CN107041470A - It is a kind of to promote feed of Lactation of Dairy Cow and preparation method thereof - Google Patents
It is a kind of to promote feed of Lactation of Dairy Cow and preparation method thereof Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The present invention relates to a kind of feed of promotion Lactation of Dairy Cow and preparation method thereof.The feed of the present invention includes the raw material of following parts by weight：20 25 parts of monkshood, 20 25 parts of radix glycyrrhizae, 23 parts of rhizoma atractylodis, 23 parts of dried orange peel, 23 parts of the bark of official magnolia, 23 parts of the tuber of pinellia, 23 parts of balloonflower root, king do not stay 28 parts, 23 parts of Yun Ling, 10 15 parts of soybean, 15 20 parts of clover stalk powder, 0.1 0.5 parts of vitamin E, 0.4 0.5 parts of lactic acid bacteria culture solution, 0.3 0.5 parts of kluyveromyces marxianus bacteria culture fluid；The feed is by preparing the mixture A of monkshood and radix glycyrrhizae, preparing nutrient solution a and nutrient solution b, lactic fermentation, prepare mixture D, secondary fermentation, addition soybean, clover step and be prepared from.The present invention passes through monkshood and radix glycyrrhizae ensiling, lactic fermentation, effect of monkshood and radix glycyrrhizae is mutually merged auxiliary, reduces monkshood toxicity, and make feed color and luster, mouthfeel more preferable；Using the secondary fermentation of kluyveromyces marxianus, each medicine property of medicine is harmonious strengthens its effect, and the quality of feed is effectively improved again.
Description
Technical field
The invention belongs to animal feed processing technique field, and in particular to a kind of feed of promotion Lactation of Dairy Cow and its preparation
Method.
Background technology
With the fast development of China's dairy industry, cow total amount is continuously increased, but due to the relatively low of overall raise competence and life
The backwardness of production capacity power, the serious problems such as current dairy generally existing milk production is low, Incidence is high, it is impossible to fully play
Cow product potentiality, cause cowboying benefit recessive drain occur.So, necessary exploitation promotes the feed of Lactation of Dairy Cow to change
Become present situation, so as to reach higher raising dairy cattle benefit.
In the prior art, promote Lactation of Dairy Cow feed mainly in feed add trace and REE elements additive or
Traditional Chinese medicine extraction component.
As the patent of Application No. " CN201510311742.0 " discloses a kind of Lactation of Dairy Cow phase feed addictive, by with
The raw material common process of lower parts by weight is formulated：0.5-0.7 parts of vitamin A, 0.01-0.02 parts of vitamine D3, dimension life
Plain E 6-10 parts, 14-18 parts of nicotinic acid, 12-20 parts of ferrous sulfate, 3-4 parts of copper sulphate, 11-19 parts of zinc sulfate, manganese sulfate 11-19
Part, 15-25 parts of 1% KI, 3-7 parts of 1% sodium selenite, 2-10 parts of 1% cobalt chloride, 895-910 parts of stone flour.
Due to lactating cow to micro first rope the need for measure be not fixed, become with growth with giving milk the stage (output of milk)
Change, and deficiency disease also seldom occurs for some trace elements such as molybdenum, fluorine etc., it is not necessary to extra supplement.But in production, due to milk cow
Raiser is not enough to the understanding of trace element or businessman misleading, excessive in dairy cow diet, cross and use micro member for a long time
The phenomenon of element happens occasionally, and so not only causes trace element largely to be accumulated in Contents in Cows, enters people by milk or beef
Body, and then health is caused harm；On the other hand, this kind of trace element is excreted in environment with fecaluria etc., can be caused again
Environmental pollution, influences human health.Such as copper, the zinc of high dose, excreted by excrement, soil hardening, contaminant water can be caused
Source, makes water quality deterioration.High-copper can also cause animal's liver copper to accumulate, so that its edibility declines, or even human body be produced
Toxic action, is also a potential threat to ecological environment.In addition, being measured and toxic dose the need for some trace elements of milk cow
Scope differs very little, if addition reaches toxic dose, frequently can lead to milk cow and disease occurs.Therefore, in milk cattle cultivating, absolutely
Trace mineral supplement can not be abused.
It is comparatively safe and reliable using traditional Chinese medicine extraction active ingredient as cow feed additive.Such as Application No.
The patent of " CN200710143609.4 " discloses a kind of feed addictive and its production method, includes the group of following parts by weight
Part：Modified montmorillonite used 400-1000, eucommia ulmoides extracts 50, bletilla striata extract 50, licorice 50, glucose oxidase 5-
50, sodium Diacetate 40-100,10-60 grams of crospovidone (PVP)；Production technology：(A) raw material prepares, with standby：Press
Material modification montmorillonite 600, sodium Diacetate 50, glucose oxidase 20, eucommia ulmoides extracts 50, the bletilla striata are weighed according to following parts by weight
Extract 50, licorice 50, crospovidone (PVP) 20；Prepare modified montmorillonite used, with standby：(1) take illiteracy de-
Stone 30g, crosses mesh size after drying and is sieved for 0.250mm.(2) montmorillonite of sieving is added to 100ml 1mol/L CX quaternary ammonium salt
In solution, it is put into 60 DEG C of thermostatted waters and vibrates 1h, then centrifuge, inclines supernatant.(3) 100ml 1mol/L CX quaternary ammoniums are added
Salting liquid, repeats the process of above-mentioned (2).(4) water washing precipitate is distilled 1 time with 100ml, centrifugation, incline supernatant, by product
In 105 DEG C of drying, grind, cross aperture and sieved for 0.250mm, is just made modified montmorillonite used.(B) eucommia ulmoides extracts are extracted with the bletilla striata
Thing, licorice carry out first time mixing, obtain once mixture；(C) glucose oxidase is added in once mixture
Carry out second to mix, obtain secondary mixture；(D) sodium Diacetate third time is added in secondary mixture to mix, and obtains three
Secondary mixture；(E) crospovidone (PVP) is added in three mixtures, four mixtures are obtained：(F) the 4th
Add modified montmorillonite used in secondary mixture, that is, obtain finished product.
But medicine effective component extracting is in extraction process, the loss of mass efficient composition can be caused, while also there is life
The problem of production. art is complicated, cost increases.
The content of the invention
For in lactating cow feed add trace element exist series of problems, and addition traditional Chinese medicine extraction effectively into
The problem of point loss of mass efficient composition existed, complex manufacturing, cost increase, in being utilized the invention provides one kind
Herbal medicine silage fermentation and secondary fermentation, with lactagogue effect, while there is invigorating the spleen, qi-regulating, strengthen the low of milk cow immunity effect
Cost green feed.
The main component of the feed is the byproduct of monkshood.Monkshood is Ranunculaceae, aconitum plant Aconitum
Carmichaeli Debx root, usually as recuperating depleted yang, mends fire supporing yang, the Chinese medicine of eliminating cold to stop pain.And in reality production
In, the sub- root of well-grown rhizome of Chinese monkshood is generally selected as Chinese medicine, its byproduct cauline leaf and fibrous root is all discarded, this is resulted in
The waste of resource.Find according to the study, a certain amount of effective component is also contained in the cauline leaf and fibrous root of monkshood.So, present invention profit
Monkshood is replaced with the monkshood cauline leaf and fibrous root of ensiling as feed, so as to reduce cost and reach the mesh that waste resource is recycled
's.
