CN102366023A - 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 PDF

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CN102366023A
CN102366023A CN2011102763780A CN201110276378A CN102366023A CN 102366023 A CN102366023 A CN 102366023A CN 2011102763780 A CN2011102763780 A CN 2011102763780A CN 201110276378 A CN201110276378 A CN 201110276378A CN 102366023 A CN102366023 A CN 102366023A
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chinese medicine
seed
inoculated
fermentation
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CN102366023B (en
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汪松林
李羽
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Liaoning Back Gold Biotechnology Co Ltd
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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

A kind of manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic
Technical field
The present invention relates to the animal and fowl fodder field, relate in particular to a kind of manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic.
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 adopted mostly and is prevented cow subclinical mastitis to the antibiotic mode of the long-term feeding of milk cow.Antibiotic has kill bacteria and the effect that suppresses pathogenic microorganisms, and therefore this feeding patterns has played the effect of certain prevention cow subclinical mastitis in actual production.But; These antibiotic can Along with people's eat milk and various milk product entering human body; 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 processing 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 under the prerequisite that has no side effect, has no drug resistance, do not have pathogenicity, preventing cow subclinical mastitis, 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, bifid lactobacillus, streptococcus fecalis etc.), bacillus preparation (comprising hay bacillus, bacillus cereus, lichens bacillus etc.), fungi preparation (comprising aspergillus oryzae etc.), saccharomycete preparation (comprising brewer's yeast etc.) etc.Existing ox with the microbial forage additive production process in; Only use protein substance and/or glucide culture medium mostly as active probiotic; Though bacterial strain quantitatively can satisfy the demands; But the activity of most of bacterial strains is not high, the field planting performance in enteron aisle a little less than, so active probiotic as milk cow forage the effect that should bring into play receive very big restriction; 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 manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic; 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 are effectively treated and are prevented cow subclinical mastitis, and then replace antibiotic use fully.
The objective of the invention is to realize through following technical scheme:
A kind of manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic comprises:
Steps A: choose the multiple milk cow that equates bacterial strain quantity former bacterial strain, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity expansion cultivation with the former bacterial strain of active probiotic, thereby obtain the primary seed solution of every kind of milk cow with active probiotic with active probiotic;
Wherein, described multiple milk cow with active probiotic comprise bacillus subtilis, bafillus natto, bacillus licheniformis, brewer's yeast 、 Chan ?in Candida, aspergillus niger, Rhizopus oryzae, Lactobacillus casei, lactobacillus acidophilus and the Bifidobacterium at least two kinds;
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 Chinese medicine fermentation first in the Chinese medicine fluid nutrient medium, thereby obtain many bacterial classification mixing Chinese medicine fermented liquids;
Wherein, described Chinese medicine fluid nutrient 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 secondary Chinese medicine fermentation in the Chinese medicine solid medium, thereby make the cow feed additive that contains the Chinese medicine active probiotic.
Technical scheme by the invention described above provides can be found out; The manufacturing approach of the cow feed additive that contains the Chinese medicine active probiotic that the embodiment of the invention provides adopts kinds of traditional Chinese medicines is cultivated has 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 being turned out is turned out far above ordinary culture medium has also strengthened the field planting ability of active probiotic in enteron aisle simultaneously; Because Chinese medicine and active probiotic have synergy to immune system, therefore will use feed addictive as milk cow with the active probiotic that Chinese medicine breeds, can strengthen the immunocompetence of milk cow greatly.We cultivate active probiotic through modern microecology principle is combined with the traditional Chinese medical science with traditional Chinese medicinal components, use feed addictive thereby produce a kind of antibiotic no drug residue, prophyiaxis and promoting growth, milk cow 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 is done to introduce simply in will describing embodiment below; Obviously, the accompanying drawing in describing below only is some embodiments of the present invention, for those of ordinary skill in the art; Do not paying under the capable prerequisite of creative work, can also obtain other accompanying drawings according to these accompanying drawings.
Fig. 1 is the schematic flow sheet of the manufacturing approach of the cow feed additive that contains the Chinese medicine active probiotic that the embodiment of the invention provided.
The specific embodiment
Following combines the accompanying drawing in the embodiment of the invention, and the technical scheme in the embodiment of the invention is carried out clear, intactly description, 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 is obtained under the creative work prerequisite.
At first, need to prove that part alleged in the present specification all is meant weight portion, make a detailed description in the face of the manufacturing approach of the cow feed additive that contains the Chinese medicine active probiotic that the embodiment of the invention provided down.
