CN102366023B - Method for preparing cow feed additive containing traditional Chinese medicine active probiotics - Google Patents
Method for preparing cow feed additive containing traditional Chinese medicine active probiotics Download PDFInfo
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Abstract
The invention discloses a method for preparing a cow feed additive containing traditional Chinese medicine active probiotics, comprising the following steps: respectively carrying out amplification culture of the strain number of a single bacterial specie on original strains of active probiotics for cows; mixing primary seed liquors of active probiotics for cows with same weight, inoculating the mixture in a seed tank medium to carry out multiple-strain mixed culture, then inoculating in a traditional Chinese medicine liquid medium to carry out a primary traditional Chinese medicine fermentation; and mixing the multiple-strain mixed traditional Chinese medicine fermentation liquor with isometric cob powder to obtain solid fermentation seeds, and then inoculating in a traditional Chinese medicine solid medium to carry out a secondary traditional Chinese medicine fermentation. According to the invention, the strain activity and colonization performance in the intestinal tracts of active probiotics in the cow feed additive are improved greatly, the prepared cow feed additive is rich in traditional Chinese medicine components, the active components with pharmacological effects in the traditional Chinese medicine components fully play the role, thus the performances of strengthening the body, treating diseases and promoting growth of the cow feed additive are improved, and furthermore completely displace antibiotics.
Description
Technical field
The present invention relates to the animal and fowl fodder field, relate in particular to a kind of manufacture method that contains the cow feed additive of active Chinese drug component probio.
Background technology
Cow subclinical mastitis is a kind of serious disease common, complicated, that can cause the dairy heavy losses, and what not only have a strong impact on cow utilizes the time limit, galactopoiesis quantity and dairy food quality, and is harmful to food hygiene and human health.For a long time, the cowboying family is mostly adopted and is prevented cow subclinical mastitis to the antibiotic mode of the long-term feeding of milk cow.Antibiotic has kill bacteria and suppresses the effect of pathogenic microorganisms, and therefore this feeding patterns has played the effect of certain prevention cow subclinical mastitis in actual production.But, these antibiotic can enter human body along with the edible milk of people and various milk product, this has just caused antibiotic abuse indirectly, might cause multiple drug induced disease, even can produce new " super drug-fast bacteria ", thereby human life and health in this feeding patterns serious threat.
For fear of or reduce milk cow and feed the medium-term and long-term antibiotic that uses of process, microecological feed additive for milk cows arises at the historic moment.So-called microecological feed additive for milk cows is the active probiotic preparation of making through special process to the useful and harmless active probiotic of milk cow.Microecological feed additive for milk cows adds auxiliary material as a kind of functional ox feed, realized having no side effect, having no drug resistance, preventing cow subclinical mastitis under without pathogenic prerequisite, not only can regulate milk cow intestinal microecology balance, improve milk cow growth rate, improve efficiency of feed utilization and can suppress the harmful intestinal tract microorganism breeding, improve the milk cow health level, strengthen milk cow immunity.At present, in existing milk cattle cultivating, the kind of common microecological feed additive for milk cows has nearly hundred kinds in lactobacillus preparation (comprising lactobacterium acidophilus, Bacillus bifidus, streptococcus fecalis etc.), bacillus preparation (comprising hay bacillus, bacillus cereus, B.lieheniformis etc.), fungi preparation (comprising aspergillus oryzae etc.), saccharomycete preparation (comprising brewer's yeast etc.) etc.Use in the microbial forage additive production process existing ox, mostly only use protein substance and/or glucide as the culture medium of active probiotic, although bacterial strain quantitatively can satisfy the demands, but the activity of most of bacterial strains is not high, field planting performance in enteron aisle a little less than, so the effect that active probiotic should be brought into play as milk cow forage is very restricted; Simultaneously, active probiotic only can strengthen the physique of milk cow to a certain extent, and the effect of its treatment and prevention cow subclinical mastitis is not fairly obvious, and therefore, existing milk cow microbial forage additive can not replace antibiotic use fully.
Summary of the invention
The embodiment of the invention provides a kind of manufacture method that contains the cow feed additive of active Chinese drug component probio, so that significantly improve the active and field planting performance in enteron aisle of the bacterial strain of active probiotic in the cow feed additive, and make the cow feed additive that makes be rich in traditional Chinese medicine ingredients, the active component that plays pharmacological action in the traditional Chinese medicine ingredients is given full play to, performance and growth promotion performance thereby the strong body that improves cow feed additive is cured the disease, effectively treat and the prevention cow subclinical mastitis, and then replace antibiotic use fully.
The objective of the invention is to be achieved through the following technical solutions:
A kind of manufacture method that contains the cow feed additive of active Chinese drug component probio comprises:
Steps A: choose the multiple milk cow that equates bacterial strain quantity with the former bacterial strain of active probiotic, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity with the former bacterial strain of active probiotic and expand and cultivate, thereby obtain the primary seed solution that every kind of milk cow is used active probiotic;
Wherein, described multiple milk cow comprises in bacillus subtilis, bafillus natto, bacillus licheniformis, brewer's yeast, candida utili, aspergillus niger, Rhizopus oryzae, Lactobacillus casei, lactobacillus acidophilus and the Bifidobacterium at least two kinds with active probiotic;
Step B: every kind of milk cow of equal weight is mixed with the primary seed solution of active probiotic, thereby obtain many bacterial classifications Hybrid TierI seed liquor;
Step C: many bacterial classifications Hybrid TierI seed liquor is inoculated into carries out many bacterial classifications Mixed culture in the seed tank culture base, mix secondary seed solution thereby obtain many bacterial classifications;
Step D: many bacterial classifications are mixed secondary seed solution be inoculated into and carry out first herb fermenting in the Chinese medicinal liquor culture medium, thereby obtain many bacterial classification mixing Chinese medicine fermented liquids;
Wherein, described Chinese medicinal liquor culture medium comprises 5%～8% brown sugar, 2%～3% traditional Chinese medicinal components, 0.2%～0.3% potassium dihydrogen phosphate, 0.3%～0.5% yeast extract and the water of surplus; Described traditional Chinese medicinal components by the Radix Astragali, Radix Codonopsis, dandelion, the root of large-flowered skullcap, rhizome of chuanxiong, the seed of cowherb, play both sides of the street, radix bupleuri, Radix Angelicae Sinensis, motherwort, kelp and Radix Glycyrrhizae form, the parts by weight that its each composition accounts for the traditional Chinese medicinal components total amount are:
The Radix Astragali: 60～80 parts, Radix Codonopsis: 60～80 parts, dandelion: 60～80 parts, the root of large-flowered skullcap: 40～60 parts,
Rhizome of chuanxiong: 10～20 parts, the seed of cowherb: 40～60 parts, play both sides of the street: 40～60 parts, radix bupleuri: 50～80 parts,
Radix Angelicae Sinensis: 40～60 parts, motherwort: 50～80 parts, kelp: 80-120 part, Radix Glycyrrhizae: 30～50 parts
Step e: with many bacterial classification mixing Chinese medicine fermented liquids with etc. the maize cob meal of weight be mixed and made into the solid state fermentation seed;
Step F: the solid state fermentation seed is inoculated into carries out the secondary herb fermenting in the Chinese medicine solid medium, thereby make the cow feed additive that contains the active Chinese drug component probio.
