subtilis 2.60 × 105cfu/
g。
Embodiment 2, feeding type probiotics are for treating white Rumen acid poisoning
Select the male Anhui White Goats at or so healthy 4 monthly ages, weight 20kg or so as experimental animal, using cereal mistake
Food method modeling according to 40g/kg stomach-filling maize flour, and carries out cud fluid inspection, blood test, fecaluria pH respectively before and after modeling
Value and infusorian count.It chooses 20 successful Anhui White Goats of modeling and is divided into control group and feeding type probiotics at random
Group (hereinafter referred experimental group).Control group uses fluid infusion [venoclysis physiological saline or 5% (mass percentage) glucose
Water], increase blood volume, promote blood circulation, alleviates acidosis symptom；Experimental group is daily on the basis of control group remedy measures
Using feeding type probiotics stomach-filling, the feeding type probiotics of embodiment 1 are suspended in distilled water and obtain feeding type
Probiotics liquid, the content of feeding type probiotics is that 5% (quality percentage contains in the feeding type probiotics liquid
Amount), feeding type probiotics liquid given low be 5mL/10kg goat, continuous one week.
Rumen content is acquired before daily stomach-filling feeding type probiotics liquid during test, using 2 layers of filtered through gauze,
Discard feed, swill obtains rumen fluid.With suction pipe by the cud drop of mixing on blood cell counting plate, close the lid glass
Piece, under an optical microscope microscopy；Part rumen fluid is separately taken to save with -80 DEG C of refrigerators for volatile fatty acid in rumen fluid
Measurement.
Data processing is carried out using 17.0 software of SPSS, measurement data uses means standard deviationForm table
Show, comparison among groups are examined using t, indicate that difference is statistically significant with P < 0.05.
One, feeding type probiotics are on influence ciliophoran in cud
It is as shown in table 1 using microscopic examination result of the optical microscopy to rumen fluid, two groups of Rumens of control group and experimental group
The trend gradually increased is presented in interior infusorian quantity, but infusorian quantity speedup is very fast in experimental group Rumen.It is testing
Group is after stomach-filling 2 days, and ciliophoran quantity has been significantly more than control group (t=10.613, P < 0.05) in experimental group rumen fluid；Later
Ciliophoran quantity significant difference (P < 0.05) always in two groups of Rumen liquid.
Table 1, feeding type probiotics are to infusorian (× 10 in cud5Item) influence
Two, influence of the feeding type probiotics to content of fatty acid in cud
Using volatile fatty acid in HPLC method measurement cud: the changes of contents of acetic acid, propionic acid and butyric acid.
1, liquid phase chromatogram condition
Chromatographic column: ZOBAX (250mm × 4.6mm, 5 μm)；
Mobile phase: methanol: potassium dihydrogen phosphate buffer solution (concentration 20mmol/L, pH=2.50)=2.5:97.5 (v/
v)；
Flow velocity: 0.6mL/min；
Column temperature: 25 DEG C；
Detection wavelength: 210nm；
Sample volume: 10 μ L.
Acetic acid standard items, propionic acid standard items and butyric acid standard items are mixed to get hybrid standard liquid sample solution, are used for HPLC
Measurement；Rumen liquid is centrifuged at 4 DEG C with 4000rpm, Rumen liquid sample solution is obtained, is measured for HPLC.
HPLC survey is carried out to acetic acid, propionic acid and the butyric acid in hybrid standard liquid and Rumen liquid according to liquid phase chromatogram condition
It is fixed, as a result as shown in Figure 1: in Fig. 1 A be acetic acid, propionic acid, three kinds of volatile fatty acid hybrid standard liquids of butyric acid chromatogram,
In, acetic acid retention time is 4.05min, and propionic acid retention time is 7.72min, and butyric acid retention time is 19.27min；B in Fig. 1
It is the chromatogram of Rumen liquid, it can be seen that being interfered around each substance peak without miscellaneous peak, separating degree in corresponding retention time
Well, it can satisfy measurement to require.
