CN108850537A - The preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation - Google Patents
The preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation Download PDFInfo
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- CN108850537A CN108850537A CN201810499883.3A CN201810499883A CN108850537A CN 108850537 A CN108850537 A CN 108850537A CN 201810499883 A CN201810499883 A CN 201810499883A CN 108850537 A CN108850537 A CN 108850537A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The present invention relates to feed additive fields, disclose a kind of preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation, using by eupatorium lindleynun var. trifoliolatum, motherwort, four kinds of prepared from traditional Chinese medicines of Herba Andrographitis and trichosanthes fruit at Chinese herb compound prepare Chinese herbal medicine digestion liquid and fermentation medium respectively；Fungi isolation medium is prepared using Chinese herbal medicine digestion liquid；The Fresh Plants of four kinds of Chinese herbal medicines of chopping are placed in culture to incision on fungi isolation medium and grow mycelia；Mycelia is forwarded on the fungi isolation medium added with 20% Chinese herbal medicine digestion liquid and turns out endogenetic fungal bacterial strain, and filters out well-grown endogenetic fungal bacterial strain；The endogenetic fungal bacterial strain filtered out is inoculated on liquid seed culture medium and turns out seed liquor；Seed liquor is inoculated on fermentation medium and is fermented, Chinese herbal feed additive is obtained.Compared with prior art, the bacteriostatic activity and enzymatic activity for the Chinese herbal feed additive that the present invention prepares improve a lot.
Description
Technical field
The present invention relates to feed additive field, in particular to a kind of Chinese herbal feed of Chinese herbal medicine endogenetic fungus fermentation adds
Add the preparation method of agent.
Background technique
Beasts, birds and aquatic products product quality requirement is continuously improved in the food-safe great attention of modern people, wide in feed
Although general addition antibiotic agents can achieve the effect for promoting growth of animal and improving animal immunizing power, but due to livestock and poultry water
Produce cultivation in illegally addition and excess addition antibiotic the case where it is commonplace, negatively affect it is outstanding day by day, pathogenic bacteria it is resistance to
The problems such as pharmacological property enhancing and medicament residue, has attracted more and more attention from people, and therefore, researches and develops safe and efficient, green novel feeding
The key subjects of animal husbandry and fishery industry are had become with Substitutes For Antibiotic.Guaranteeing that feed addictive meets the basis of national regulation
On, natural Chinese medicinal herb feed addictive has gradually embodied the advantage of oneself.
Eupatorium lindleynun var. trifoliolatum, motherwort, Herba Andrographitis and trichosanthes fruit are traditional Chinese medicine, are rich in flavone compound, terpene lactones, green original
Acid is polymerized to the compound that enol etc. has notable biological activity to people and livestock and poultry.These active materials can effectively enhance animal machine
The immune function of body promotes the development of Immune Organs of Body, and can inhibit Escherichia coli, staphylococcus aureus, common change
The common various pathogenic microorganisms such as shape bacillus and salmonella typhimurium promote cultivated animals health.Recent studies indicate that
Using eupatorium lindleynun var. trifoliolatum, motherwort, Herba Andrographitis and trichosanthes fruit and its extract as poultry, domestic animal, aquatic products feed addictive, not only
It can prevent the generation of Animal diseases, adjust Immune Function In Animals, reduce prophylactic, increase edible safety, moreover it is possible to improve meat
Product edible quality and mouthfeel.But there are medication complicated component, dosage for the application and research due to Chinese herbal feed additive greatly,
The problems such as nutrient concentrations are low, palatability is poor, effect is unstable, the large-scale promotion for hindering Chinese herbal feed additive is answered
With.
Summary of the invention
Goal of the invention：Aiming at the problems existing in the prior art, the present invention provides a kind of fermentation of Chinese herbal medicine endogenetic fungus
The preparation method of Chinese herbal feed additive, the bacteriostatic activity and enzymatic activity for the Chinese herbal feed additive prepared have very big
It improves.
Technical solution：The present invention provides a kind of preparation sides of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation
Method includes the following steps：S1：Using by eupatorium lindleynun var. trifoliolatum, motherwort, four kinds of prepared from traditional Chinese medicines of Herba Andrographitis and trichosanthes fruit at Chinese herbal medicine answer
Side prepares Chinese herbal medicine digestion liquid and fermentation medium respectively；S2：Fungi isolation medium is prepared using the Chinese herbal medicine digestion liquid；
S3：The Fresh Plants of four kinds of Chinese herbal medicines of chopping are placed in culture to incision on the fungi isolation medium and grow mycelia；
S4：The mycelia is forwarded on the fungi isolation medium added with the 20% Chinese herbal medicine digestion liquid and turns out interior life
Fungal bacterial strain, and filter out well-grown endogenetic fungal bacterial strain；S5：The endogenetic fungal bacterial strain filtered out is inoculated in liquid strain
Seed liquor is turned out on sub- culture medium；S6：The seed liquor is inoculated on the fermentation medium and is fermented, Chinese herbal medicine is obtained
Feed addictive.