As shown in Figure 1, a kind of manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic specifically can may further comprise the steps:
Steps A: choose the multiple milk cow that equates bacterial strain quantity former bacterial strain, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity expansion cultivation with the former bacterial strain of active probiotic, thereby obtain the primary seed solution of every kind of milk cow with active probiotic with active probiotic;
Wherein, described multiple milk cow with active probiotic comprise bacillus subtilis, bafillus natto, bacillus licheniformis, brewer's yeast 、 Chan ?in Candida, aspergillus niger, Rhizopus oryzae, Lactobacillus casei, lactobacillus acidophilus and the Bifidobacterium at least two kinds;
Particularly; Choose the former bacterial strain of the multiple milk cow of equal bacterial strain quantity accordingly with 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:
The former bacterial strain seed inclined-plane that (1) will separate the bacillus subtilis after purifying is inoculated in the triangular flask of bacillus subtilis bacterium culture medium, and shaken cultivation 24 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane that (2) will separate the bafillus natto after purifying is inoculated in the triangular flask of bafillus natto culture medium, and shaken cultivation 24 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane that (3) will separate the bacillus licheniformis after purifying is inoculated in the triangular flask of bacillus licheniformis culture medium, and shaken cultivation 24 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane that (4) will separate the brewer's yeast after purifying is inoculated in the triangular flask of brewer's yeast culture medium, and shaken cultivation 48 hours, thereby obtains 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) will separate purify after De Chan ?the former bacterial strain seed inclined-plane of Candida be inoculated into Chan ?in the triangular flask of Candida culture medium, and shaken cultivation 48 hours, thus Huo De Chan ?the primary seed solution of Candida; Chan ?the Candida culture medium comprise 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 ， Chan ?the inoculative proportion of Candida be every 500mL De Chan ?Candida inoculation of medium 100mL De Chan ?Candida.
And/or,
The former bacterial strain seed inclined-plane that (6) will separate the aspergillus niger after purifying is inoculated in the triangular flask of aspergillus niger culture medium, and shaken cultivation 48 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane that (7) will separate the Rhizopus oryzae after purifying is inoculated in the triangular flask of Rhizopus oryzae culture medium, and shaken cultivation 48 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane that (8) will separate the Lactobacillus casei after purifying is inoculated in the Kolle flask of Lactobacillus casei culture medium, and cultivates 48 hours down at 37 ℃, thereby obtains 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 64.H. 124.O. 26.; English name is POLYSORBATE 80, and Chinese synonym is polyoxyethylene sorbitan monooleate or emulsifying agent T80, soluble in water, ethanol, methyl alcohol and ethyl acetate; Be mainly used in solubilizer or the emulsifying agent of doing 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 bigger 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,
The former bacterial strain seed inclined-plane that (9) will separate the lactobacillus acidophilus after purifying is inoculated in the Kolle flask of lactobacillus acidophilus culture medium, and cultivates 48 hours down at 37 ℃, thereby obtains 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 bigger 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,
The former bacterial strain seed inclined-plane that (10) will separate the Bifidobacterium after purifying is inoculated in the Kolle flask of Bifidobacterium culture medium, and cultivates 48 hours down at 37 ℃, thereby obtains the primary seed solution of Bifidobacterium; The Bifidobacterium 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 bigger 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 the Bifidobacterium inoculation of medium 100mL 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 practical application; 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, mix then, 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-sided factors such as comprehensive cost of manufacture and serviceabilities, each composition of this seed tank culture base accounts for total mass ratio can select the scheme in the following table 1 for use, and the embodiment one in wherein should showing 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 ℃,, be cooled to 30 ℃ again to seed tank culture base sterilization 25 minutes;
Particularly, in practical application, preferably select for use under 121 ℃ the seed tank culture base was sterilized 25 minutes, can be killed fully to guarantee the harmful bacterium 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% here is meant 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 to having inoculated carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, and the pH value in the seed tank culture base reaches fermentation in 4.8 o'clock to be accomplished, and promptly obtains many bacterial classifications and mixes secondary seed solution.Generally speaking, the anaerobic fermentation of seed tank culture base generally can be accomplished in 48～72 hours, and after fermentation was accomplished, the number of active probiotic can reach every milliliter 1,500,000,000 strain.