A kind of feed of promotion Lactation of Dairy Cow, includes the raw material of following parts by weight：20-25 parts of monkshood, 20-25 parts of radix glycyrrhizae,
2-3 parts of rhizoma atractylodis, 2-3 parts of dried orange peel, 2-3 parts of the bark of official magnolia, 2-3 parts of the tuber of pinellia, 2-3 parts of balloonflower root, king do not stay 2-8 parts, 2-3 parts of Yun Ling, lactic acid
0.4-0.5 parts of bacteria culture fluid, 0.3-0.5 parts of kluyveromyces marxianus bacteria culture fluid, 10-15 parts of soybean, clover stalk powder 15-
20 parts, 0.1-0.5 parts of vitamin E.
Preferably, the raw material of following parts by weight is included：25 parts of monkshood, 25 parts of radix glycyrrhizae, 2 parts of rhizoma atractylodis, 2 parts of dried orange peel, the bark of official magnolia 2
Part, 2 parts of the tuber of pinellia, 2 parts of balloonflower root, king do not stay 2 parts, 2 parts of Yun Ling, 0.5 part of lactic acid bacteria culture solution, the culture of kluyveromyces marxianus bacterium
0.3 part of liquid, 10 parts of soybean, 15 parts of clover stalk powder, 0.1 part of vitamin E.
Preferably, the lactic acid bacteria culture solution is to be inoculated in Pediococcus acidilactici in liquid MRS culture mediums by 2% inoculum concentration
Culture, it is 0.6 × 10 that culture, which is obtained to exponential phase, under the conditions of 30-37 DEG C8CFU/ML-1.0×108CFU/ML lactic acid
Bacteria culture fluid；The kluyveromyces marxianus bacteria culture fluid is to be inoculated in kluyveromyces marxianus bacterium by 2% inoculum concentration
Cultivated in liquid potato culture, it is 0.6 × 10 that culture, which is obtained to exponential phase, under the conditions of 35-38 DEG C8CFU/ML-
1.0×108CFU/ML kluyveromyces marxianus bacteria culture fluid.
Preferably, the monkshood is the cauline leaf or fibrous root of monkshood, and radix glycyrrhizae used is the stem or fibrous root of radix glycyrrhizae.
The preparation method of the feed of promotion Lactation of Dairy Cow described in any of the above-described kind, comprises the following steps：
Step 1, monkshood and the mixture A of radix glycyrrhizae are prepared：Monkshood and radix glycyrrhizae are dried to moisture respectively and reach 50-60%,
Then guillotine cutting is well mixed into fragment, obtains mixture A；
Step 2, lactic acid bacteria culture solution and kluyveromyces marxianus bacteria culture fluid are prepared：Extracting lactic acid piece coccus, connects by 2%
Kind of amount is inoculated in liquid MRS culture mediums and cultivated, under the conditions of 30-37 DEG C culture obtain to exponential phase be 0.6 ×
108CFU/ML-1.0×108CFU/ML lactic acid bacteria culture solution；Kluyveromyces marxianus bacterium is taken, is inoculated in by 2% inoculum concentration
Cultivated in liquid potato culture, it is 0.6 × 10 that culture, which is obtained to exponential phase, under the conditions of 35-38 DEG C8CFU/ML-
1.0×108CFU/ML kluyveromyces marxianus bacteria culture fluid；
Step 3, lactic fermentation：The mixture that the lactic acid bacteria culture solution that step 2 is prepared is obtained with step 1 is by 1：100
It is well mixed, tight, ferment at constant temperature then is wrapped up with film, fermentate B is obtained；
Step 4, mixture D is prepared：The rhizoma atractylodis dried, dried orange peel, the bark of official magnolia, the tuber of pinellia, balloonflower root, Wang Buliu, Yun Ling are taken, mixing is simultaneously
Crush, cross 80-120 mesh sieves, obtain mixture D；
Step 5, secondary fermentation：The fermentate B mixing that the mixture D and step 3 that step 4 is obtained are obtained, is mixed
Thing D, kluyveromyces marxianus bacteria culture fluid is accessed by mixture D, is wrapped up tight, ferment at constant temperature with film, is obtained fermentate E；
Step 6, addition soybean, clover：Soybean, clover are crushed, 80-120 mesh sieves is crossed, is mixed with fermentate E, is stirred,
Obtain the feed of the present invention.
Preferably, length 1-3cm of the guillotine cutting into fragment in the step 1.
Preferably, the condition of ferment at constant temperature is in the step 3：Fermented 20-25 days under 35-38 DEG C of constant temperature.
Preferably, the amount ratio of mixture D and kluyveromyces marxianus bacteria culture fluid is 1 in the step 5：49；It is described
Ferment at constant temperature is fermented 10-15 days under 40-45 DEG C of constant temperature.
Preferably, mixing speed is 1000-3000r/min in the step 6, and mixing time is 10-15min
Compared with prior art, beneficial effects of the present invention：
(1) present invention passes through monkshood and radix glycyrrhizae ensiling, lactic fermentation, effect of monkshood and radix glycyrrhizae is mutually merged auxiliary,
So as to so that monkshood toxicity is reduced, and make feed color and luster, mouthfeel more preferable；
(2) present invention is using the secondary fermentation of kluyveromyces marxianus, and each medicine property of medicine is harmonious strengthens its effect, together
Shi Caiyong saccharomycete biologies take oxygen by force, promote lactic acid bacteria fast-growth to breed and the generation metabolite that fermented together with yeast, that is, press down
The growth of other miscellaneous bacterias has been made, the quality of feed is effectively improved again, while reducing the loss of dry, egg has been also add
Bai Hanliang, so as to improve palatability and digestibility；
(3) addition soybean, clover are to have covered its feed medium-height grass flavour of a drug in the present invention, so that feed mouthfeel is more preferable, fragrant
Taste is more sufficient；Soybean, clover are nutritious simultaneously, improve milk quality, improve milk production；
(4) present invention, so as to reach the purpose that discarded object is recycled, is produced into it using monkshood, the byproduct of radix glycyrrhizae
Sheet is lower, be easier to be received by dairy farmer.
Brief description of the drawings
Fig. 1 is active ingredient comparative result one before and after secondary fermentation of the present invention；
Fig. 2 is active ingredient comparative result two before and after secondary fermentation of the present invention；
Fig. 3 is embodiment 2-4 different formulations feed quality Basic Evaluation comparative results.
Embodiment
Further detailed description is done to the present invention with reference to specific embodiment, but embodiments of the present invention are not limited to
This.
Embodiment 1：
A kind of feed of promotion Lactation of Dairy Cow, includes the raw material of following parts by weight：22 parts of monkshood, 22 parts of radix glycyrrhizae, rhizoma atractylodis 3.5
Part, 3.5 parts of dried orange peel, 3.5 parts of the bark of official magnolia, 3.5 parts of the tuber of pinellia, 3.5 parts of balloonflower root, king do not stay 8 parts, 2.5 parts of Yun Ling, lactic acid bacteria culture solution
0.4 part, 0.5 part of kluyveromyces marxianus bacteria culture fluid, 12 parts of soybean, 18 parts of clover stalk powder, 0.5 part of vitamin E.
The preparation method of the feed of the promotion Lactation of Dairy Cow of above-mentioned raw materials proportioning, comprises the following steps：
(1) fibrous root or cauline leaf of monkshood and radix glycyrrhizae are dried to moisture respectively and reaches 50%-60%, guillotine cutting is into 1-3cm
Fragment, be well mixed, obtain mixture A.