Preferably, in the S1, in the Chinese herb compound, four kinds of eupatorium lindleynun var. trifoliolatum, motherwort, Herba Andrographitis and trichosanthes fruit medium-height grass
The mass ratio of medicine is 1:1:1:1.
Preferably, in the S1, the preparation method of the Chinese herbal medicine digestion liquid is as follows：The Chinese herb compound is crushed
To 40 mesh, 20 times of distilled water is added, decocts coarse filtration after 30 min, takes filtrate to be centrifuged 15min in 8000r/min, takes supernatant
Liquid obtains the Chinese herbal medicine digestion liquid after being concentrated under reduced pressure 5 times.
Preferably, in the S1, the group of the fermentation medium becomes：Chinese herb compound 70%, wheat bran 25%, glucose
3%,（NH4）2SO41.5%, KH2PO40.3%, MgSO40.2% and water 50-60%.
Preferably, in the S2, the preparation method of the fungi isolation medium is as follows：In Martin solid medium
The middle fungi isolation medium that the 5% Chinese herbal medicine digestion liquid is added, pH are natural.
Preferably, in the S5, the group of the liquid seed culture medium becomes：Glucose 5%, beancake powder 1.5%, yeast
Powder 0.5%, KH2PO40.2%, MgSO4·7H2O 0.1%, CaCO3 0.4%。
Preferably, in the S5, the condition of culture of the seed liquor is：28 DEG C, 180 r/min, 72h.
Preferably, in the S6, the water content of the fermentation medium is 50 ~ 60%, pH naturally, after 30 min of sterilizing,
The seed liquor is inoculated on the fermentation medium by volume mass than the inoculum concentration for 10%, after 32 DEG C of fermentation 72h, 50
DEG C drying, obtain the Chinese herbal feed additive.
Beneficial effect：The interior life isolated from four kinds of eupatorium lindleynun var. trifoliolatum, motherwort, Herba Andrographitis and trichosanthes fruit Chinese herbal medicines in the present invention
Fungi can produce a variety of different enzymes during growth metabolism, by macromolecular substances in Chinese herbal feed additive and resist
Trophic factors decomposes or is converted to small-molecule substance, can also generate the secondary metabolite that diseases prevention helps growth, can effectively enhance
Inhibit the work of the various pathogenic microorganisms such as Escherichia coli, staphylococcus aureus, proteus vulgaris and salmonella typhimurium
Property, to improve feed palatability and feed conversion rate, enhancing drug effect and reduction feeding cost promote animal growth, mention
The fertility of high animal and growth performance etc..
Specific embodiment
The present invention is described in detail With reference to embodiment.
It the use of mass ratio is 1:1:1:1 eupatorium lindleynun var. trifoliolatum, motherwort, four kinds of prepared from traditional Chinese medicines of Herba Andrographitis and trichosanthes fruit at medium-height grass
Medicine compound weighs the Chinese herb compound that 20g is crushed to 40 mesh, and 20 times of distilled water is added, and decocts coarse filtration after 30 min, takes filter
Liquid is centrifuged 15min in 8000r/min, takes supernatant, is concentrated under reduced pressure 5 times, obtains Chinese herbal medicine digestion liquid, save backup in 4 DEG C of refrigerators；
Fermentation medium is prepared, wherein Chinese herb compound accounts for 70%, and wheat bran accounts for 25%, and glucose accounts for 3%,（NH4）2SO4Account for 1.5%, KH2PO4
Account for 0.3%, MgSO40.2% is accounted for, water content accounts for 50%；The preparation of 5% Chinese herbal medicine digestion liquid is added in Martin solid medium to come true
Bacterium isolation medium, pH are natural.
Four kinds of fresh medicinal clear water of medium-height grass are rinsed well surface dirt, with deionized water sonic oscillation 10min.?