As seen from the above technical solution provided by the invention, the manufacture method of the cow feed additive that contains the active Chinese drug component probio that the embodiment of the invention provides adopts kinds of traditional Chinese medicines is cultivated good facilitation to milk cow growth and resistance against diseases active probiotic, thereby make the active component in the Chinese medicine give full play to pharmacological action, and the active probiotic that the activity that makes the active probiotic of turning out is turned out far above ordinary culture medium has also strengthened the colonization ability of active probiotic in enteron aisle simultaneously; Because Chinese medicine and active probiotic have synergy to immune system, the active probiotic that therefore will breed with Chinese medicine can strengthen the immunocompetence of milk cow greatly as feed additive for dairy cows.We cultivate active probiotic by modern microecology principle is combined with the traditional Chinese medical science with traditional Chinese medicinal components, thereby produce a kind of antibiotic no drug residue, prophyiaxis and promoting growth, feed additive for dairy cows with low cost of replacing fully.
Description of drawings
In order to be illustrated more clearly in the technical scheme of the embodiment of the invention, the accompanying drawing of required use was done to introduce simply during the below will describe embodiment, apparently, accompanying drawing in the following describes only is some embodiments of the present invention, for those of ordinary skill in the art, not paying under the capable prerequisite of creative work, can also obtain other accompanying drawings according to these accompanying drawings.
The schematic flow sheet of the manufacture method of the cow feed additive that contains the active Chinese drug component probio that Fig. 1 provides for the embodiment of the invention.
The specific embodiment
Following is clearly and completely described the technical scheme in the embodiment of the invention in conjunction with the accompanying drawing in the embodiment of the invention, and obviously, described embodiment only is the present invention's part embodiment, rather than whole embodiment.Based on embodiments of the invention, those of ordinary skills belong to protection scope of the present invention not making the every other embodiment that obtains under the creative work prerequisite.
At first, need to prove that part alleged in the present specification all refers to weight portion, the below makes a detailed description to the manufacture method of the cow feed additive that contains the active Chinese drug component probio that the embodiment of the invention provides.
As shown in Figure 1, a kind of manufacture method that contains the cow feed additive of active Chinese drug component probio specifically can may further comprise the steps:
Steps A: choose the multiple milk cow that equates bacterial strain quantity with the former bacterial strain of active probiotic, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity with the former bacterial strain of active probiotic and expand and cultivate, thereby obtain the primary seed solution that every kind of milk cow is used active probiotic;
Wherein, described multiple milk cow comprises in bacillus subtilis, bafillus natto, bacillus licheniformis, brewer's yeast, candida utili, aspergillus niger, Rhizopus oryzae, Lactobacillus casei, lactobacillus acidophilus and the Bifidobacterium at least two kinds with active probiotic;
Particularly, choose accordingly the multiple milk cow that the equates bacterial strain quantity former bacterial strain of active probiotic, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity with the former bacterial strain of active probiotic and expand and cultivate, can comprise with the primary seed solution of active probiotic thereby obtain every kind of milk cow:
(1) with the former bacterial strain seed inclined plane inoculating of the bacillus subtilis behind the separating-purifying in the triangular flask of bacillus subtilis bacterium culture medium, and shaken cultivation 24 hours, thus obtain the primary seed solution of bacillus subtilis; The bacillus subtilis bacterium culture medium comprises 3 parts beef extract, 10 parts peptone, 5 parts sodium chloride, 20 parts agar and 1000 parts water; Wherein, the pH value of this culture medium should just can be inoculated in 7.0～7.2 scope, to guarantee that bacterial strain activity and bacterial strain survival condition can not destroyed.Need to prove that the inoculative proportion of bacillus subtilis is the bacillus subtilis of the bacillus subtilis inoculation of medium 100mL of every 500mL.
And/or,
(2) with the former bacterial strain seed inclined plane inoculating of the bafillus natto behind the separating-purifying in the triangular flask of bafillus natto culture medium, and shaken cultivation 24 hours, thus obtain the primary seed solution of bafillus natto; The bafillus natto culture medium comprises 3 parts beef extract, 10 parts peptone, 5 parts sodium chloride, 20 parts agar and 1000 parts water; The pH value of this culture medium should just can be inoculated in 7.0～7.2 scope, to guarantee that bacterial strain activity and bacterial strain survival condition can not destroyed.Need to prove that the inoculative proportion of bafillus natto is the bafillus natto of the bafillus natto inoculation of medium 100mL of every 500mL.