(2) extracting lactic acid piece coccus, is inoculated in liquid MRS culture mediums by 2% inoculum concentration and cultivated, and is cultivated under the conditions of 35 DEG C
It is 0.6 × 10 to obtain to exponential phase8CFU/ML-1.0×108CFU/ML lactic acid bacteria culture solution；Marx's Crewe is taken to tie up ferment
Female bacterium, is inoculated in liquid potato culture by 2% inoculum concentration and cultivated, cultivated and obtained to exponential phase under the conditions of 37 DEG C
For 0.6 × 108CFU/ML-1.0×108CFU/ML kluyveromyces marxianus bacteria culture fluid；
(3) mixture A that the lactic acid bacteria culture solution that step (2) prepares obtains with step (1) is weighed by 1：100 mixing are equal
It is even, tight, 35 DEG C of constant temperature are wrapped up with film, ferments 22 days, obtains fermentate B.
(4) rhizoma atractylodis dried, dried orange peel, the bark of official magnolia, the tuber of pinellia, balloonflower root, Wang Buliu, Yun Ling are crushed, crosses 100 mesh sieves, must mix
Thing D.
(5) mixture D and fermentate B are mixed, obtains mixture D, mixture D is pressed 1:49 access Marx Crewe dimensions
Yeast bacteria culture fluid, wraps up tight with film, in 42 DEG C of constant temperature, ferments 13 days, obtains fermentate E.
(6) soybean, clover are crushed, crosses 120 mesh sieves, mixed with fermentate E, mixing speed is 2000r/min, during stirring
Between be 12min, obtain the present invention feed.
In order to be better described the synergistic action effect between each raw material of the invention, and preparation method of the present invention technique
Feature, is verified below by the method for experiment.
Experiment 1：Before and after lactic fermentation, the detection of mixture A and fermentate B active ingredients and toxicity
1.1 experimental method：
1.1.1 effective liquorice assay method
The preparation of reference substance solution
Extracting liquorice glycosides reference substance, ammonium glycyrrhetate reference substance are appropriate, accurately weighed, plus 70% ethanol is respectively prepared every lmL and contained
The μ g of liquiritin 20, ammonium glycyrrhetate 0.2mg solution, are produced (glycyrrhizic acid weight=ammonium glycyrrhetate weight/1.0207).
The preparation of need testing solution
Mixture A and fermentate B is taken to distinguish and (cross No. three sieves) about 0.2g, it is accurately weighed, put in conical flask with cover, precision adds
Enter 70% ethanol 100mL, close plug, weighed weight, ultrasonically treated (power 250W, frequency 40kHz) 30 minutes is let cool, then weighed
Weight, the weight of less loss is supplied with 70% ethanol, is shaken up, filtration, is taken subsequent filtrate, is produced.
Determination method
It is accurate respectively to draw reference substance solution and each l0 μ L of need testing solution, liquid chromatograph is injected, determines, produces.
1.1.2 monkshood active ingredient and toxicity test method
The preparation of reference substance solution
Take mesaconine reference substance, Hypaconitine reference substance, aconitine reference substance, benzoylmesaconine reference substance,
Benzoyl aconine reference substance, benzoyl time aconine reference substance are appropriate, accurately weighed, plus isopropanol-dichloromethane (1：
1) mixed solution that every lmL respectively contains 10 μ g is made in mixed solution, produces.
The preparation of need testing solution
Mixture A and fermentate B is taken to distinguish and (cross No. three sieves) about 2g, it is accurately weighed, put in conical flask with cover, ammonification test solution
3mL, precision adds isopropanol-ethyl acetate (1：1) mixed solution 50mL, weighed weight, ultrasonically treated (power 300W, frequency
40kHz, water temperature is below 25 DEG C) 30 minutes, let cool, then weighed weight, with isopropanol-ethyl acetate (1:1) mixed solution is mended
The weight of sufficient less loss, shakes up, filtration.Precision measures subsequent filtrate 25mL, and less than 40 DEG C are recovered under reduced pressure solvent to doing, and residue precision adds
Enter isopropanol-dichloromethane (1:1) mixed solution 3mL dissolves, filtering, takes subsequent filtrate, produces.
Determination method
It is accurate respectively to draw reference substance solution and need testing solution each 10 μ L injection liquid chromatographs, determine, produce.
1.2 experimental result：
Table 1 ferments front and rear active ingredient comparative result
| Content before fermentation | Content after fermentation | |
| Liquiritin (mg/g) | 14.3±1.1 | 17.4±1.3* |
| Glycyrrhizic acid (mg/g) | 25.1±1.5 | 27.8±2.1 |
| Mesaconine (μ g/g) | 812.1±12.9 | 91±6.4** |
| Hypaconitine (μ g/g) | 215.2±9.8 | 46±1.2** |
| Aconitine (μ g/g) | 316.4±10.2 | 52±12.1** |
| Benzoylmesaconine (μ g/g) | 23±1.4 | 95±5.8** |
| Benzoyl aconine (μ g/g) | 27±1.6 | 98±6.7** |
| Benzoyl time aconine (μ g/g) | 16±1.1 | 79±4.5** |
" * " represents that experimental group is in P compared with before fermentation in table<0.05 has significant difference；" before * * " are represented and fermented
Compared to experimental group in P<0.01 has pole significant difference.
The main diester-type alkaloids of toxic component (mesaconine, Hypaconitine, aconitine) of monkshood, diester-type alkaloids
Toxicity to human body is mainly shown as suppression breathing and arrhythmogenic effect, and the mechanism of alkaloid proarrhythmia is to promote the heart
Myocyte's Na+ channel openers, accelerate to flow in Na+, promote cell depolarization, so as to cause arrhythmia cordis.
Generally with removing toxic substances synergist of the radix glycyrrhizae as monkshood.Radix glycyrrhizae has the effect of solution monkshood toxicity.From theory of traditional Chinese medical science
For, radix glycyrrhizae it is sweet can and, reconcile the monkshood property of medicine to detoxify, can reconcile the property of monkshood partial temperature, the anti-pungent too that dissipates is allowed to Gui Yiping
With；Its sweet energy is slow, relaxes monkshood high violent with removing toxic substances；Its sweet energy is mended, and is strengthened the body resistance to consolidate the constitution, radix glycyrrhizae gives compatibility acridness and sweetness activating yang with attached,
Yang-energy is sufficient, strengthens the beautiful removing toxic substances of body tolerance：Radix glycyrrhizae for can the moon can sun medicine, sweet temperature is generally to help sun, and emergency is generally benefit the moon
As a result.For doctor trained in Western medicine is theoretical, the composition of radix glycyrrhizae reduction monkshood toxicity is considered glycyrrhizic acid mostly.Containing multiple in glycyrrhizic acid molecule
Carboxyl, with stronger acidity, can occur precipitation reaction with a variety of alkaloids in monkshood, generate water insoluble macromolecular network
Compound, so as to reduce ester alkaloid amount in decoction.Meanwhile, the hydrolysate glucuronic acid of glycyrrhizic acid in vivo is body solution
One of important substance of poison.Glucuronic acid can be combined with internal hydroxyl or carboxyl compound, generate glucuronic acid
Glycoside derivates are external through urine ejection.It can be with Aconitum alkaloids hydroxy combining, generation low toxicity or nontoxic glucuronic acid network
Compound and by urine ejection.The synergistic effect of Radix Glycyrrhizae on Radix Aconiti Preparata is mainly manifested in the synergy between both each compositions.