Under aseptic condition, surface sterilization is carried out to Chinese herbal medicine, specific step is as follows：Aseptic water washing 3 times, 0.1% mercuric chloride solution impregnates
10min, 8% liquor natrii hypochloritis impregnate 5min, aseptic water washing 3 times, 75% ethyl alcohol impregnate 5min, aseptic water washing 3 times.Disinfection
The medicinal aseptic operation of medium-height grass afterwards is cut into leaf (2-5) mm × (2-5) mm, and the segment of root and stem 5-10mm is placed in fungi separation
On culture medium, culture dish is sealed with sealed membrane, 28 DEG C of constant temperature incubation 3-5d；Every ware connects 4-5 sample, observes at material cut
Mycelia (bacterium colony) is grown, incision mycelia is taken, is forwarded on the fungi isolation medium added with 20% Chinese herbal medicine digestion liquid and continues
Culture, filters out well-grown bacterial strain using Tip Splitting picking method in the endogenetic fungal bacterial strain turned out.
All endogenetic fungal bacterial strains filtered out are inoculated in liquid seed culture medium respectively（Glucose 5%, beancake powder
1.5%, yeast powder 0.5%, KH2PO40.2%, MgSO4·7H2O0.1%, CaCO30.4%.）In, 28 DEG C, under the conditions of 180 r/min
72h is cultivated, seed liquor is obtained.Fermentation medium 30g is weighed in 250 mL triangular flasks by formula, and pH is naturally, tap water adjusting contains
Water 50%, after 110 DEG C of 30 min of sterilizing, by 10%（V/m）The good seed liquor of inoculum concentration inoculated and cultured, while it is sterile to be inoculated with
The Chinese herbal medicine fermentation material of water is control group.After 32 DEG C of culture 72h, 50 DEG C of drying obtain the Chinese herbal medicine of Chinese herbal medicine endogenetic fungus fermentation
Feed addictive.
By tissue block method, 6 plants are filtered out altogether from Chinese herb compound in the fungi separation training being gradually incremented by containing concentration
Support well-grown endogenetic fungal bacterial strain on base.
The endogenetic fungus seed liquor isolated is subjected to the fermentation material leaching liquor that ferments to obtain to Chinese herb compound, it is sterile to be inoculated with
The Chinese herbal medicine fermentation material of water is control group, studies fermentation material leaching liquor to indicator strain Escherichia coli（E. coli）, golden yellow Portugal
Grape coccus（S. aureus）, proteus vulgaris（P. vulgaris）And salmonella typhimurium（S. typhimurium）Four
The bacteriostatic activity of strain indicator bacteria.The following Tables 1 and 2 of experimental result.
1 fermentation material leaching liquor bacteriostatic experiment of table
From 1 bacteriostatic experiment data of table it is found that the suppression of the fermentation material leaching liquor for the 6 plants of endogenetic fungus isolated from Chinese herb compound
Bacterium activity is enhanced compared with control group bacteriostatic activity.Wherein number is FG-B and the bacteriostatic activity of two plants of bacterial strains of FY-B is more right
It is the most obvious according to group bacteriostatic activity enhancing.
Bacterial strain FG-B fermentation material leaching liquor is husky to Escherichia coli, staphylococcus aureus, proteus vulgaris and mouse typhus
The bacteriostatic activity of door Salmonella improves 60.91%, 47.42%, 48.84%, 40.34% compared with control group respectively；The leaching of bacterial strain FY-B fermentation material
Extract is to the bacteriostatic activity of Escherichia coli, staphylococcus aureus, proteus vulgaris and salmonella typhimurium compared with control group
It is respectively increased 62.73%, 52.58%, 54.65%, 38.66%.
The experiment of 2 fermentation material leaching liquor enzyme activity of table
From 2 enzyme activity experimental data of table it is found that the enzyme of the fermentation material leaching liquor for the 6 plants of endogenetic fungus isolated from Chinese herb compound
Active is enhanced compared with control group enzymatic activity.The enzymatic activity for the bacterial strain that wherein number is FY-B increases compared with control group enzymatic activity
It is the most obvious by force.CNCase, FPA, protease enzymatic activity respectively reach 2397U/mL, 821U/mL, 1657U/mL.
Comprehensive analysis bacteriostatic experiment and enzyme activity experimental data can obtain, six filtered out from Chinese herbal feed additive compound
The bacteriostatic activity of the fermentation material leaching liquor of the well-grown strain of strain and enzymatic activity compared with control group bacteriostatic activity and enzymatic activity all
It is greatly improved, wherein the bacteriostatic activity for the bacterial strain that number is FY-B and enzymatic activity increase compared with control group bacteriostatic activity and enzymatic activity
It is the most obvious by force.Bacterial strain FY-B fermentation material leaching liquor is to Escherichia coli, staphylococcus aureus, proteus vulgaris and mouse typhus
The bacteriostatic activity of salmonella is respectively increased 62.73%, 52.58%, 54.65%, 38.66% compared with control group；CNCase, FPA, albumen
Enzyme enzymatic activity respectively reaches 2397U/mL, 821U/mL, 1657U/mL.