Step D: many bacterial classifications are mixed secondary seed solution be inoculated into and carry out Chinese medicine fermentation first in the Chinese medicine fluid nutrient medium, thereby obtain many bacterial classification mixing Chinese medicine fermented liquids;
Wherein, Described Chinese medicine fluid nutrient 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 medicine fluid nutrient 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-sided factors such as comprehensive cost of manufacture and serviceabilities, each composition of this Chinese medicine fluid nutrient medium accounts for total mass ratio can select the preferred version in the following table 2 for use, and the embodiment 1 in wherein should showing is a 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 traditional Chinese medicinal components, and each composition accordingly accounts for the parts by weight of traditional Chinese medicinal components total amount can select the preferred version in the following table 3 for use, and wherein the unit in this table is weight portion, and embodiment a is a most preferred embodiment:
Table 3:
The traditional Chinese medicine ingredients title Instance a Instance b Instance c Instance 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 Chinese medicine fermentation first in the Chinese medicine fluid nutrient medium accordingly, can may further comprise the steps thereby obtain many bacterial classification mixing Chinese medicine fermented liquids:
Step D1: preparation Chinese medicine fluid nutrient medium;
Step D2: under 115 ℃～121 ℃,, be cooled to 30 ℃ again to herb liquid body medium sterilization 20 minutes;
Particularly, in practical application, preferably select for use under 121 ℃, can be killed fully to guarantee the harmful bacterium in the Chinese medicine fluid nutrient medium to herb liquid body medium sterilization 20 minutes.
Step D3: many bacterial classifications are mixed secondary seed solution be inoculated into (the inoculative proportion according to 5%～10% here is meant that many bacterial classifications of 5 to 10 unit volumes of Chinese medicine liquid inoculation of medium of per 100 unit volumes mix secondary seed solution) in the Chinese medicine fluid nutrient medium according to 5%～10% inoculative proportion;
Step D4: the Chinese medicine fluid nutrient medium to having inoculated carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, and the pH value in the Chinese medicine fluid nutrient medium reaches fermentation in 5.0 o'clock to be accomplished, and promptly obtains many bacterial classification mixing Chinese medicine fermented liquids; Generally speaking, the anaerobic fermentation of Chinese medicine fluid nutrient medium generally can be accomplished in 48～72 hours, and after fermentation was accomplished, the number of active probiotic can reach every milliliter 2,000,000,000 strain.
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 (the microwaves corresponding sterilization is meant that adopting frequency 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, do not have harmful bacteria in the active probiotic solid state fermentation seed that is synthesized with assurance.
Step F: the solid state fermentation seed is inoculated into carries out secondary Chinese medicine fermentation in the Chinese medicine solid medium, thereby make the cow feed additive that contains the Chinese medicine active probiotic.
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, (winnofil is claimed but also claimed to precipitated calcium carbonate not only 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 commercial production at home; Precipitated calcium carbonate adopts carbonizatin method to make mostly---the calcining of raw materials such as lime stone is generated lime and carbon dioxide; Add entry again and make lime become milk of lime, feed carbon dioxide then and make 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 is formed; 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-sided factors such as comprehensive cost of manufacture and serviceabilities, each composition of this Chinese medicine solid medium accounts for total mass ratio can select the preferred version in the following table 4 for use, and wherein the embodiment A in this table is a 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 for use, and wherein the unit in the table 3 is weight portion, and the embodiment a in the table 3 is a 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 fermentation of secondary Chinese medicine in the Chinese medicine solid medium accordingly, may further comprise the steps thereby make the cow feed additive that contains the Chinese medicine active probiotic:
Step F 1: preparation Chinese medicine solid medium;
Step F 2: under 115 ℃～121 ℃,, be cooled to 30 ℃ again to Chinese medicine solid medium sterilization 20 minutes;
Particularly, in practical application, preferably select for use under 121 ℃ the Chinese medicine solid medium was sterilized 25 minutes, can be killed fully to guarantee the harmful bacterium 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% here is meant 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, fermentation is accomplished when the temperature of Chinese medicine solid medium no longer rises, and promptly obtains to contain the cow feed additive of Chinese medicine active probiotic.Generally speaking, the sealing and fermenting of Chinese medicine solid medium generally can be accomplished in 5～7 days, and after fermentation was accomplished, the number of active probiotic can reach every milliliter 4,500,000,000 strain.After sieving, material after fermentation accomplished can be packaged into the cow feed additive that contains the Chinese medicine active probiotic.