2, Specification Curve of Increasing
Accurate acetic acid standard items, propionic acid standard items and each 100 μ L of butyric acid standard items of measuring is in 10mL volumetric flask respectively, to go
Ionized water is settled to 10mL, obtains hybrid standard liquid stoste；Hybrid standard liquid stoste is used into deionized water doubling dilution, respectively
Obtain the hybrid standard product solution of following 8 concentration gradients: acetic acid is respectively 16.25,32.5,65,130,260,520,1040,
2080μg/mL；Propionic acid is respectively 15.51,31.03,62.06,124.1,248.2,496.5,993,1986 μ g/mL；Butyric acid point
Not Wei 14.84,29.7,59.38,118.75,237.5,475,950,1900 μ g/mL, successively surveyed under the chromatographic condition of step 1
The content of acetic acid, propionic acid and butyric acid in the hybrid standard liquid of fixed each concentration.It is horizontal seat by ordinate, peak area of concentration
Mark carries out the standard curve (table 2) that linear regression obtains each standard substance.
Table 2, acetic acid, propionic acid, three kinds of volatile fatty acids of butyric acid standard curve
3, Precision Experiment
It is accurate respectively to measure acetic acid standard items, propionic acid standard items and butyric acid standard items in right amount in 10mL volumetric flask, with go from
Sub- water is settled to 10mL, obtains mixed liquor, by mixed liquor.A certain amount of mixed liquor is accurately drawn, under the chromatographic condition of step 1
Continuous sample introduction 6 times, more each substance peak area, and RSD value is calculated, whether complied with standard with this method of inspection precision.
The results are shown in Table 3, and the RSD value of acetic acid, propionic acid and butyric acid is respectively 1.1%, 0.9% and 1.3% in mixed liquor,
Illustrate that instrument precision is good.
Table 3, precision experiment result
4, the rate of recovery is tested
Accurate acetic acid standard items, propionic acid standard items and the butyric acid standard items of measuring are appropriate respectively, are placed in 10mL volumetric flask, point
Not Pei Zhi known concentration acetic acid, propionic acid and butyric acid mixed liquor (original amount), every kind of substance is three concentration (table 4).Upwards
State in the acetic acid, propionic acid and butyric acid mixed liquor of known concentration be added known quantity acetic acid, propionic acid and butyric acid (additional amount), obtain to
The acetic acid of survey, propionic acid and to butyric acid mixed liquor.Acetic acid to be measured, propionic acid and butyric acid mixed liquor sample introduction are measured, standard song is utilized
The content (measured amount) of three kinds of substances of line equation calculation, the rate of recovery=measured amount/(original amount+additional amount).In the chromatography of step 1
Under the conditions of measure peak area, each content of material is calculated according to the standard curve that step 2 is established, calculates the rate of recovery.
The results are shown in Table 4, and each substance sample recovery rate is 96% or more, the method for measuring three kinds of volatile fatty acids
Accurate Determining can be carried out to the content of acetic acid, propionic acid and butyric acid.
Table 4, rate of recovery experimental result
5, in Rumen liquid acetic acid, propionic acid and butyric acid content measurement
By Rumen liquid at 4 DEG C, it is centrifuged 10min under the conditions of 4000rpm, Rumen liquid sample solution is obtained, in step 1
Chromatographic condition under during sequentially determining test control group and acetic acid in the Rumen liquid of experimental group, propionic acid and butyric acid contain
Amount.
Table 5 is the acetic acid content measurement result in the Rumen liquid of control group and experimental group, in two groups of Rumen liquid
The trend gradually increased is presented in acetic acid content, but the acetic acid content speedup in experimental group Rumen liquid is very fast.In Tiny ecosystem
After preparation stomach-filling 4d, the content of acetic acid has been significantly more than control group (t=9.281, P < 0.05) in experimental group rumen fluid；Later two
Content significant difference (P < 0.05) always of acetic acid in group Rumen liquid.
Table 5, feeding type probiotics to acetic acid content in Rumen liquid influence (μg/mL)
Table 6 is the propionic acid content measurement result in the Rumen liquid of control group and experimental group, in two groups of Rumen liquid
The trend gradually increased is presented in propionic acid content, but the propionic acid content speedup in experimental group Rumen liquid is very fast.In Tiny ecosystem
After preparation stomach-filling 2d, the propionic acid content in experimental group Rumen liquid has been significantly more than control group (t=5.574, P < 0.05)；It
Propionic acid content in two groups of Rumen liquid significant difference (P < 0.05) always afterwards.
Table 6, feeding type probiotics to propionic acid content in Rumen liquid influence (μg/mL)
Table 7 is the butyric acid assay in control group and the Rumen liquid of experimental group as a result, in two groups of Rumen liquid
The trend gradually increased is presented in butyric acid content, but the butyric acid content speedup in experimental group Rumen liquid is very fast.In Tiny ecosystem
After preparation stomach-filling 2d, the butyric acid content in experimental group Rumen liquid has been significantly more than control group (t=5.568, P < 0.05)；It
Butyric acid content in two groups of Rumen liquid significant difference (P < 0.05) always afterwards.