Morphologic observation is carried out to the preferable endogenetic fungus of the bacteriostatic activity and enzymatic activity that filter out and preliminary classification is identified.It is logical
Bacterium colony observation and conventional microscopy are crossed, is illustrated according to Fungal identification handbook and Dendrochium, is initially identified as aspergillus oryzae, is named as rice
Aspergillus FY-B.As a result such as table 3.
3 endogenetic fungus morphological feature of table
Above embodiment only technical concepts and features to illustrate the invention, its object is to allow person skilled in the art's energy
The solution contents of the present invention much of that are simultaneously implemented accordingly, and it is not intended to limit the scope of the present invention.It is all spiritual according to the present invention
The equivalent transformation or modification that essence is done, should be covered by the protection scope of the present invention.
Claims (8)
1. a kind of preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation, which is characterized in that including following
Step：
S1：Using by eupatorium lindleynun var. trifoliolatum, motherwort, Herba Andrographitis and trichosanthes fruit four kinds of prepared from traditional Chinese medicines at Chinese herb compound prepare respectively
Herbal medicine digestion liquid and fermentation medium；
S2：Fungi isolation medium is prepared using the Chinese herbal medicine digestion liquid；
S3：The Fresh Plants of four kinds of Chinese herbal medicines of chopping are placed in culture to incision on the fungi isolation medium and grow bacterium
Silk；
S4：The mycelia is forwarded on the fungi isolation medium added with the 20% Chinese herbal medicine digestion liquid and is turned out
Endogenetic fungal bacterial strain, and filter out well-grown endogenetic fungal bacterial strain；
S5：The endogenetic fungal bacterial strain filtered out is inoculated on liquid seed culture medium and turns out seed liquor；
S6：The seed liquor is inoculated on the fermentation medium and is fermented, Chinese herbal feed additive is obtained.
2. the preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation according to claim 1, special
Sign is, in the S1, in the Chinese herb compound, eupatorium lindleynun var. trifoliolatum, motherwort, four kinds of Chinese herbal medicines of Herba Andrographitis and trichosanthes fruit quality
Than being 1:1:1:1.
3. the preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation according to claim 1, special
Sign is, in the S1, the preparation method of the Chinese herbal medicine digestion liquid is as follows：
The Chinese herb compound is crushed to 40 mesh, 20 times of distilled water is added, decocts coarse filtration after 30 min, take filtrate in
8000r/min is centrifuged 15min, takes supernatant, obtains the Chinese herbal medicine digestion liquid after being concentrated under reduced pressure 5 times.
4. the preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation according to claim 1, special
Sign is, in the S1, the group of the fermentation medium becomes：Chinese herb compound 70%, wheat bran 25%, glucose 3%,（NH4）2SO41.5%, KH2PO4 0.3%, MgSO40.2% and water 50-60%.
5. the preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation according to claim 1, special
Sign is, in the S2, the preparation method of the fungi isolation medium is as follows：5% is added in Martin solid medium
The fungi isolation medium of the Chinese herbal medicine digestion liquid, pH are natural.
6. the preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation according to claim 1, special
Sign is, in the S5, the group of the liquid seed culture medium becomes：Glucose 5%, beancake powder 1.5%, yeast powder 0.5%,
KH2PO40.2%, MgSO4·7H2O 0.1%, CaCO3 0.4%。
7. the preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation according to claim 1, special
Sign is, in the S5, the condition of culture of the seed liquor is：28 DEG C, 180 r/min, 72h.
8. the preparation method of the Chinese herbal feed additive of Chinese herbal medicine endogenetic fungus fermentation according to claim 1, special
Sign is, in the S6, the water content of the fermentation medium is 50 ~ 60%, pH naturally, after 30 min of sterilizing, by described kind
Sub- liquid is inoculated on the fermentation medium by volume mass than the inoculum concentration for 10%, and after 32 DEG C of fermentation 72h, 50 DEG C of drying are obtained
The Chinese herbal feed additive.
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2018
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| CN101480226A (en) * | 2009-01-23 | 2009-07-15 | 熊忠辉 | Fungal fermentate feed addictive as well as preparation method and use thereof |
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