In the technical aspect, the invention is described in traditional Chinese medicine composition for the efficacy of each component: Astragalus - sweet and warm, qi Sun, the active ingredient of Astragalus polysaccharides can improve immunity; Codonopsis - sweet nature, complement in Qi, and the stomach, the active ingredient saponins can improve immunity; dandelion - sweet natured can detoxify, and reducing swelling with dandelion sterols, choline, inulin and other common ingredients and skullcap suppress harmful mastitis bacteria (Staphylococcus aureus and Streptococcus pyogenes); skullcap - heat and dryness, cooling blood detoxification; Sichuan dome - blood circulation of qi, yin, circulation, Sichuan dome of blood gas medicine, let Blood complement rather than stagnation; Vaccaria - promoting blood circulation longitude, under the breast carbuncle, which contains triterpenoid saponins, glycosides; Passepartout - blood circulation, Qufeng, diuresis through longitude; Bupleurum - bitter, slightly cold, hepatobiliary classics, there Shugan and stomach, Sun's functional, antibacterial and antiviral and immunomodulatory functions; Angelica - sweet and warm, not only blood, but also blood, its active ingredient SF can activate macrophages, immune function play; Motherwort - stasis, containing a variety of alkaloids and organic selenium, can improve the body's immune system; Khumbu - Pinggan, suppressing gas relieves congestion, high iodine content and rich in organic Tongru factors can increase milk yield; licorice - with detoxification, to reconcile the various drugs, their active ingredient glycyrrhizin in cell immunomodulator.The mentioned traditional Chinese medicinal components of the present invention is the growth characteristic according to milk cow, the treatment of cow subclinical mastitis and the breed demand in prevention mechanism and the milk cattle cultivating process; The drug matching principle of adopt monarch, minister, helping, make; It is formulated to choose a few flavor Chinese medicines that can promote milk cow digestion and growth, can strengthen the milk cow immunity of organisms, can improve the milk cow resistance against diseases respectively---and at first select for use the Radix Astragali, Radix Codonopsis, Radix Angelicae Sinensis as monarch drug in a prescription; Improve animal body immunity, reach the purpose of benefiting qi and nourishing blood: select for use dandelion, the root of large-flowered skullcap, radix bupleuri to improve its anti-bacteria and anti-virus ability simultaneously; And select rhizome of chuanxiong, the seed of cowherb for use, play both sides of the street, motherwort, kelp breast promoting blood circulation and removing blood stasis, logical and improve the output of milk; Select for use the Radix Glycyrrhizae not only can be clearing heat and detoxicating and can also be in harmonious proportion all the other all medicines at last.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 technology of not describing in detail all can adopt strain culturing equipment general in the prior art and strain culturing technology, do not remake detailed description in this application.Various Chinese medicines described in the present invention all can be buied in each big Chinese medicine pharmacy easily, therefore this are no longer detailed.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.
It is thus clear that; 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 of raising cow feed additive is cured the disease; Effectively treat and the prevention cow subclinical mastitis, and then replace antibiotic use fully.
The above; Be merely the preferable specific embodiment of the present invention, but protection scope of the present invention is not limited thereto, any technical staff who is familiar with the present technique field is 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 manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic is characterized in that, comprising:
Steps A: choose the multiple milk cow that equates bacterial strain quantity former bacterial strain, and respectively every kind of milk cow is carried out single bacterial classification bacterial strain quantity expansion cultivation with the former bacterial strain of active probiotic, thereby obtain the primary seed solution of every kind of milk cow with active probiotic with active probiotic;
Wherein, described multiple milk cow with active probiotic comprise bacillus subtilis, bafillus natto, bacillus licheniformis, brewer's yeast 、 Chan ?in Candida, aspergillus niger, Rhizopus oryzae, Lactobacillus casei, lactobacillus acidophilus and the Bifidobacterium at least two kinds;
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 Chinese medicine fermentation first in the Chinese medicine fluid nutrient medium, thereby obtain many bacterial classification mixing Chinese medicine fermented liquids;
Wherein, described Chinese medicine fluid nutrient 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 secondary Chinese medicine fermentation in the Chinese medicine solid medium, thereby make the cow feed additive that contains the Chinese medicine active probiotic.