Fermentation can preserve feed and its nutriment for a long time, meanwhile, the activity of microorganism makes ensilage with fragrance
Sour-sweet taste, improves the palatability of domestic animal.Also, by reference to document, it is found that fermentation can give birth to the diester-type of monkshood
Alkaloids (mesaconine, Hypaconitine, aconitine) are converted into monoester alkaloid (benzoyl mesaconine, benzoyl time rhizome of Chinese monkshood
Alkali, benzoyl aconite alkali) so that the toxicity reduction of monkshood, and fermenting can be such that effect of monkshood and radix glycyrrhizae mutually merges
Auxiliary.
So, the present invention first uses lactobacillus-fermented radix glycyrrhizae and monkshood.
Active ingredient comparative result before and after fermentation：In table 1, effective liquorice (liquiritin, glycyrrhizic acid) is substantially
Increase, diester-type alkaloids (mesaconine, Hypaconitine, aconitine) content of monkshood is substantially reduced, monoester alkaloid (benzene
The new aconine of formyl, benzoyl aconine, benzoyl time aconine) showed increased, its content meets《Middle traditional Chinese medicines
Allusion quotation》Standard.
Experiment 2：Before and after secondary fermentation, active ingredient compares
2.1 experimental method：
2.1.1 radix glycyrrhizae, monkshood effective constituent determination be ibid
2.1.2 Atisine chloride Atractydin assay
The preparation of reference substance solution
Take Atisine chloride Atractydin reference substance appropriate, it is accurately weighed, plus the solution that every 1mL contains 20 μ g is made in methanol, produces.
The preparation of need testing solution
Mixture D and fermentate E is taken to distinguish and (cross No. three sieves) about 0.2g, it is accurately weighed, put in conical flask with cover, precision adds
Enter methanol 50mL, close plug, weighed weight, ultrasonically treated (power 250W, frequency 40kHz) 1 hour is let cool, then weighed weight, is used
Methanol supplies the weight of less loss, shakes up, filtration, takes subsequent filtrate, produces.
Determination method
It is accurate respectively to draw reference substance solution and each 10 μ L of need testing solution, liquid chromatograph is injected, determines, produces.
2.1.3 content of hesperidin is determined
The preparation of reference substance solution
Take aurantiamarin reference substance appropriate, it is accurately weighed, plus solution of every lmL containing 0.4mg is made in methanol, produces.
The preparation of need testing solution
Mixture D and fermentate E about lg respectively is taken, it is accurately weighed, put in apparatus,Soxhlet's, plus petroleum ether (60
90mL), 2 are heated to reflux3 hours, petroleum ether is discarded, the dregs of a decoction are volatilized, plus methanol 80mL is reheated and is back to extracting liquid colourless,
Let cool, filter, filtrate is put in 100mL measuring bottles, with a small amount of methanol fraction time washing container, washing lotion is filtered in the same measuring bottle of people, plus first
Alcohol shakes up to scale, produces.
Determination method
It is accurate respectively to draw reference substance solution and need testing solution each 5 μ L injection liquid chromatographs, determine, produce.
2.1.4 magnolol and honokiol assay
The preparation of reference substance solution
Take magnolol reference substance, honokiol reference substance appropriate, it is accurately weighed, plus every lmL is respectively prepared containing the bark of official magnolia in methanol
The μ g of 40 μ g honokiols of phenol 24 solution, is produced.
The preparation of need testing solution
Mixture D and fermentate E is taken to distinguish and (cross No. three sieves) about 0.2g, it is accurately weighed, put in conical flask with cover, precision adds
Enter methanol 25mL, shake up, close plug impregnates 24 hours, filtration, and precision measures subsequent filtrate 5mL, put in 25mL measuring bottles, plus methanol is extremely
Scale, shakes up, and produces.
Determination method
It is accurate respectively to draw each 4 μ L of the above two reference substance solution and μ L of need testing solution 4, liquid chromatograph is injected, is surveyed
It is fixed, produce.
2.1.5 the assay of Platycodin D
The preparation of reference substance solution
Take Platycodin-D for Reference Substance appropriate, it is accurately weighed, plus solution of every lmL containing 0.5mg is made in methanol, produces.
The preparation of need testing solution
Mixture D and fermentate E is taken to distinguish and (cross No. two sieves) about 2g, accurately weighed, precision adds alcohol 50mL in 50%, claims
Determine weight, ultrasonically treated (power 250W, frequency 40kHz) 30 minutes is let cool, then weighed weight, is evaporated with 50% methanol, residual
Slag adds water 20mL, and low-grade fever makes dissolving, is extracted 3 times with the shaking of water saturated n-butanol, and each 20mL merges n-butanol liquid, uses ammonia
Test solution 50mL is washed, and discards ammoniacal liquor, then is washed with the water 50mL of n-butanol saturation, discards aqueous, n-butanol liquid is evaporated, and residue adds
Methanol 3mL mixes dissolving, plus silica gel 0.5g thoroughly, puts and is evaporated in water-bath, is added on silicagel column 100120 mesh, 10g, internal diameter is 2cm,
- methanol (9 is burnt with dichloromethane：1) in mixed solution wet method dress post, with chloroform-methanol (9：1) mixed solution 50mL is eluted,
Discard eluent, then with chloroform-methanol-water (60：20：3) mixed solution 100mL is eluted, and discards eluent, after with trichlorine
Methane-methanol-water (60：29：6) mixed solution 100mL is eluted, and is collected eluent, is evaporated, residue adds methanol to dissolve, is transferred to
In 5mL measuring bottles, plus methanol is to scale, shakes up, filtration, produces.
Determination method
It is accurate respectively to draw the μ L of reference substance solution 10, the μ L of need testing solution 10, liquid chromatograph is injected, determines, uses external standard
Two-point method logarithmic equation is calculated, and is produced.
2.1.6 Vaccarin assay
The preparation of reference substance solution
Take Vaccarin reference substance appropriate, it is accurately weighed, plus solution of every lmL containing 0.lmg is made in 70% methanol,
Produce.
The preparation of need testing solution
Mixture D and fermentate E is taken to distinguish and (cross No. three sieves) about 1.2g, it is accurately weighed, put in conical flask with cover, precision adds
Enter 70% methanol 50mL, weighed weight, ultrasonically treated (power 250W, frequency 33kHz) 30 minutes is let cool, then weighed weight, is used
70% methanol supplies the weight of less loss, shakes up, filtration, takes subsequent filtrate, produces.
Determination method
It is accurate respectively to draw reference substance solution and each l0 μ L of need testing solution, liquid chromatograph is injected, determines, produces.
2.1.7 the measure of lactic acid content
The drafting of standard curve
The μ gmL of lactate standard liquid 1,2,3,4,5,6 are respectively configured-1, be separately added into colorimetric cylinder lactate standard liquid 1mL,
4.0% copper-bath 0.05m L, add concentrated sulfuric acid 6mL, react 5min, are cooled to after less than 20 DEG C, then respectively at each ratio
1% alkali solubility xenol solution 0.05mL is added in colour tube, is well mixed, 6-8h is placed at room temperature, overnight, in 570nm
Lower carry out colorimetric estimation, draws out standard curve.
Sample is determined
1g fermented feeds are dissolved in 100mL distilled water, 4000rpm centrifugation 10min, supernatant 1mL are taken, by standard curve
Lactic acid content (%) in determination sample.
2.1.8 the measure of oligomeric polysaccharide content
The preparation of reference substance solution
Mannose, rhamnose, glucose, galactolipin, arabinose, xylose standard product 0.5g accurately are weighed, 50% second is used
Alcoholic solution dissolves and is settled to 10mL, is configured to the standard reserving solution that concentration is 50mg/mL, produces.