Table 7, feeding type probiotics to butyric acid content in Rumen liquid influence (μg/mL)
The optimization of the preparation condition of the feeding type probiotics of embodiment 3, embodiment 1
Microballoon characteristic is investigated: respectively when each bacterial strain microballoon prepares completion, room temperature 1 month after preparation, 2 months laggard
Row carries bacterium amount measurement.1g microballoon is weighed, sets in sterile purified water and stirs, is filtered with filter paper, supernatant is taken to carry out viable count measurement,
As surface bacteria containing amount.Take equivalent microballoon, be put in sterile purified water be crushed, 37 DEG C of constant temperature oscillations for 24 hours after, filtered with filter paper,
Supernatant is taken to carry out viable count measurement, as total bacterium amount carries bacterium amount=total bacterium amount-surface bacterium amount.
One, single factor exploration carries the influence of bacterium amount to each bacterial strain microballoon
Select Lactobacillus casei (Lactobacillus casei) ACCC10640, candida tropicalis (Candida
Tropicalis) ACCC21161 and bacillus subtilis (Bacillus subtilis) ACCC02157 is used for probiotics
Preparation.
The liquid spawn of Lactobacillus casei in following experiments with fermentation medium A at 35-37 DEG C the preparation method is as follows: trained
The culture (all substances in culture vessel) that feeding 46h-54h is obtained.Specifically the preparation method is as follows: by activated cheese cream
Bacillus is inoculated in the 500mL triangular flask of the fermentation medium A equipped with 100mL, 35-37 DEG C, shaking table 160rpm (radius of turn
It 20mm) cultivates 48 hours, all substances (culture) in triangular flask are the liquid spawn of Lactobacillus casei.The cheese cream bar
Lactobacillus casei content is 8.6 × 10 in the liquid spawn of bacterium9cfu/mL。
The liquid spawn of candida tropicalis in following experiments is the preparation method is as follows: with fermentation medium A at 28-30 DEG C
The culture (all substances in culture vessel) that culture 32h-40h is obtained.Specifically the preparation method is as follows: by the activated torrid zone
Candida is inoculated in the 500mL triangular flask of the fermentation medium A equipped with 100mL, 28-30 DEG C, 180rpm (radius of turn
20mm) shaken cultivation 36 hours, all substances (culture) in triangular flask are the liquid spawn of candida tropicalis.The heat
Candida tropicalis content is 7.8 × 10 in liquid spawn with Candida10cfu/mL。
The liquid spawn of bacillus subtilis in following experiments is the preparation method is as follows: with fermentation medium A at 30-32 DEG C
The culture (all substances in culture vessel) that culture 18h-24h is obtained.Specifically the preparation method is as follows: by activated withered grass
Bacillus is inoculated in the 500mL triangular flask of the fermentation medium A equipped with 100mL, 30-32 DEG C, 180rpm (radius of turn
20mm) shaken cultivation 20 hours, all substances (culture) in triangular flask are bacillus subtilis bacterium solution.The withered grass gemma
Bacillus subtilis bacterial content is 9.6 × 10 in bacillus bacterium solution9cfu/mL。
Every liter of fermentation medium A is prepared as follows: 10g glucose, 10g peptone, 1.5g KH2PO4、0.80g
MnSO4、1.00g MgSO4, 1.0g NaCl, be settled to l000mL, pH value 6.5 with distilled water；It sterilizes 20 points under the conditions of 121 DEG C
Clock.
Sodium alginate aqueous solution: 10g sodium alginate is added in deionized water, is configured to the alginic acid that concentration is 10g/L
Sodium water solution；15g sodium alginate is added in deionized water, the sodium alginate aqueous solution that concentration is 15g/L is configured to；By 20g
Sodium alginate is added in deionized water, is configured to the sodium alginate aqueous solution that concentration is 20g/L；The addition of 30g sodium alginate is gone
In ionized water, it is configured to the sodium alginate aqueous solution that concentration is 30g/L.
Calcium chloride-chitosan liquid: appropriate calcium chloride dissolution is added to the acetum that pH value is 4, then weighs certain
The chitosan dissolution of amount obtains calcium chloride-chitosan liquid.Calcium chloride concentration in calcium chloride-chitosan liquid can be 40g/
L, 50g/L, 60g/L, 80g/L or 100g/L；The concentration of chitosan is 10g/L.