And/or,
(3) with the former bacterial strain seed inclined plane inoculating of the bacillus licheniformis behind the separating-purifying in the triangular flask of bacillus licheniformis culture medium, and shaken cultivation 24 hours, thus obtain the primary seed solution of bacillus licheniformis; The bacillus licheniformis culture medium comprises 3 parts beef extract, 10 parts peptone, 5 parts sodium chloride, 20 parts agar and 1000 parts water; The pH value of this culture medium should just can be inoculated in 7.0～7.2 scope, to guarantee that bacterial strain activity and bacterial strain survival condition can not destroyed.Need to prove that the inoculative proportion of bacillus licheniformis is the bacillus licheniformis of the bacillus licheniformis inoculation of medium 100mL of every 500mL.
And/or,
(4) with the former bacterial strain seed inclined plane inoculating of the brewer's yeast behind the separating-purifying in the triangular flask of brewer's yeast culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of brewer's yeast; The brewer's yeast culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water.Need to prove that the inoculative proportion of brewer's yeast is the brewer's yeast of the brewer's yeast inoculation of medium 100mL of every 500mL.
And/or,
(5) with the former bacterial strain seed inclined plane inoculating of the candida utili behind the separating-purifying in the triangular flask of candida utili culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of candida utili; The candida utili culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water.Need to prove that the inoculative proportion of candida utili is the candida utili of the candida utili inoculation of medium 100mL of every 500mL.
And/or,
(6) with the former bacterial strain seed inclined plane inoculating of the aspergillus niger behind the separating-purifying in the triangular flask of aspergillus niger culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of aspergillus niger; The aspergillus niger culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water.Need to prove that the inoculative proportion of aspergillus niger is the aspergillus niger of the aspergillus niger inoculation of medium 100mL of every 500mL.
And/or,
(7) with the former bacterial strain seed inclined plane inoculating of the Rhizopus oryzae behind the separating-purifying in the triangular flask of Rhizopus oryzae culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of Rhizopus oryzae; The Rhizopus oryzae culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water.Need to prove that the inoculative proportion of Rhizopus oryzae is the Rhizopus oryzae of the Rhizopus oryzae inoculation of medium 100mL of every 500mL.
And/or,
(8) with the former bacterial strain seed inclined plane inoculating of the Lactobacillus casei behind the separating-purifying in the Kolle flask of Lactobacillus casei culture medium, and 37 ℃ of lower cultivations 48 hours, thereby obtain the primary seed solution of Lactobacillus casei; The Lactobacillus casei culture medium comprise 20 parts glucose, 10 parts peptone, 10 parts beef extract, 5 parts yeast extract, 5 parts natrium citricum, 2 parts sodium dihydrogen phosphate, 5 parts sodium acetate, 0.2 part epsom salt, 0.25 part four water manganese sulfates, (molecular formula of Tween 80 is C to 1 part Tween 80
64H
124O
26English name is POLYSORBATE 80, Chinese synonym is polyoxyethylene sorbitan monooleate or emulsifying agent T80, soluble in water, ethanol, methyl alcohol and ethyl acetate, be mainly used in doing solubilizer or the emulsifying agent of parenteral solution and oral liquid, the dispersant of capsule, the emulsifying agent of ointment and matrix, the matrix of suppository or the emulsifying agent in the food industry) and 1000 parts water; The pH value of this culture medium should just can be inoculated in 6.2～6.4 scope, thereby guarantees that bacterial strain can have larger activity, and guarantees that the bacterial strain existing state can not destroyed.Need to prove that the inoculative proportion of Lactobacillus casei is the Lactobacillus casei of the Lactobacillus casei inoculation of medium 100mL of every 500mL.
And/or,
(9) with the former bacterial strain seed inclined plane inoculating of the lactobacillus acidophilus behind the separating-purifying in the Kolle flask of lactobacillus acidophilus culture medium, and 37 ℃ of lower cultivations 48 hours, thereby obtain the primary seed solution of lactobacillus acidophilus; The lactobacillus acidophilus culture medium comprises 20 parts glucose, 10 parts peptone, 10 parts beef extract, 5 parts yeast extract, 5 parts natrium citricum, 2 parts sodium dihydrogen phosphate, 5 parts sodium acetate, 0.2 part epsom salt, 0.25 part four water manganese sulfates, 1 part Tween 80 and 1000 parts water; The pH value of this culture medium should just can be inoculated in 6.2～6.4 scope, thereby guarantees that bacterial strain can have larger activity, and guarantees that the bacterial strain existing state can not destroyed.Need to prove that the inoculative proportion of lactobacillus acidophilus is the lactobacillus acidophilus of the lactobacillus acidophilus inoculation of medium 100mL of every 500mL.
And/or,
(10) with the former bacterial strain seed inclined plane inoculating of the Bifidobacterium behind the separating-purifying in the Kolle flask of Medium of Bifidobacterium, and 37 ℃ of lower cultivations 48 hours, thereby obtain the primary seed solution of Bifidobacterium; Medium of Bifidobacterium comprises 20 parts glucose, 10 parts peptone, 10 parts beef extract, 5 parts yeast extract, 5 parts natrium citricum, 2 parts sodium dihydrogen phosphate, 5 parts sodium acetate, 0.2 part epsom salt, 0.25 part four water manganese sulfates, 1 part Tween 80 and 1000 parts water; The pH value of this culture medium should just can be inoculated in 6.2～6.4 scope, thereby guarantees that bacterial strain can have larger activity, and guarantees that the bacterial strain existing state can not destroyed.Need to prove that the inoculative proportion of Bifidobacterium is the Bifidobacterium of inoculation 100mL in the Medium of Bifidobacterium of every 500mL.
Step B: every kind of milk cow of equal weight is mixed with the primary seed solution of active probiotic, thereby obtain many bacterial classifications Hybrid TierI seed liquor;
Particularly, in actual applications, preferably from the described 10 kinds of milk cows of steps A with all selecting the identical weight primary seed solution the primary seed solution of active probiotic, then mix, thereby make the cow feed additive that finally makes with these 10 kinds of probios.