Need testing solution
Feed Sample 1g is weighed in 100mL beaker, the ethanol solutions of 25mL 50% are added, in ultrasonic oscillator
After ultrasonic extraction 20min, it is transferred in 50mL volumetric flasks, 50% ethanol solution is settled to scale, mixes, take supernatant, with
0.45 μm of membrane filtration, filtrate is for analysis.
Determination method
It is accurate respectively to draw reference substance solution and each 20 μ L of need testing solution, liquid chromatograph is injected, determines, produces.
2.1.9 the measure of protein content
Sample treatment
1g samples are weighed, accurately to 0.0002g, are put into kjeldahl flask, copper sulphate 0.4g, anhydrous potassium sulfate 6g is added,
Selenium powder 0.004g, with sample mixed, then enriching sulfuric acid 12.5mL, carefully heats on stove disappearing to boil, treats sample coking, foam disappears
Lose, strengthen firepower, to solution clarification, reheat at least 1h.By the above-mentioned boil liquid cooling that disappears, 100mL capacity is transferred to without loss
In bottle, constant volume is sample decomposed solution after cooling, takes the boric acid solutions of 20mL 2%, plus mixed indicator 2 to drip, and makes semimicro distillation dress
This solution is immersed in the end put, and accurately pipettes in sample decomposed solution 10mL injection reative cells, is stoppered entrance glass stopper, adds
10mL 40%NaoH solution, carefully lifts glass stopper and is allowed to flow into reative cell, is stoppered glass stopper, and the sealing that adds water in porch
It is good, gas leakage is prevented, is distilled, self-absorption liquid is changed into blueness and starts timing, after 4 minutes, condensation pipe end is left absorption liquid level, then
Distillation 1 minute, condensation pipe end is cleaned with distilled water, and washing lotion flows into absorbing liquid.
Titration
Absorbing liquid after absorbing ammonia is titrated with 0.02mol/L standard liquid immediately, and solution becomes bois de rose by blueness and is
Terminal, record consumes the volume of standard liquid, calculates protein content.
2.2 experimental result
Microorganism produces a variety of enzymes such as cellulase, lignoenzyme, lipase during growth metabolism.Enzyme is that have height
The biocatalyst of catalytic efficiency is spent, it makes to quickly complete under complicated biochemical reaction.Microorganism has powerful decomposition
Conversion capability, and abundant secondary metabolite can be produced.
Fermented tcm has the advantage that：(1) content of effective ingredient, Ministry of Public Health's institute for drug control experiment are fully improved
Show, fermented tcm only needs to 1/28 amount, just can be played with a amount of common water extract and be equal to drug effect；(2) attenuation is played
The effect of synergy；(3 reduce pharmaceutical molecular weight.Medicine enters the effective active component that some can not be used directly after body, leads to
Microbial degradation is crossed to be absorbed and used into small-molecule active substance.Small-molecule active substance is due to being easier to by blood-brain barrier
With body cell protein binding, so having more high activity than macromolecular substances.Pharmacokinetic proves that methods of glycosides is in intestines
It is difficult to absorb in road, it is most of that the smaller aglycon of molecular weight need to be decomposed into through enteron aisle bacterial enzyme and curative effect is played.
During Fig. 1 is active ingredient comparative result one before and after secondary fermentation, Fig. 1, " * " represents the experimental group compared with control group
In P<0.05 has significant difference, and " * * " represent that experimental group is in P compared with control group<0.01 has pole significant difference.Can by Fig. 1
Know：Before and after secondary fermentation, diester-type alkaloids (mesaconine, Hypaconitine, aconitine) content is extremely significantly reduced, Arabic
Sugar and Xylose Content extremely significantly increase, monoester alkaloid (benzoyl Hypaconitine), aurantiamarin, magnolol, rhamnose and half
Lactose content significantly increases.
During Fig. 2 is active ingredient comparative result two before and after secondary fermentation, Fig. 2, " * " represents the experimental group compared with control group
In P<0.05 has significant difference, and " * * " represent that experimental group is in P compared with control group<0.01 has pole significant difference.Can by Fig. 2
Know：Before and after secondary fermentation, lactic acid, protein, glycyrrhizic acid, liquiritin have all increased.Wherein lactic acid content has before and after fermentation
Significant difference, protein content has pole significant difference before and after fermentation.
Control group is the feed of cattle farm, is formulated (parts by weight)：Corn 48；Soya-bean cake 20；Wheat bran 22；Yeast 5；Food
Salt 2；Calcium bicarbonate 1；Additive 2；Wherein additive is the combination of nicotinic acid, zinc methionine and vitamin b.
Embodiment 2：
A kind of feed of promotion Lactation of Dairy Cow, includes the raw material of following parts by weight：25 parts of monkshood, 25 parts of radix glycyrrhizae, rhizoma atractylodis 2
Part, 2 parts of dried orange peel, 2 parts of the bark of official magnolia, 2 parts of the tuber of pinellia, 2 parts of balloonflower root, king do not stay 2 parts, 2 parts of Yun Ling, 0.5 part of lactic acid bacteria culture solution, Marx
0.3 part of kluyveromyces, 10 parts of soybean, 15 parts of clover stalk powder, 0.1 part of vitamin E.
The preparation method of the feed of the promotion Lactation of Dairy Cow of above-mentioned raw materials proportioning, comprises the following steps：
(1) fibrous root or cauline leaf of monkshood and radix glycyrrhizae are dried to moisture respectively and reach 60%, guillotine cutting into 1cm fragment,
It is well mixed, obtain mixture A.
(2) extracting lactic acid piece coccus activation bacterium solution, is inoculated in MRS culture mediums by 2% inoculum concentration, 24 is cultivated under the conditions of 37 DEG C
Hour, it is 0.6 × 10 to obtain exponential phase8CFU/ML-1.0×108CFU/ML lactic acid bacteria culture solution；Take Marx's Crewe
Saccharomycete activating solution is tieed up, is inoculated in by 2% inoculum concentration in potato culture, is cultivated 28 hours under the conditions of 38 DEG C, obtains logarithm
Growth period is 0.6 × 108CFU/ML-1.0×108CFU/ML kluyveromyces marxianus bacteria culture fluids.
(3) mixture that the lactic acid bacteria culture solution that step (2) prepares obtains with step (1) is weighed by 1:100 mixing are equal
It is even, tight, 37 DEG C of constant temperature are wrapped up with film, ferments 25 days, obtains fermentate B.
(4) rhizoma atractylodis dried, dried orange peel, the bark of official magnolia, the tuber of pinellia, balloonflower root, Wang Buliu, Yun Ling are crushed, crosses 120 mesh sieves, must mix
Thing D.
(5) mixture D and fermentate B are mixed, obtains mixture D, mixture D is pressed 1：49 access Marx Crewe dimensions
Yeast bacteria culture fluid, wraps up tight with film, in 45 DEG C of constant temperature, ferments 15 days, obtains fermentate E.
(6) soybean, clover are crushed, crosses 120 mesh sieves, mixed with fermentate E, mixing speed is 3000r/min, during stirring
Between be 15min, obtain the present invention feed.
Embodiment 3：
A kind of feed of promotion Lactation of Dairy Cow, is made up of the raw material of following parts by weight：20 parts of monkshood, 20 parts of radix glycyrrhizae, rhizoma atractylodis 3
Part, 3 parts of dried orange peel, 3 parts of the bark of official magnolia, 3 parts of the tuber of pinellia, 3 parts of balloonflower root, king do not stay 5 parts, 3 parts of Yun Ling, 0.4 part of lactic acid bacteria culture solution, Marx
0.46 part of kluyveromyces, 15 parts of soybean, 20 parts of clover stalk powder, 0.2 part of vitamin E.