Liquid spawn is added into sodium alginate colloidal solution, obtains strain-sodium alginate liquid (hereinafter referred to as A liquid),
Strain -4 DEG C of sodium alginate liquid standing is removed into bubble for 24 hours.Strain-sodium alginate liquid is added drop-wise to chlorination with syringe
In calcium-chitosan liquid (hereinafter referred to as B liquid), and with 600rmin-1Speed stirring certain time makes chitosan and sodium alginate
Glue nucleus forming reaction microreactor ball occurs.When the active constituent of liquid spawn is Lactobacillus casei, it is (dry to obtain Lactobacillus casei microballoon
Lactobacillus paracasei microbial inoculum)；When the active constituent of liquid spawn is candida tropicalis, the candida tropicalis microballoon (torrid zone is obtained
Candida microbial inoculum)；When the active constituent of liquid spawn is bacillus subtilis, bacillus subtilis microballoon (withered grass is obtained
Gemma bacillus agent).
1, sodium alginate concentration carries the influence of bacterium amount to each bacterial strain microballoon
The volume ratio of 5%, A liquid and B liquid that every kind of bacterium solution additional amount is fixed as sodium alginate colloidal solution volume is fixed as
Calcium chloride concentration is fixed as 50g/L, reaction time 20min in 2:1, B liquid, investigates seaweed in sodium alginate colloidal solution respectively
The influence that bacterium amount is formed is carried to each bacterial strain microballoon when sour na concn is respectively 10g/L, 15g/L, 20g/L and 30g/L.
The results are shown in Table 8, with the increase of sodium alginate concentration in sodium alginate colloidal solution, three kinds of bacterial strain microballoons
It carries bacterium amount and shows the trend for first increasing and reducing afterwards, three kinds of bacterial strain microballoons carry bacterium amount when sodium alginate concentration is 20g/L
Maximum, Lactobacillus casei microballoon are 3.8 × 105Cfu/g microballoon, candida tropicalis microballoon are 7.2 × 104Cfu/g microballoon, it is withered
Careless bacillus microballoon is 5.2 × 105Cfu/g microballoon.When sodium alginate concentration is low, microballoon mechanical strength is poor, stirred
It is easily broken in journey and is lost the bacterial strain of package；And when sodium alginate concentration is excessively high, viscosity is consequently increased, whipping process
Middle difference microballoon mutually collides adhesion, causes microballoon to rupture when external force carries out separated, also will affect load bacterium amount.
Table 8, sodium alginate concentration carry the influence of bacterium amount to each bacterial strain microballoon
2, calcium chloride concentration carries the influence of bacterium amount to each bacterial strain microballoon
Sodium alginate concentration is fixed on 20g/L in sodium alginate colloidal solution, and every kind of bacterium solution additional amount is fixed as alginic acid
5%, the A liquid of sodium colloidal solution volume and the volume ratio of B liquid are fixed as 2:1, reaction time 20min, investigate chlorine in B liquid respectively
Change calcium concentration is 40g/L, carries the influence that bacterium amount is formed to each bacterial strain microballoon when 60g/L, 80g/L and 100g/L.
The results are shown in Table 9, and the load bacterium amount of three kinds of bacterial strain microballoons first increases with the increase presentation of calcium chloride concentration to be reduced afterwards
Trend, it is 4.1 × 10 that it is maximum that Lactobacillus casei microballoon carries bacterium amount when calcium chloride concentration is 60g/L5Cfu/g microballoon；The torrid zone
Candida microballoon carries bacterium amount maximum when calcium chloride concentration is 40g/L, is 6.7 × 104Cfu/g microballoon；Bacillus subtilis
Microballoon carries bacterium amount maximum when calcium chloride concentration is 80g/L, is 7.5 × 105Cfu/g microballoon.Calcium chloride concentration and microsphere surface
Compactness it is directly related, when calcium chloride concentration is smaller, microballoon is not fine and close enough, thallus can be leaked out from gap cause carry bacterium
Amount reduces；When calcium chloride concentration is excessive, compactness is too strong, so that most of thallus can not be from densification when measurement carries bacterium amount
It is leaked out in microballoon, and the content of the load bacterium amount of practical measurement is caused to reduce.