Step C: many bacterial classifications Hybrid TierI seed liquor is inoculated into carries out many bacterial classifications Mixed culture in the seed tank culture base, mix secondary seed solution thereby obtain many bacterial classifications;
Wherein, corresponding seed tank culture base comprises 3%～8% brown sugar, 0.2%～0.5% potassium dihydrogen phosphate, 0.3%～0.5% yeast extract, 0.2%～0.5% peptone and the water of surplus, and each constituent content of seed tank culture base described herein is mass percent;
Particularly, in actual production was used, for many factors such as comprehensive cost of manufacture and serviceabilities, each composition of this seed tank culture base accounts for total mass ratio can select scheme in the following table 1, and the embodiment one in wherein should table is scheme most preferably:
Table 1:
| Embodiment | Brown sugar | Potassium dihydrogen phosphate | Yeast extract | Peptone | Water |
| Embodiment one | 8％ | 0.22％ | 0.5％ | 0.3％ | Surplus |
| Embodiment two | 6％ | 0.2％ | 0.4％ | 0.4％ | Surplus |
| Embodiment three | 7％ | 0.25 | 0.4％ | 0.4％ | Surplus |
| Embodiment four | 5％ | 0.3％ | 0.5％ | 0.2％ | Surplus |
Further, many bacterial classifications Hybrid TierI seed liquor is inoculated into carries out many bacterial classifications Mixed culture in the seed tank culture base accordingly, can may further comprise the steps thereby obtain many bacterial classifications mixing secondary seed solution:
Step C1: preparation seed tank culture base;
Step C2: under 115 ℃～121 ℃, to seed tank culture base sterilization 25 minutes, be cooled to again 30 ℃;
Particularly, in actual applications, preferably select under 121 ℃ the seed tank culture base was sterilized 25 minutes, can be killed fully to guarantee the harmful bacteria in the seed tank culture base.
Step C3: many bacterial classifications Hybrid TierI seed liquor is inoculated in the seed tank culture base (many bacterial classifications Hybrid TierI seed liquor that the inoculative proportion according to 3%～5% herein refers to 3 to 5 unit volumes of inoculation in the seed tank culture base of per 100 unit volumes) according to 3%～5% inoculative proportion;
Step C4: the seed tank culture base that docking finishes carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, finishes until the pH value in the seed tank culture base reaches fermentation in 4.8 o'clock, namely obtains many bacterial classifications and mixes secondary seed solution.Generally speaking, the anaerobic fermentation of seed tank culture base generally can be finished in 48～72 hours, and after fermentation was finished, the number of active probiotic can reach every milliliter of 1,500,000,000 strains.
Step D: many bacterial classifications are mixed secondary seed solution be inoculated into and carry out first herb fermenting in the Chinese medicinal liquor culture medium, thereby obtain many bacterial classification mixing Chinese medicine fermented liquids;
Wherein, described Chinese medicinal liquor culture medium comprises 5%～8% brown sugar, 2%～3% traditional Chinese medicinal components, 0.2%～0.3% potassium dihydrogen phosphate, 0.3%～0.5% yeast extract and the water of surplus, and each constituent content of Chinese medicinal liquor culture medium described herein is mass percent; Described traditional Chinese medicinal components by the Radix Astragali, Radix Codonopsis, dandelion, the root of large-flowered skullcap, rhizome of chuanxiong, the seed of cowherb, play both sides of the street, radix bupleuri, Radix Angelicae Sinensis, motherwort, kelp and Radix Glycyrrhizae form, the parts by weight that its each composition accounts for the traditional Chinese medicinal components total amount are:
The Radix Astragali: 60～80 parts, Radix Codonopsis: 60～80 parts, dandelion: 60～80 parts, the root of large-flowered skullcap: 40～60 parts,
Rhizome of chuanxiong: 10～20 parts, the seed of cowherb: 40～60 parts, play both sides of the street: 40～60 parts, radix bupleuri: 50～80 parts,
Radix Angelicae Sinensis: 40～60 parts, motherwort: 50～80 parts, kelp: 80-120 part, Radix Glycyrrhizae: 30～50 parts.
Particularly, in actual production is used, for many factors such as comprehensive cost of manufacture and serviceabilities, each composition of this Chinese medicinal liquor culture medium accounts for total mass ratio can select preferred version in the following table 2, and the embodiment 1 in wherein should table is most preferred embodiment:
Table 2:
| Embodiment | Brown sugar | Traditional Chinese medicinal components | Potassium dihydrogen phosphate | Yeast extract | Water |
| Embodiment 1 | 6％ | 3％ | 0.2％ | 0.5％ | Surplus |
| Embodiment 2 | 7％ | 2.5％ | 0.3％ | 0.3％ | Surplus |
| Embodiment 3 | 5.5％ | 2.5％ | 0.25％ | 0.4％ | Surplus |
| Embodiment 4 | 8％ | 2％ | 0.25％ | 0.45％ | Surplus |
Corresponding with it, the composition of corresponding traditional Chinese medicinal components, and each composition parts by weight of accounting for the traditional Chinese medicinal components total amount can select the preferred version in the following table 3, and wherein the unit in this table is weight portion, and embodiment a is most preferred embodiment:
Table 3:
| The traditional Chinese medicine ingredients title | Example a | Example b | Example c | Example d |
| The Radix Astragali | 80 | 70 | 75 | 60 |
| Radix Codonopsis | 80 | 60 | 65 | 75 |
| Dandelion | 80 | 75 | 70 | 60 |
| The root of large-flowered skullcap | 50 | 40 | 55 | 60 |
| Rhizome of chuanxiong | 20 | 15 | 15 | 10 |
| The seed of cowherb | 50 | 40 | 55 | 60 |
| Play both sides of the street | 50 | 60 | 55 | 40 |
| Radix bupleuri | 50 | 60 | 70 | 80 |
| Radix Angelicae Sinensis | 60 | 55 | 50 | 40 |
| Motherwort | 60 | 50 | 55 | 70 |
| Kelp | 120 | 110 | 100 | 90 |
| Radix Glycyrrhizae | 50 | 30 | 40 | 45 |
Further, many bacterial classifications mixing secondary seed solution are inoculated into carry out first herb fermenting in the Chinese medicinal liquor culture medium accordingly, can may further comprise the steps thereby obtain many bacterial classification mixing Chinese medicine fermented liquids:
Step D1: preparation Chinese medicinal liquor culture medium;
Step D2: under 115 ℃～121 ℃, to Chinese medicinal liquor medium sterilization 20 minutes, be cooled to again 30 ℃;
Particularly, in actual applications, preferably select under 121 ℃ Chinese medicinal liquor medium sterilization 20 minutes, can be killed fully to guarantee the harmful bacteria in the Chinese medicinal liquor culture medium.