The feed of the promotion Lactation of Dairy Cow of above-mentioned raw materials proportioning, comprises the following steps：
(1) fibrous root or cauline leaf of monkshood and radix glycyrrhizae are dried to moisture respectively and reach 50%, guillotine cutting into 3cm fragment,
It is well mixed, obtain mixture A.
(2) extracting lactic acid piece coccus activation bacterium solution, is inoculated in MRS culture mediums by 2% inoculum concentration, 24 is cultivated under the conditions of 30 DEG C
Hour, it is 0.6 × 10 to obtain exponential phase8CFU/ML-1.0×108CFU/ML lactic acid bacteria culture solution；Marx's Crewe is tieed up
Saccharomycete activating solution, is inoculated in potato culture by 2% inoculum concentration, is cultivated 28 hours under the conditions of 35 DEG C, obtains logarithm life
Long-term is 0.6 × 108CFU/ML-1.0×108CFU/ML kluyveromyces marxianus bacteria culture fluid.
(3) mixture that the lactic acid bacteria culture solution that step (2) prepares obtains with step (1) is weighed by 1:100 mixing are equal
It is even, tight, 38 DEG C of constant temperature are wrapped up with film, ferments 20 days, obtains fermentate B.
(4) rhizoma atractylodis dried, dried orange peel, the bark of official magnolia, the tuber of pinellia, balloonflower root, Wang Buliu, Yun Ling are crushed, crosses 80 mesh sieves, obtain mixture
D。
(5) mixture D and fermentate B are mixed, obtains mixture D, mixture D is pressed 1:49 access Marx Crewe dimensions
Yeast bacteria culture fluid, wraps up tight with film, in 40 DEG C of constant temperature, ferments 10 days, obtains fermentate E.
(6) soybean, clover are crushed, crosses 120 mesh sieves, mixed with fermentate E, mixing speed is 1000r/min, during stirring
Between be 10min, obtain the present invention feed.
Embodiment 4：
The present embodiment is the comparative example as embodiment 2, will use kluyveromyces marxianus in the feedstuff of embodiment 2
Nutrient solution is substituted, and the amount of other raw material is than same as Example 2；The mistake of lactic acid bacteria culture solution preparation is eliminated in preparation method
Journey, the process of lactic acid bacteria culture solution fermentation is fermented with kluyveromyces marxianus nutrient solution to be replaced, other steps and the phase of embodiment 2
Together.Purpose is with carrying out Contrast on effect with two kinds of bacterium fermented feeds of embodiment 2 with single bacterium fermented feed.It is specific as follows：
A kind of feed of promotion Lactation of Dairy Cow, includes the raw material of following parts by weight：25 parts of monkshood, 25 parts of radix glycyrrhizae, rhizoma atractylodis 2
Part, 2 parts of dried orange peel, 2 parts of the bark of official magnolia, 2 parts of the tuber of pinellia, 2 parts of balloonflower root, king do not stay 2 parts, 2 parts of Yun Ling, 0.8 part of kluyveromyces marxianus, big
10 parts of beans, 15 parts of clover stalk powder, 0.1 part of vitamin E.
The preparation method of the feed of the promotion Lactation of Dairy Cow of above-mentioned raw materials proportioning, comprises the following steps：
(1) fibrous root or cauline leaf of monkshood and radix glycyrrhizae are dried to moisture respectively and reach 60%, guillotine cutting into 1cm fragment,
It is well mixed, obtain mixture A.
(2) kluyveromyces marxianus bacterium is taken, is inoculated in by 2% inoculum concentration in potato culture, is trained under the conditions of 38 DEG C
Support 28 hours, it is 0.6 × 10 to obtain exponential phase8CFU/ML-1.0×108CFU/ML kluyveromyces marxianus bacteria culture fluids.
(3) mixture that the kluyveromyces marxianus bacteria culture fluid that step (2) prepares obtains with step (1) is weighed to press
1:100 are well mixed, and tight, 37 DEG C of constant temperature are wrapped up with film, are fermented 25 days, are obtained fermentate B.
(4) rhizoma atractylodis dried, dried orange peel, the bark of official magnolia, the tuber of pinellia, balloonflower root, Wang Buliu, Yun Ling are crushed, crosses 120 mesh sieves, must mix
Thing D.
(5) mixture D and fermentate B are mixed, obtains mixture D, mixture D is pressed 1:49 access Marx Crewe dimensions
Yeast bacteria culture fluid, wraps up tight with film, in 45 DEG C of constant temperature, ferments 15 days, obtains fermentate E.
(6) soybean, clover are crushed, crosses 120 mesh sieves, mixed with fermentate E, mixing speed is 3000r/min, during stirring
Between be 15min, be promoted the feed of Lactation of Dairy Cow.
Embodiment 5：
The present embodiment is the comparative example as embodiment 2, and the kluyveromyces marxianus in the feedstuff of embodiment 2 are trained
Nutrient solution is substituted with lactic acid bacteria culture solution, and the amount of other raw material is than same as Example 2；Marx gram is eliminated in preparation method
Process prepared by Shandong dimension Yeast Cultivation liquid, the process of kluyveromyces marxianus nutrient solution fermentation is replaced with lactobacillus-fermented, its
Its step is same as Example 2.Purpose is with being imitated with two kinds of bacterium fermented feeds of embodiment 2 with single bacterium fermented feed
Fruit contrasts.It is specific as follows：
A kind of feed of promotion Lactation of Dairy Cow, includes the raw material of following parts by weight：25 parts of monkshood, 25 parts of radix glycyrrhizae, rhizoma atractylodis 2
Part, 2 parts of dried orange peel, 2 parts of the bark of official magnolia, 2 parts of the tuber of pinellia, 2 parts of balloonflower root, king do not stay 2 parts, 2 parts of Yun Ling, 0.8 part of lactic acid bacteria culture solution, soybean 10
Part, 15 parts of clover stalk powder, 0.1 part of vitamin E.
The preparation method of the feed of the promotion Lactation of Dairy Cow of above-mentioned raw materials proportioning, comprises the following steps：
(1) fibrous root or cauline leaf of monkshood and radix glycyrrhizae are dried to moisture respectively and reach 60%, guillotine cutting into 1cm fragment,
It is well mixed, obtain mixture A.
(2) extracting lactic acid piece coccus, is inoculated in MRS culture mediums by 2% inoculum concentration, is cultivated 24 hours, is obtained under the conditions of 37 DEG C
It is 0.6 × 10 to exponential phase8CFU/ML-1.0×108CFU/ML lactic acid bacteria culture solution.
(3) mixture that the lactic acid bacteria culture solution that step (2) prepares obtains with step (1) is weighed by 1:100 mixing are equal
It is even, tight, 37 DEG C of constant temperature are wrapped up with film, ferments 25 days, obtains fermentate B.
(4) rhizoma atractylodis dried, dried orange peel, the bark of official magnolia, the tuber of pinellia, balloonflower root, Wang Buliu, Yun Ling are crushed, crosses 120 mesh sieves, must mix
Thing D.
(5) mixture D and fermentate B are mixed, obtains mixture D, mixture D is pressed 1:49 access lactic acid bacteria cultures
Liquid, wraps up tight with film, in 45 DEG C of constant temperature, ferments 15 days, obtains fermentate E.