Table 9, calcium chloride concentration carry the influence of bacterium amount to each bacterial strain microballoon
3, the volume ratio of A liquid and B liquid carries the influence of bacterium amount to each bacterial strain microballoon
Sodium alginate concentration is fixed on 20g/L in sodium alginate colloidal solution, and every kind of bacterium solution additional amount is fixed as alginic acid
Calcium chloride concentration is fixed as 50g/L, reaction time 20min in 5%, the B liquid of sodium colloidal solution volume, investigates A liquid and B respectively
The volume ratio of liquid is 2:1,1:1,1:2, carries the influence that bacterium amount is formed to each bacterial strain microballoon when 1:3.
The results are shown in Table 10, and Lactobacillus casei microballoon Lactobacillus casei when the volume ratio of A liquid and B liquid is 1:2 carries bacterium
Amount is maximum, is 3.6 × 105Cfu/g microballoon；Candida tropicalis microballoon carries bacterium amount most when the volume ratio of A liquid and B liquid is 1:1
It greatly, is 6.4 × 104Cfu/g microballoon；Bacillus subtilis microballoon carries bacterium amount maximum when the volume ratio of A liquid and B liquid is 2:1, is
6.3×105Cfu/g microballoon.Liquor capacity compares load bacterium amount and has no direct effect, may be by shadow to the influence for carrying bacterium amount
The viscosity of mixed solution system is rung to realize.
The volume ratio of table 10, A liquid and B liquid carries the influence of bacterium amount to each bacterial strain microballoon
4, the glue nuclear reaction time carries the influence of bacterium amount to each bacterial strain microballoon
Sodium alginate concentration is fixed on 20g/L in sodium alginate colloidal solution, and every kind of bacterium solution additional amount is fixed as alginic acid
Calcium chloride concentration is fixed as 50g/L in 5%, the B liquid of sodium colloidal solution volume, and the volume ratio of A liquid and B liquid is fixed as 2:1, respectively
The glue nuclear reaction time is investigated to carry the influence that bacterium amount is formed to each bacterial strain microballoon when 10min, 30min, 60min and 120min.
As a result as shown in table 11, with the increase in reaction time, the load bacterium amount of three groups of bacterial strain microballoons, which shows, gradually to drop
Low trend, this is because forming initial stage in microballoon, there are many not completely enclosed gaps in microsphere surface, for a long time in liquid
Impregnating in system can cause bacterial strain to be lost from these gaps to reduce the load bacterium amount of microballoon.
Table 11, glue nuclear reaction time carry the influence of bacterium amount to each bacterial strain microballoon
Two, orthogonal experiment determines the optimum preparating condition of each bacterial strain microballoon
Orthogonal experiment selects four factor three horizontal, to carry bacterium amount as evaluation index, evaluates microspheres, orthogonal arrage
As shown in table 12.
Table 12, orthogonal experiment condition
Orthogonal experiment results are as shown in table 13, and according to very poor size, the factor sequence for influencing bacterial strain microballoon is alginic acid
Na concn > calcium chloride concentration > A, B volume ratio > reaction time, the best production technology of Lactobacillus casei microballoon are as follows: sodium alginate
Concentration is 15g/L, and the volume ratio of calcium chloride concentration 60g/L, A liquid and B liquid is 1:1, reaction time 10min；Tropical vacation silk
The best production technology of yeast microballoon are as follows: sodium alginate concentration 15g/L, the volume of calcium chloride concentration 40g/L, A liquid and B liquid
Than for 1:1, the glue nuclear reaction time is 10min；The best production technology of bacillus subtilis microballoon are as follows: sodium alginate concentration is
The volume ratio of 15g/L, calcium chloride concentration 60g/L, A liquid and B liquid is 1:1, and the glue nuclear reaction time is 10min.
Table 13, Orthogonal experiment results
Three, the optimum condition verifying of each bacterial strain microballoon preparation
The best production technology of each bacterial strain obtained according to step 2 prepares the microballoon of each bacterial strain respectively, and investigates each bacterial strain
The load bacterium amount of the microballoon 30d after preparation, 60d change.The result shows that Lactobacillus casei microballoon, candida tropicalis microballoon and withered grass
Three kinds of probiotics bacterial balls of bacillus microballoon contain bacterium amount variation less in 60d Storage period, do not substantially reduce, illustrate herein
The probiotics micro-sphere structure prepared under process conditions stablizes (table 14).