Step D3: many bacterial classifications are mixed secondary seed solution be inoculated into (inoculative proportion according to 5%～10% herein refers to that many bacterial classifications of 5 to 10 unit volumes of Chinese medicinal liquor inoculation of medium of per 100 unit volumes mix secondary seed solution) in the Chinese medicinal liquor culture medium according to 5%～10% inoculative proportion;
Step D4: the Chinese medicinal liquor culture medium that docking finishes carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, finishes until the pH value in the Chinese medicinal liquor culture medium reaches fermentation in 5.0 o'clock, namely obtains many bacterial classification mixing Chinese medicine fermented liquids; Generally speaking, the anaerobic fermentation of Chinese medicinal liquor culture medium generally can be finished in 48～72 hours, and after fermentation was finished, the number of active probiotic can reach every milliliter of 2,000,000,000 strains.
Step e: with many bacterial classification mixing Chinese medicine fermented liquids with etc. the maize cob meal of weight be mixed and made into the solid state fermentation seed;
Particularly, corresponding maize cob meal preferably passes through microwave sterilization (corresponding microwave sterilization refers to that proportion is the heat energy killing microorganisms that produces of the microwave irradiation of 300MHz～300000MHz and the method for gemma.This method is fit to the sterilization of liquid and solid material, and solid material is had desiccation) after maize cob meal, not have harmful bacteria in the active probiotic solid state fermentation seed that guarantees to be synthesized.
Step F: the solid state fermentation seed is inoculated into carries out the secondary herb fermenting in the Chinese medicine solid medium, thereby make the cow feed additive that contains the active Chinese drug component probio.
Wherein, corresponding Chinese medicine solid medium is by 60～70 parts wheat bran, 15～20 parts of dregs of beans, 10～15 parts corn flour, 2～3 parts brown sugar, 2～3 parts traditional Chinese medicinal components, (precipitated calcium carbonate not only claims but also claims winnofil to 2～4 parts precipitated calcium carbonate, is the trickle calcium carbonate of the particle of settling volume between 2.4～2.8mL/g that makes with the chemical process method; In the industrial production at home, precipitated calcium carbonate adopts carbonizatin method to make mostly---the calcining of the raw materials such as lime stone is generated lime and carbon dioxide, adding entry makes lime become milk of lime again, then passing into carbon dioxide makes the milk of lime carbonization generate precipitation of calcium carbonate, at last to precipitation of calcium carbonate dewater, drying and pulverization process get final product), 0.5～0.8 part yeast extract, 0.2～0.5 part fulvic acid forms; Described traditional Chinese medicinal components comprises the Radix Astragali, Radix Codonopsis, dandelion, the root of large-flowered skullcap, rhizome of chuanxiong, the seed of cowherb, plays both sides of the street, radix bupleuri, Radix Angelicae Sinensis, motherwort, kelp and Radix Glycyrrhizae, and the parts by weight that its each composition accounts for the traditional Chinese medicinal components total amount are:
The Radix Astragali: 60～80 parts, Radix Codonopsis: 60～80 parts, dandelion: 60～80 parts, the root of large-flowered skullcap: 40～60 parts,
Rhizome of chuanxiong: 10～20 parts, the seed of cowherb: 40～60 parts, play both sides of the street: 40～60 parts, radix bupleuri: 50～80 parts,
Radix Angelicae Sinensis: 40～60 parts, motherwort: 50～80 parts, kelp: 80-120 part, Radix Glycyrrhizae: 30～50 parts.
Particularly, in actual production is used, for many factors such as comprehensive cost of manufacture and serviceabilities, each composition of this Chinese medicine solid medium accounts for total mass ratio can select preferred version in the following table 4, and wherein the embodiment A in this table is most preferred embodiment; Corresponding with it, the composition of corresponding traditional Chinese medicinal components, and each composition parts by weight of accounting for the traditional Chinese medicinal components total amount can select the preferred version in the as above table 3, and wherein the unit in the table 3 is weight portion, and the embodiment a in the table 3 is most preferred embodiment.
Table 4:
| Composition | Embodiment one | Embodiment two | Embodiment three | Embodiment four |
| Wheat bran | 66.8 | 61.8 | 63.8 | 64.8 |
| Dregs of beans | 15 | 18 | 17 | 16 |
| Corn flour | 10 | 12 | 10 | 12 |
| Brown sugar | 2 | 3 | 2.5 | 2 |
| Traditional Chinese medicinal components | 2.5 | 2.5 | 2.5 | 2.5 |
| Precipitated calcium carbonate | 2 | 2 | 3 | 2 |
| Yeast extract | 0.5 | 0.5 | 0.7 | 0.5 |
| Fulvic acid | 0.2 | 0.2 | 0.5 | 0.2 |
Further, the solid state fermentation seed is inoculated into carries out the secondary herb fermenting in the Chinese medicine solid medium accordingly, may further comprise the steps thereby make the cow feed additive that contains the active Chinese drug component probio:
Step F 1: preparation Chinese medicine solid medium;
Step F 2: under 115 ℃～121 ℃, to Chinese medicine solid medium sterilization 20 minutes, be cooled to again 30 ℃;
Particularly, in actual applications, preferably select under 121 ℃ the Chinese medicine solid medium was sterilized 25 minutes, can be killed fully to guarantee the harmful bacteria in the Chinese medicine solid medium.