(6) soybean, clover are crushed, crosses 120 mesh sieves, mixed with fermentate E, mixing speed is 3000r/min, during stirring
Between be 15min, be promoted the feed of Lactation of Dairy Cow.
The Basic Evaluation of the feed prepared below by 3 pairs of embodiment 2-5 different formulations of experiment, different preparation conditions, reality
Apply effect and the comparison to ox immunity.
Test 3 different formulations feed Basic Evaluations, implementation result and the comparison to ox immunity
3.1 different formulations feed quality Basic Evaluations
3.1.1 experimental method：
3.1.1.1 the measure of dry matter content
Determined according to GOST R 52838-2007；
3.1.1.2 the measure of acid content of lignin
Determined according to GB/T20805-2006；
3.1.1.3 the measure of acid detergent fiber (ADF) content
Determined according to GB/T20805-2006；
Crude protein 3.1.1.4 (CP) assay
Determined according to GB/T6432-1994；
Crude fibre 3.1.1.5 (CF) assay
Determined according to GB/T 6434-2006；
3.1.1.6 the measure of neutral detergent fiber (ADF) content
Determined according to GB/T20806-2006；
3.1.2 experimental result：
Conventional milk cow roughage nutrient composition content number be evaluate roughage quality most basic index, mainly include
Dry (DM), crude fibre (CF), neutral detergent fiber (NDF), acid detergent fiber (ADF), acidic cleaning lignin
(ADL), crude protein (CP) etc..NDF is relevant with cud volume degree of filling and daily feedings, and its content and energy concentration are into negative
Close, high NDF contents can limit the feed intake and its energy utilization efficiency to roughage of ox in roughage, and NDF contents are higher,
Roughage quality is lower.Roughage ADF contents and its organic matter digestibility (OMD) are negatively correlated, and ADF contents are higher, roughage
Quality is lower.The change of CP contents can reflect the damaed cordition of roughage nutrient in preparation process, but in view of the micro- life of cud
Effect of the thing to protein, it is necessary to consider degraded situation of the diet protein in cud.
Fig. 3 is embodiment 2-5 different formulations feed quality Basic Evaluation comparative results.In Fig. 3, shoulder mark difference lowercase
Represent significant difference (P ＜ 0.05)；Shoulder mark difference capitalization represents difference extremely significantly (P ＜ 0.01)；Shoulder mark same letter or
Represent difference not significantly (P ＞ 0.05) without letter.As can be known from Fig. 3：Different formulations feed quality Basic Evaluation mutually compares
It is each compared with, the present invention to implement each content of material of formula and control has notable difference, while dry in the feed of embodiment 2, thick
At most, crude fibre, neutral detergent fiber, acid detergent fiber, acidic cleaning content of lignin are minimum for protein content.
3.2 implementation results are tested：
100 healthy milk cows are selected, five groups are randomly divided into, are 300 grams/head daily, experimental period is 60 days.The output of milk is adopted
With the actual method weighed, i.e., daily weigh for 3 times in the morning, afternoon and evening and keeps a record at the output of milk.
Experimental result is shown in Table 2：
Table 2 tests milk crop statistical result
In table 2, milk is purchased by 4 yuan/kilogram prices.
As can be known from Table 2：The formula of embodiment 2 is optimal, and the output of milk is most.
Influence of the 3.3 different formulations feeds to ox immunity
In just examination morning the 50th day phase, collection test ox venous blood 20mL, stands 30min, treats serum at room temperature on an empty stomach
After precipitation, it is stand-by that blood serum sample is made in low temperature 3000r/min centrifugation 15min.Using kit measurement total protein (BCA methods), in vain
Albumen (Bromocresol green), glutamic-pyruvic transaminase/glutamic-oxalacetic transaminease (reitman-frankel method)；RNA isolation kit determines immunoglobulin A, G, M
(IgA, IgG, IgM), TAC, total number born and concentration of malondialdehyde.All operations are in strict accordance with examination
Agent box operation instructions are carried out.
Experimental result is shown in Table 3：
Influence of the different formulations feed of table 3 to cow's serum immunologic function parameter
In table, the different lowercase letter indication differences of colleague's data shoulder mark are significantly (P ＜ 0.05)；Shoulder mark difference capital letter matrix
Show difference extremely significantly (P ＜ 0.01)；Shoulder mark same letter represents difference not significantly (P ＞ 0.05) without letter.
As can be known from Table 3：Compared with other groups, eat total protein in the ox that embodiment 2 is formulated matched somebody with somebody feed, serum,
At most, glutamic-oxalacetic transaminease, glutamic-pyruvic transaminase content are minimum, illustrate that embodiment 2 is matched somebody with somebody for albumin, IgA, IgG, IgM content
Feed to improve ox immunity best results.
Influence of the different formulations feed of table 4 to the anti-oxidant parameter of cow's serum
In table, the different lowercase letter indication differences of colleague's data shoulder mark significantly (P<0.05)；Shoulder mark difference capital letter matrix
Show difference extremely significantly (P<0.01)；Shoulder mark same letter represents difference not significantly (P without letter>0.05).
Total protein content has reacted Absorption of the body to albumen, and the relation with humoral immunity in serum.White egg
It is the transport agent of water-soluble relatively low material in the main body and blood for constitute plasma colloid osmotic pressure in vain, to liver in protein metabolism
In play an important role.Glutamic-pyruvic transaminase and glutamic-oxalacetic transaminease are the important symbols for reflecting liver and cardiac function, if active
It is too high, illustrate that liver and heart are possible to be compromised.Immunoglobulin is that the mankind and higher mammal are produced in vivo by after antigenic stimulus
Raw class protein that can be with antigen generation specific effect, also known as antibody, play an important roll to immunity of organisms, are immunized
The height of level reflects resistivity of the body to disease indirectly.TAC is to weigh body Function of Antioxidant System
The composite target of situation, can represent and reflect body enzyme and the comprehensive effect and body free radical of non-enzyme Antioxidation Mechanism
Metabolism status, can be used to evaluate the height of oxidation resistance.
Influence of the different formulations feed to ox immunity：
As can be known from Table 3：Eat total protein, white egg in the ox that the embodiment of the present invention 2 and 3 is formulated matched somebody with somebody feed, its serum
In vain, IgA, IgG, IgM content are apparently higher than eating control group farm now with the corresponding index of the ox of feed；Its glutamic-oxalacetic transaminease, paddy
Pyruvic transaminase content is significantly lower than the corresponding index for eating the ox that feed is now used on control group farm.
Compared with other groups, eat total protein in the ox that embodiment 2 is formulated matched somebody with somebody feed, its serum, albumin, IgA,
At most, glutamic-oxalacetic transaminease, glutamic-pyruvic transaminase content are minimum for IgG, IgM content, illustrate that embodiment 2 is formulated matched somebody with somebody feed to improving
Ox immunity best results.
It is also possible to find out, the ox of the embodiment 4 fermented in composition of raw materials using single bacterium and embodiment 5 is eaten, its
Total protein, albumin, IgA, IgG, IgM content are higher than the corresponding index for eating the ox that feed is now used on control group farm in serum, but
It is less than the corresponding index for the ox for eating the feed that the formula of embodiment 2 and embodiment 3 is matched somebody with somebody；Its glutamic-oxalacetic transaminease, third turn of ammonia of paddy
Enzyme content is higher than the corresponding index for eating the ox that feed is now used on control group farm, but is below eating the formula of embodiment 2 and embodiment 3
The corresponding index of the ox for the feed matched somebody with somebody.