Table 14, each bacterial strain microballoon study on the stability
Claims (7)
1. a kind of feeding type probiotics, active constituent Cheesecake lactobacillus, candida tropicalis and bacillus subtilis
Composition；The Lactobacillus casei is Lactobacillus casei (Lactobacillus casei) ACCC10640, the false silk ferment in the torrid zone
Mother is candida tropicalis (Candida tropicalis) ACCC21161, and the bacillus subtilis is bacillus subtilis
(Bacillus subtilis)ACCC02157；In the active constituent, the Lactobacillus casei, the candida tropicalis and
The Colony Forming Unit ratio of the bacillus subtilis is (3-7): (1-4): (5-11)；In the feeding type probiotics
Active component content is (3-9) × 105cfu/g；
The feeding type probiotics are prepared as follows: by Lactobacillus casei microbial inoculum, Candida tropicalis agent and
Bacillus subtilis microbial agent carries out being mixed to get the feeding type probiotics；The active constituent of the Lactobacillus casei microbial inoculum
For the Lactobacillus casei；The active constituent of the Candida tropicalis agent is the candida tropicalis；The withered grass bud
The active constituent of spore bacillus microbial inoculum is the bacillus subtilis.
2. feeding type probiotics according to claim 1, it is characterised in that: the feeding type probiotics are by institute
State active constituent and auxiliary material composition；The auxiliary material is made of sodium alginate, calcium chloride and chitosan.
3. feeding type probiotics according to claim 1, it is characterised in that: the Lactobacillus casei microbial inoculum, described
The mixed proportion of Candida tropicalis agent and the bacillus subtilis microbial agent meets in the feeding type probiotics
The Colony Forming Unit ratio of the Lactobacillus casei, the candida tropicalis and the bacillus subtilis is (3-7): (1-
4): (5-11).
4. feeding type probiotics described in any claim in -3 according to claim 1, it is characterised in that: the cheese
Lactobacillus microbial inoculum is prepared by following methods: Lactobacillus casei-sodium alginate liquid being added drop-wise in calcium chloride-chitosan liquid and is obtained
To the Lactobacillus casei microbial inoculum, the Lactobacillus casei-sodium alginate liquid is by sodium alginate aqueous solution and the cheese
The liquid that the bacterium solution of lactobacillus is mixed to get；The calcium chloride-chitosan liquid is by 40g-100g calcium chloride and 10g-20g shell
Glycan is dissolved in liquid obtained in the acetum that the pH value of 1L is 4；
The Candida tropicalis agent is prepared by following methods: candida tropicalis-sodium alginate liquid being added drop-wise to described
Obtain the Candida tropicalis agent in calcium chloride-chitosan liquid, the candida tropicalis-sodium alginate liquid be by
The liquid that the sodium alginate aqueous solution and the bacterium solution of the candida tropicalis are mixed to get；
The bacillus subtilis microbial agent is prepared by following methods: bacillus subtilis-sodium alginate liquid being added drop-wise to described
Obtain the bacillus subtilis microbial agent in calcium chloride-chitosan liquid, the bacillus subtilis-sodium alginate liquid be by
The liquid that the sodium alginate aqueous solution and the bacterium solution of the bacillus subtilis are mixed to get.
5. the product for the treatment of or adjuvant treatment animal rumens acid poisoning, active constituent is that any right is wanted in claim 1-4
Seek the feeding type probiotics.
6. product according to claim 5, it is characterised in that: the product has the function of following 4 kinds:
1) increase the content of acetic acid in rumen fluid；
2) increase the content of propionic acid in rumen fluid；
3) increase the content of butyric acid in rumen fluid；
4) increase ciliophoran quantity in rumen fluid.
7. feeding type probiotics described in any claim have following 4 kinds of functions in preparation in claim 1-4
Application in product:
1) increase the content of acetic acid in rumen fluid；
2) increase the content of propionic acid in rumen fluid；
3) increase the content of butyric acid in rumen fluid；
4) increase ciliophoran quantity in rumen fluid.
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| CN103689218A (en) * | 2013-12-05 | 2014-04-02 | 中国农业大学 | Complex fungicide for improving feed fiber degradation rate and positively regulating rumen fermentation propionic acid generation as well as application thereof |
| CN104664169A (en) * | 2015-03-18 | 2015-06-03 | 内蒙古粮食科学研究设计院有限责任公司 | Biological enhancer for ruminant feeds, and preparation method for biological enhancer |
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| CN103689218A (en) * | 2013-12-05 | 2014-04-02 | 中国农业大学 | Complex fungicide for improving feed fiber degradation rate and positively regulating rumen fermentation propionic acid generation as well as application thereof |
| CN104664169A (en) * | 2015-03-18 | 2015-06-03 | 内蒙古粮食科学研究设计院有限责任公司 | Biological enhancer for ruminant feeds, and preparation method for biological enhancer |
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