Step F 3: the solid state fermentation seed liquor is inoculated in the Chinese medicine solid medium (the solid state fermentation seed that the inoculative proportion according to 20%～30% herein refers to 20 to 30 unit volumes of inoculation in the Chinese medicine solid medium of per 100 unit volumes) according to 20%～30% inoculative proportion;
Step F 4: the Chinese medicine solid medium that will inoculate places and carries out sealing and fermenting under 37 ℃, when the temperature of Chinese medicine solid medium reaches 40 ℃ the Chinese medicine solid medium is overturn; Continuing fermentation, time fermentation is finished until the temperature of Chinese medicine solid medium no longer rises, and namely obtains to contain the cow feed additive of active Chinese drug component probio.Generally speaking, the sealing and fermenting of Chinese medicine solid medium generally can be finished in 5～7 days, and after fermentation was finished, the number of active probiotic can reach every milliliter of 4,500,000,000 strains.After sieving, material after fermentation finished can be packaged into the cow feed additive that contains the active Chinese drug component probio.
In technique scheme, the effect of each composition of traditional Chinese medicinal components of the present invention is: the Radix Astragali---sweet warm in nature, the tonifying Qi and lifting yang of distinguishing the flavor of, and its active component astragalus polyose can improve immunity of organisms; Radix Codonopsis---the sweet property of distinguishing the flavor of is flat, tonifying middle-Jiao and Qi and taste, and its active component saponin can improve immunity of organisms; The flat flavor of dandelion---property is sweet can be clearing heat and detoxicating, and dispersing swelling and dissipating binds contains the composition such as taraxasterol, choline, synanthrin and the root of large-flowered skullcap suppresses mammitis harmful bacteria (staphylococcus aureus and micrococcus scarlatinae) jointly; The root of large-flowered skullcap---clearing heat and moistening dryness, the effects such as removing pattogenic heat from the blood and toxic material from the body; Rhizome of chuanxiong---blood-activating and qi-promoting, yin-nourishing, stagnation resolvation, rhizome of chuanxiong are the gas medicine in the blood, allow the blood tonify without causing stagnation; The seed of cowherb---Huo Xue Tong Longitude, the lactogenesis carbuncle that disappears, it contains triterpenoid saponin, glycosides displayed; Play both sides of the street---promoting blood circulation and removing blood stasis, Qu Feng Tong Network, Li Shui Tong Longitude; Radix bupleuri---bitter, cold nature are returned the liver and gall warp, and easypro liver and stomach are arranged, and rise the function of sun, and anti-bacteria and anti-virus also has immunoloregulation function; Radix Angelicae Sinensis---it is sweet warm in nature to distinguish the flavor of, and can enrich blood, and can invigorate blood circulation again, and its active component sodium ferulate energy activating macrophage plays immunologic function; Motherwort---promoting blood circulation contains multiple alkaloid and Organic Selenium, can improve the immunity of body; Kelp---nourish yin and calm the liver, press down the gas dissipating bind, organic iodine content is high and be rich in logical Bittner milk factor and can improve milk production of cow; Radix Glycyrrhizae---have clearing heat and detoxicating, coordinating the drug actions of a prescription, its active component glycyrrhizic acid is cell immunomodulator.The mentioned traditional Chinese medicinal components of the present invention is the growth characteristic according to milk cow, cultivation demand in the treatment of cow subclinical mastitis and prevention mechanism and the milk cattle cultivating process, adopt monarch, minister, assistant, the drug matching principle that makes, choose respectively and can promote milk cow digestion and growth, can strengthen the milk cow immunity of organisms, the a few herbs that can improve the milk cow resistance against diseases are formulated---at first select the Radix Astragali, Radix Codonopsis, Radix Angelicae Sinensis is as monarch drug in a prescription, improve animal body immunity, reach the purpose of benefiting qi and nourishing blood: select simultaneously dandelion, the root of large-flowered skullcap, radix bupleuri improves its anti-bacteria and anti-virus ability; And select rhizome of chuanxiong, the seed of cowherb, play both sides of the street, motherwort, kelp breast promoting blood circulation and removing blood stasis, logical and improve the output of milk; Select at last the Radix Glycyrrhizae not only can be clearing heat and detoxicating and can also be in harmonious proportion all the other all medicines.This shows that traditional Chinese medicinal components provided by the present invention not only can promote the speed of growth of milk cow, and can strengthen the disease resistance ability of milk cow, thereby can effectively treat and prevent cow subclinical mastitis.
Need to prove that strain culturing equipment used in the present invention and the strain culturing technique of not describing in detail all can adopt strain culturing equipment general in the prior art and strain culturing technique, no longer are described in detail in this application.Various Chinese medicines described in the present invention all can be buied in each large Chinese medicine pharmacy easily, therefore this are no longer described in detail.The former bacterial strain of various active probiotic bacterium described in the present invention all can be introduced from Chinese common micro-organisms culture presevation administrative center (CGMCC) or agriculture microorganism fungus kind preservation administrative center (ACCC), with existing state and the survival quality of guaranteeing each bacterial classification.
As seen, the enforcement of the embodiment of the invention can significantly improve the active and field planting performance in enteron aisle of bacterial strain of active probiotic in the cow feed additive, and make the cow feed additive that makes be rich in traditional Chinese medicine ingredients, the active component that plays pharmacological action in the traditional Chinese medicine ingredients is given full play to, performance and growth promotion performance thereby the strong body that improves cow feed additive is cured the disease, effectively treat and the prevention cow subclinical mastitis, and then replace antibiotic use fully.
The above; only for the better specific embodiment of the present invention, but protection scope of the present invention is not limited to this, anyly is familiar with those skilled in the art in the technical scope that the present invention discloses; the variation that can expect easily or replacement all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain of claims.