As can be known from Table 4：Eat total antioxidation energy in the ox that the embodiment of the present invention 2 and 3 is formulated matched somebody with somebody feed, its serum
Power, total Superoxide dismutase ability are apparently higher than eating control group farm now with the corresponding index of the ox of feed；Its mda content is obvious
Less than eating control group farm now with the corresponding index of the ox of feed.
Compared with other groups, TAC, total super oxygen in the ox that embodiment 2 is formulated matched somebody with somebody feed, serum are eaten
Change enzyme ability is most strong, and mda content is minimum, illustrates that embodiment 2 is formulated matched somebody with somebody feed to improving ox immunity best results.
It is also possible to find out, the ox of the embodiment 4 fermented in composition of raw materials using single bacterium and embodiment 5 is eaten, its
TAC, total Superoxide dismutase ability are higher than the corresponding index for eating the ox that feed is now used on control group farm in serum, still
The corresponding index of the ox for the feed matched somebody with somebody less than the formula for eating embodiment 2 and embodiment 3；Its mda content, which is less than, eats control group
Farm is above eating the phase of the ox for the feed that the formula of embodiment 2 and embodiment 3 is matched somebody with somebody now with the corresponding index of the ox of feed
Answer index.
The substantial amounts of the results show more than, the raw material proportioning of the feed of promotion Lactation of Dairy Cow of the invention and its preparation
Feed to promote Lactation of Dairy Cow have obvious effect.
Above content is to combine specific preferred embodiment further description made for the present invention, it is impossible to assert
The specific implementation of the present invention is confined to these explanations.For general technical staff of the technical field of the invention,
On the premise of not departing from present inventive concept, some simple deduction or replace can also be made, should all be considered as belonging to the present invention's
Protection domain.
Claims (9)
1. a kind of feed of promotion Lactation of Dairy Cow, it is characterised in that：Include the raw material of following parts by weight：20-25 parts of monkshood, radix glycyrrhizae
20-25 parts, 2-3 parts of rhizoma atractylodis, 2-3 parts of dried orange peel, 2-3 parts of the bark of official magnolia, 2-3 parts of the tuber of pinellia, 2-3 parts of balloonflower root, king do not stay 2-8 parts, Yun Ling 2-3
Part, 0.4-0.5 parts of lactic acid bacteria culture solution, 0.3-0.5 parts of kluyveromyces marxianus bacteria culture fluid, 10-15 parts of soybean, clover straw
15-20 parts of stalk powder, 0.1-0.5 parts of vitamin E.
2. the feed of promotion Lactation of Dairy Cow according to claim 1, it is characterised in that：Include the raw material of following parts by weight：
25 parts of monkshood, 25 parts of radix glycyrrhizae, 2 parts of rhizoma atractylodis, 2 parts of dried orange peel, 2 parts of the bark of official magnolia, 2 parts of the tuber of pinellia, 2 parts of balloonflower root, king do not stay 2 parts, 2 parts of Yun Ling,
0.5 part of lactic acid bacteria culture solution, 0.3 part of kluyveromyces marxianus bacteria culture fluid, 10 parts of soybean, 15 parts of clover stalk powder, dimension life
Plain 0.1 part of E.
3. the feed of promotion Lactation of Dairy Cow according to claim 2, it is characterised in that：The lactic acid bacteria culture solution is by breast
Sour piece coccus is inoculated in liquid MRS culture mediums by 2% inoculum concentration and cultivated, and culture obtains to logarithm giving birth under the conditions of 30-37 DEG C
Long-term is 0.6 × 108CFU/ML-1.0×108CFU/ML lactic acid bacteria culture solution；The kluyveromyces marxianus bacteria culture fluid
It is to be inoculated in kluyveromyces marxianus bacterium in liquid potato culture by 2% inoculum concentration to cultivate, under the conditions of 35-38 DEG C
It is 0.6 × 10 that culture, which is obtained to exponential phase,8CFU/ML-1.0×108CFU/ML kluyveromyces marxianus bacteria culture fluid.
4. the feed of promotion Lactation of Dairy Cow according to claim 3, it is characterised in that：The monkshood for monkshood cauline leaf or
Person's fibrous root, radix glycyrrhizae used is the stem or fibrous root of radix glycyrrhizae, and water content is 50-60%.
5. the preparation method of the feed of the promotion Lactation of Dairy Cow described in claim any one of 1-4, it is characterised in that：Including following
Step：
Step 1, monkshood and the mixture A of radix glycyrrhizae are prepared：Monkshood and radix glycyrrhizae are dried to moisture respectively and reach 50-60%, then
Guillotine cutting is well mixed into fragment, obtains mixture A；
Step 2, lactic acid bacteria culture solution and kluyveromyces marxianus bacteria culture fluid are prepared：Extracting lactic acid piece coccus, by 2% inoculum concentration
It is inoculated in liquid MRS culture mediums and cultivates, it is 0.6 × 10 that culture, which is obtained to exponential phase, under the conditions of 30-37 DEG C8CFU/
ML-1.0×108CFU/ML lactic acid bacteria culture solution；Kluyveromyces marxianus bacterium is taken, liquid horse is inoculated in by 2% inoculum concentration
Cultivated in bell potato culture medium, it is 0.6 × 10 that culture, which is obtained to exponential phase, under the conditions of 35-38 DEG C8CFU/ML-1.0×
108CFU/ML kluyveromyces marxianus bacteria culture fluid；
Step 3, lactic fermentation：The mixture A that the lactic acid bacteria culture solution that step 2 is prepared is obtained with step 1 is by 1：100 mixing
Uniformly, tight, ferment at constant temperature then is wrapped up with film, obtains fermentate B；
Step 4, mixture D is prepared：The rhizoma atractylodis dried, dried orange peel, the bark of official magnolia, the tuber of pinellia, balloonflower root, Wang Buliu, Yun Ling are taken, is mixed and powder
It is broken, 80-120 mesh sieves are crossed, mixture D is obtained；
Step 5, secondary fermentation：The fermentate B mixing that the mixture D and step 3 that step 4 is obtained are obtained, obtains mixture D,
Mixture D is accessed into kluyveromyces marxianus bacteria culture fluid, tight, ferment at constant temperature is wrapped up with film, obtains fermentate E；
Step 6, addition soybean, clover：Soybean, clover are crushed, 80-120 mesh sieves is crossed, is mixed with fermentate E, stirs, obtains
The feed of the present invention.
6. preparation method according to claim 5, it is characterised in that：Length 1- of the guillotine cutting into fragment in the step 1
3cm。
7. preparation method according to claim 5, it is characterised in that：The condition of ferment at constant temperature is in the step 3：35-
Fermented 20-25 days under 38 DEG C of constant temperatures.
8. preparation method according to claim 5, it is characterised in that：Mixture D is tieed up with Marx's Crewe in the step 5
The amount ratio of yeast bacteria culture fluid is 49：1；The ferment at constant temperature is fermented 10-15 days under 40-45 DEG C of constant temperature.
9. preparation method according to claim 5, it is characterised in that：Mixing speed is 1000-3000r/ in the step 6
Min, mixing time is 10-15min.
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| CN107594074A (en) * | 2017-09-27 | 2018-01-19 | 东川区隆兴肉牛养殖专业合作社 | A kind of cattle feed of hygrometric state fermentation protein and preparation method thereof |
| CN109043135A (en) * | 2018-09-18 | 2018-12-21 | 广西壮族自治区农业科学院农产品加工研究所 | A kind of lactagogue forage containing banana flowers |
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