2. the manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic according to claim 1; It is characterized in that; The described former bacterial strain of choosing the multiple milk cow of equal bacterial strain quantity with 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:
The former bacterial strain seed inclined-plane of bacillus subtilis after separate purifying is inoculated in the triangular flask of bacillus subtilis bacterium culture medium, and shaken cultivation 24 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane of the bafillus natto after separate purifying is inoculated in the triangular flask of bafillus natto culture medium, and shaken cultivation 24 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane of bacillus licheniformis after separate purifying is inoculated in the triangular flask of bacillus licheniformis culture medium, and shaken cultivation 24 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane of the brewer's yeast after separate purifying is inoculated in the triangular flask of brewer's yeast culture medium, and shaken cultivation 48 hours, thereby obtains 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 De Chan after separate purifying ?the former bacterial strain seed inclined-plane of Candida be inoculated into Chan ?in the triangular flask of Candida culture medium, and shaken cultivation 48 hours, thus Huo De Chan ?the primary seed solution of Candida; Chan ?the Candida culture medium comprise 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 of the aspergillus niger after separate purifying is inoculated in the triangular flask of aspergillus niger culture medium, and shaken cultivation 48 hours, thereby obtains 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,
The former bacterial strain seed inclined-plane of the Rhizopus oryzae after separate purifying is inoculated in the triangular flask of Rhizopus oryzae culture medium, and shaken cultivation 48 hours, thereby obtains 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 that separates the Lactobacillus casei after purifying is inoculated in the Kolle flask of Lactobacillus casei culture medium, and cultivated 48 hours down at 37 ℃, thus the primary seed solution of acquisition 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 that separates the lactobacillus acidophilus after purifying is inoculated in the Kolle flask of lactobacillus acidophilus culture medium, and cultivated 48 hours down at 37 ℃, thus the primary seed solution of acquisition 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 that separates the Bifidobacterium after purifying is inoculated in the Kolle flask of Bifidobacterium culture medium, and cultivated 48 hours down at 37 ℃, thus the primary seed solution of acquisition Bifidobacterium; The Bifidobacterium 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.
3. according to claim 1 and the 2 described manufacturing approaches that contain the cow feed additive of Chinese medicine active probiotic; 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 ℃,, be cooled to 30 ℃ again to seed tank culture base sterilization 25 minutes;
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 to having inoculated carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, and the pH value in the seed tank culture base reaches fermentation in 4.8 o'clock to be accomplished, and promptly obtains many bacterial classifications and mixes secondary seed solution.
4. the manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic according to claim 1 and 2; It is characterized in that; Described many bacterial classifications mixing secondary seed solution are inoculated into carried out Chinese medicine fermentation first in the Chinese medicine fluid nutrient medium, may further comprise the steps thereby obtain many bacterial classification mixing Chinese medicine fermented liquids:
Step D1: preparation Chinese medicine fluid nutrient medium;
Step D2: under 115 ℃～121 ℃,, be cooled to 30 ℃ again to herb liquid body medium sterilization 20 minutes;
Step D3: many bacterial classifications are mixed secondary seed solution be inoculated in the Chinese medicine fluid nutrient medium according to 5%～10% inoculative proportion;
Step D4: the Chinese medicine fluid nutrient medium to having inoculated carries out anaerobic fermentation, and the rotating speed of installation for fermenting is 180r/min, and fermentation temperature is 32 ℃, and the pH value in the Chinese medicine fluid nutrient medium reaches fermentation in 5.0 o'clock to be accomplished, and promptly obtains many bacterial classification mixing Chinese medicine fermented liquids.
5. the manufacturing approach that contains the cow feed additive of Chinese medicine active probiotic according to claim 1 and 2; It is characterized in that; The described solid state fermentation seed is inoculated into carried out the fermentation of secondary Chinese medicine in the Chinese medicine solid medium, may further comprise the steps thereby make the cow feed additive that contains the Chinese medicine active probiotic:
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 is formed; 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 ℃,, be cooled to 30 ℃ again to Chinese medicine solid medium sterilization 20 minutes;
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, fermentation is accomplished when the temperature of Chinese medicine solid medium no longer rises, and promptly obtains to contain the cow feed additive of Chinese medicine active probiotic.
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CN106260542A (en) * 2016-07-29 2017-01-04 河南德邻生物制品有限公司 A kind of lactating sow Chinese medicine microbial fermentation preparation and preparation method thereof
CN107080079A (en) * 2017-06-17 2017-08-22 哈尔滨伟平科技开发有限公司 The preparation method of one boar Chinese herbal feed additive
CN107080080A (en) * 2017-06-17 2017-08-22 哈尔滨伟平科技开发有限公司 A kind of preparation method of pig feed additive
CN107319051A (en) * 2017-07-27 2017-11-07 贵州源熙生物研发有限公司 A kind of stem of noble dendrobium red yeast rice and preparation method and application
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CN108653487A (en) * 2018-06-27 2018-10-16 张家口万全区科泰生物科技有限公司 It is a kind of enhancing milk cow immunity active peptide and its application
CN110637789A (en) * 2019-11-15 2020-01-03 陈武茂 Ecological breeding method for cattle
CN110679545A (en) * 2019-11-15 2020-01-14 陈武茂 Beef cattle ecological breeding method
CN112753889A (en) * 2020-12-31 2021-05-07 内江师范学院 Motherwort mildew preventive, preparation method and application in fish feed
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