Claims (5)
1. a manufacture method that contains the cow feed additive of active Chinese drug component probio is characterized in that, comprising:
Steps A: choose the multiple milk cow that equates bacterial strain quantity with the former bacterial strain of active probiotic, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity with the former bacterial strain of active probiotic and expand and cultivate, thereby obtain the primary seed solution that every kind of milk cow is used active probiotic;
Wherein, described multiple milk cow comprises in bacillus subtilis, bafillus natto, bacillus licheniformis, brewer's yeast, candida utili, aspergillus niger, Rhizopus oryzae, Lactobacillus casei, lactobacillus acidophilus and the Bifidobacterium at least two kinds with active probiotic;
Step B: every kind of milk cow of equal weight is mixed with the primary seed solution of active probiotic, thereby obtain many bacterial classifications Hybrid TierI seed liquor;
Step C: many bacterial classifications Hybrid TierI seed liquor is inoculated into carries out many bacterial classifications Mixed culture in the seed tank culture base, mix secondary seed solution thereby obtain many bacterial classifications;
Step D: many bacterial classifications are mixed secondary seed solution be inoculated into and carry out first herb fermenting in the Chinese medicinal liquor culture medium, thereby obtain many bacterial classification mixing Chinese medicine fermented liquids;
Wherein, described Chinese medicinal liquor culture medium comprises 5%～8% brown sugar, 2%～3% traditional Chinese medicinal components, 0.2%～0.3% potassium dihydrogen phosphate, 0.3%～0.5% yeast extract and the water of surplus; Described traditional Chinese medicinal components by the Radix Astragali, Radix Codonopsis, dandelion, the root of large-flowered skullcap, rhizome of chuanxiong, the seed of cowherb, play both sides of the street, radix bupleuri, Radix Angelicae Sinensis, motherwort, kelp and Radix Glycyrrhizae form, the parts by weight that its each composition accounts for the traditional Chinese medicinal components total amount are:
The Radix Astragali: 60～80 parts, Radix Codonopsis: 60～80 parts, dandelion: 60～80 parts, the root of large-flowered skullcap: 40～60 parts,
Rhizome of chuanxiong: 10～20 parts, the seed of cowherb: 40～60 parts, play both sides of the street: 40～60 parts, radix bupleuri: 50～80 parts,
Radix Angelicae Sinensis: 40～60 parts, motherwort: 50～80 parts, kelp: 80-120 part, Radix Glycyrrhizae: 30～50 parts;
Step e: with many bacterial classification mixing Chinese medicine fermented liquids with etc. the maize cob meal of weight be mixed and made into the solid state fermentation seed;
Step F: the solid state fermentation seed is inoculated into carries out the secondary herb fermenting in the Chinese medicine solid medium, thereby make the cow feed additive that contains the active Chinese drug component probio.
2. the manufacture method that contains the cow feed additive of active Chinese drug component probio according to claim 1, it is characterized in that, the described multiple milk cow of the choosing equal bacterial strain quantity former bacterial strain of active probiotic, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity with the former bacterial strain of active probiotic and expand and cultivate, comprise with the primary seed solution of active probiotic thereby obtain every kind of milk cow:
With the former bacterial strain seed inclined plane inoculating of the bacillus subtilis behind the separating-purifying in the triangular flask of bacillus subtilis bacterium culture medium, and shaken cultivation 24 hours, thus obtain the primary seed solution of bacillus subtilis; The bacillus subtilis bacterium culture medium comprises 3 parts beef extract, 10 parts peptone, 5 parts sodium chloride, 20 parts agar and 1000 parts water;
And/or,
With the former bacterial strain seed inclined plane inoculating of the bafillus natto behind the separating-purifying in the triangular flask of bafillus natto culture medium, and shaken cultivation 24 hours, thus obtain the primary seed solution of bafillus natto; The bafillus natto culture medium comprises 3 parts beef extract, 10 parts peptone, 5 parts sodium chloride, 20 parts agar and 1000 parts water;
And/or,
With the former bacterial strain seed inclined plane inoculating of the bacillus licheniformis behind the separating-purifying in the triangular flask of bacillus licheniformis culture medium, and shaken cultivation 24 hours, thus obtain the primary seed solution of bacillus licheniformis; The bacillus licheniformis culture medium comprises 3 parts beef extract, 10 parts peptone, 5 parts sodium chloride, 20 parts agar and 1000 parts water;
And/or,
With the former bacterial strain seed inclined plane inoculating of the brewer's yeast behind the separating-purifying in the triangular flask of brewer's yeast culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of brewer's yeast; The brewer's yeast culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water;
And/or,
With the former bacterial strain seed inclined plane inoculating of the candida utili behind the separating-purifying in the triangular flask of candida utili culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of candida utili; The candida utili culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water;
And/or,
With the former bacterial strain seed inclined plane inoculating of the aspergillus niger behind the separating-purifying in the triangular flask of aspergillus niger culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of aspergillus niger; The aspergillus niger culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water;
And/or,
With the former bacterial strain seed inclined plane inoculating of the Rhizopus oryzae behind the separating-purifying in the triangular flask of Rhizopus oryzae culture medium, and shaken cultivation 48 hours, thus obtain the primary seed solution of Rhizopus oryzae; The Rhizopus oryzae culture medium comprises 200 parts potato, 20 parts sucrose, 3 parts potassium dihydrogen phosphate, 1.5 parts magnesium sulfate, 20 parts agar and 1000 parts water;
And/or,
The former bacterial strain seed inclined plane inoculating of the Lactobacillus casei behind the separating-purifying in the Kolle flask of Lactobacillus casei culture medium, and 37 ℃ of lower cultivations 48 hours, thereby is obtained the primary seed solution of Lactobacillus casei; The Lactobacillus casei culture medium comprises 20 parts glucose, 10 parts peptone, 10 parts beef extract, 5 parts yeast extract, 5 parts natrium citricum, 2 parts sodium dihydrogen phosphate, 5 parts sodium acetate, 0.2 part epsom salt, 0.25 part four water manganese sulfates, 1 part Tween 80 and 1000 parts water;
And/or,
The former bacterial strain seed inclined plane inoculating of lactobacillus acidophilus behind the separating-purifying in the Kolle flask of lactobacillus acidophilus culture medium, and 37 ℃ of lower cultivations 48 hours, thereby is obtained the primary seed solution of lactobacillus acidophilus; The lactobacillus acidophilus culture medium comprises 20 parts glucose, 10 parts peptone, 10 parts beef extract, 5 parts yeast extract, 5 parts natrium citricum, 2 parts sodium dihydrogen phosphate, 5 parts sodium acetate, 0.2 part epsom salt, 0.25 part four water manganese sulfates, 1 part Tween 80 and 1000 parts water;
And/or,
The former bacterial strain seed inclined plane inoculating of Bifidobacterium behind the separating-purifying in the Kolle flask of Medium of Bifidobacterium, and 37 ℃ of lower cultivations 48 hours, thereby is obtained the primary seed solution of Bifidobacterium; Medium of Bifidobacterium comprises 20 parts glucose, 10 parts peptone, 10 parts beef extract, 5 parts yeast extract, 5 parts natrium citricum, 2 parts sodium dihydrogen phosphate, 5 parts sodium acetate, 0.2 part epsom salt, 0.25 part four water manganese sulfates, 1 part Tween 80 and 1000 parts water.
3. the manufacture method that contains the cow feed additive of active Chinese drug component probio according to claim 1 and 2, it is characterized in that, described many bacterial classifications Hybrid TierI seed liquor is inoculated into carried out many bacterial classifications Mixed culture in the seed tank culture base, may further comprise the steps thereby obtain many bacterial classifications mixing secondary seed solution:
Step C1: preparation seed tank culture base;
Wherein, described seed tank culture base comprises 3%～8% brown sugar, 0.2%～0.5% potassium dihydrogen phosphate, 0.3%～0.5% yeast extract, 0.2%～0.5% peptone and the water of surplus;
Step C2: under 115 ℃～121 ℃, to seed tank culture base sterilization 25 minutes, be cooled to again 30 ℃;
Step C3: many bacterial classifications Hybrid TierI seed liquor is inoculated in the seed tank culture base according to 3%～5% inoculative proportion;
Step C4: the seed tank culture base that docking finishes carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, finishes until the pH value in the seed tank culture base reaches fermentation in 4.8 o'clock, namely obtains many bacterial classifications and mixes secondary seed solution.
4. the manufacture method that contains the cow feed additive of active Chinese drug component probio according to claim 1 and 2, it is characterized in that, described many bacterial classifications mixing secondary seed solution are inoculated into carried out first herb fermenting in the Chinese medicinal liquor culture medium, may further comprise the steps thereby obtain many bacterial classification mixing Chinese medicine fermented liquids:
Step D1: preparation Chinese medicinal liquor culture medium;
Step D2: under 115 ℃～121 ℃, to Chinese medicinal liquor medium sterilization 20 minutes, be cooled to again 30 ℃;
Step D3: many bacterial classifications are mixed secondary seed solution be inoculated in the Chinese medicinal liquor culture medium according to 5%～10% inoculative proportion;
Step D4: the Chinese medicinal liquor culture medium that docking finishes carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, finishes until the pH value in the Chinese medicinal liquor culture medium reaches fermentation in 5.0 o'clock, namely obtains many bacterial classification mixing Chinese medicine fermented liquids.
5. the manufacture method that contains the cow feed additive of active Chinese drug component probio according to claim 1 and 2, it is characterized in that, the described solid state fermentation seed is inoculated into carried out the secondary herb fermenting in the Chinese medicine solid medium, may further comprise the steps thereby make the cow feed additive that contains the active Chinese drug component probio:
Step F 1: preparation Chinese medicine solid medium;
Wherein, described Chinese medicine solid medium is by 60～70 parts wheat bran, 15～20 parts of dregs of beans, 10～15 parts corn flour, 2～3 parts brown sugar, 2～3 parts traditional Chinese medicinal components, 2～4 parts precipitated calcium carbonate, 0.5～0.8 part yeast extract, 1～2 part sea-tangle, 0.2～0.5 part fulvic acid forms; Described traditional Chinese medicinal components comprises the Radix Astragali, Radix Codonopsis, dandelion, the root of large-flowered skullcap, rhizome of chuanxiong, the seed of cowherb, plays both sides of the street, radix bupleuri, Radix Angelicae Sinensis, motherwort, kelp and Radix Glycyrrhizae, and the parts by weight that its each composition accounts for the traditional Chinese medicinal components total amount are:
The Radix Astragali: 60～80 parts, Radix Codonopsis: 60～80 parts, dandelion: 60～80 parts, the root of large-flowered skullcap: 40～60 parts,
Rhizome of chuanxiong: 10～20 parts, the seed of cowherb: 40～60 parts, play both sides of the street: 40～60 parts, radix bupleuri: 50～80 parts,
Radix Angelicae Sinensis: 40～60 parts, motherwort: 50～80 parts, kelp: 80-120 part, Radix Glycyrrhizae: 30～50 parts;
Step F 2: under 115 ℃～121 ℃, to Chinese medicine solid medium sterilization 20 minutes, be cooled to again 30 ℃;
Step F 3: the solid state fermentation seed liquor is inoculated in the Chinese medicine solid medium according to 20%～30% inoculative proportion;
Step F 4: the Chinese medicine solid medium that will inoculate places and carries out sealing and fermenting under 37 ℃, when the temperature of Chinese medicine solid medium reaches 40 ℃ the Chinese medicine solid medium is overturn; Continuing fermentation, time fermentation is finished until the temperature of Chinese medicine solid medium no longer rises, and namely obtains to contain the cow feed additive of active Chinese drug component probio.
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